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Sample records for 86rb vblizi porogov

  1. Effect of selenite on 86Rb uptake by rat lens

    International Nuclear Information System (INIS)

    The effect was observed in vitro of selenite on the uptake of 86Rb by the lens in two weeks old and adult rats. Also measured was the uptake of 86Rb by the lens in vitro in 30-days old rats with cataracts induced by the administration of selenite at day 14 after birth and in control animals of the same age. Selenite in a concentration of 0.4 mM and more caused an insignificant decrease in 86Rb uptake by the lens in adult rats while in the lens of young rats the uptake decreased significantly already at concentrations lower by two orders. Lenses with cataracts took up significantly less 86Rb than transparent lenses; body weight, the weight of the fresh lens and its dry mass decreased while the water content in the lens was higher as compared with control groups. (author). 2 tabs., 26 refs

  2. Loss of receptor-mediated 86Rb efflux from pig aortic endothelial cells in culture.

    OpenAIRE

    Ager, A.; Martin, W

    1983-01-01

    The responsiveness of freshly-isolated and subcultured pig aortic endothelial cells to adenosine triphosphate (ATP), bradykinin and ionophore A23187 was compared by monitoring agonist-induced 86Rb efflux. ATP, bradykinin and ionophore A23187 stimulated 86Rb efflux from freshly-isolated cells. ATP and bradykinin, which act via specific receptors, were less effective at inducing 86Rb efflux from subcultured cells but ionophore A23187 was as effective on subcultured as on freshly-isolated cells....

  3. 86Rb Distribution in the Lung of the Rabbit with Pneumothorax

    International Nuclear Information System (INIS)

    86Rb uptake of some organs and tissues, eg. both lungs, both renal cortices. small intestine, liver and skeletal muscle were studied in the control and the rabbit subjected to pneumothorax. 86Rb in the form of chloride mixed with physiological saline was intravenously injected. The doses were 100 μc for a rabbit. The rabbits were sacrificed at intervals of 10, 20, 40, and 60 seconds after the injection of 86Rb, by the injection of saturated KCI solution. After scarification, the organ and tissue sample were quickly removed. 86Rb uptake in gm of the organs and tissues were measured. On the basis of uptake value, administered doses and body weight, % dose/gm tissues per 200 gm body weight was calculated. Followings were the results: 1. Pneumothorax resulted in a marked elevation in 86Rb uptake value of collapsed lung and returned to normal level lately. 2. Contralateral lung of pneumothorax also showed marked elevation in 86Rb uptake value and recovered to normal level. 3. Initial 86Rb uptake value of liver, small intestine of the rabbit with pneumothorax showed some elevation as compared to control, but that of late stage were similar with control. 4. Local blood flow determination by means of 86Rb uptake were inadequate in the collapsed lung of pneumothorax. 5. It was suggested that the mechanism for the initial elevation of 86Rb uptake value in each organs and tissue were different from each other.

  4. Leiurus quinquestriatus venom inhibits BRL 34915-induced 86Rb+ efflux from the rat portal vein

    International Nuclear Information System (INIS)

    The effect of the crude venom of the Israeli scorpion Leiurus quinquestriatus hebraeus on the 86Rb+ efflux stimulated by the K+ channel opener BRL 34915 in the rat portal vein was examined. Applied alone, the venom greatly increased the spontaneous mechanical activity of and the concomitant 86Rb+ efflux from the vessel. When the excitability of the vein was suppressed by the dihydropyridine calcium antagonist, PN 200-110, the 86Rb+ efflux stimulated by BRL 34915 could be shown to be inhibited by the venom. From the concentration dependence of this inhibition an IC50 value of 0.17 +/- 0.01 mg/ml was estimated. This venom is thus the most potent blocker of BRL 34915-evoked 86Rb+ efflux reported so far. 17 references, 2 figures

  5. Burn-up cross sections of 51Cr, 59Fe, 65Zn, 86Rb, 103Ru

    International Nuclear Information System (INIS)

    Targets of Cr, Fe, Zn, Rb, and Ru were irradiated in the hydraulic tube of the Oak Ridge HFIR reactor at a neutron flux of 2.6 x 1015 n/cm2sec for 1 day and 20 days. The reactor burn-up cross sections (in barns) of the radioactive product nuclides are: 51Cr, 59Fe, 65Zn, 60 +- 30; 86Rb, 103Ru, <20

  6. Characterization of Ca(2+)-activated 86Rb+ fluxes in rat C6 glioma cells: a system for identifying novel IKCa-channel toxins.

    OpenAIRE

    de-Allie, F. A.; Bolsover, S. R.; Nowicky, A. V.; Strong, P N

    1996-01-01

    1. The pharmacological characteristics of a putative Ca2+ activated K+ channel (IKCa channel) in rat glioma C6 cells were studied in the presence of the Ca2+ ionophore, ionomycin and various K+ channel blockers, 86Rb+ being used as a radioisotopic tracer for K+. 2. The resting 86Rb+ influx into C6 cells was 318 +/- 20 pmol s-1. The threshold for ionomycin activation of 86Rb+ influx was approx. 100 nM. At ionomycin concentrations above the activation threshold, the initial rate of 86Rb+ influx...

  7. Aging: ouabain-sensitive /sup 86/Rb+ uptake rate and responsiveness to digoxin in rat left atrial muscle

    Energy Technology Data Exchange (ETDEWEB)

    Kennedy, R.H.; Seifen, E.

    1989-01-01

    Previous work in anesthetized rats has demonstrated that the sensitivity to cardiotoxic actions of cardiotonic steroids is increased in senescence, and studies in crude homogenates and partially purified membrane preparations have suggested that this altered responsiveness is related to an aging-associated reduction in the sarcolemmal content of Na,K-adenosine triphosphatase. This decrease in Na,K-adenosine triphosphatase could enhance the sensitivity to digitalis-like compounds by reducing the reserve capacity of the Na+-pump and thus the extent of digitalis-induced pump inhibition required before the onset of toxicity. Current experiments examined dose-dependent actions of digoxin in atrial muscle isolated from 3-, 12- and 24- to 25-month-old rats and determined if alterations in responsiveness correlated with changes in ouabain-sensitive 86Rb+ uptake rate, an estimate of Na+-pump activity. Atrial preparations from aged rats were more sensitive to the cardiotoxic actions of digoxin; however, the inotropic efficacy before the onset of toxicity was not affected by age. Both 1) the maximum attainable ouabain-sensitive 86Rb+ uptake rate and 2) the difference between maximum uptake rate and that monitored in preparations stimulated at 4.0 Hz decreased progressively with age. These results indicate that atrial muscle from aged rats is more sensitive to direct toxic effects of digoxin and suggest that this lower tolerance is mediated, at least in part, by a reduction in Na+-pump reserve capacity.

  8. Fractionated irradiation and early changes in noradrenaline induced potassium efflux ([sup 86]Rb[sup +]) in rat parotid gland

    Energy Technology Data Exchange (ETDEWEB)

    Franzen, L.; Littbrand, B.; Henriksson, R. (Univ. Hospital, Dept. of Oncology, Umeaa (Sweden)); Sundstroem, S. (Univ. Hospital, Dept. of Histology and Cell Biology, Umeaa (Sweden)); Karlsson, M. (Univ. Hospital, Dept. of Radiation Physics, Umeaa (Sweden)); Gustafsson, H. (Univ. Hospital, Dept. of Oto-Laryngology, Umeaa (Sweden))

    1992-01-01

    The effects of fractionated irradiation on the electrolyte fluid secretion from rat parotid gland were studied. Secretion was measured as noradrenaline stimulated potassium efflux in vitro with [sup 86]Rb[sup +] as tracer for potassium. The irradiation was delivered either as a five-day schedule (total dose 20, 25, 30, 35, 40, 45 Gy) or a two-day schedule (total dose 24, 32 Gy). The noradrenaline stimulated efflux was decreased in comparison with contralateral controls 10 days after the last irradiation. The effect was dose-dependent. Based on the data available, [alpha]/[beta] ratio of the used system was calculated to about 20 Gy, which corresponds to other results regarding early radiation effects. (orig.).

  9. Evaluation of crop residues on potassium kinetics in an acid soil and potassium use efficiency in potato-garlic sequence using tracer 86Rb

    International Nuclear Information System (INIS)

    Greenhouse and laboratory studies were conducted on an acid soil in order to evaluate the role of two crop residues i.e. paddy and wheat along with farmyard manure on potassium kinetics and its availability in the potato-garlic sequence using tracer 86Rb. Under rapid equilibrium, application of crop residues of paddy, wheat straw and FYM were able to enhance soil pH and organic carbon content. In addition, their application helped in enhancing soil K availability indices like water soluble, available and non-exchangeable -K. This was further augmented by the Q/I studies using 86Rb where application of organic residues helped in lowering the potassium buffering capacity of the soil. Greenhouse study supplemented the results obtained from laboratory study where application of crop residues/FYM were able to improve the potato yield significantly and maintained higher concentration of K in potato leaf at early growth stages. A significant correlation was obtained between leaf K and haulms-K with that of 86Rb activities in potato leaf at 35 days and 86Rb absorbed in the haulms, respectively. Residues/ FYM and PK application to potato left sufficient residual effect on succeeding garlic crop. In potato-garlic sequence, K recovery was highest with FYM while N and P recoveries were higher with wheat residues. The nutrient recoveries with PK application followed law of diminishing returns. (author)

  10. sup 86 Rb(K) influx and ( sup 3 H)ouabain binding by human platelets: Evidence for beta-adrenergic stimulation of Na-K ATPase activity

    Energy Technology Data Exchange (ETDEWEB)

    Turaihi, K.; Khokher, M.A.; Barradas, M.A.; Mikhailidis, D.P.; Dandona, P. (Royal Free Hospital and School of Medicine, London (England))

    1989-08-01

    Although active transport of potassium into human platelets has been demonstrated previously, there is hitherto no evidence that human platelets have an ouabain-inhibitable Na-K ATPase in their membrane. The present study demonstrates active rubidium (used as an index of potassium influx), {sup 86}Rb(K), influx into platelets, inhibitable by ouabain, and also demonstrates the presence of specific ({sup 3}H)ouabain binding by the human platelet. This {sup 86}Rb(K) influx was stimulated by adrenaline, isoprenaline, and salbutamol, but noradrenaline caused a mild inhibition. Active {sup 86}Rb(K) influx by platelets was inhibited markedly by timolol, mildly by atenolol, but not by phentolamine. Therefore, active {sup 86}Rb(K) influx in human platelets is enhanced by stimulation of beta adrenoceptors of the beta 2 subtype. The platelet may therefore replace the leukocyte in future studies of Na-K ATPase activity. This would be a considerable advantage in view of the ease and rapidity of preparation of platelets.

  11. Concentration-response relationship of {alpha}{sub 1}-adrenoceptor-stimulated increase of {sup 86}Rb{sup +} efflux in rat heart

    Energy Technology Data Exchange (ETDEWEB)

    Andersen, G.Oe.; Enger, M.; Skomedal, T.; Osnes, J.B. [Univ. of Oslo, Dept. of Pharmacology (Norway)

    1996-10-01

    The aim of the present study was to establish a concentration-response relationship for the {alpha}{sub 1}-adrenoceptor mediated increase of{sup 86}Rb{sup +} efflux, and to characterize the sensitivity of this response to the selective {alpha}{sub 1}-adrenoceptor antagonist prazosin. Isolated rat hearts were perfused retrogradely at constant flow and at 31 deg. Timolol (10{sup -}26 mol/l) was used to block {beta}-adrenoceptors. After a loading period with {sup 86}Rb{sup +} and 55 min. washout, the hearts were exposed to phenylephrine in a concentration range from 3x10{sup -8} mol/l to 10{sup -4} mol/l. Control experiments comparing the effects of {alpha}{sub 1}-adrenoceptor stimulation on {sup 86}Rb{sup +} efflux and {sup 42}K{sup +} efflux were performed. {alpha}{sub 1}-Adrenoceptor stimulation increased the {sup 86}Rb{sup +} efflux with a pD{sub 2}=6.35{+-}0.20 (mean{+-}S.E.M.) The maximal response to phenylephrine was 22.5{+-}2.0% (mean{+-}S.E.M.) of the control values. The concentration-response curve was shifted to higher concentration of agonist in the presence of the {alpha}{sub 1}-adrenoceptor antagonist prazosin (3x10{sup -10} mol/l). The calculated inhibition constant for prazosin was 6.1x10{sup -11} mol/l. {sup 86}Rb{sup +} was found to be a suitable K{sup +} analogue in the study of relative changes in K{sup +} efflux concentration-dependently. A high sensitivity to prazosin confirmed the involvement of the {alpha}{sub 1}-adrenoceptor population. (au) 37 refs.

  12. Progesterone, some progesterone derivatives and urinary digoxin-like substances from pregnant women in radioimmuno- and /sup 86/Rb-Uptake assays of digoxin

    Energy Technology Data Exchange (ETDEWEB)

    Vinge, E.; Helgesen-Rosendal, S.; Baeckstroem, T.

    1988-01-01

    Progesterone and some derivatives were tested in a radioimmunoassay (RIA) of digoxin and in a bioassay measuring the /sup 86/Rb-uptake into red blood cells as in index of Na/sup +/, K/sup +/-ATPase activity. The digitalis-like activity of the hormones was compared with that found in chromatographic fractions of material extracted from the urines of pregnant women at term. Progesterone at concentrations > 10/sup -6/ fM cross-reacted in the RIA, and at 10/sup -3/ M it decreased /sup 86/Rb-uptake by 18%. The anaesthetic progesterone derivates 5..cap alpha..-pregnance-3..cap alpha..-ol-20-one and 5..cap alpha..-pregnane-3,20-dione crossreacted to a lesser degree in the RIA and lacked effect in the bioassay. Similar results were obtained with pregnandiol-glucuronide, the major urinary metabolite of progesterone. In contrast, several fractions of the urinary material had significant effects in both assays. It is concluded that the digitalis-like activity of progesterone is not coupled to properties associated with its anaesthetic effects. Furthermore, although progesterone may account for a part of the endogenous digoxin-like substances in serum of neonates and pregnant women, neither progesterone proper nor pregnandiol-glucuronide explains the great amount of digoxin-like substances found in the urines.

  13. ON Cajanus cajan's AND Dalbergia bulansae's Absor ptivitr OF 32p AND 86Rb%木豆、南岭黄檀对磷钾元素吸收效应的探讨

    Institute of Scientific and Technical Information of China (English)

    侯开卫; 陈玉德

    1983-01-01

    @@ 本文就木豆对磷钾(用铷代替钾)元素的吸收及其生长效应,磷素在木豆各器官的分布以及不同营养元素和不同含氮量对南岭黄檀吸收磷素的影响等方面进行了分析论述.

  14. Ouabain enhancement of compound 48/80 induced histamine secretion from rat peritoneal mast cells: dependence on extracellular sodium

    DEFF Research Database (Denmark)

    Knudsen, T; Bertelsen, Niels Haldor; Johansen, Torben

    1992-01-01

    Purified populations of rat peritoneal mast cells were used to study the effect of ouabain on compound 48/80-induced histamine secretion and on 86Rb+ uptake. 86Rb+ was used as a tracer for extracellular K+. The calculated value of the ouabain-sensitive uptake of K+ and 86Rb+ was considered a meas...

  15. Blood flow measurements in selected oral tissues in dogs using radiolabelled microspheres and rubidium-86

    International Nuclear Information System (INIS)

    Radiolabelled microspheres and 86Rb were used in dogs to measure oral tissue blood flow. The tissues studied were mandibular alveolar and basal bone, oral mucosa, gingiva, lip, tongue and soft tissue of hard palate. A suspension of 6 x 106 cerium-141 labelled microspheres, 15 + - 5 μm in diameter was injected into the left ventricle through an arterial catheter in 9 healthy adult dogs. Cardiovascular stability was assessed during the catheterization and injection of microspheres. Following the microsphere injection, 500 μCi of 86Rb was injected intravenously and the animal was killed within 60s. Blood flow measured using 86Rb was not significantly different (p > 0.05) from radiolabelled microsphere flow in mandibular basal bone, oral mucosa and gingiva, suggesting that in these oral tissues nutritional or capillary exchange flow is similar to total blood flow. In mandibular alveolar bone, lip, tongue and soft tissue of hard palate, however, 86Rb measurements were significantly greater (p 86Rb shows promise for continued studies of blood flow in healthy and pathological oral tissues. (author)

  16. GABA/sub B/ receptor activation inhibits Ca2+-activated potassium channels in synaptosomes: involvement of G-proteins

    International Nuclear Information System (INIS)

    86Rb-efflux assay from preloaded synaptosomes of rat cerebral cortex was developed to study the effect of GABA/sub B/ receptor agonist baclofen on Ca2+-activated K+-channels. Depolarization of 86Rb-loaded synaptosomes in physiological buffer increased Ca2+-activated 86Rb-efflux by 400%. The 86Rb-efflux was blocked by quinine sulfate, tetraethylammonium, and La3+ indicating the involvement of Ca2+-activated K+-channels. (-)Baclofen inhibited Ca2+-activated 86Rb-efflux in a stereospecific manner. The inhibitory effect of (-)baclofen was mediated by GABA/sub B/ receptor activation, since it was blocked by GABA/sub B/ antagonist phaclofen, but not by bicuculline. Further, pertussis toxin also blocked the ability of baclofen or depolarizing action to affect Ca2+-activated K+-channels. These results suggest that baclofen inhibits Ca2+-activated K+-channels in synaptosomes and these channels are regulated by G-proteins. This assay may provide an ideal in vitro model to study GABA/sub B/ receptor pharmacology

  17. GABA/sub B/ receptor activation inhibits Ca/sup 2 +/-activated potassium channels in synaptosomes: involvement of G-proteins

    Energy Technology Data Exchange (ETDEWEB)

    Ticku, M.K.; Delgado, A.

    1989-01-01

    /sup 86/Rb-efflux assay from preloaded synaptosomes of rat cerebral cortex was developed to study the effect of GABA/sub B/ receptor agonist baclofen on Ca/sup 2 +/-activated K/sup +/-channels. Depolarization of /sup 86/Rb-loaded synaptosomes in physiological buffer increased Ca/sup 2 +/-activated /sup 86/Rb-efflux by 400%. The /sup 86/Rb-efflux was blocked by quinine sulfate, tetraethylammonium, and La/sup 3 +/ indicating the involvement of Ca/sup 2 +/-activated K/sup +/-channels. (-)Baclofen inhibited Ca/sup 2 +/-activated /sup 86/Rb-efflux in a stereospecific manner. The inhibitory effect of (-)baclofen was mediated by GABA/sub B/ receptor activation, since it was blocked by GABA/sub B/ antagonist phaclofen, but not by bicuculline. Further, pertussis toxin also blocked the ability of baclofen or depolarizing action to affect Ca/sup 2 +/-activated K/sup +/-channels. These results suggest that baclofen inhibits Ca/sup 2 +/-activated K/sup +/-channels in synaptosomes and these channels are regulated by G-proteins. This assay may provide an ideal in vitro model to study GABA/sub B/ receptor pharmacology.

  18. Changes in cardiac glycoside receptor sites 86 rubidium uptake and intracellular sodium concentrations in the erythrocytes of patients receiving digoxin during the early phases of treatment of cardiac failure in regular rhythm and of atrial fibrillation

    International Nuclear Information System (INIS)

    Measurements of the binding of 12-α-[3H]-digoxin to the membranes of intact erythrocytes, erythrocytic 86Rb uptake and intraerythrocytic sodium concentrations have been made in the red cells of patients receiving digoxin in the short-term for atrial fibrillation or cardiac failure in regular rhythm. During the first few days of treatment [3H]-digoxin binding and 86Rb uptake fall and intraerythrocytic sodium concentrations rise. Subsequently parallel fluctuations occur in [3H]-digoxin binding and 86Rb uptake but not in intraerythrocytic sodium concentrations and the significance of the fluctuations is discussed. The values of all three measurements correlate significantly with the response of the heart in sinus rhythm as measured by QS2I. Plasma digoxin concentrations do not correlate with QS2I. (author)

  19. Effects of VA mycorrhizas fungus on phosphorus and potassium uptake in tea seedlings

    International Nuclear Information System (INIS)

    Tea (Camellia sinensis) seeds were sown on sterilized acidic yellow soil (pH 5.6) in a pot experiment and treated as follows: 1) inoculated with VA mycorrhizas fungus (Glomus citricolum), 2) nonmycorrhizal as control, top dressed with 32P-single superphosphate (M-32P) and 86Rb-rubidium chloride (M-86Rb). The results showed that the percentage of VA mycorrhizas infection was 52.6% for M-32P and 56.7% for M-86Rb. Plant height, dry weight and the uptake of phosphorus and potassium were 2.1 and 1.8 times, 2.4 and 2.5 times, 5.6 and 4.1 times as that of control respectively. The utilization rate of phosphorus and potassium were raised by 14.10% and 17.13% respectively

  20. Racial differences in red cell cation transport and their relationship to essential hypertension

    International Nuclear Information System (INIS)

    Red cell cation transport has been studied in normotensive and essential hypertensive groups of white and black (West Indian) subjects. In vitro uptake of the potassium analogue 86Rb was measured during short-term incubation of erythrocytes in the presence and absence of ouabain. Sodium pump activity was significantly greater (p less than 0.0005) in white hypertensives than in white normotensives. No such difference was observed between black hypertensive and normotensives. 86Rb uptake was significantly lower in black than in white normotensive individuals; this racial differences was not due to a difference in sodium pump activity

  1. Bradykinin and vasopressin stimulate Na/sup +/-K/sup +/-Cl/sup -/ cotransport in cultured endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Brock, T.A.; Brugnara, C.; Canessa, M.; Gimbrone, M.A. Jr.

    1986-06-01

    The authors have characterized a Na/sup +/-K/sup +/-Cl/sup -/ cotransporter in vascular endothelial cells (EC) cultured from different blood vessels and species that is inhibited by the diuretics furosemide and bumentanide. Inward /sup 86/Rb influx transported by the Na/sup +/-K/sup +/ pump in cultured EC from bovine and pig aorta, bovine vena cava, and baboon cephalic vein but not in human umbilical or saphenous vein EC. External Na/sup +/ or Cl/sup -/-stimulated, ouabain-insensitive /sup 86/Rb influx is equal to furosemide or bumetanide-sensitive /sup 86/Rb influx. Ouabain-insensitive /sup 22/Na influx is also partially inhibited by these drugs and stimulated by increasing external K/sup +/ or Cl/sup -/. Net Na/sup +/ extrusion occurs via the Na/sup +/-K/sup +/-Cl/sup -/ cotransporter in the absence of external K/sup +/, whereas net Na/sup +/ influx occurs at higher external K/sup +/. Maximal concentrations (100 nM) of bradykinin and vasopressin increase the initial rate of bumetanide-sensitive /sup 86/Rb influx by approx.60 and 70%. Addition of either ethyleneglycol-bis(..beta..-aminotethylether)-N,N'-tetraacetic acid or LaCl/sub 3/ (to block calcium influx) prevents bradykinin-stimulated /sup 86/Rb influx. When intracellular calcium is elevated using ionomycin (100 nM), a Ca/sup 2 +/ionophore, bumetanide-sensitive /sup 86/Rb influx increases approx.twofold. In contrast, isoproterenol (100 ..mu..M) and forskolin (50 /sup +/M), adenylate cyclase stimulators, decrease furosemide-sensitive /sup 86/Rb influx. Thus in certain types of cultured EC, a Na/sup +/-K/sup +/-Cl/sup -/ cotransporter mediates a fraction of K/sup +/ influx quantitatively as important as the Na/sup +/-K/sup +/ pump (ouabain-sensitive /sup 86/Rb influx) and appears to be modulated by Ca/sup 2 +/ and cyclic nucleotides.

  2. ATP-sensitive K+ channels that are blocked by hypoglycemia-inducing sulfonylureas in insulin-secreting cells are activated by galanin, a hyperglycemia-inducing hormone

    International Nuclear Information System (INIS)

    The action of the hyperglycemia-inducing hormone galanin, a 29-amino acid peptide names from its N-terminal glycine and C-terminal amidated alanine, was studied in rat insulinoma (RINm5F) cells using electrophysiological and 86Rb+ flux techniques. Galanin hyperpolarizes and reduces spontaneous electrical activity by activating a population of APT-sensitive K+ channels with a single-channel conductance of 30 pS (at -60 mV). Galanin-induced hyperpolarization and reduction of spike activity are reversed by the hypoglycemia-inducing sulfonylurea glibenclamine. Glibenclamide blocks the galanin-activated ATP-sensitive K+ channel. 86Rb+ efflux from insulinoma cells is stimulated by galanin in a dose-dependent manner. The half-maximum value of activation is found at 1.6 nM. Galanin-induced 86Rb+ efflux is abolished by glibenclamide. The half-maximum value of inhibition is found at 0.3 nM, which is close to the half-maximum value of inhibition of the ATP-dependent K+ channel reported earlier. 86Rb+ efflux studies confirm the electrophysiological demonstration that galanin activates and ATP-dependent K+ channel

  3. ATP-sensitive K/sup +/ channels that are blocked by hypoglycemia-inducing sulfonylureas in insulin-secreting cells are activated by galanin, a hyperglycemia-inducing hormone

    Energy Technology Data Exchange (ETDEWEB)

    de Weille, J.; Schmid-Antomarchi, H.; Fosset, M.; Lazdunski, M.

    1988-02-01

    The action of the hyperglycemia-inducing hormone galanin, a 29-amino acid peptide names from its N-terminal glycine and C-terminal amidated alanine, was studied in rat insulinoma (RINm5F) cells using electrophysiological and /sup 86/Rb/sup +/ flux techniques. Galanin hyperpolarizes and reduces spontaneous electrical activity by activating a population of APT-sensitive K/sup +/ channels with a single-channel conductance of 30 pS (at -60 mV). Galanin-induced hyperpolarization and reduction of spike activity are reversed by the hypoglycemia-inducing sulfonylurea glibenclamine. Glibenclamide blocks the galanin-activated ATP-sensitive K/sup +/ channel. /sup 86/Rb/sup +/ efflux from insulinoma cells is stimulated by galanin in a dose-dependent manner. The half-maximum value of activation is found at 1.6 nM. Galanin-induced /sup 86/Rb/sup +/ efflux is abolished by glibenclamide. The half-maximum value of inhibition is found at 0.3 nM, which is close to the half-maximum value of inhibition of the ATP-dependent K/sup +/ channel reported earlier. /sup 86/Rb/sup +/ efflux studies confirm the electrophysiological demonstration that galanin activates and ATP-dependent K/sup +/ channel.

  4. Cell interactions in concanavalin A activated cation flux and DNA synthesis of mouse lymphocytes

    DEFF Research Database (Denmark)

    Owens, T; Kaplan, J G

    1980-01-01

    Co-culture at constant cell density of nude mouse spleen cells (by themselves unresponsive to the T-cell mitogen concanavalin A (Con A)), with congenic T-enriched lymphocyte suspensions and Con A caused anomalously high activation of K+ transport (measured by 86Rb uptake) and of incorporation...

  5. Cotransport of water by Na¿-K¿-2Cl¿ cotransporters expressed in Xenopus oocytes

    DEFF Research Database (Denmark)

    Zeuthen, Thomas; Macaulay, Nanna

    2012-01-01

    The NKCC1 and NKCC2 isoforms of the mammalian Na¿–K¿–2Cl¿ cotransporter were expressed in Xenopus oocytes and the relation between external ion concentration and water fluxes determined.Water fluxes were determined from changes in the oocytes volume and ion fluxes from 86Rb+ uptake. Isotonic...

  6. Regulation of the sodium/potassium/chloride cotransporter by calcium and cyclic AMP in cultured vascular smooth muscle cells

    Energy Technology Data Exchange (ETDEWEB)

    Higgins, B.L.; Smith, L.; Smith, J.B.

    1987-05-01

    The activity of the Na/K/Cl cotransporter in smooth muscle cells cultured from rat aorta was assayed by measuring the initial rate of furosemide-inhibitable /sup 86/Rb influx or efflux. Five uM furosemide or 0.2 uM bumetanide inhibited influx by 50%. Furosemide-inhibitable /sup 86/Rb influx depended on the presence of all 3 ions in the external medium. The dependence on Na and K was hyperbolic with apparent Km values of 45 and 5 mM, respectively. The dependence on Cl was sigmoidal. Assuming a stoichiometry of 1:1:2 for Na:K:Cl, a Km for Cl of 60 mM was obtained from a Hofstee plot of the data. Rapidly growing cells had 3 fold higher cotransport activity than quiescent cells. Angiotensin II (ANG) stimulated furosemide-inhibitable /sup 86/Rb efflux by 2 fold. An ANG receptor antagonist prevented ANG from increasing cotransport activity. Two calcium ionophores, A23187 and ionomycin, increased cotransport activity by 2 fold. Phorbol myristate acetate had no effect on cotransport activity. Isoproterenol, dibutyryl cyclic AMP, cholera toxin, or methylisobutylxanthine inhibited furosemide-sensitive /sup 86/Rb influx by 35 to 50%. From these findings they conclude that increasing cytoplasmic free calcium stimulates cotransport activity, whereas increasing cellular cyclic AMP inhibits the cotransporter.

  7. Cationic and secretory effects of glimepiride and glibenclamide in perifused rat islets

    Energy Technology Data Exchange (ETDEWEB)

    Lebrun, P.; Malaisse, W.J. (Laboratories of Pharmacology and Experimental Medicine, Brussels Free University, Brussels (Belgium))

    1992-01-01

    The effects of glimepiride and glibenclamide upon [sup 86]Rb outflow, [sup 45]Ca outflow and insulin release were examined in rat islets perfused at low (zero to 2.8 mM) or close-to-normal (8.3 mM) D- glucose concentrations. At the low hexose concentrations, a marked and not reversible decrease in [sup 86]Rb outflow contrasted with a rapid and reversible increase in [sup 45]Ca outflow. The latter increase was abolished in the absence of extracellular Ca[sup 2+], and was not associated with any pronounced stimulation of insulin release. Inversely, in the presence of 8.3 mM D-glucose, a comparable increase in [sup 45]Ca efflux now coincided with an increase in [sup 86]Rb efflux and a marked and not reversible stimulation of insulin release. Whether in terms of the time course or glucose dependency of the cationic and secretory responses, a coupled increase in both [sup 40]Ca inflow and [sup 45]Ca outflow thus coincided with either negative or positive changes in [sup 86]Rb outflow and either minimal or marked changes in insulin output. Such dissociated behaviors suggest that the insulinotropic action of hypoglycemic sulfonylureas is not necessarily attributable solely to a primary decrease in K[sup +] conductance. (au).

  8. K+ transport across the lamprey erythrocyte membrane: characteristics of a Ba(2+)- and amiloride-sensitive pathway.

    Science.gov (United States)

    Kirk, K

    1991-09-01

    The characteristics of K+ transport in erythrocytes from the river lamprey (Lampetra fluviatilis) were investigated using standard radioisotope flux techniques. The cells were shown to have a ouabain-sensitive transport pathway that carried 43K+ and 86Rb+ into the cell at similar rates. Most of the ouabain-resistant 43K+ and 86Rb+ influx was via a pathway that was insensitive to cotransport inhibitors and to the replacement of extracellular Cl- or Na+. This pathway showed a strong selectivity for 43K+ over 86Rb+. It was inhibited fully by Ba2+ (I50 approximately 2.8 mumol l-1), amiloride (I50 approximately 150 mumol l-1) and ethylisopropylamiloride (I50 approximately 3.3 mumol l-1) and less effectively by quinine and by the tetraethylammonium ion. Inhibition by Ba2+ took full effect within a few minutes whereas the full inhibitory effect of amiloride took more than 1 h to develop. Experiments with the membrane potential probe [14C]tetraphenylphosphonium ion gave results consistent with the lamprey erythrocyte membrane having a Ba(2+)-sensitive K+ conductance that was significantly greater than the membrane Na+ conductance and which gave rise to a marked dependence of the membrane potential on the extracellular K+ concentration. The rate constants for Ba(2+)-sensitive 43K+ and 86Rb+ influx decreased (proportionally) with increasing extracellular K+ concentration in a manner that was consistent with the transport being via a conductive pathway. The decrease was attributed to a depolarisation of the membrane (in response to the increasing extracellular K+ concentration) and a consequent decrease in the driving force for the conductive movement of 43K+ and 86Rb+ into the cells. Ba(2+)-sensitive 86Rb+ influx increased significantly with decreasing cell volume and with increasing intracellular pH (at a constant extracellular pH) but increased only slightly with increasing extracellular pH. The pathway operated normally in the complete absence of extracellular Ca2+ but

  9. Comparative phloem Mobility of nickel in nonsenescent plants. [Pisum sativa L. ; Pelargonium zonale L

    Energy Technology Data Exchange (ETDEWEB)

    Neumann, P.M.; Chamel, A.

    1986-06-01

    /sup 63/Ni was applied to nonsenescent source leaves and found to be transported to sink tissues in pea (Pisum saativum L.) and geranium plants (Pelargonium zonale L.). The comparative mobilities (percent tracer transported out of source leaf division % /sup 86/Rb transported) for /sup 63/Ni in peas was 2.12 and in geranium 0.25. The value for the phloem mobile /sup 86/Rb was 1.00. By contrast, the comparative mobility of /sup 45/Ca, which is relatively immobile in the phloem, was low (0.05 in peas, 0.00 in geranium). Interruption of the phloem pathway between source and sink leaves by steam girdling almost completely inhibited /sup 63/Ni accumulation in the sink leaves of both species. The authors conclude that Ni is transported from nonsenescent source leaves to sink tissues via the phloem of leguminous and nonleguminous plants.

  10. Evidence for differential action of indoleacetic acid upon ion fluxes in single cells of Petroselinum sativum.

    Science.gov (United States)

    Bentrup, F W; Pfrüner, H; Wagner, G

    1973-12-01

    The apparent influx of (36)Cl(-) and (86)Rb(+)/K(+) into cells from the higher plant Petroselinum sativum has been measured during the presence and absence in the culture medium of indolacetic acid (IAA) which is an essential auxin of these cells. While 10(-5) M IAA did not significantly affect the influx of (86)Rb(+)/K(+), it substantially reduced that of (36)Cl(-), i.e. by a factor 0.25 within 30 min. This differential action of IAA, which holds for a reasonable range of external pH, is assumed to bear on current hypotheses that the primary events of auxin action involve plasmalemma functions. PMID:24474466

  11. Regulation of ATP-sensitive K+ channels in insulinoma cells: Activation by somatostatin and protein kinase C and the role of cAMP

    International Nuclear Information System (INIS)

    The actions of somatostatin and of the phorbol ester 4β-phorbol 12-myristate 13-acetate (PMA) were studied in rat insulinoma (RINm5F) cells by electrophysiological and 86Rb+ flux techniques. Both PMA and somatostatin hyperpolarize insulinoma cells by activating ATP-sensitive K+ channels. The presence of intracellular GTP is required for the somatostatin effects. PMA- and somatostatin-induced hyperpolarization and channel activity are inhibited by the sulfonylurea glibenclamide. Glibenclamide-sensitive 86Rb+ efflux from insulinoma cells is stimulated by somatostatin in a dose-dependent manner (half maximal effect at 0.7 nM) and abolished by pertussis toxin pretreatment. Mutual roles of a GTP-binding protein, of protein kinase C, and of cAMP in the regulation of ATP-sensitive K+ channels are discussed

  12. Na/K ATPase inhibition by digitalis-like factors in neonates

    Energy Technology Data Exchange (ETDEWEB)

    Bottorff, M.B.; Songu-Mize, E.; Hoon, T.J.; Phelps, S.J.; Kamper, C.A.

    1986-03-01

    At the authors institution, 48% of neonates < 1 month of age had false-positive digoxin immunoassay determinations while not receiving digoxin, presumably due to an endogenous digoxin-like immunoreactive substance (DLIS) in the plasma. Plasma from 3 neonates positive for DLIS by fluorescence polarization immunoassay (FPIA) was evaluated for inhibitory activity on human red blood cell (RBC) Na/K ATPase. Neonatal plasma aliquots containing DLIS concentrations (conc) of 0.24, 0.37, 0.43, 0.49 and 0.61 ng/ml (3.07 - 7.81 x 10/sup -10/M) were incubated with human RBC and /sup 86/Rb in order to measure /sup 86/Rb uptake inhibition with respect to DLIS negative neonatal plasma. /sup 86/Rb uptake inhibition by digoxin-spiked human serum (1.07 x 10/sup -10/ - 4.57 x 10/sup -6/M) was also measured. Percent inhibition vs. log molar conc plots for DLIS and digoxin were compared. DLIS inhibited Na/K ATPase in a linear fashion over the range studied. Comparing the linear portions of the conc-inhibition curves for digoxin and DLIS, the molar conc of digoxin producing 40% inhibition of /sup 86/Rb uptake is 333 times greater than the molar conc of DLIS producing similar inhibition. Therefore, DLIS in neonatal serum as measured by FPIA has approximately 300 times greater inhibitory activity than digoxin. The presence of circulating DLIS may reflect an adaptive or maladaptive response to some, as yet unknown, process early in life.

  13. Partial purification of endogenous digitalis-like compound(s) in cord blood

    Energy Technology Data Exchange (ETDEWEB)

    Balzan, S.; Ghione, S.; Biver, P.; Gazzetti, P.; Montali, U. (C.N.R. Institute of Clinical Physiology, Pisa (Italy))

    1991-02-01

    Increasing evidence indicates the presence of endogenous digitalis-like compound(s) in human body fluids. In this preliminary report, we describe a study of the partial purification by HPLC of these compounds in the plasma of neonates (who have particularly high concentrations of this substance) and adults. Plasma samples from neonates (cord blood) and adults, lyophilized and extracted with methanol, were applied on a 300 x 3.9 mm C18 Nova Pak column and eluted with a mobile phase of acetonitrile/methanol/water (17/17/66 or 14/14/72 by vol) and, after 30 min, with 100% methanol. We assayed eluted fractions for inhibitory activity of 86Rb uptake and for digoxin-like immunoreactivity. The elution profile revealed a first peak of inhibitory activity of 86Rb uptake at the beginning of the chromatography; another peak was eluted with the 100% methanol. The two peaks also cross-reacted with antidigoxin antibodies. Because the second peak could possibly reflect the nonspecific interference of various lipophilic compounds, we focused our attention on the first peak. For these fractions dose-response curves for 86Rb uptake and for displacement of digoxin were parallel, respectively, to those of ouabain and digoxin, suggesting similarities of digoxin-like immunoreactive substance to cardiac glycosides. Similar chromatographic profiles were also obtained for plasma from adults, suggesting that the endogenous glycoside-like compound(s) in the neonate may be the same as those in the adult.

  14. Oligosaccharide composition of the neurotoxin responsive Na+ channel and the requirement of sialic acid for activity

    International Nuclear Information System (INIS)

    The neurotoxin responsive Na+ channel was purified to homogeneity in an 18% yield from a clonal cell line of mouse neuroblastoma, N-18, metabolically labeled with L-[3H]fucose. The Na+ channel, a glycoprotein, M/sub r/=200,000 (gradient 7-14% PAGE) was digested with Pronase and the glycopeptides were characterized by serial lectin affinity chromatography. greater than 90% of the oligosaccharides contained sialic acid and 18% were biantennary, 39% were triantennary and 30% tetraantennary. The glycoprotein was reconstituted into artificial phospholipid vesicles and 86Rb flux was stimulated (65%) by 200 μM veratridine and 1.2 μg of scorpion venom and was inhibited (95%) by 5 μM tetrodotoxin. The requirement of sialic acid for Na+ channel activity was demonstrated since neuraminidase (0.01 U) treatment of the reconstituted glycoprotein eliminated the response of 86Rb flux to the stimulating neurotoxins. In other experiments, treatment of N-18 cells with 10 μM swainsonine, an inhibitor of glycoprotein processing, altered the oligosaccharide composition of the Na+ channel. When the abnormally glycosylated Na+ channel was reconstituted into artificial phospholipid vesicles, 86Rb flux in response to neurotoxins was impaired. Thus, glycosylation of the polypeptide with oligosaccharides of specific composition and structure is essential for expression of the biological activity of the neurotoxin responsive Na+ channel

  15. A functional assay to measure postsynaptic gamma-aminobutyric acidB responses in cultured spinal cord neurons: Heterologous regulation of the same K+ channel

    Energy Technology Data Exchange (ETDEWEB)

    Kamatchi, G.L.; Ticku, M.K. (Univ. of Texas Health Science Center, San Antonio (USA))

    1991-02-01

    The stimulation of postsynaptic gamma-aminobutyric acid (GABA)B receptors leads to slow inhibitory postsynaptic potentials due to the influx of K(+)-ions. This was studied biochemically, in vitro in mammalian cultured spinal cord neurons by using 86Rb as a substitute for K+. (-)-Baclofen, a GABAB receptor agonist, produced a concentration-dependent increase in the 86Rb-influx. This effect was stereospecific and blocked by GABAB receptor antagonists like CGP 35 348 (3-aminopropyl-diethoxymethyl-phosphonic acid) and phaclofen. Apart from the GABAB receptors, both adenosine via adenosine1 receptors and 5-hydroxytryptamine (5-HT) via 5-HT1 alpha agonists also increased the 86Rb-influx. These agonists failed to show any additivity between them when they were combined in their maximal concentration. In addition, their effect was antagonized specifically by their respective antagonists without influencing the others. These findings suggest the presence of GABAB, adenosine1 and 5-HT1 alpha receptors in the cultured spinal cord neurons, which exhibit a heterologous regulation of the same K(+)-channel. The effect of these agonists were antagonized by phorbol 12,13-didecanoate, an activator of protein kinase C, and pretreatment with pertussis toxin. This suggests that these agonists by acting on their own receptors converge on the same K(+)-channel through the Gi/Go proteins. In summary, we have developed a biochemical functional assay for studying and characterizing GABAB synaptic pharmacology in vitro, using spinal cord neurons.

  16. A functional assay to measure postsynaptic gamma-aminobutyric acidB responses in cultured spinal cord neurons: Heterologous regulation of the same K+ channel

    International Nuclear Information System (INIS)

    The stimulation of postsynaptic gamma-aminobutyric acid (GABA)B receptors leads to slow inhibitory postsynaptic potentials due to the influx of K(+)-ions. This was studied biochemically, in vitro in mammalian cultured spinal cord neurons by using 86Rb as a substitute for K+. (-)-Baclofen, a GABAB receptor agonist, produced a concentration-dependent increase in the 86Rb-influx. This effect was stereospecific and blocked by GABAB receptor antagonists like CGP 35 348 (3-aminopropyl-diethoxymethyl-phosphonic acid) and phaclofen. Apart from the GABAB receptors, both adenosine via adenosine1 receptors and 5-hydroxytryptamine (5-HT) via 5-HT1 alpha agonists also increased the 86Rb-influx. These agonists failed to show any additivity between them when they were combined in their maximal concentration. In addition, their effect was antagonized specifically by their respective antagonists without influencing the others. These findings suggest the presence of GABAB, adenosine1 and 5-HT1 alpha receptors in the cultured spinal cord neurons, which exhibit a heterologous regulation of the same K(+)-channel. The effect of these agonists were antagonized by phorbol 12,13-didecanoate, an activator of protein kinase C, and pretreatment with pertussis toxin. This suggests that these agonists by acting on their own receptors converge on the same K(+)-channel through the Gi/Go proteins. In summary, we have developed a biochemical functional assay for studying and characterizing GABAB synaptic pharmacology in vitro, using spinal cord neurons

  17. A 28,000 mol. wt toxin from Bacillus thuringiensis israelensis induces cation transport in rat muscle cultures.

    Science.gov (United States)

    Cahan, R; Shainberg, A; Pechatnikov, I; Nitzan, Y

    1995-07-01

    The mechanism by which the Bacillus thuringiensis israelensis (Bti) 28,000 mol. wt toxin exerts its effect on mature muscle cultures was examined. The toxin inhibited Na+/K(+)-ATPase activity as revealed by 86Rb influx. A 50% inhibition of Na+/K(+)-ATPase activity was obtained with 0.2 microgram/ml of the toxin. The inhibition was time and dose dependent, and it was reversible with low doses of the toxin (up to 0.2 microgram/ml. A considerable release of 86Rb was obtained by doses greater than 0.2 microgram/ml. The 86Rb release was also time and dose dependent. This effect is probably non-specific, since 45Ca influx is also accelerated by toxin-treated cultures. Pre-incubation of the toxin with phosphotidylserine (PS) antagonized the toxin. It is concluded that the toxin is a hydrophobic protein which interacts with the membrane. In low doses this interaction reduces the activity of the sodium pump and in high doses it causes non-specific permeability of the sarcolemma. PMID:8588218

  18. The mechanism of patulin's cytotoxicity and the antioxidant activity of indole tetramic acids

    Energy Technology Data Exchange (ETDEWEB)

    Riley, R.T.; Showker, J.L. (Toxicology and Mycotoxins Research Unit, U.S. Department of Agriculture/Agricultural Research Service, Athens, GA (USA))

    1991-06-01

    In LLC-PK1 cells exposed to patulin (50 microM), lipid peroxidation, abrupt calcium influx, extensive blebbing, and total LDH release appeared to be serially connected events with each representing a step in the loss of structural integrity of the plasma membrane. The aforementioned patulin-induced events were prevented by concurrent incubation with butylated hydroxytoluene, deferoxamine, and cyclopiazonic acid, a fungal metabolite. Patulin also caused depletion of nonprotein sulfhydryls, increased 86Rb+ efflux, dome collapse, and eventually the loss of cell viability. These events were not prevented by antioxidants, results consistent with the hypothesis that they were also serially connected but occurring parallel to those previously mentioned. The earliest events observed in patulin-treated cells were the decrease in nonprotein sulfhydryls and increase in 86Rb+ efflux (5 min) which occurred before statistically significant alterations in protein-bound sulfhydryls. The increased potassium efflux (86Rb+ efflux) occurred via a pathway distinct from BaCl2, quinine, or tetraethylammonium sensitive potassium channels. This is the first published report of the antioxidant activity of indole tetramic acids (cyclopiazonic acid and cyclopiazonic acid imine). The protective effect of tetramic acids in LLC-PK1 cells was restricted to indole tetramic acids, and their prevention of lipid peroxidation did not involve iron chelation. The results of this study demonstrate that cyclopiazonic acid is a potent inhibitor of azide-insensitive, ATP-dependent, a23187-sensitive calcium uptake by the lysate of LLC-PK1 cells. This result is consistent with the hypothesis that the endoplasmic reticulum calcium transport ATPase is a sensitive target for cyclopiazonic acid in LLC-PK1 cells.

  19. Perfusion changes in the RIF-1 tumour and normal tissues after carbogen and nicotinamide, individually and combined.

    OpenAIRE

    Honess, D. J.; Bleehen, N.M.

    1995-01-01

    The strategy of combining carbogen breathing and nicotinamide to overcome chronic and acute hypoxia respectively is being evaluated clinically. The effects of both agents individually and in combination on relative perfusion of 400-700 mm3 RIF-1 tumours and normal tissues were measured by 86Rb extraction. Carbogen breathing alone for 6 min increased relative tumour perfusion by 50-70% compared with control at flow rates of 50 to 200 ml min-1, but the effect was lost at 300 ml min-1. All flow ...

  20. Blood flow modification by nicotinamide and metoclopramide in mouse tumours growing in different sites.

    OpenAIRE

    Hirst, D G; Joiner, B.; Hirst, V. K.

    1993-01-01

    Nicotinamide (NA) and metoclopramide (MCA) have been shown to be sensitisers of the effects of radiation and drugs in experimental rodent tumours growing in skin and muscle. We have used 86Rb uptake to investigate the effects of these two drugs on the distribution of blood to a mouse carcinoma (NT) growing in skin, muscle or the gut wall, as well as to the host normal tissue. NA caused an increase in cardiac output distribution (COD) of between 17 and 92% to tumours in the three sites. When t...

  1. Radionuclides for investigating the accumulation of toxic elements in algae and fish

    International Nuclear Information System (INIS)

    Radionuclides are very suitable for investigating the accumulation of toxic elements in algae and fish. The most important results obtained from using 51Cr(III) (VI), 65Zn, 74As(III) (V), CH374AsO(OH)2, (CH3)274AsO(OH), (CH3)374As, 85Sr, 86Rb, /sup 115m/Cd, 133Ba, 137Cs, 203Hg(II), CH3203HgCl and C6H5203HgCl for the study of the accumulation, release and chemical transformation of the above species in algae and fish are summarized. (author)

  2. Radionuclides in the investigation of the cumulation of toxic elements on alga and fish

    International Nuclear Information System (INIS)

    The present paper surveys the most important results obtained using 51Cr(III)(VI), 65Zn, 74As(III)(V), CH374AsO(OH)2, (CH3)274AsO(OH), (CH3)374As, 85Sr, 86Rb, /sup 115m/Cd, 133Ba, 137Cs, 203Hg(II), CH3203HgCl and C6H5203HgCl for studying the cumulation, release and chemical transformation of the above species on alga and fish. (author)

  3. The use of radiotracers in natural rubber research in Malaysia

    International Nuclear Information System (INIS)

    Five main areas where radiotracers have been used in research on natural rubber are on studies on the uptake of plant nutrients by Hevea brasiliensis using 32P, 86Rb and 45Ca as tracers; biosynthesis of Hevea rubber using 14C or 3H labelled intermediates; translocation and metabolism of 14C. Ethephon in Hevea brasiliensis; use of radiotracers as analytical tools and adsorption of labelled fatty acid soaps on natural rubber lattices. These studies are discussed to show the powerful tool that radiotracers provide in agricultural, biochemical and chemical research on natural rubber. (author)

  4. Ouabain inhibition of the sodium-potassium pump: estimation of ED50 in different types of human leucocytes in vitro

    DEFF Research Database (Denmark)

    Møller, B; Vaag, Allan; Johansen, Torben

    1990-01-01

    uptake of K+ (86Rb+) that is sensitive to ouabain, 10(-5) mol l-1. 2. Dose-response curves for inhibition of the Na(+)-K+ pump were obtained after exposure of the cells to various concentrations of ouabain for 2.5 h when the level of pump inhibition was considered to be at steady state. 3. The ED50...... but somewhat higher when determined in an artificial medium (Ringer solution) than when determined in autologous plasma. 4. The ED50 values observed in the present study were very similar to KD values reported in the literature on ligand binding of tritiated ouabain to the same types of human leucocytes....

  5. Ouabain-binding and 86rubidium-uptake in lymphocytes of normal and borderline hypertensive subjects

    DEFF Research Database (Denmark)

    Nielsen, J R; Pedersen, K E; Johansen, Torben;

    1983-01-01

    In borderline hypertensives cellular sodium concentration seems to be increased, indicating that cellular abnormalities are present in the early course of essential hypertension. In order to study the mechanisms underlying this finding the number of sodium/potassium pump sites and the cation pump...... activity were studied in lymphocytes of nine borderline hypertensives (27 (20-36) years) and nine controls (28 (20-36) years). Maximum 3H-ouabain binding and 86Rb-uptake were taken as measures of the number of pump sites and cation pump activity, respectively. The median number of sodium/potassium pump...

  6. Intestinal excretion of metals by rats

    International Nuclear Information System (INIS)

    The excretion of 65Zn, sup(115m)Cd, 203Hg, 207Bi, 210Pb, 60Co, 64Cu, 85Sr and 86Rb in the perfused sections of the intestinal tract in vivo was investigated by the pendular perfusion method. After intravenous administration the excretion of metals was investigated in the jejunum, in the colon and in some experiments also in the ileum. The fluid net movement in the jejunum and colon was measured in dependency on the energy spectrum of the applied metal isotope by means of 14C or 3H-polyethylene glycol 2000. (orig./MG)

  7. A quantitative radiochemical study of ionic and molecular transport in bovine dental enamel

    International Nuclear Information System (INIS)

    A radiochemical method was developed to determine quantitatively and simultaneously the transport of up to four different compounds through dental enamel. The compounds chosen were [3H]-sorbitol, [14C]-glycerol, 36Cl- and 86Rb+. Effective diffusion coefficients of these compounds determined at 40C were considerably different for different specimens of enamel. Thus values for [3H]-sorbitol varied for 0.04 x 10-8 to 2.5 x 10-8 cm2s-1. Most enamel membranes showed an ion selective behaviour by which the cations were more mobile than the anions. A molecular sieve effect was observed for glycerol and sorbitol. (author)

  8. The effects of digoxin and β-methyldigoxin on the heart rate of decompensated patients with atrial fibrillation

    International Nuclear Information System (INIS)

    Eighteen patients with atrial fibrillation were given digoxin 0.13 mg twice daily for 3 weeks and β-methyldigoxin 0.10 mg twice daily for another 3 weeks. At the end of each 3 week period an exercise test was performed and the effects on the heart rate of the two drugs were compared. No difference in heart rate was obtained at rest, wheareas the heart rate after 6 min of exercise was higher during treatment with digoxin (131 beats/min) than when the patients were taking β-methyldigoxin (124 beats/min). There were no significant differences between digoxin and β-methyldigoxin in their effects on the ECT (R-R intervals, T-wave, Q-T duration). The plasma concentrations of the two glycosides were determined by radioimmunoassay and by 86Rb-uptake inhibition assay. Comparable plasma concentration values (1.0 ng/ml for digoxin, 1.1 ng/ml for β-methyldigoxin, mean values) were obtained by radioimmunoassay, but the 86Rb-technique gave significantly higher values (mean 1.5 ng/ml) for β-methyldigoxin. It is concluded that β-methyldigoxin is equal to digoxin for producing slowing of the heart rate in patients with atrial fibrillation. (orig.)

  9. Effect of temperature on the formation and inactivation of syringomycin E pores in human red blood cells and bimolecular lipid membranes.

    Science.gov (United States)

    Agner, G; Kaulin, Y A; Schagina, L V; Takemoto, J Y; Blasko, K

    2000-06-01

    The effects of temperature on the formation and inactivation of syringomycin E (SRE) pores were investigated with human red blood cells (RBCs) and lipid bilayer membranes (BLMs). SRE enhanced the RBC membrane permeability of 86Rb and monomeric hemoglobin in a temperature dependent manner. The kinetics of 86Rb and hemoglobin effluxes were measured at different temperatures and pore formation was found to be only slightly affected, while inactivation was strongly influenced by temperature. At 37 degrees C, SRE pore inactivation began 15 min after and at 20 degrees C, 40 min after SRE addition. At 6 degrees C, below the phase transition temperature of the major lipid components of the RBC membrane, no inactivation occurred for as long as 90 min. With BLMs, SRE induced a large current that remained stable at 14 degrees C, but at 23 degrees C it decreased over time while the single channel conductance and dwell time did not change. The results show that the temperature dependent inactivation of SRE pores is due to a decrease in the number of open pores.

  10. Actions of ionomycin in rat parotid gland

    Energy Technology Data Exchange (ETDEWEB)

    Poggioli, J.; Leslie, B.A.; McKinney, J.S.; Weiss, S.J.; Putney, J.W. Jr.

    1982-04-01

    The effects of ionomycin (SQ 23,377), a carboxylic acid Ca-ionophore, on the rat parotid acinar cell were investigated. Ionomycin stimulated 86Rb efflux from parotid slices and was substantially more potent and efficacious than the Ca-ionophore, A-23187. The release of 86Rb was dependent on the concentration of ionomycin and of Ca. Ionomycin also stimulated 22Na uptake and 3H-protein secretion, but did not stimulate the incorporation of 32PO4 into phosphatidylinositol. These observations are consistent with an action of ionomycin in increasing cytosolic Ca by acting as an ionophore and not involving endogenous receptors. Pretreatment with ionomycin inhibited the transient, Ca-independent responses to carbachol or physalaemin. When ionomycin was added to parotid cells pre-equilibrated with 45Ca, a net loss of radiocalcium was observed. These observations suggest that ionomycin can release the receptor-regulated cellular Ca pool. Morphological studies did not reveal any nonspecific deleterious effects in the cells after incubation with 2.67 microM ionomycin.

  11. Rho family GTP binding proteins are involved in the regulatory volume decrease process in NIH3T3 mouse fibroblasts

    DEFF Research Database (Denmark)

    Pedersen, Stine F; Beisner, Kristine H; Willumsen, Berthe M;

    2002-01-01

    kinase inhibitor Y-27632 and the phosphatidyl-inositol 3 kinase (PI3K) inhibitor wortmannin. The maximal rates of swelling-activated K+ (86 Rb+ as tracer) and taurine ([3H]taurine as tracer) efflux after a 30 % reduction in extracellular osmolarity were increased about twofold in cells with maximal Rho......AV14 expression compared to wild-type cells, but were unaffected by Y-27632. The volume set points for activation of release of both osmolytes appeared to be reduced by RhoAV14 expression. The maximal taurine efflux rate constant was potentiated by the tyrosine phosphatase inhibitor Na(3)VO(4), and...... inhibited by the tyrosine kinase inhibitor genistein. The magnitude of the swelling-activated Cl- current (I(Cl,swell) ) was higher in RhoAV14 than in wild-type cells after a 7.5 % reduction in extracellular osmolarity, but, in contrast to 86Rb+ and [3H]taurine efflux, similar in both strains after a 30...

  12. Lysophosphatidylcholines containing polyunsaturated fatty acids were found as Na+,K+-ATPase inhibitors in acutely volume-expanded hog

    International Nuclear Information System (INIS)

    Na+,K+-ATPase inhibitors activities against the specific binding of ouabain to Na+,K+-ATPase and 86Rb uptake into hog erythrocytes have been purified from the plasma of acutely saline-infused hog. The purifications were performed by a combination of Amberlite XAD-2 adsorption chromatography and four steps of high-performance liquid chromatography with four different types of columns. Fast atom bombardment (FAB) mass and proton NMR spectrometric studies identified the purified substances as γ-arachidoyl- [LPCA(γ), 34%], β-arachidoyl- [LPCA(β), 4%], γ-linoleoyl- (LPCL, 33%), and γ-oleoyl- (LPCO, 25%) lysophosphatidylcholine, expressed in molar ratio in the plasma. Small amounts of γ-docosapentaenoyl-, γ-eicosatrienoyl-, and γpalmitoyllysophosphatidylcholine were also detected by both FAB mass and 1H NMR spectrometric studies. The inhibition of Na+,K+-ATPase activity due to these compounds was always more sensitive than that of both ouabain-binding and 86Rb uptake activities. The ouabain-displacing activity in plasma due to these compounds increased with time during saline infusion. The maximal plasma level was approximately 10 times higher than that in the preinfusion plasma sample. Although these results suggest that γ-acyl-LPC's with long-chain polyunsaturated fatty acids are not simple competitive inhibitors to Na+,K+-ATPase, these compounds could be implicated in the pathogenesis of the circulation abnormality through the modulation of membrane enzyme

  13. Inhibition of Na(+) -K+ pump activity by divalent cations in intact peritoneal mast cells of the rat

    DEFF Research Database (Denmark)

    Knudsen, T; Berthelsen, Carsten; Johansen, Torben

    1990-01-01

    1. The inhibition by the divalent cations magnesium, barium and strontium and the trivalent ion lanthanum of the Na(+) -K+ pump in the plasma membrane of rat peritoneal mast cells was studied in pure mast cell populations by measurement of the ouabain-sensitive uptake of the radioactive potassium......-resistant uptake was not changed. Half maximum decrease in the ouabain-sensitive K+(86Rb+)-uptake was observed with 1.8 mM magnesium, 1.2mM barium and 0.7 mM strontium. 4. The trivalent ion lanthanum blocked almost completely the ouabain-sensitive K+(86Rb+)-uptake at a concentration of 1 microM as does 1 m......M calcium. Combining either of these ions with magnesium had no further inhibitory effect on the ouabain-sensitive uptake. 5. In conclusion, in addition to the previously suggested modulation by calcium of the activity of the Na+ (-)K+ pump, evidence is provided in this investigation that the modulation may...

  14. Potassium fluxes in Chlamydomonas reinhardtii. II. Compartmental analysis

    International Nuclear Information System (INIS)

    42K+ and 86Rb+ were used to determine the subcellular distribution of potassium in Chlamydomonas reinhardtii by compartmental analysis. In both wild type and a mutant strain, three distinct compartments (referred to as I, II, and III) were apparent. Using 42K+, we found that these had half-lives for K+ exchange of 1.07 min, 12.8 min, and 2.9 h, respectively, in wild-type cells and 0.93 min, 14.7 min, and 9.8 h, respectively, for the mutants. Half-lives were not significantly different when 86Rb+ was used to trace K+. Compartments I and II probably correspond to the cell wall and cytoplasm, respectively. Based on the lack of a large central vacuole in Chlamydomonas, the effect of a dark pretreatment on the kinetic properties of compartment III and the similarity between the [K+] of compartment III and that of isolated chloroplasts, this slowly exchanging compartment was identified as the chloroplast. Growth of wild-type cells at 100 micromolars (instead of 10 mM K+) caused no change of cytoplasmic [K+] but reduced chloroplast [K+] very substantially. The mutants failed to grow at 100 micromolars K+

  15. Studies on the methods of inorganic nutrient application in coconut

    International Nuclear Information System (INIS)

    Using carrier free 32P, tagged single superphosphate and 86Rb, the efficiency of different methods of plant injection and soil placement techniques for fertilizer applications was examined. In the plant injection techniques the radioactivity was fed to the palms through growing roots tips, cut ends of roots, stem injection and leaf axils. The application of radioactivity through the cut ends of roots was most efficient since 32P was detected in 10 m tall palms, four hours after application. In stem, leaf axil and growing roots tips injection the 32P was detected after 8, 12 and 18 h. Out of four methods of soil application, the quickest recovery of 32P in the palms was detected after 7 days of placement when applied by the hole method. The 32P activity in the palms through circular trenches, strips and basin methods was recorded after 8, 8 and 11 days of application respectively. The accumulation of 86Rb was significantly higher than 32P. With plant injection technique the accumulation of activity was found to be significantly higher than with soil placement methods. The rate of radioactivity absorption was 10 to 60 time faster in the former technique as compared to that of the latter. The application of radioactivity through cut ends of roots and circular trench methods, were found to be better and may recommended for nutrient application in coconut. (orig.)

  16. Potassium transport across rat alveolar epithelium: evidence for an apical Na+-K+ pump.

    Science.gov (United States)

    Basset, G; Bouchonnet, F; Crone, C; Saumon, G

    1988-06-01

    1. Experiments were performed on rat lungs into which various solutions were instilled whilst the lungs were perfused with either whole blood or Ringer solution. Instillation of ion-free glucose solution led to a net flux of fluid and ions into the alveolar spaces. K+ ions entered faster than Na+ ions and reached a concentration about twice that in the perfusate. Ouabain in the perfusate (basolateral side) prevented the rise in alveolar K+ concentration above that in the perfusate, indicating a transcellular pathway. Ba2+ in the instillate (apical side) hindered the entry of K+ into alveoli, suggesting the presence of apical K+ channels. 2. When Ringer solution was instilled, K+ was continuously removed from the alveoli and the K+ concentration in the instillate remained constant or decreased slightly depending on the rate of fluid absorption. The net K+ efflux from alveoli to blood was 0.23 pmol/(cm2 s). When Ba2+ was added to the instillate the net K+ efflux increased to 0.36 pmol/(cm2 s). Apical ouabain reversed the K+ flux resulting in a net K+ flux of 0.19 pmol/(cm2 s) into the alveoli. This suggests the presence of an Na+-K+-ATPase located in the apical membrane of some alveolar cells. 3. The K+ transport from instillate (Ringer solution) to perfusate was traced by means of 86Rb which was added to the instillate. Ouabain in the instillate did not affect fluid absorption but reduced the apparent 86Rb permeability by 50% although the paracellular permeability (estimated with [3H]mannitol) was unaffected. This also indicates the presence of an apical Na+-K+-ATPase. When ouabain was added to the perfusate, the apparent 86Rb permeability doubled. These findings indicate that recirculation of 86Rb (and K+) occurs due to the activity of both apical and basolateral Na+-K+-ATPases. 4. When ouabain was placed on both sides of the epithelium, preventing transcellular transport, the passive 86Rb permeability was 10.3 x 10(-8) cm/s (assuming an alveolar surface area of

  17. Chromatographic purification of neutron capture molybdenum-99 from cross-contaminant radionuclides

    International Nuclear Information System (INIS)

    Technetium-99m is called the work horse, for many reasons, in nuclear medicine diagnostic purposes. It is produced as the β-decay of 99Mo radionuclide. Molybdenum-99 gel type generators are considered as a suitable alternative of the conventional chromatographic alumina columns loaded with fission molybdenum-99. 99Mo neutron-capture is cross-contaminated with radionuclides originated from activation of chemical impurities in the Mo target such 60C0, 65Zn, 95Zr, 175Hf, 181Hf, 86Rb, 134Cs, 141Ce, 152Eu, 140La,51Cr, 124Sb,46Sc, 54Mn, 59Fe and / or fast neutrons interactions with the stable isotopes of molybdenum such as 92mNb, 95Nb and 95Zr. To prevent contamination of the eluted 99mTc, successive purification methods were made. After complete dissolution of the irradiated target wrapped with thin Al foil in 5 M NaOH solution, hydrogen peroxide was added to start precipitation of Fe(OH)3. The formed Fe (III) minerals allow complete elimination of some radio contaminants from the molybdate solute such as 152Eu, 140La,141Ce, 45Mn and 92mNb in addition to partial elimination of 46Sc, 60Co and 59Fe radionuclides. The remaining supernatant was acidified by concentrated nitric acid to ph 9.5 for precipitation of Al(OH)3 with complete elimination of radio contaminants such as 95Zr 175Hf, 181Hf, 65Zn, 124Sb, 51Cr, 46Sc, 60Co and 59Fe. 134Cs and 86Rb radionuclides were not affected by precipitation of Fe(OH)3 or Al(OH)3. Chromatographic column of potassium nickel hexacyanoferrate (II) (KNHCF) has high affinity towards elimination of 134Cs and 86Rb radionuclides. Highly pure molybdate-99Mo solution was processed for preparation of zirconium molybdate gel generator with 99mTc eluate of high radionuclidic, radiochemical and chemical purity suitable for use in medical purposes.

  18. Early events elicited by bombesin and structurally related peptides in quiescent Swiss 3T3 cells. II. Changes in Na/sup +/ and Ca/sup 2 +/ fluxes, Na/sup +//K/sup +/ pump activity, and intracellular pH

    Energy Technology Data Exchange (ETDEWEB)

    Mendoza, S.A.; Schneider, J.A.; Lopez-Rivas, A.; Sinnett-Smith, J.W.; Rozengurt, E.

    1986-06-01

    The amphibian tetradecapeptide, bombesin, and structurally related peptides caused a marked increase in ouabain-sensitive /sup 86/Rb/sup +/ uptake (a measure of Na/sup +//K/sup +/ pump activity) in quiescent Swiss 3T3 cells. This effect occurred within seconds after the addition of the peptide and appeared to be mediated by an increase in Na/sup +/ entry into the cells. The effect of bombesin on Na/sup +/ entry and Na/sup +//K/sup +/ pump activity was concentration dependent with half-maximal stimulation occurring at 0.3-0.4 nM. The structurally related peptides litorin, gastrin-releasing peptide, and neuromedin B also stimulated ouabain-sensitive /sup 86/Rb/sup +/ uptake; the relative potencies of these peptides in stimulating the Na/sup +//K/sup +/ pump were comparable to their potencies in increasing DNA synthesis. Bombesin increased Na/sup +/ influx, at least in part, through an Na/sup +//H/sup +/ antiport. The peptide augmented intracellular pH and this effect was abolished in the absence of extracellular Na/sup +/. In addition to monovalent ion transport, bombesin and the structurally related peptides rapidly increased the efflux of /sup 45/Ca/sup 2 +/ from quiescent Swiss 3T3 cells. This Ca/sup 2 +/ came from an intracellular pool and the efflux was associated with a 50% decrease in total intracellular Ca/sup 2 +/. The peptides also caused a rapid increase in cytosolic free calcium concentration. Prolonged pretreatment of Swiss 3T3 cells with phorbol dibutyrate, which causes a loss of protein kinase C activity, greatly decreased the stimulation of /sup 86/Rb/sup +/ uptake and Na/sup +/ entry by bombesin implicating this phosphotransferase system in the mediation of part of these responses to bombesin. Since some activation of monovalent ion transport by bombesin was seen in phorbol dibutyrate-pretreated cells, it is likely that the peptide also stimulates monovalent ion transport by a second mechanism.

  19. Purification of alpha-toxin from Staphylococcus aureus and application to cell permeabilization

    Energy Technology Data Exchange (ETDEWEB)

    Lind, I.; Ahnert-Hilger, G.; Fuchs, G.; Gratzl, M.

    1987-07-01

    Crude alpha-toxin was produced by Staphylococcus aureus, strain Wood 46. The amount of exotoxin was monitored during growth and all subsequent purification steps by determination of its hemolytic activity against rabbit erythrocytes. The culture supernatant was treated with ammonium sulfate (75% saturation). The resulting precipitate was dialyzed and subjected to cation-exchange chromatography. The fractions containing the hemolytic activity were further purified by gel chromatography. The final product was enriched by a factor of 8.5 compared to the crude toxin. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified toxin exhibited one major band. It caused the release of /sup 86/Rb+ and ATP from rat insulinoma (RIN A2) as well as pheochromocytoma cells (PC12) in culture, indicating efficient permeabilization of their plasma membranes for small molecules.

  20. Effect of temperature and light on foliar absorption of P and Rb by Chrysanthemum and Pilea

    Directory of Open Access Journals (Sweden)

    Szczepan Marczyński

    2013-12-01

    Full Text Available Young plants of Pilea cadierei Gagnep Guillaum and Chrysanthemum morifolium Ramat. 'Giant # 4 Indianapolis White' were grown in Hoagland's solution in growth chambers. Their leaves were treated with rubidium phosphate double labelled with 33P and 86Rb. Light intensity, period of pretreatment in light or dark, daylength, and air temperature had different influences on foliar uptake of each ion, as did plant species and leaf surface. With all variables tested, uptake and translocation of Rb was much greater than of P. Absorption of both P and Rb through the lower surface was as much as 8 times greater than through upper surface, especially with Pilea. Light had a greater effect upon uptake of both P and Rb by Chrysanthemum than by Pilea, but did not influence uptake as much as previously reported.

  1. Partial characterization of endogenous digoxinlike substance in human urine

    Energy Technology Data Exchange (ETDEWEB)

    Vinge, E.; Ekman, R.

    1988-01-01

    Urinary samples were collected from individuals not taking cardiac glycosides. Aliquots of 30 ml were passed through preparative octadecylsilane-bonded phase columns and eluted in fractions by stepwise increasing concentrations of acetonitrile. Eluted fractions were analysed for their contents of endogenous digoxinlike substance (EDLS) by radioimmunoassay of digoxin and by a bioassay of cardiac glycosides, which measures the uptake of rubidium (/sup 86/Rb) by erythrocytes as an index of Na+, K+-ATPase activity. In both assays, digoxinlike activity was found in several fractions, but the highest values were consistently measured in the fractions eluted with 40% acetonitrile. Greater amounts of EDLS were recovered from the urine of pregnant women than from the urine of men and nonpregnant women.

  2. Application of neutron activation analysis for the determination of wines originating from different vineyards

    International Nuclear Information System (INIS)

    Neutron activation analysis has been used for the determination of trace elements in different wines originating from various french vineyards. Non-destructive technologies are used for short and middle half-life radionuclides (28Al - 76As - 49Ca - 38Cl - 42K - 27Mg - 56Mn - 24Na - 52V). A radiochemical separation is necessary for longer half-life radionuclides (60Co - 52Cr - 134Cs - 59Fe - 86Rb - 65Zn). The results of the study show that the identification of vineyards based on the determination of specific oligo-elements is feasible. However, more data are needed to demonstrate that the knowledge of the amounts of specific oligo-elements in a wine corresponding to a given vineyard can be used to disclose frauds, particularly in the cases of wine watering or mixtures of wines originating from different vineyards. (author)

  3. Decontamination by shotblasting of radioactivity deposited on an asphalt road

    International Nuclear Information System (INIS)

    Long-lived fission products may be deposited in the environment after a serious reactor accident. From previous experiments it is known that if firehosing is to be used for decontamination it has to be done soon after the deposition. It is therefore worthwhile to study another decontamination method. An experimental study has been conducted of how well shotblasting can remove contamination from an asphalt road. In shotblasting a thin layer of the surface is loosened by the impact of small steel balls, and in the same procedure the surface dust is vacuumed up and the steel balls recovered. The contaminant was 86Rb, which behaves as caesium. As reference, the weathering of identical contamination on an asphalt road, a concrete road and a road covered with small concrete stones was studied concurrently. (author)

  4. Use of radioisotopes in study of herbicide effect mechanism

    International Nuclear Information System (INIS)

    The degradation of 35S-prometryne in 8 agricultural crops and 3 woody species is explained. S released from the -SCH3 group was incorporated into cystine, a sulphate anion, and in plants from the family Brassicaceae into sinalbin and glucobrassicin. The main cause of resistance of Convolvulus arvensis to atrazine (labelled on rings with 14C) was its ability to degrade atrazine into non-toxic metabolites (hydroxyatrazine, conjugate with glutathione). Bidisin, Isobarnon and Suffix decreased the uptake, transport, and metabolism of phosphorus (32P), calcium (45Ca) and potassium (86Rb) in Avena fatua; they are used for its killing. In barley and wheat no decrease was observed with the exception of a limited transport of potassium in the youngest leaf. Bidisin and Isobarnon inhibited the fixation of 14CO2 in A. fatua within 2 hours after application. (author)

  5. Diphenyleneiodonium, an inhibitor of NOXes and DUOXes, is also an iodide-specific transporter

    Directory of Open Access Journals (Sweden)

    C. Massart

    2014-01-01

    Full Text Available NADPH oxidases (NOXes and dual oxidases (DUOXes generate O2.− and H2O2. Diphenyleneiodonium (DPI inhibits the activity of these enzymes and is often used as a specific inhibitor. It is shown here that DPI, at concentrations similar to those which inhibit the generation of O2 derivatives, activated the efflux of radioiodide but not of its analog 99mTcO4− nor of the K+ cation mimic 86Rb+ in thyroid cells, in the PCCl3 rat thyroid cell line and in COS cell lines expressing the iodide transporter NIS. Effects obtained with DPI, especially in thyroid cells, should therefore be interpreted with caution.

  6. Counting efficiency for radionuclides decaying by beta and gamma-ray emission

    International Nuclear Information System (INIS)

    In this paper, counting efficiency vs figure of merit for beta and gamma-ray emitters has been computed. It is assumed that the decay scheme has only a gamma level and the beta-ray emission may be coincident with the gamma-rays or the internal-conversion electrons. The radionuclides tabulated are: 20O, 20F, 28Al, 35P,41Ar, 42K, 47Se, 62Fe, 66Cu, 81Ge, 86Rb, 104Rh, 108Ru, 112Pd, 121Sn(m), 122In, 129I, 141Ce, 142Pr, 151Sm, 170Tm, 171Tm, 194Os, 203Hg, 205Hg, 210Pb, 225Ra, 244Am(m). It has been assumed that the liquid is a toluene based scintillator solution in standard glass vials containing 10 cm3. (Author)

  7. Shrinkage insensitivity of NKCC1 in myosin II-depleted cytoplasts from Ehrlich ascites tumor cells

    DEFF Research Database (Denmark)

    Hoffmann, Else K; Pedersen, Stine F

    2007-01-01

    Protein phosphorylation/dephosphorylation and cytoskeletal reorganization regulate the Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) during osmotic shrinkage; however, the mechanisms involved are unclear. We show that in cytoplasts, plasma membrane vesicles detached from Ehrlich ascites tumor cells (EATC......) by cytochalasin treatment, NKCC1 activity evaluated as bumetanide-sensitive (86)Rb influx was increased compared with the basal level in intact cells yet could not be further increased by osmotic shrinkage. Accordingly, cytoplasts exhibited no regulatory volume increase after shrinkage. In cytoplasts......, cortical F-actin organization was disrupted, and myosin II, which in shrunken EATC translocates to the cortical region, was absent. Moreover, NKCC1 activity was essentially insensitive to the myosin light chain kinase (MLCK) inhibitor ML-7, a potent blocker of shrinkage-induced NKCC1 activity in intact...

  8. Water permeability of Na+-K+-2C1- cotransporters in mammalian epithelial cells

    DEFF Research Database (Denmark)

    Hammann, Steffen; Herrera-Perez, J.J.; Bundgaard, Magnus;

    2005-01-01

    Water transport properties of the Na+-K+-2Cl- cotransporter (NKCC) were studied in cultures of pigmented epithelial cells (PE) from the ciliary body of the eye. Here, the membrane that faces upwards contains NKCCs and can be subjected to rapid changes in bathing solution composition and osmolarity....... The anatomy of the cultured cell layer was investigated by light and electron microscopy. The transport rate of the cotransporter was determined from the bumetanide-sensitive component of 86Rb+ uptake, and volume changes were derived from quenching of the fluorescent dye calcein. The water permeability (Lp......) of the membrane was halved by the specific inhibitor bumetanide. The bumetanide-sensitive component of the water transport exhibited apparent saturation at osmotic gradients higher than 200 mosmol l-1. Cell shrinkages produced by NaCl or KCl were smaller than those elicited by equi-osmolar applications...

  9. Chiral recognition of pinacidil and its 3-pyridyl isomer by canine cardiac and smooth muscle: Antagonism by sulfonylureas

    International Nuclear Information System (INIS)

    Pinacidil, a potassium channel opener (PCO), relaxes vascular smooth muscle by increasing potassium ion membrane conductance, thereby causing membrane hyperpolarization. PCOs also act on cardiac muscle to decrease action potential duration (APD) selectively. To examine the enantiomeric selectivity of pinacidil, the stereoisomers of pinacidil (a 4-pyridylcyanoguanidine) and its 3-pyridyl isomer (LY222675) were synthesized and studied in canine Purkinje fibers and cephalic veins. The (-)-enantiomers of both pinacidil and LY222675 were more potent in relaxing phenylephrine-contracted cephalic veins and decreasing APD than were their corresponding (+)-enantiomers. The EC50 values for (-)-pinacidil and (-)-LY222675 in relaxing cephalic veins were 0.44 and 0.09 microM, respectively. In decreasing APD, the EC50 values were 3.2 microM for (-)-pinacidil and 0.43 microM for (-)-LY222675. The eudismic ratio was greater for the 3-pyridyl isomer than for pinacidil in both cardiac (71 vs. 22) and vascular (53 vs. 17) tissues. (-)-LY222675 and (-)-pinacidil (0.1-30 microM) also increased 86Rb efflux from cephalic veins to a greater extent than did their respective optical antipodes. The antidiabetic sulfonylurea, glyburide (1-30 microM), shifted the vascular concentration-response curve of (-)-pinacidil to the right by a similar extent at each inhibitor concentration. Glipizide also antagonized the response to (-)-pinacidil, but was about 1/10 as potent with a maximal shift occurring at 10 and 30 microM. Glyburide antagonized the vascular relaxant effects of 0.3 microM (-)-LY222675 (EC50, 2.3 microM) and reversed the decrease in APD caused by 3 microM (-)-LY222675 (EC50, 1.9 microM). Nitroprusside did not alter 86Rb efflux, and vascular relaxation induced by sodium nitroprusside was unaffected by sulfonylureas

  10. Glucose homeostasis in pregnant rats submitted to dietary protein restriction.

    Science.gov (United States)

    de Mello, Maria Alice Rostom; Luciano, Eliete; Carneiro, Everardo Magalhães; Latorraca, Márcia Queiroz; Machado de Oliveira, Camnila Aparecida; Boschero, Antonio Carlos

    2003-01-01

    In the present work, we examined the effects of feeding a low protein diet during pregnancy on glucose-induced insulin secretion and glucose homeostasis in rats. Young (60 days), pregnant (P) or non-pregnant (NP) rats were fed during pregnancy or for 21 days (the NP) a normal (17%) or a low (6%) protein diet. Serum glucose and insulin levels and pancreas insulin content in the fed state; total area under serum glucose curve (AG) after a glucose load and serum glucose disappearance rate (Kitt) after insulin administration; as well as 86Rb outflow, 45Ca uptake and insulin secretion by isolated pancreatic islets in response to glucose were evaluated. Serum glucose was lower in 17%-P (12%) and 6%-P (27%) than in corresponding NP-rats. Serum insulin was higher in 17%-P (153%) and 6%-P (77%) compared to the corresponding NP-rats. Pancreatic insulin was higher in 6%-rats (55%) than in 17%-rats. No differences were found in AG among the groups whereas Kitt was lower in 6%-NP and higher in 6%-P than in the equivalent 17% rats. Increasing glucose concentration from 2.8 to 16.7 mmol/l, reduced 86Rb outflow from isolated islets from all groups. Increasing glucose concentration from 2.8 to 16.7 mmol/l elevated 45Ca uptake by 17%-NP (47%), 17%-P (40%) and 6%-P (214%) islets but not by 6%-NP ones. The increase in 45Ca uptake was followed by an increase in insulin release by the 17%-NP (2767%), 17%-P (2850%) and 6%-P (1200%) islets. In conclusion, 6%-P rats show impaired glucose induced insulin secretion related to reduced calcium uptake by pancreatic islets. However, the poor insulin secretion did not fully compensate the high peripheral sensitivity to the hormone, resulting in hypoglycemia. PMID:15686122

  11. C-peptide increases Na,K-ATPase expression via PKC- and MAP kinase-dependent activation of transcription factor ZEB in human renal tubular cells.

    Directory of Open Access Journals (Sweden)

    Dana Galuska

    Full Text Available BACKGROUND: Replacement of proinsulin C-peptide in type 1 diabetes ameliorates nerve and kidney dysfunction, conditions which are associated with a decrease in Na,K-ATPase activity. We determined the molecular mechanism by which long term exposure to C-peptide stimulates Na,K-ATPase expression and activity in primary human renal tubular cells (HRTC in control and hyperglycemic conditions. METHODOLOGY/PRINCIPAL FINDINGS: HRTC were cultured from the outer cortex obtained from patients undergoing elective nephrectomy. Ouabain-sensitive rubidium ((86Rb(+ uptake and Na,K-ATPase activity were determined. Abundance of Na,K-ATPase was determined by Western blotting in intact cells or isolated basolateral membranes (BLM. DNA binding activity was determined by electrical mobility shift assay (EMSA. Culturing of HRTCs for 5 days with 1 nM, but not 10 nM of human C-peptide leads to increase in Na,K-ATPase α(1-subunit protein expression, accompanied with increase in (86Rb(+ uptake, both in normal- and hyperglycemic conditions. Na,K-ATPase α(1-subunit expression and Na,K-ATPase activity were reduced in BLM isolated from cells cultured in presence of high glucose. Exposure to1 nM, but not 10 nM of C-peptide increased PKCε phosphorylation as well as phosphorylation and abundance of nuclear ERK1/2 regardless of glucose concentration. Exposure to 1 nM of C-peptide increased DNA binding activity of transcription factor ZEB (AREB6, concomitant with Na,K-ATPase α(1-subunit mRNA expression. Effects of 1 nM C-peptide on Na,K-ATPase α(1-subunit expression and/or ZEB DNA binding activity in HRTC were abolished by incubation with PKC or MEK1/2 inhibitors and ZEB siRNA silencing. CONCLUSIONS/SIGNIFICANCE: Despite activation of ERK1/2 and PKC by hyperglycemia, a distinct pool of PKCs and ERK1/2 is involved in regulation of Na,K-ATPase expression and activity by C-peptide. Most likely C-peptide stimulates sodium pump expression via activation of ZEB, a transcription

  12. Correlation between catecholamine release and sodium pump inhibition in the perfused adrenal gland of the cat

    Science.gov (United States)

    Garcia, A.G.; Garcia-Lopez, E.; Montiel, C.; Nicolas, G.P.; Sanchez-Garcia, P.

    1981-01-01

    1 Ca2+ reintroduction to retrogradely perfused and ouabain (10-4 M)-treated cat adrenal glands caused a catecholamine secretory response which was greater the longer the time of exposure to the cardiac glycoside. Such a response was proportional to the external Na+ concentration [Na+]o. 2 A qualitatively similar, yet smaller response was observed when glands were perfused with Krebs solution lacking K+ ions; thus, K+ deprivation mimicked the secretory effects of ouabain. Catecholamine secretion evoked by Ca2+ reintroduction in K+-free solution (0-K+) was also proportional to [Na+]o and greater the longer the time of exposure of the gland to 0-K+ solution. 3 The ionophore X537A also mimicked the ouabain effects, since Ca2+ reintroduction to glands treated with this agent (25 μM) caused a sharp secretory response. When added together with X537A, ouabain (10-4 M) did not modify the response to the ionophore. 4 N-ethylmaleimide (NEM), another Na+, K+-ATPase inhibitor, did not evoke the release of catecholamines; on the contrary, NEM (10-4 M) inhibited the catecholamine secretory response to high [K+]o, acetylcholine, Ca2+ reintroduction and ouabain. 5 Ouabain (10-4 M) inhibited the uptake of 86Rb into adreno-medullary tissue by 60%. Maximal inhibition had already occurred 2 min after adding the drug, indicating a lack of temporal correlation between ATPase inhibition and the ouabain secretory response, which took longer (about 30-40 min) to reach its peak. NEM (10-4 M) blocked 86Rb uptake in a similar manner. 6 The results are further evidence in favour of the presence of a Na+-Ca2+ exchange system in the chromaffin cell membrane, probably involved in the control of [Ca2+]i and in the modulation of catecholamine secretion. This system is activated by increasing [Na+]i, either directly (ionophore X537A, increased [Na+]o) or indirectly (Na+ pump inhibition). However, the simple inhibition of Na+ pumping does not always lead to a catecholamine secretory response; such is

  13. Late radiation damage in bone, bone marrow and brain vasculature, with particular emphasis upon fractionation models

    International Nuclear Information System (INIS)

    X-ray induced changes in rat and human bone and bone marrow vasculature and in rat brain vasculature were measured as a function of time after irradiation and absorbed dose. The absorbed dose in the organ varied from 5 to 25 Gy for single dose irradiations and from 19 to 58 Gy for fractionated irradiations.The number of fractions varied from 3 to 10 for the rats and from 12 to 25 for the human. Blood flow changes were measured using an ''1''2''5I antipyrine or ''8''6RbCl extraction technique. The red blood cell (RBC) volume was examined by ''5''1Cr labelled red cells. Different fractionation models have been compared. Radiation induced reduction of bone and bone marrow blood flow were both time and dose dependent. Reduced blood flow 3 months after irradiation would seem to be an important factor in the subsequent atrophy of bones. With a single dose of 10 Gy the bone marrow blood flow returned to the control level by 7 months after irradiation. In the irradiated bone the RBC volume was about same as that in the control side but in bone marrow the reduction was from 32 to 59%. The dose levels predicted by the nominal standard dose (NSD) formula produced about the same damage to the rat femur seven months after irradiation when the extraction of ''8''6Rb chloride and the dry weight were concerned as the end points. However, the results suggest that the NSB formula underestimates the late radiation damage in bone marrow when a small number of large fractions are used. In the irradiated brains of the rats the blood flow was on average 20.4% higher compared to that in the control group. There was no significant difference in brain blood flow between different fractionation schemes. The value of 0.42 for the exponent of N corresponds to the average value for central nervous system tolerance in the literature. The model used may be sufficiently accurate for clinical work provided the treatment schemes used do not depart too radically from standard practice

  14. Effects of radiosensitising agent nicotinamide on relative tissue perfusion and kidney junction in C3H mice

    International Nuclear Information System (INIS)

    Nicotinamide is an effective radiosensitiser of murine tumours, functioning by improving tumour perfusion by decreasing the proportion of intermittently closed capillaries. The effect of nicotinamide on relative tissue perfusion of RIF-1 tumour and normal skin, muscle, lung, liver, kidney and spleen were investigated using the 86Rb extraction technique. A dose of 1000 mg/kg was shown to have transient effects on tumour, skin and lung perfusion but to have sustained effects on muscle (a drop to 80% of control), liver, kidney and spleen (with increased ranging from 165% to 280% of control) from 0.5 to 4 h after treatment i.e. during the period of maximum radiosensitisation. These increases were evident at doses as low as 100 mg/kg. The data suggest that the radiosensitisation induced by nicotinamide in the mouse may be associated with these perfusion changes. Nicotinamide was also shown to have a substantial inhibitory effect on renal function, inhibiting 51CrEDTA clearance by a factor (± 2 SE) of 2.56 ± 0.19 and 125I-iodohippurate clearance by a factor of 2.07 ± 0.45 at 1000 mg/kg. These effects were shown to be dose-related, and to be evident at doses from 400 mg/kg upwards. This suggests that nicotinamide potentiation of co-administered cytotoxic agents may be mediated by reduced renal clearance of the cytotoxic drug, thus increasing the plasma half-life. (author)

  15. Altered erythrocyte Na-K pump in anorectic patients

    International Nuclear Information System (INIS)

    The status of the erythrocyte sodium pump was evaluated in a group of patients suffering from anorexia nervosa and a group of healthy female control subjects. Anorectic patients showed significantly higher mean values of digoxin-binding sites/cell (ie, the number of Na-K-ATPase units) with respect to control subjects while no differences were found in the specific 86Rb uptake (which reflects the Na-K-ATPase activity) between the two groups. A significant correlation was found between relative weight and the number of Na-K-ATPase pump units (r = -0.66; P less than 0.0001). Anorectic patients showed lower serum T3 concentrations (71.3 +/- 53 ng/dL) with respect to control subjects (100.8 +/- 4.7 ng/dL; P less than 0.0005) and a significant negative correlation between T3 levels and the number of pump units (r = -0.52; P less than 0.003) was found. This study therefore shows that the erythrocyte Na-K pump may be altered in several anorectic patients. The authors suggest that this feature could be interrelated with the degree of underweight and/or malnutrition

  16. Innovations in isotope techniques to enhance the evaluation and management of nutrient sources

    International Nuclear Information System (INIS)

    The increasing world population and the need to produce more food is putting increasing pressures on soil and water resources. Increasingly, studies of interactions between nutrients rather than single nutrient studies are becoming important as systems intensify and a wider range of nutrients is added. The incorporation of isotopes into these studies will greatly assist in understanding the driving forces that determine system productivity and sustainability. Studies of N dynamics have been greatly assisted by the use of the stable 15N, and studies of water, other nutrients, and C have also been assisted by the use of radioactive isotopes such as 3H, 14C, 32P, 35S and 86Rb. However, increasing restrictions on the use of radioactive substances are beginning to severely limit their availability for such studies. Stable isotopes such as 13C, 15N and 34S offer the prospect of replacing radioactive isotopes, or of their being used in combination with radioactive isotopes, to minimize the perceived risks and/or to allow the tracing of two components in the system. The innovative use of isotopes in plant-nutrient studies utilizing direct labelling with stable isotopes, multiple direct labelling with stable and/or radioactive isotopes and/or utilizing natural abundance, reverse dilution, or a combination of direct labelling and reverse dilution, are presented. (author)

  17. Combined radioactivation methods in determining trace elements

    International Nuclear Information System (INIS)

    The authors have devised a method of radioactivation analysis (RAA) for determining 32 elements by NAA, proton-activation analysis (PAA), and emission spectral analysis (ESA). Here they examine element distributions in certain plants by NAA, PAA, and DAA (deuteron activation analysis) as described elsewhere. The results are compared with those from activation analysis (AA) and ESA. The authors used five species of medicinal plant: Plantago Major L, Salvia officinalis L., Artemisia absinthium L., Alhagi Persarum, and Eremurus. They used referenced methods for preparing the samples, irradiating them in the reactor or cyclotron, and measuring the radioactivity. The γ-ray spectra for the activated samples from all the plants gave peaks representing 24Na, 42K, 56Mn, 140La, 82Br, 124Sb, 46Sc, 59Fe, 198Au, 139Ce, 153Sm, 86Rb, 65Zn, 60Co, and 147MSn. The concentrations of Fe, Sb, Sn, Zn, Rb, Co were determined when the irradiated samples have been kept for 10 days. To determine Ca, Fe, Ti, Cu, Zn, and Sr, ash disks were irradiated by proton and deuteron beams in a cyclotron, where they recorded radiation from the 48Sc, 56Co, 48V, 65Zn, 67Ga, 88Y. The conclusions are that all these plants and the parts of them accumulate the elements in different ways

  18. The physiological significance of HKT1, a Na{sup +} - coupled high affinity K{sup +} transporter in `Triticum aestivum`

    Energy Technology Data Exchange (ETDEWEB)

    Box, S.; Schachtman, D.P. [University of Adelaide, SA (Australia). Department of Botany

    1997-12-31

    Full text: Several mechanisms for high affinity K{sup +} uptake by higher plants have been proposed:-an ATP-energised K:+ pump, a K{sup +}/H{sup +} antiport and a H{sup +}coupled carrier. Recently, a Na{sup +}--coupled high affinity K{sup +} transporter, HKT1, was isolated from wheat roots. Whilst Na{sup +}K{sup +} symports have been described in charophyte algae, the cloning of HKT1 from wheat is the first, evidence that this type d transport mechanism may function in higher plants. Is the activity of HKT1 an important mechanism involved in K{sup +} acquisition by wheat? The aim of this study was to assess the physiological significance of Na{sup +}- coupled high affinity K{sup +} uptake in T. aestivum. To determine whether HKT1 plays a significant role in wheat growth, we measured the dry weights and ion content of plants grown in a range of [K{sup +}], with and without Na{sup +}. To directly assess the activity of Na{sup +}- coupled K{sup +} transport, {sup 86}Rb{sup +} and {sup 22}Na{sup +} flux analyses were performed on the elongation zones and whole roots of intact seedlings, expressing a high affinity K{sup +} uptake system. The results of these growth and tracer flux studies will be discussed in relation to the expression of the gene encoding HKT1 in T. aestivum

  19. Syringotoxin pore formation and inactivation in human red blood cell and model bilayer lipid membranes.

    Science.gov (United States)

    Szabó, Zsófia; Gróf, Pál; Schagina, Ludmila V; Gurnev, Philip A; Takemoto, Jon Y; Mátyus, Edit; Blaskó, Katalin

    2002-12-23

    The effect of syringotoxin (ST), a member of the cyclic lipodepsipeptides family (CLPs) produced by Pseudomonas syringae pv. syringae on the membrane permeability of human red blood cells (RBCs) and model bilayer lipid membranes (BLMs) was studied and compared to that of two recently investigated CLPs, syringomycin E (SRE) and syringopeptin 22A (SP22A) [Biochim. Biophys. Acta 1466 (2000) 79 and Bioelectrochemistry 52 (2000) 161]. The permeability-increasing effect of ST on RBCs was the least among the three CLPs. A time-dependent ST pore inactivation was observed on RBCs at 20 and 37 degrees C but not at 8 degrees C. From the kinetic model worked out parameters as permeability coefficient of RBC membrane for 86Rb(+) and pores mean lifetime were calculated. A shorter pores mean lifetime was calculated at 37 degrees C then at 20 degrees C, which gave us an explanation for the unusual slower rate of tracer efflux measured at 37 degrees C then that at 20 degrees C. The results obtained on BLM showed that the pore inactivation was due to a decrease in the number of pores but not to a change of their dwell time or conductance.

  20. Altered erythrocyte Na-K pump in anorectic patients

    Energy Technology Data Exchange (ETDEWEB)

    Pasquali, R.; Strocchi, E.; Malini, P.; Casimirri, F.; Ambrosioni, E.; Melchionda, N.; Labo, G.

    1985-07-01

    The status of the erythrocyte sodium pump was evaluated in a group of patients suffering from anorexia nervosa and a group of healthy female control subjects. Anorectic patients showed significantly higher mean values of digoxin-binding sites/cell (ie, the number of Na-K-ATPase units) with respect to control subjects while no differences were found in the specific /sup 86/Rb uptake (which reflects the Na-K-ATPase activity) between the two groups. A significant correlation was found between relative weight and the number of Na-K-ATPase pump units (r = -0.66; P less than 0.0001). Anorectic patients showed lower serum T3 concentrations (71.3 +/- 53 ng/dL) with respect to control subjects (100.8 +/- 4.7 ng/dL; P less than 0.0005) and a significant negative correlation between T3 levels and the number of pump units (r = -0.52; P less than 0.003) was found. This study therefore shows that the erythrocyte Na-K pump may be altered in several anorectic patients. The authors suggest that this feature could be interrelated with the degree of underweight and/or malnutrition.

  1. Isolation and characterization of a specific endogenous Na/sup +/, K/sup +/-ATPase inhibitor from bovine adrenal

    Energy Technology Data Exchange (ETDEWEB)

    Tamura, M.; Lam, T.T.; Inagami, T.

    1988-06-14

    In order to identify a specific endogenous Na/sup +/,K/sup +/-ATPase inhibitor which could possibly be related to salt-dependent hypertension, the authors looked for substances in the methanol extract of bovine whole adrenal which show all of the following properties: (i) inhibitory activity for Na/sup +/,K/sup +/-ATPase; (ii) competitive displacing activity against (/sup 3/H)ouabain binding to the enzyme; (iii) inhibitory activity for /sup 86/Rb uptake into intact human erythrocytes; and (iv) cross-reactivity with sheep anti-digoxin-specific antibody. After stepwise fractionation of the methanol extract of bovine adrenal glands by chromatography on a C/sub 18/ open column, a 0-15% acetonitrile fraction was fractionated by high-performance liquid chromatography on a Zorbax octadecylsilane column. One of the most active fractions in 0-15% acetonitrile was found to exhibit all of the four types of the activities. It was soluble in water and was distinct from various substances which have been known to inhibit Na/sup +/,K/sup +/-ATPase. These results strongly suggest that this water-soluble nonpeptidic Na/sup +/,K/sup +/-ATPase inhibitor may be a specific endogenous regulator for the ATPase.

  2. Endogenous inotropic substance from heart tissue has digitalis-like properties

    Energy Technology Data Exchange (ETDEWEB)

    Khatter, J.C.; Agbanyo, M.; Navaratnam, S. (Univ. of Manitoba, Winnipeg (Canada))

    1991-01-01

    In the past few years, we developed an extraction procedure which we successfully used to isolate a crude fraction containing digitalis-like substance (DLS) from porcine left ventricular tissue. In this study, the crude fraction was found to cross-react with digoxin antibodies and showed immunoreactivity of 4.25 {plus minus} 0.6 ng digoxin equivalent/ml. On further purification of the crude fraction using silica gel G column chromatography, a fraction C was obtained, which was highly positive inotropic on canine trabeculae and it dose-dependently inhibited ouabain sensitive {sup 86}Rb{sup +} uptake in rate heart slices. A 50% inhibition of uptake was obtained by 25 ul of fraction C. Fraction C also inhibited canine kidney Na{sup +}, K{sup +}-ATPase dose-dependently and a 50% inhibition of this enzyme required 17 ul of fraction C. Ashing of the fraction C at 500{degree}C resulted in loss of inotropic and enzyme inhibitory activities, indicating an organic nature of the unknown digitalis-like substance.

  3. Effect of ADH on rubidium transport in isolated perfused rat cortical collecting tubules

    Energy Technology Data Exchange (ETDEWEB)

    Schafer, J.A.; Troutman, S.L.

    1986-06-01

    Unidirectional fluxes of 86Rb+ were measured as an indicator of potassium transport in isolated rat cortical collecting tubules perfused and bathed at 38 degrees C with isotonic solutions in which Rb+ replaced K+. Under control conditions the lumen-to-bath flux (Jl----b) was significantly less than the bath-to-lumen flux (Jb----l), indicating net Rb+ secretion. Net secretion increased approximately 180% after addition of 100 microU/ml of arginine vasopressin (ADH) to the bathing solution, due to a rapid and reversible increase in Jb----l from 4.6 +/- 0.8 to 9.0 +/- 1.9 pmol X min-1 X mm-1 with no significant change in Jl----b. The ADH effect was completely inhibited by 2 mM luminal Ba2+. The average transepithelial voltage (Ve) was not significantly different from zero in the control period but became lumen negative (-5 to -10 mV) after ADH. With 10(-5) M amiloride in the lumen Ve was lumen positive (+2 to +4 mV) and was unaltered by ADH or Ba2+, yet ADH produced a significant but attentuated increase in Jb----l with no change in Jl----b. The results indicate that ADH augments net K+ secretion either by an increase in the Ba2+-sensitive conductance of the apical membrane or by an increase in the electrochemical potential driving force for net Rb+ secretion through this pathway.

  4. Characterization of ultraviolet light-induced ouabain-resistant mutations in Chinese hamster cells

    International Nuclear Information System (INIS)

    Ouabain-resistant mutations in Chinese hamster cells have been quantitatively characterized. The mutation frequencies were found to be induced curvilinearly with treatments of increasing doses of ultraviolet light (UV). For the range of UV doses tested (5-20 J/m2), the observed mutation frequency, Y, as a function of UV dose X, follows a curvilinear function, Y=(-28+13.37X - 1.52X2+0.08 X3).10-6. The frequencies of UV-induced mutations were directly correlated with cell survival, indicating a similar causal relationship between cell killing and mutation induction. Under the same experimental conditions, X-rays induced 6-thioguanine-, but not ouabain-, resistant mutations, UV-induced ouabain-resistant (ouasup(r)) mutants exhibit a selection disadvantage. Their phenotypic expressions are modifiable by various agents. Wild type and 16 ouasup(r) mutants were compared with respect to their sensitivity to ouabain inhibition of 86Rb uptake by whole cells. All the ouasup(r) mutants assayed are less sensitive to the drug than are wild-type cells. In the absence of ouabain, the Na+-K+-ATPase activities can be significantly higher or lower than that of the wild-type cells. (Auth.)

  5. Membrane potential and surface potential in mitochondria: uptake and binding of lipophilic cations.

    Science.gov (United States)

    Rottenberg, H

    1984-01-01

    The uptake and binding of the lipophilic cations ethidium+, tetraphenylphosphonium+ (TPP+), triphenylmethylphosphonium+ (TPMP+), and tetraphenylarsonium+ (TPA+) in rat liver mitochondria and submitochondrial particles were investigated. The effects of membrane potential, surface potentials and cation concentration on the uptake and binding were elucidated. The accumulation of these cations by mitochondria is described by an uptake and binding to the matrix face of the inner membrane in addition to the binding to the cytosolic face of the inner membrane. The apparent partition coefficients between the external medium and the cytosolic surface of the inner membrane (K'o) and the internal matrix volume and matrix face of the inner membrane (K'i) were determined and were utilized to estimate the membrane potential delta psi from the cation accumulation factor Rc according to the relation delta psi = RT/ZF ln [(RcVo - K'o)/(Vi + K'i)] where Vo and Vi are the volume of the external medium and the mitochondrial matrix, respectively, and Rc is the ratio of the cation content of the mitochondria and the medium. The values of delta psi estimated from this equation are in remarkably good agreement with those estimated from the distribution of 86Rb in the presence of valinomycin. The results are discussed in relation to studies in which the membrane potential in mitochondria and bacterial cells was estimated from the distribution of lipophilic cations. PMID:6492133

  6. Ionic and secretory response of pancreatic islet cells to minoxidil sulfate

    Energy Technology Data Exchange (ETDEWEB)

    Antoine, M.H.; Hermann, M.; Herchuelz, A.; Lebrun, P. (Laboratory of Pharmacology, Brussels Free University School of Medicine (Belgium))

    1991-07-01

    Minoxidil sulfate is an antihypertensive agent belonging to the new class of vasodilators, the K+ channel openers. The present study was undertaken to characterize the effects of minoxidil sulfate on ionic and secretory events in rat pancreatic islets. The drug unexpectedly provoked a concentration-dependent decrease in 86Rb outflow. This inhibitory effect was reduced in a concentration-dependent manner by glucose and tolbutamide. Minoxidil sulfate did not affect 45Ca outflow from islets perfused in the presence of extracellular Ca++ and absence or presence of glucose. However, in islets exposed to a medium deprived of extracellular Ca++, the drug provoked a rise in 45Ca outflow. Whether in the absence or presence of extracellular Ca++, minoxidil sulfate increased the cytosolic free Ca++ concentration of islet cells. Lastly, minoxidil sulfate increased the release of insulin from glucose-stimulated pancreatic islets. These results suggest that minoxidil sulfate reduces the activity of the ATP-sensitive K+ channels and promotes an intracellular translocation of Ca++. The latter change might account for the effect of the drug on the insulin-releasing process. However, the secretory response to minoxidil sulfate could also be mediated, at least in part, by a modest Ca++ entry.

  7. Limulus test for pyrogens and radiometric sterility tests on radiopharmaceuticals. Part of a coordinated programme

    International Nuclear Information System (INIS)

    Sterility testing of radiopharmaceuticals prepared at BARC were carried out using the radiometric technique (Radiometric detection of the metabolic product 14Co2). Batches of different radiopharmaceuticals were tested for pyrogen using the limulus lysate method and the results were compared with the rabbit method. The results of sterility test on 202 batches of 19 different radiopharmaceuticals show that the radiometric method can be used for sterility testing of radiopharmaceuticals labelled with 35S, 51Cr, 57Co, 59Fe, 82Br, 86Rb, sup(99m)Tc, sup(113m)In, 125I and 169Yb. The radiometric test proves to be more rapid than the conventional one for the sterility testing of such radiopharmaceuticals. Detection time is between 6-21 hours. In the case of 131I-labelled radiopharmaceuticals and in the case of chlormerodrin-Hg-203, it was found an interference due to volatile species (sup(131m)Xe in the case of 131I and some volatile mercury form in the case of chlormerodrin). In these cases it would be possible to carry out the radiometric sterility test after separation of the microorganisms from the radioactive material (by filtration). The limulus lysate method can be employed for control of various pyrogen-prone raw materials and radiopharmaceuticals. Such method is the only method at present available for detecting the low level pyrogen contamination in intrathecal injections. The limulus test is more rapid than the rabbit test

  8. Measurement of the hyperfine splitting of {sup 133}Cs atoms in superfluid helium

    Energy Technology Data Exchange (ETDEWEB)

    Imamura, K., E-mail: kimamura@riken.jp [RIKEN Nishina Center (Japan); Furukawa, T. [Tokyo Metropolitan University, Department of Physics (Japan); Yang, X. F. [Peking University, School of Physics (China); Mitsuya, Y. [Meiji University, Department of Physics (Japan); Fujita, T. [Osaka University, Department of Physics (Japan); Hayasaka, M. [Tokyo Gakugei University, Department of Physics (Japan); Kobayashi, T. [RIKEN Center for Advanced Photonics (Japan); Hatakeyama, A. [Tokyo University of Agriculture and Technology, Department of Applied Physics (Japan); Ueno, H. [RIKEN Nishina Center (Japan); Odashima, H. [Meiji University, Department of Physics (Japan); Matsuo, Y. [Hosei University, Department of Advanced Sciences (Japan)

    2015-04-15

    We have been developing a new nuclear laser spectroscopy method named “OROCHI” (Optical RI-atom Observation in Condensed Helium as Ion-catcher). OROCHI utilizes superfluid helium (He II) not only as an efficient stopping medium of highly energetic ions but also as a host matrix of in-situ atomic laser spectroscopy. Using these characteristic of He II, we produce atomic spin polarization and measure Zeeman and hyperfine structure (HFS) splitting using laser-RF (radio frequency) / MW (microwave) double resonance method. From the measured energy splittings, we can deduce nuclear spins and moments. So far, we have conducted a series of experiments using both stable ({sup 85,87}Rb, {sup 133}Cs, {sup 197}Au, {sup 107,109}Ag) and unstable isotopes ({sup 84,86}Rb) to confirm the feasibility of OROCHI method, especially observing Zeeman resonance and determining nuclear spins. The measurement of HFS splitting of atoms introduced into He II is indispensable to clarify the nuclear properties by deducing nuclear moments as well as the study of nuclear spins. For this purpose, we perform a precision measurement of HFS of {sup 133}Cs atoms immersed in He II using laser ablation technique. In this paper, we describe the result of the experiment.

  9. Upper limit of 83Rb release into the gas phase from a 83mKr calibration source for the XENON project

    International Nuclear Information System (INIS)

    The isomer 83mKr with its half-life of 1.83 h is an ideal calibration source for a liquid noble gas dark matter experiment like the XENON project. For such a low counting experiment the possibility that traces of the much longer living mother isotop 83Rb (t1/2 = 86.2 d) contaminate the detector must be avoided. In this work the 83Rb release of a 1.8 MBq strong 83Rb source embedded in zeolite spheres has been investigated by searching for the characteristic 83Rb γ lines with the ultra-sensitive germanium detector Gator at LNGS after collecting a possible 83Rb release in a cryogenic trap for about 10 days. No signal has been found. The corresponding upper limit for the 83Rb release of 200μBq means, that such a 83Rb source as 83mKr generator can be used at the XENON project as well as for the KATRIN experiment. % without the danger of contaminating the detector with 83Rb. The germanium detector also allowed to set upper limits on the possible release of the isotopes 84Rb and 86Rb, which were produced during the 83Rb production at the Rez cyclotron to some amount.

  10. Insulin regulation of Na/K pump activity in rat hepatoma cells

    International Nuclear Information System (INIS)

    Insulin rapidly increases Na/K pump activity in HTC rat hepatoma cells in tissue culture, as measured by the ouabain-sensitive influx of the potassium analogue 86Rb+. Increased influx is observed within minutes and is maximal (70% above control) within 1-2 h. The effect appears to be mediated by the insulin receptors, as: the concentration dependence on insulin is identical to that for insulin induction of tyrosine aminotransferase and stimulation of 2-aminoisobutyric acid transport, proinsulin is 6% as potent as insulin, and the effect is blocked by anti-receptor antibodies. The early stimulation of potassium influx is not blocked by cycloheximide and is not associated with an increased number of pump sites as measured by 3H-ouabain binding. The insulin effect is blocked by amiloride, which blocks sodium influx, and is mimicked by the sodium ionophore monensin, which increases sodium influx and intracellular accumulation. Insulin also rapidly increases the initial rate of 22Na+ influx, suggesting that insulin may enhance Na/K pump activity, in part, by increasing intracellular sodium concentration. Incubation of HTC cells with insulin for 24 h causes complete unresponsiveness to the insulin induction of transaminase and stimulation of amino acid transport, a phenomenon mediated by postbinding mechanisms. In contrast, similar incubation with insulin does not cause unresponsiveness to the insulin stimulation of Na/K pump activity. Therefore, the site of regulation of responsiveness to insulin must be distal to, or separate from, those events causing stimulation of ion fluxes

  11. Insulin regulation of Na/K pump activity in rat hepatoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Gelehrter, T.D.; Shreve, P.D.; Dilworth, V.M.

    1984-05-01

    Insulin rapidly increases Na/K pump activity in HTC rat hepatoma cells in tissue culture, as measured by the ouabain-sensitive influx of the potassium analogue 86Rb+. Increased influx is observed within minutes and is maximal (70% above control) within 1-2 h. The effect appears to be mediated by the insulin receptors, as: the concentration dependence on insulin is identical to that for insulin induction of tyrosine aminotransferase and stimulation of 2-aminoisobutyric acid transport, proinsulin is 6% as potent as insulin, and the effect is blocked by anti-receptor antibodies. The early stimulation of potassium influx is not blocked by cycloheximide and is not associated with an increased number of pump sites as measured by /sup 3/H-ouabain binding. The insulin effect is blocked by amiloride, which blocks sodium influx, and is mimicked by the sodium ionophore monensin, which increases sodium influx and intracellular accumulation. Insulin also rapidly increases the initial rate of /sup 22/Na+ influx, suggesting that insulin may enhance Na/K pump activity, in part, by increasing intracellular sodium concentration. Incubation of HTC cells with insulin for 24 h causes complete unresponsiveness to the insulin induction of transaminase and stimulation of amino acid transport, a phenomenon mediated by postbinding mechanisms. In contrast, similar incubation with insulin does not cause unresponsiveness to the insulin stimulation of Na/K pump activity. Therefore, the site of regulation of responsiveness to insulin must be distal to, or separate from, those events causing stimulation of ion fluxes.

  12. Theoretical Study of the Molecular and Electronic Structures of CPDT-TCNQ and Its Difluoro and Dimethyl Derivatives

    Institute of Scientific and Technical Information of China (English)

    GONG Zhi-Jun; LI Qian-Shu

    2006-01-01

    CPDT-TCNQ and its derivatives are good candidates for charge-transfer acceptors.In this work, the electronic ground and excited states of CPDT-TCNQ as well as its difluoro and dimethyl derivatives are studied. The ground state optimized structures and energies were obtained using a restricted (closed-shell) density functional theory (DFT) as approximated by the various hybrid functionals (RB3LYP, RB3P86, RB3PW91). The 6-31G** and 6-31+G** basis sets were employed in calculations. All derivatives are planar and exhibit a quinoid structure in their electronic ground states. The energy and oscillator strength of the first 15 singlet-singlet electronic transitions have been investigated by applying the time-dependent density functional theory (TD-DFT) approximations to the correspondingly optimized ground state geometries. The results show the strongest absorption in electronic spectra of molecules due to the HOMO-LUMO electronic transition of the thiophene backbone.

  13. Lack of interaction between digoxin and quinidine in cultured heart cells

    International Nuclear Information System (INIS)

    Previous investigations have raised the possibility that the digoxin-quinidine interaction is associated with a reduction in the positive inotropic effect of digoxin due to displacement of digoxin from cardiac as well as skeletal muscle. To circumvent some of the complexities presented by intact animal models, this interaction was investigated in cultured chick embryo ventricular cells. Quinidine, even at relatively high concentrations (10(-4)--2 x 10(-3) M), did not significantly affect positive inotropic effects of digoxin and did not protect against cellular contracture induced by toxic digoxin concentrations, despite preincubation of cells with quinidine for 60 min. The effects of digoxin on monovalent cation transport, as judged by active uptake of the K analog 86Rb, were also not altered by 10(-4) M to 2 x 10(-3) M quinidine. These data suggest that quinidine does not displace digoxin from Na, K adenosine triphosphatase binding sites in this preparation. Although these data must be extrapolated to the intact animal with caution, our findings suggest that changes in digoxin clearance are more likely of primary importance in the digoxin-quinidine interaction, and indicate that the approximately 2-fold increase in serum digoxin concentration observed after addition of quinidine would be expected to have direct effects on myocardial cells comparable with those seen with increased digoxin concentration in the absence of quinidine

  14. Development of NIRS models to predict protein and amylose content of brown rice and proximate compositions of rice bran.

    Science.gov (United States)

    Bagchi, Torit Baran; Sharma, Srigopal; Chattopadhyay, Krishnendu

    2016-01-15

    With the escalating persuasion of economic and nutritional importance of rice grain protein and nutritional components of rice bran (RB), NIRS can be an effective tool for high throughput screening in rice breeding programme. Optimization of NIRS is prerequisite for accurate prediction of grain quality parameters. In the present study, 173 brown rice (BR) and 86 RB samples with a wide range of values were used to compare the calibration models generated by different chemometrics for grain protein (GPC) and amylose content (AC) of BR and proximate compositions (protein, crude oil, moisture, ash and fiber content) of RB. Various modified partial least square (mPLSs) models corresponding with the best mathematical treatments were identified for all components. Another set of 29 genotypes derived from the breeding programme were employed for the external validation of these calibration models. High accuracy of all these calibration and prediction models was ensured through pair t-test and correlation regression analysis between reference and predicted values. PMID:26258697

  15. Mineral cycling in a young Douglas fir forest stand

    International Nuclear Information System (INIS)

    Radiotracers for phosphorus (32P), potassium (substitute 86Rb), and calcium (45Ca) were used to follow the turnover of nutrients entering the forest floor with rainwash. Phosphorus moved readily through the forest floor to be strongly retained by the acid mineral soil. Considerable phosphorus absorbed by trees was correlated with forest-floor leachate contents and became concentrated in physiologically active tissues. Losses with litterfall and rainwash were nondetectable; this indicated high retention and internal redistribution. Rapid movement toward sinks appeared to be characteristic. In contrast, the larger amounts of calcium added were absorbed by the exchange capacity of the forest floor, with subsequent small but relatively unrestricted movement through the soil and trees. Both calcium and potassium absorption by trees was correlated with mineral-soil leachate contents. Annual potassium mobility fell between that of phosphorus and calcium and appeared to be more dependent on temperature and moisture conditions of the forest ecosystem, with potassium moving rapidly in subcycles during wet periods of the growing season

  16. Measurement of the hyperfine splitting of 133Cs atoms in superfluid helium

    International Nuclear Information System (INIS)

    We have been developing a new nuclear laser spectroscopy method named “OROCHI” (Optical RI-atom Observation in Condensed Helium as Ion-catcher). OROCHI utilizes superfluid helium (He II) not only as an efficient stopping medium of highly energetic ions but also as a host matrix of in-situ atomic laser spectroscopy. Using these characteristic of He II, we produce atomic spin polarization and measure Zeeman and hyperfine structure (HFS) splitting using laser-RF (radio frequency) / MW (microwave) double resonance method. From the measured energy splittings, we can deduce nuclear spins and moments. So far, we have conducted a series of experiments using both stable (85,87Rb, 133Cs, 197Au, 107,109Ag) and unstable isotopes (84,86Rb) to confirm the feasibility of OROCHI method, especially observing Zeeman resonance and determining nuclear spins. The measurement of HFS splitting of atoms introduced into He II is indispensable to clarify the nuclear properties by deducing nuclear moments as well as the study of nuclear spins. For this purpose, we perform a precision measurement of HFS of 133Cs atoms immersed in He II using laser ablation technique. In this paper, we describe the result of the experiment

  17. Increased activity of digoxin-like substance in low-renin hypertension in acromegaly

    Energy Technology Data Exchange (ETDEWEB)

    Soszynski, P.; Slowinska-Srzednicka, J.; Zgliczynski, S. (Medical Center for Postgraduate Education, Warsaw (Poland))

    1990-01-01

    Arterial hypertension is common in acromegaly, but the pathogenesis of this complication remains unknown. To determine the role of an endogenous Na,K pump inhibitor/digoxin-like substance (DLS) in the pathogenesis of hypertension in acromegaly 76 subjects: 28 with acromegaly, 20 with essential hypertension and 28 healthy controls were studied. Serum DLS was measured with the use of radioimmunoassay and bioassay by the inhibition of digoxin-sensitive erythrocyte 86-Rb uptake. In acromegaly, the activity of DLS was significantly increased and plasma renin activity decreased in the hypertensive group, as compared with that of the normotensive group and controls. Moreover, DLS was elevated in the low-renin group of essential hypertension, as compared with that of the normal/high-renin group or controls. The activity of DLS correlated positively with mean arterial pressure and negatively with plasma renin activity, but not with growth hormone levels. In conclusion, an endogenous sodium pump inhibitor/digoxin-like substance may play a role in the pathogenesis of low-renin hypertension in acromegaly.

  18. Limits on the release of Rb isotopes from a zeolite based 83mKr calibration source for the XENON project

    CERN Document Server

    Hannen, V; Arneodo, F; Baudis, L; Beck, M; Bokeloh, K; Ferella, A D; Giboni, K; Lang, R F; Lebeda, O; Ortjohann, H -W; Schumann, M; Spalek, A; Venos, D; Weinheimer, C

    2011-01-01

    The isomer 83mKr with its half-life of 1.83 h is an ideal calibration source for a liquid noble gas dark matter experiment like the XENON project. However, the risk of contamination of the detector with traces of the much longer lived mother isotop 83Rb (86.2 d half-life) has to be ruled out. In this work the release of 83Rb atoms from a 1.8 MBq 83Rb source embedded in zeolite beads has been investigated. To do so, a cryogenic trap has been connected to the source for about 10 days, after which it was removed and probed for the strongest 83Rb gamma-rays with an ultra-sensitive Germanium detector. No signal has been found. The corresponding upper limit on the released 83Rb activity means that the investigated type of source can be used in the XENON project and similar low-background experiments as 83mKr generator without a significant risk of contaminating the detector. The measurements also allow to set upper limits on the possible release of the isotopes 84Rb and 86Rb, traces of which were created alongside ...

  19. Phorbol ester-induced protein secretion in rat parotid gland. Relationship to the role of inositol lipid breakdown and protein kinase C activation in stimulus-secretion coupling

    Energy Technology Data Exchange (ETDEWEB)

    Putney, J.W. Jr.; McKinney, J.S.; Aub, D.L.; Leslie, B.A.

    1984-09-01

    When added to rat parotid gland slices incubated in vitro, 4 alpha-phorbol-dibutyrate (PDBu) induced a dose-dependent increase in protein secretion, but did not affect membrane permeability to K+ (as determined by /sup 86/Rb efflux). The response to PDBu was unaffected by the removal of extracellular Ca2+ and was not markedly potentiated by incubation with the phosphodiesterase inhibitor, methylisobutylxanthine. PDBu did not activate phospholipase C breakdown of inositol lipids as shown by a failure to increase formation of soluble inositol phosphates. When applied in combination with the Ca2+ ionophore, ionomycin, a secretory rate was obtained that was greater than the predicted sum of rates obtained when the two drugs were given alone. These results, when taken with the reported results of others, are consistent with an action of PDBu in activating protein kinase C and suggest that this enzyme plays an important role in the pathway linking receptor activation to protein secretion, but not K+ flux, in the parotid gland.

  20. A simple assay for agonist-regulated Cl and K conductances in salt-secreting epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Venglarik, C.J.; Bridges, R.J.; Frizzell, R.A. (Univ. of Alabama, Birmingham (USA))

    1990-08-01

    We developed a convenient flux assay that permits simultaneous measurement of Cl and K conductance pathways in Cl-secreting epithelial cells. Monolayers of the colonic tumor cell line T84 were preloaded with 125I and 86Rb, and isotope effluxes were monitored by a sample-replace procedure. The adenosine 3',5'-cyclic monophosphate (cAMP)-mediated agonists forskolin and prostaglandin E2 increased I efflux with little effect on Rb efflux, whereas the Ca-mediated agonists ionomycin, A23187, and carbachol increased both I and Rb effluxes. Simultaneous determinations of I and Cl or Rb and K effluxes indicated that I and Rb provide good measures of the effluxes of Cl and K, respectively. Forskolin- and ionomycin-stimulated I effluxes were inhibited by the Cl-channel blockers diphenylamine-2-dicarboxylate (DPC), 5-nitro-2-(3-phenylpropyl-amino)benzoic acid (NPPB), and 2-(cyclopentyl-6,7-dichloro-2,3-dihydro-2-methyl-1-oxo-1H- inden-5-yl-oxy)acetic acid (IAA-94) and by high external K. The Rb efflux evoked by ionomycin was inhibited by the K-channel blockers Ba and charybdotoxin. These findings suggest that I and Rb effluxes provide qualitative estimates of agonist-stimulated Cl and K conductance pathways. Thus this method can provide a simple and relatively inexpensive screening assay for Cl and K conductances in cultured cells to assess the effects of agonist, blockers, or genetic manipulations.

  1. Feasibility of the instrumental neutron activation analysis of entire archaeological pottery. Part 1: Precision of the results and radiological safety of the process

    International Nuclear Information System (INIS)

    The feasibility of the instrumental neutron activation analysis of entire pieces of archaeological pottery, using low thermal neutron fluxes, is examined. The study takes into account the chemical elements relevant for archaeological investigations, as well as the degree of accuracy required for such kind of research. It is shown that after irradiation of a typical pottery sample of about 1 kg during 45 minutes, at a thermal flux of about 109 n.cm-2.s-1, analytical signals are obtained, by gamma spectrometry, with counting statistics better than 1%, for 76As, 131Ba, 141Ce, 60Co, 134Cs, 181Hf, 140La, 24Na, 122Sb, 46Sc, 153Sm and 233Pa, whereas 51Cr, 152Eu, 42K, 86Rb, 175Yb and 65Zn can be detected with counting statistics within 1% and 2%. On the other hand, the statistics of measurement are relatively poor (orders of 3% - 10%) for 177Lu, 147Nd, 239Np, 160Tb and 181Ta. The feasibility of accomplishment reliable quantitative determinations, taking into account the complexity of the analysis of entire pieces of archaeological pottery is discussed, which involves factors such as high masses, as well as asymmetric and variable shapes. (orig.)

  2. Activity identification of peptide containing rat sodium pump α2 subunit M1-M2 extramembrane fragment in vitro%大鼠钠泵α2亚单位M1~M2膜外区多肽体外活性鉴定

    Institute of Scientific and Technical Information of China (English)

    张明娟; 杨军; 强磊; 王蓉; 宋亚燔; 牛小麟

    2008-01-01

    本文旨在研究含大鼠钠泵α2亚单位M1~M2膜外区的多肽片段(peptide contmning rat sodium pump α2 subunit M1-M21extramembrane fragment,RES2衍生物)是否具有与内源性钠泵抑制因子结合以及改善钠泵α亚单位活性的作用.采用Fmoc固相合成法合成多肽(Leu-Ala-Ala-Met-Glu-Asp-Glu-Pro-Ser-Asn-Asp-Asn-Gly-Gly-Gly-Ser),产物经高效液相色谱技术纯化,并进行质谱分析.采用放射性配体.受体结合法检测其与哇巴因的结合能力,进而采用人红细胞86Rb摄取实验检测其生物学活性.结果显示,RES2衍生物与3H-哇巴因具有一定的结合能力,其平衡解离常数(Kd)为38.46 nmol/L,IC50为6.353nmol/L.RES2衍生物可以阻断哇巴因对红细胞膜Na+,K+-ATPase的抑制作用,使红细胞86Rb摄取率从38.53%上升到83.69%左右,并呈一定的剂量依赖关系.因此,RES2衍生物不仅具有体外结合活性,而且具有改善钠泵活性的生物学效应,为其成为有效的抗高血压药物提供科学依据.%In order to explore the activity of a peptide containing rat sodium pump α2 subunit M I-M2 extramembrane fragment (RES2derivative) in vitro, the peptide (Len-Ala-Ala-Met-Glu-Asp-Glu-Pro-Ser-Asn-Asp-Asn-Gly-Gly-Gly-Ser) was synthesized by pep-tide synthesizer with Fmoc method and purified by high performance liquid chromatography (HPLC). Its binding activity wasidentified by radioligand-receptor binding assay (RRA) and its bioactivity was measured by erythrocyte 86Rb uptake. The results ofsaturation binding experiment and competitive binding experiment showed that the synthesized RES2 derivative had the capability tobind to 3H-ouabain. The dissociation constant (Kd) was 38.46 nmol/L and IC50 was 6.353 nmol/L. Erythrocyte 86Rb uptake experimentshowed that the RES2 derivative blocked the inhibitory effect of ouabain on the sodium pump on erythrocyte membrane in a dose-dependent manner. The results showed that the RES2 derivative is capable of binding to ouabain and

  3. Dextromethorphan and its metabolite dextrorphan block alpha3beta4 neuronal nicotinic receptors.

    Science.gov (United States)

    Hernandez, S C; Bertolino, M; Xiao, Y; Pringle, K E; Caruso, F S; Kellar, K J

    2000-06-01

    Dextromethorphan (DM), a structural analog of morphine and codeine, has been widely used as a cough suppressant for more than 40 years. DM is not itself a potent analgesic, but it has been reported to enhance analgesia produced by morphine and nonsteroidal anti-inflammatory drugs. Although DM is considered to be nonaddictive, it has been reported to reduce morphine tolerance in rats and to be useful in helping addicted subjects to withdraw from heroin. Here we studied the effects of DM on neuronal nicotinic receptors stably expressed in human embryonic kidney cells. Studies were carried out to examine the effects of DM on nicotine-stimulated whole cell currents and nicotine-stimulated (86)Rb(+) efflux. We found that both DM and its metabolite dextrorphan block nicotinic receptor function in a noncompetitive but reversible manner, suggesting that both drugs block the receptor channel. Consistent with blockade of the receptor channel, neither drug competed for the nicotinic agonist binding sites labeled by [(3)H]epibatidine. Although DM is approximately 9-fold less potent than the widely used noncompetitive nicotinic antagonist mecamylamine in blocking nicotinic receptor function, the block by DM appears to reverse more slowly than that by mecamylamine. These data indicate that DM is a useful antagonist for studying nicotinic receptor function and suggest that it might prove to be a clinically useful neuronal nicotinic receptor antagonist, possibly helpful as an aid for helping people addicted to nicotine to refrain from smoking, as well as in other conditions where blockade of neuronal nicotinic receptors would be helpful. PMID:10869398

  4. The Na{sup +}/K{sup +} -pump in rat peritoneal mast cells: Some aspects of regulatio of activity and cellular fusion

    Energy Technology Data Exchange (ETDEWEB)

    Knudsen, T. [Odense Univ., Dept. of Pharmacology, Inst. of Medical Biology, The Faculty of Health Scineces (Denmark)

    1995-12-31

    The mast cell contains potent mediators of inflammation which are released after IgE-directed and non-IgE-directed stimulation of the cell. This highly specialized cell is therefore ascribed a role in the pathogenesis of disease states in which the inflammatory response plays a role for the development of the clinical symptoms. Thus, besides being of interest in basic research, studies of the cellular processes leading to release of inflammatory mediators from the mast cell also also have important clinical implications. The aim of the present work has been to document the existence of the Na{sup +}/K{sup +}-pump in rat peritoneal mast cells, to investigate the regulation of the pump activity and to explore whether modulation of the pump activity interferes with the cellular stimulus/secretion coupling mechanism. The Na{sup +}/K{sup +}-pump activity following stimulation of the mast cell was also investigated. The pump activity was assessed as the ouabain-sensitive cellular potassium uptake with {sup 86}Rb{sup +} as a tracer for potassium. The histamine release from the mast cell following IgE-directed and non-IgE-directed stimulation of the cell was used as a parameter of cellular degranulation. Histamine was measured by spectrofluorometry. Besides describing aspects of the function and regulation of the Na{sup +}/K{sup +}-pump in the rat peritoneal mast cell, this thesis points to the potential role of sodium transport mechanisms in mast cell physiology. Pharmacological manipulations of such transport mechanisms might in the future add to the treatment of allergic diseases. (au) 253 refs.

  5. Effects of Ouabain on Proliferation of Human Endothelial Cells Correlate with Na+,K+-ATPase Activity and Intracellular Ratio of Na+ and K.

    Science.gov (United States)

    Tverskoi, A M; Sidorenko, S V; Klimanova, E A; Akimova, O A; Smolyaninova, L V; Lopina, O D; Orlov, S N

    2016-08-01

    Side-by-side with inhibition of the Na+,K+-ATPase ouabain and other cardiotonic steroids (CTS) can affect cell functions by mechanisms other than regulation of the intracellular Na+ and K+ ratio ([Na+]i/[K+]i). Thus, we compared the dose- and time-dependences of the effect of ouabain on intracellular [Na+]i/[K+]i ratio, Na+,K+-ATPase activity, and proliferation of human umbilical vein endothelial cells (HUVEC). Treatment of the cells with 1-3 nM ouabain for 24-72 h decreased the [Na+]i/[K+]i ratio and increased cell proliferation by 20-50%. We discovered that the same ouabain concentrations increased Na+,K+-ATPase activity by 25-30%, as measured by the rate of (86)Rb(+) influx. Higher ouabain concentrations inhibited Na+,K+-ATPase, increased [Na+]i/[K+]i ratio, suppressed cell growth, and caused cell death. When cells were treated with low ouabain concentrations for 48 or 72 h, a negative correlation between [Na+]i/[K+]i ratio and cell growth activation was observed. In cells treated with high ouabain concentrations for 24 h, the [Na+]i/[K+]i ratio correlated positively with proliferation inhibition. These data demonstrate that inhibition of HUVEC proliferation at high CTS concentrations correlates with dissipation of the Na+ and K+ concentration gradients, whereas cell growth stimulation by low CTS doses results from activation of Na+,K+-ATPase and decrease in the [Na+]i/[K+]i ratio.

  6. Effects of atrial and brain natriuretic peptides upon cyclic GMP levels, potassium transport, and receptor binding in rat astrocytes

    International Nuclear Information System (INIS)

    The ability of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) to alter cyclic GMP levels and NaKCl cotransport in rat neocortical astrocytes was determined. At concentrations of 10(-9)-10(-6) M, rat ANP99-126 (rANF), rat ANP102-126 (auriculin B), and rat ANP103-126 (atriopeptin III) stimulated 6- to 100-fold increases in cyclic GMP levels. Porcine BNP (pBNP) and rat BNP (rBNP) were 20%-90% as effective as rANF over most of this concentration range, although 10(-6) M pBNP produced a greater effect than rANF. NaKCl cotransport as measured by bumetanide-sensitive 86Rb+ influx was not altered by exposure of astrocytes to 10(-6)M rANF, pBNP, or rBNP. Both pBNP and rBNP, as well as rat ANP103-123 (atriopeptin I) and des[gl18, ser19, gly20, leu21, gly22] ANF4-23-NH2 (C-ANF4-23) strongly competed for specific 125I-rANF binding sites in astrocyte membranes with affinities ranging from 0.03 to 0.4 nM, suggesting that virtually all binding sites measured at subnanomolar concentrations of 125I-rANF were of the ANP-C (ANF-R2) receptor subtype. These receptors are thought to serve a clearance function and may be linked to a guanylate cyclase activity that is chemically and pharmacologically distinct from that coupled to ANP-A (ANF-R1) receptors. ANP receptors on astrocytes may function in limiting the access of ANP and BNP to neurons involved in body fluid and cardiovascular regulation

  7. Mechanisms of mercurial and arsenical inhibition of tyrosine absorption in intestine of the winter flounder Pseudopleuronectus americanus

    Energy Technology Data Exchange (ETDEWEB)

    Musch, M.W.; Chauncey, B.; Schmid, E.C.; Kinne, R.K.; Goldstein, L. (Mount Desert Island Biological Laboratory, Salsbury Cove, ME (USA))

    1990-06-01

    Effects of HgCl2 (100 microM) para-chloromercuribenzene sulfonate (PCMBS) (1 mM), and oxophenylarsine (OPA) (250 microM) were determined on (a) the rate of Na pump activity in intact winter flounder intestine; (b) activity of Na-K-ATPase in tissue homogenates; and (c) Na-dependent and Na-independent uptake of tyrosine in brush border membrane vesicles. Initial rate of uptake (influx) of 86Rb from the serosal solution of tissues mounted in Ussing chambers, a measure of Na-K-ATPase activity in the intact cell, was inhibited by all three agents with differing time courses. Rapidly permeating HgCl2 inhibited influx to the same degree as ouabain at 30 min, whereas the effects of PCMBS and OPA required 90 min. Cell potassium was also measured as an indirect indicator of ATPase activity and cell membrane permeability. All three agents decreased cell K, although effects on cell K lagged behind those for inhibition of the ATPase. At the concentrations used in the Ussing chamber (or at one-tenth concentration), all agents completely inhibited Na-K-ATPase activity in enzyme assays performed with tissue homogenates. In contrast, only HgCl2 decreased Na-dependent uptake of tyrosine by brush border membrane vesicles. These results suggest that mercurial and arsenical effects on tyrosine absorption are due to inhibition of the Na-K-ATPase thus decreasing the driving force for the cellular uptake by the Na-tyrosine cotransport system. Direct effects on Na-tyrosine cotransport may play a role in the inhibition observed with HgCl2, but not for PCMBS or OPA.

  8. Membrane-Mediated Decrease in Root Exudation Responsible for Phorphorus Inhibition of Vesicular-Arbuscular Mycorrhiza Formation

    Science.gov (United States)

    Graham, James H.; Leonard, Robert T.; Menge, John A.

    1981-01-01

    The mechanism responsible for phosphorus inhibition of vesicular-arbuscular mycorrhiza formation in sudangrass (Sorghum vulgare Pers.) was investigated in a phosphorus-deficient sandy soil (0.5 micrograms phosphorus per gram soil) amended with increasing levels of phosphorus as superphosphate (0, 28, 56, 228 micrograms per gram soil). The root phosphorus content of 4-week-old plants was correlated with the amount of phosphorus added to the soil. Root exudation of amino acids and reducing sugars was greater for plants grown in phosphorus-deficient soil than for those grown in the phosphorus-treated soils. The increase in exudation corresponded with changes in membrane permeability of phosphorus-deficient roots, as measured by K+ (86Rb) efflux, rather than with changes in root content of reducing sugars and amino acids. The roots of phosphorus-deficient plants inoculated at 4 weeks with Glomus fasciculatus were 88% infected after 9 weeks as compared to less than 25% infection in phosphorus-sufficient roots; these differences were correlated with root exudation at the time of inoculation. For plants grown in phosphorus-deficient soil, infection by vesicular-arbuscular mycorrhizae increased root phosphorus which resulted in a decrease in root membrane permeability and exudation compared to nonmycorrhizal plants. It is proposed that, under low phosphorus nutrition, increased root membrane permeability leads to net loss of metabolites at sufficient levels to sustain the germination and growth of the mycorrhizal fungus during pre- and postinfection. Subsequently, mycorrhizal infection leads to improvement of root phosphorus nutrition and a reduction in membrane-mediated loss of root metabolites. PMID:16661955

  9. Influence of fentanyl and morphine on intestinal circulation

    Energy Technology Data Exchange (ETDEWEB)

    Tverskoy, M.; Gelman, S.; Fowler, K.C.; Bradley, E.L.

    1985-06-01

    The influence of fentanyl and morphine on the intestinal circulation was evaluated in an isolated loop preparation in 37 dogs anesthetized with pentobarbital intravenously. Selected intestinal segments were pumped with aortic blood at a constant pressure of 100 mm Hg. A mixture of /sup 86/Rb and 9-micron spheres labeled with /sup 141/Ce was injected into the arterial cannula supplying the intestinal loop, while mesenteric venous blood was collected for activity counting. A strong correlation was found between the clearances of rubidium and microspheres (r = 0.97, P less than 0.0001), suggesting that the shunting of 9-micron spheres through the intestines reflects the shunting of blood through nonnutritive vessels. Intravenous fentanyl decreased oxygen uptake (O/sub 2/up), and vascular resistance (VR), and increased blood flow (BF), rubidium and microsphere clearances (Cl-Rb, Cl-Sph, respectively), and permeability--surface area product (PS) in a dose-related fashion. Intravenous morphine in a dose of 1 mg X kg-1 increased Cl-Rb (nutritive BF) without changes in total (nutritive and nonnutritive) BF. This increase in nutritive BF is probably related to morphine-induced histamine release. Morphine in a dose of 5 mg X kg-1 was accompanied by vasoconstriction that was completely abolished by alpha-adrenoceptor blockade. The data suggest that morphine-induced intestinal vasoconstriction is mediated via a release of epinephrine, apparently from the adrenal medulla. It is concluded that changes in the intestinal circulation during anesthesia with narcotics might play a certain role in the cardiovascular homeostasis during anesthesia and surgery. An increase in oxygen content in portal venous blood, resulting from a decrease in intestinal oxygen uptake, should facilitate hepatic oxygenation.

  10. Cellular effects of beta-adrenergic and of cAMP stimulation on potassium transport in rat alveolar epithelium.

    Science.gov (United States)

    Saumon, G; Basset, G; Bouchonnet, F; Crone, C

    1989-07-01

    Alveolar fluid absorption is greatly enhanced by cAMP and by beta-adrenergic agonists via an increase in Na+ transport. Little is known about K+ homeostasis under these circumstances. We studied K+ transport across alveolar epithelium in isolated perfused rat lungs stimulated either by dibutyryl-cAMP or isoproterenol. K+ fluxes and the apparent permeability of 86Rb across the epithelium (alveoli to plasma) were interpreted according to a model involving two types of cells, B and L, distinguished by the location of Na+-K+-ATPases (basal and luminal). Water is considered to be absorbed by B cells in a solute-coupled process energized by a basolateral Na+-K+-ATPase that is stimulated by isoproterenol and cAMP. K+ transport out of the alveoli is due to the activity of a Na+-K+-ATPase located in the apical membrane of L cells. In the present study net transport rate of K+ was -0.5 +/- 0.15 nmol/s, n = 20 (out of alveoli) in control conditions. When the epithelium was stimulated by dibutyryl-cAMP (10(-4) mol/l) net absorption of K+ reversed to net 'secretion' into alveoli (3.2 +/- 0.31 nmol/s), fluid absorption was not stimulated. K+ 'secretion' was abolished by apical Ba2+, indicating it was due to opening of apical K+ channels. Basolateral ouabain reversed net K+ 'secretion' to net absorption indicating that K+ entry into alveoli was dependent on activity of B cell basolateral Na+-K+-ATPase (masking simultaneous K+ removal by apical L cell Na+-K+-pump). When larger concentrations of dibutyryl-cAMP (10(-3) mol/l) or when isoproterenol were used to stimulate the epithelium there was a tripling of fluid absorption.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2571117

  11. Cation transport in oxidant-stressed human erythrocytes: heightened N-ethylmaleimide activation of passive K+ influx after mild peroxidation.

    Science.gov (United States)

    Sheerin, H E; Snyder, L M; Fairbanks, G

    1989-07-24

    Normal and chronically dehydrated (hereditary xerocytosis) human red cells were subjected to mild peroxidative treatment (315 microM hydrogen peroxide (H2O2), 15 min) in the presence of azide. The subsequent expression of passive (ouabain-resistant) K+ transport activities was analyzed by measurement of 86Rb+ influx. Peroxidation of normal red cells did not affect basal K+ transport activity, but the increment in K+ influx elicited by 0.5 mM N-ethylmaleimide (NEM) was increased 3-fold. The enhanced K+ influx was chloride-dependent, but only partially inhibited by 0.1 mM furosemide. Stimulated activity declined progressively after NEM activation, but could be restored by a second NEM treatment. Prior conversion of hemoglobin to the carbonmonoxy form abolished the response to peroxide, while 200 microM butylated hydroxytoluene (BHT) exerted only partial inhibition, suggesting that the effect of H2O2 requires interaction of activated, unstable hemoglobin species with the membrane, but that lipid peroxidation is not sufficient. Peroxidation following NEM treatment also enhanced NEM activation, indicating that enhancement does not require altered NEM reactions with stimulatory or inhibitory sites. Passive K+ transport in hereditary xerocytosis red cells was not activated by NEM, with or without H2O2 pretreatment. The results demonstrate that modest peroxidative damage to red cells can heighten the activation of a transport system that is thought to be capable of mediating net K+ efflux and volume reduction in cells that express it. Models are proposed in which the effects of NEM, H2O2, cell swelling and other factors are mediated by conformational changes in a postulated subpopulation of anion channel (Band 3) molecules that bind the K+ transporter. PMID:2758051

  12. Chloride transport in a glioma cell line

    Energy Technology Data Exchange (ETDEWEB)

    Wolpaw, E.W.

    1984-01-01

    Maintenance of the extracellular environment is a major function of central nervous system astroglia. The transport of Cl/sup -/ across the cell membrane may be an integral part of this function, since Cl/sup -/ transport has been implicated in homeostasis of cell volume, pH, and extracellular K/sup +/ concentration. The work presented here investigated Cl/sup -/ transport in the glioma cell line LRM55. Results indicate that LRM55 cells are a good model for astroglia and that these cells contain three Cl/sup -/ transporters; a Cl/sup -//HCO/sub 3//sup -/ exchanger, a K/sup +//Cl/sup -/ cotransporter, and a Cl/sup -//SO/sub 4//sup 2 -/ exchanger. Ion transport studies measured the fluxes of Cl/sup -/ (as /sup 36/Cl/sup -/), K/sup +/ (as /sup 86/Rb/sup +/), and SO/sub 4//sup 2 -/ (as /sup 35/SO/sub 4//sup 2 -/). Cl/sup -/ flux was trans-simulated by Cl/sup -/ or HCO/sub 3//sup -/ and was inhibited by SITS or furosemide. External K/sup +/ stimulated Cl/sup -/ influx and external Cl/sup -/ stimulated Rb/sup +/ influx. Furosemide, but not SITS, inhibited the K/sup +//Cl/sup -/ cotransporter. High K/sup +/ medium increased cell volume and Cl/sup -/ content. Steady-state Cl/sup -/ concentration was at least twice that predicted from passive equilibration according to the Nernst equation. SO/sub 4//sup 2 -/ flux was trans-stimulated by SO/sub 4//sup 2 -/ or by Cl/sup -/. Cl/sup -/ was a competitive inhibitor of SO/sub 4//sup 2 -/ influx, but SO/sub 4//sup 2 -/ had no detectable effect on Cl/sup -/ influx or efflux. SO/sub 4//sup 2 -/ flux was inhibited by SITS or furosemide.

  13. Tectonic-and petrological interpretation of geochronological data on the basement at the Southeast border of the Quadrilatero Ferrifero, Minas Gerais, Brazil

    International Nuclear Information System (INIS)

    In an attempt to elucidate the regional geological evolution of the Ouro Preto and Mariana Districts, SE Quadrilatero Ferrifero, Minas Gerais, the available petrographical and structural data were interpreted together with 5 Rb/Sr and 7 K/Ar age determinations. Various types of metamorphic rocks were identified, and four phases of regional petrographic evolution could be distinguished: a) pre-metamorphic phase indicated by relict sedimentary and other textures; b) pre-deformation metamorphic phase, manifested by several minerals, such as microcline; c) major syntectonic crystallization phase associated with the main regional diastrophism, presumaby the Minas orogeny; d) post-deformation phase, with retrograde metamorphic minerals. With one exception, the apparent K/Ar ages obtained on biotites and amphiboles were concordant, ranging between 480 and 540 m.y., and can be referred to the terminal episodes of cooling in the Brazilian cycle. The Rb/Sr data obtained from 3 whole-rock analyses, when plotted on a Sr87/Sr86 - Rb87/Sr86 diagram, indicated an age of approximately 2.000m.y., with an initial Sr87/Sr86 ratio of 0.711. In this diagram, 2 other speciments from a single outerop plotted significantly above of the reference isochron of 2.000m.y. The apparent ages of about 2.700 m.y. are concordant with those for the Barbacena Group which occurs more toward the eastern side of the study area. From interpretation of petrographical and structural data, the following regional geological evolution can be inferred; a)2.700m.y. - Formation of rocks during the pre-Minas episode (diastrophism in Barbacena Series, or in the Bacao Complex; b) 2.000m.y. - Trans-Amazonian cycle - Main phase of syntectonic crystallization - Minas Series diastrophism; c) 500-600m.y. - Brazilian cycle - retrograde metamorphism, thrust tectonics and regional fracturing. (Author)

  14. Use of an external source (60Co) for 32P detection efficiency determination by the Cerenkov effect, in soil extracts

    International Nuclear Information System (INIS)

    The detection of 32P in aqueous extracts is usually made with the aid of a Geiger-Muller detector, with thin window and sample on a planchet. Presently the technique is being developed of detection of high energy beta particles emitters (32P, 42K, 86Rb) through the Cerenkov effect, using a commercial liquid scintillation system. This technique, despite being approximately 30 times more sensitive, has the inconvenience of varying the detection efficiency, mainly for color samples (soil extracts, for instance). From this stems the need for determining the detection efficiency for each sample. The internal standardization and channels ratio methods show a series of drawbacks, mainly the non-reutilization of the samples (1st method) and statistical uncertainty for low activity samples (2nd method). The elimination of these dreawbacks can be achieved through the utilization of the external standardization method. A 60Co source with 1,4 μCi activity has been adapted to the sample elevator of the detector system, and a comparison was made with the channels ratio method to evaluate the efficiency of 32P detection in soil extracts (P extraction and fractionation). The external standardization method showed to be more accurate, besides being influenced to a lesser degree by high voltage variation, sample volume and vial types. In the case of large samples, it is advisable to carry out detection in vials filled up to their full capacity; in the case of small samples, the whole volume should be transferred to the vials and completed up to 9 ml for nylon vials,10 ml for glass vials and 11 to 14 ml for polyethilene vials. On the other hand, plastic vials showed higher detection efficiency than ones. As to background radiation, the lowest rates were given by nylon vials and the highest by Beckman glass vials

  15. Hypotonicity and cell volume regulation in shark rectal gland: role of organic osmolytes and F-actin.

    Science.gov (United States)

    Ziyadeh, F N; Mills, J W; Kleinzeller, A

    1992-03-01

    Hypotonic stress (reduction of external tonicity from approximately 900 mosM and 295 mM NaCl to approximately 600 mosM and 135 mM NaCl) produced a relatively slow regulatory volume decrease (RVD) in dogfish shark (Squalus acanthias) rectal gland cells. During the 5-h experiment, cell K+ content remained unchanged; cell content of Na+ and Cl- dropped in the initial swelling phase by some 50% (reflecting the corresponding reduction in medium NaCl), and then remained unchanged during volume recovery phase. Also, cellular fluxes of 86Rb+ and urea were not affected by hypotonic stress. However, hypotonicity enhanced 10- to 20-fold the efflux of organic cell osmolytes taurine, betaine, and trimethyloxamine, and this accounted for the loss of osmotically obliged water during RVD. Enhancement of osmolyte efflux by hypotonic stress was abolished by readjusting the low-Na+ saline to isotonicity (approximately 900 mosM) with innocuous cations (choline+, Li+, or N-methylglucamine+). The results suggest that reduction of medium tonicity may be the determinant for the RVD response to hypotonic stress. The above properties of the observed RVD were also displayed when studying changes on cell F-actin at the basolateral cell face; hypotonic stress (medium with 135 mM NaCl) produced a rapid disappearance of fluorescence related to this cytoskeletal component, whereas no such changes were seen in low-Na+ salines made isotonic with choline or N-methylglucamine chloride nor in a saline made hyposmolar by omitting urea. Hence, hypotonicity is required to affect F-actin organization (depolymerization?). These changes of F-actin fluorescence are transient; they were completed within 5-10 min of hypotonic stress, and afterwards a gradual reconstitution of cell F-actin organization was seen. The above observations are consistent with the assumption that, in shark rectal gland cells, transient loss of cytoskeleton (F-actin) organization at the basolateral cell face, induced by hypotonicity

  16. Tumor necrosis factor-alpha induces Cl- and K+ secretion in human distal colon driven by prostaglandin E2.

    Science.gov (United States)

    Schmitz, H; Fromm, M; Bode, H; Scholz, P; Riecken, E O; Schulzke, J D

    1996-10-01

    Increased levels of tumor necrosis factor-alpha (TNF-alpha) have been found in, for example, inflammatory bowel disease (IBD) and human immunodeficiency virus (HIV) infection. To investigate a possible contribution of TNF-alpha to the pathogenesis of diarrhea in these diseases, ion transport of human distal colon was studied in the Ussing chamber in vitro. Serosal addition of TNF-alpha increased short-circuit current (Isc) of partially stripped tissues in a dose-dependent manner. Maximum Isc increase of 1.8 +/- 0.2 mumol.h-1.cm-2 was reached after 60 +/- 9 min at 200 ng/ml TNF-alpha. Bidirectional tracer flux measurements revealed that TNF-alpha induced an increase in 36 Cl serosal-to-mucosal flux, a decrease in 36Cl- mucosal-to-serosal flux, and a slight increase in K+ secretion indicated by an increased secretory 86Rb net flux. In the highly differentiated colonic epithelial cell line HT-29/B6, TNF-alpha had no effect on Isc, suggesting a mediation step located in the subepithelium. This supposition was supported by measurements on totally stripped human tissues, since removal of subepithelial layers by total stripping reduced the TNF-alpha effect by 40%. Experiments with tetrodotoxin (10(-6)M) indicated that the TNF-alpha effect was not mediated by the enteric nervous system. The specific 5-lipoxygenase blocker ICI-230487 (5 x 10(-8)M) also had no effect on TNF-alpha action. In contrast, inhibition of cyclooxygenase by indomethacin (10(-6)M inhibited the effect of TNF-alpha. Radioimmunoassay of prostaglandin E2 (PGE2) in the serosal bathing solution revealed an increase in PGE2 production/release after addition of TNF-alpha, which paralleled the Isc response. We conclude that TNF-alpha changed Cl- and K+ transport toward secretion in human colon. This effect was mediated by PGE2 produced by subepithelial cells. Thus TNF-alpha could be a mediator of diarrhea during intestinal inflammation, e.g., in IBD and HIV infection. PMID:8897887

  17. Vasopressin alters the mechanism of apical Cl- entry from Na+:Cl- to Na+:K+:2Cl- cotransport in mouse medullary thick ascending limb

    Energy Technology Data Exchange (ETDEWEB)

    Sun, A.; Grossman, E.B.; Lombardi, M.; Hebert, S.C. (Brigham and Women' s Hospital, Boston, MA (USA))

    1991-02-01

    Experiments were performed using in vitro perfused medullary thick ascending limbs of Henle (MTAL) and in suspensions of MTAL tubules isolated from mouse kidney to evaluate the effects of arginine vasopressin (AVP) on the K+ dependence of the apical, furosemide-sensitive Na{sup +}:Cl{sup {minus}} cotransporter and on transport-related oxygen consumption (QO{sub 2}). In isolated perfused MTAL segments, the rate of cell swelling induced by removing K+ from, and adding one mM ouabain to, the basolateral solution (ouabain(zero-K+)) provided an index to apical cotransporter activity and was used to evaluate the ionic requirements of the apical cotransporter in the presence and absence of AVP. In the absence of AVP cotransporter activity required Na{sup +} and Cl{sup {minus}}, but not K{sup +}, while the presence of AVP the apical cotransporter required all three ions. {sup 86}Rb{sup +} uptake into MTAL tubules in suspension was significant only after exposure of tubules to AVP. Moreover, {sup 22}Na{sup +} uptake was unaffected by extracellular K+ in the absence of AVP while after AVP exposure {sup 22}Na{sup +} uptake was strictly K{sup +}-dependent. The AVP-induced coupling of K{sup +} to the Na{sup +}:Cl{sup {minus}} cotransporter resulted in a doubling in the rate of NaCl absorption without a parallel increase in the rate of cellular {sup 22}Na{sup +} uptake or transport-related oxygen consumption. These results indicate that arginine vasopressin alters the mode of a loop diuretic-sensitive transporter from Na{sup +}:Cl{sup {minus}} cotransport to Na{sup +}:K{sup +}:2Cl{sup {minus}} cotransport in the mouse MTAL with the latter providing a distinct metabolic advantage for sodium transport. A model for AVP action on NaCl absorption by the MTAL is presented and the physiological significance of the coupling of K{sup +} to the apical Na{sup +}:Cl{sup {minus}} cotransporter in the MTAL and of the enhanced metabolic efficiency are discussed.

  18. Aespoe Hard Rock Laboratory. Final report of the first stage of the tracer retention understanding experiments

    International Nuclear Information System (INIS)

    The first stage of the Tracer Retention Understanding Experiments (TRUE) was performed as a SKB funded project. The overall objectives of TRUE are to develop the understanding of radionuclide migration and retention in fractured rock, to evaluate the realism in applied model concepts, and to assess whether the necessary input data to the models can be collected from site characterisation. Further, to evaluate the usefulness and feasibility of different model approaches, and finally to provide in situ data on radionuclide migration and retention. The strive for address with multiple approaches is facilitated through a close collaboration with the Aespoe Task Force on Modelling of Groundwater Flow and Transport of Solutes. The TRUE programme is a staged programme which addresses various scales from laboratory (22Na+ 47Ca2+ ≅ 85Sr2+ 86Rb+ ≅ 133Ba2+ The field tracer tests, using essentially the same cocktail of sorbing tracers as in the laboratory, were found to show the same relative sorbtivity as seen in the laboratory. A test using 137Cs showed that after termination of the test, some 63% of the injected activity remained sorbed in the rock. The interpretation of the in situ tests with sorbing tracers was performed using the LaSAR approach, developed as a part of the TRUE project. In this approach the studied flow path is viewed as a part of an open fracture. Key processes are spatially variable advection and mass transfer. The evaluation shows that laboratory diffusion data are not representative for in situ conditions, and that a close fit between field and modelled breakthrough is obtained only when a parameter group which includes diffusion is enhanced with a factor varying between 32-50 for all tracers and experiments (except for Cs) and 137 for Cs. Our interpretation is that the enhancement is mainly due to higher diffusivity/porosity and higher sorption in the part of the altered rim zone of the feature which is accessible over the time scales of the in

  19. ZZ MATXSLIBJ33, JENDL-3.3 based, 175 N-42 photon groups (VITAMIN-J) MATXS library for discrete ordinates multi-group

    International Nuclear Information System (INIS)

    1 - Description of program or function: JENDL-3.3 based, 175 neutron-42 photon groups (VITAMIN-J) MATXS library for discrete ordinates multi-group transport codes. Format: MATXS. Number of groups: 175 neutron, 42 gamma-ray. Nuclides: 337 nuclides contained in JENDL-3.3: H-1, H-2, He-3, He-4, Li-6, Li-7, Be-9, B-10, B-11, C-Nat, N-14, N-15, O-16, F-19, Na-23, Mg-24, Mg-25, Mg-26, Al-27, Si-28, Si-29, Si-30, P-31, S-32, S-33, S-34, S-36, Cl-35, Cl-37, Ar-40, K-39, K-40, K-41, Ca-40, Ca-42, Ca-43, Ca-44, Ca-46, Ca-48, Sc-45, Ti-46, Ti-47, Ti-48, Ti-49, Ti-50, V-Nat, Cr-50, Cr-52, Cr-53, Cr-54, Mn-55, Fe-54, Fe-56, Fe-57, Fe-58, Co-59, Ni-58, Ni-60, Ni-61, Ni-62, Ni-64, Cu-63, Cu-65, Ga-69, Ga-71, Ge-70, Ge-72, Ge-73, Ge-74, Ge-76, As-75, Se-74, Se-76, Se-77, Se-78, Se-79, Se-80, Se-82, Br-79, Br-81, Kr-78, Kr-80, Kr-82, Kr-83, Kr-84, Kr-85, Kr-86, Rb-85, Rb-87, Sr-86, Sr-87, Sr-88, Sr-89, Sr-90, Y-89, Y-91, Zr-90, Zr-91, Zr-92, Zr-93, Zr-94, Zr-95, Zr-96, Nb-93, Nb-94, Nb-95, Mo-92, Mo-94, Mo-95, Mo-96, Mo-97, Mo-98, Mo-99, Mo-100, Tc-99, Ru-96, Ru-98, Ru-99, Ru-100, Ru-101, Ru-102, Ru-103, Ru-104, Ru-106, Rh-103, Rh-105, Pd-102, Pd-104, Pd-105, Pd-106, Pd-107, Pd-108, Pd-110, Ag-107, Ag-109, Ag-110m, Cd-106, Cd-108, Cd-110, Cd-111, Cd-112, Cd-113, Cd-114, Cd-116, In-113, In-115, Sn-112, Sn-114, Sn-115, Sn-116, Sn-117, Sn-118, Sn-119, Sn-120, Sn-122, Sn-123, Sn-124, Sn-126, Sb-121, Sb-123, Sb-124, Sb-125, Te-120, Te-122, Te-123, Te-124, Te-125, Te-126, Te-127m, Te-128, Te-129m, Te-130, I-127, I-129, I-131, Xe-124, Xe-126, Xe-128, Xe-129, Xe-130, Xe-131, Xe-132, Xe-133, Xe-134, Xe-135, Xe-136, Cs-133, Cs-134, Cs-135, Cs-136, Cs-137, Ba-130, Ba-132, Ba-134, Ba-135, Ba-136, Ba-137, Ba-138, Ba-140, La-138, La-139, Ce-140, Ce-141, Ce-142, Ce-144, Pr-141, Pr-143, Nd-142, Nd-143, Nd-144, Nd-145, Nd-146, Nd-147, Nd-148, Nd-150, Pm-147, Pm-148, Pm-148m, Pm-149, Sm-144, Sm-147, Sm-148, Sm-149, Sm-150, Sm-151, Sm-152, Sm-153, Sm-154, Eu-151, Eu-152, Eu-153, Eu-154, Eu-155, Eu

  20. Aespoe Hard Rock Laboratory. Final report of the first stage of the tracer retention understanding experiments

    Energy Technology Data Exchange (ETDEWEB)

    Winberg, A. [Conterra AB, Uppsala (Sweden); Andersson, Peter [Geosigma AB, Uppsala (Sweden); Hermanson, Jan [Golder Grundteknik, Solna (Sweden); Byegaard, Johan [Chalmers Univ. of Technology, Goeteborg (Sweden). Dept. of Nuclear Chemistry; Cvetkovic, V. [Royal Inst. of Tech., Stockholm (Sweden). Dept. of Water Resources Engineering; Birgersson, Lars [Kemakta Konsult AB, Stockholm (Sweden)

    2000-03-15

    from its surrounding. The near proximity of the experimental array to the tunnel (10-15 m) implies a strong gradient (approximately 10%) in the structure, which has to be overcome and controlled during the experiments. A methodology for characterising fracture pore space using resin injection, excavation using large diameter coring and subsequent analysis with photo-microscopic and image analysis techniques was developed and tested at a separate site. The results show that epoxy resin can be injected over several hours, and that the estimated areal spread is in the order of square metres. The mean apertures of the two investigated samples were 239 and 266 microns, respectively. Assessment of spatial correlation show practical ranges in the order of a few millimetres. Performed tracer tests with conservative tracers in Feature A show that the feature is connected between its interpreted intercepts in the array. The parameters evaluated from the conservative tests; flow porosity, dispersivity and fracture conductivity are similar, indicating a relative homogeneity. Previous work has identified cationic tracers, featured by sorption through ion exchange, as the most suitable tracers for sorbing tracer experiments at ambient Aespoe conditions. Laboratory experiments on generic Aespoe material and site-specific material included batch sorption experiments on various size fractions of the geological material, and through diffusion experiments on core samples of variable length on a centimetre length scale. The sorbtivity was found to be strongly affected by the biotite content and the sorption was also found to increase with contact time. The sorbtivity was found to follow the relative order; {sup 22}Na{sup +} < {sup 47}Ca{sup 2+} {approx_equal} {sup 85}Sr{sup 2+} << {sup 86}Rb{sup +} {approx_equal} {sup 133}Ba{sup 2+} The field tracer tests, using essentially the same cocktail of sorbing tracers as in the laboratory, were found to show the same relative sorbtivity as seen

  1. ZZ FSXJ32, MCNP nuclear data library based on JENDL-3.2. ZZ FSXLIBJ33, MCNP nuclear data library based on JENDL-3.3

    International Nuclear Information System (INIS)

    1 - Description of program or function: - NEA-1424/03: JENDL-3.2 based MCNP library. Format: MCNP. Number of groups: Continuous energy cross section library. Nuclides: H, He, Li, Be, B, C, N, O, F, Na, Mg, Al, Si, P, S, Cl, Ar, K, Ca, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Ga, Ge, As, Se, Br, Kr, Rb, Sr, Y, Zr, Nb, Mo, Tc, Ru, Rh, Pd, Ag, Cd, In, Sn, Sb, Te, I, Xe, Cs, Ba, La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Hf, Ta, W, Pb, Bi, Ra, Ac, Th, Pa, U, Np, Pu, Am, Cm, Bk, Cf, Es, Fm. Temperatures: 293 K, 600 K, 900 K, 1200 K, 1500 K, 2000 K. Origin: JENDL-3.2. The temperature-dependent continuous energy cross section library for MCNP, FSXJ32, was prepared from JENDL-3.2 for a variety of applications in the field of atomic energy. - NEA-1424/06: April 2005: This is the DVD version of ZZ-FSXJ32 NEA-1424/03. - NEA-1424/07: This version differs from version NEA-1424/05 in the following: Index files xsdir.fsxlb331 and xsdir.fsxlb332 have been updated, since atomic weights were missing for 23 nuclides. JENDL-3.3 based MCNP library. Format: MCNP. Number of groups: Continuous energy cross section library. Nuclides: 337 nuclides contained in JENDL-3.3. H-1, H-2, He-3, He-4, Li-6, Li-7, Be-9, B-10, B-11, C-Nat, N-14, N-15, O-16, F-19, Na-23, Mg-24, Mg-25, Mg-26, Al-27, Si-28, Si-29, Si-30, P-31, S-32, S-33, S-34, S-36, Cl-35, Cl-37, Ar-40, K-39, K-40, K-41, Ca-40, Ca-42, Ca-43, Ca-44, Ca-46, Ca-48, Sc-45, Ti-46, Ti-47, Ti-48, Ti-49, Ti-50, V-Nat, Cr-50, Cr-52, Cr-53, Cr-54, Mn-55, Fe-54, Fe-56, Fe-57, Fe-58, Co-59, Ni-58, Ni-60, Ni-61, Ni-62, Ni-64, Cu-63, Cu-65, Ga-69, Ga-71, Ge-70, Ge-72, Ge-73, Ge-74, Ge-76, As-75, Se-74, Se-76, Se-77, Se-78, Se-79, Se-80, Se-82, Br-79, Br-81, Kr-78, Kr-80, Kr-82, Kr-83, Kr-84, Kr-85, Kr-86, Rb-85, Rb-87, Sr-86, Sr-87, Sr-88, Sr-89, Sr-90, Y-89, Y-91, Zr-90, Zr-91, Zr-92, Zr-93, Zr-94, Zr-95, Zr-96, Nb-93, Nb-94, Nb-95, Mo-92, Mo-94, Mo-95, Mo-96, Mo-97, Mo-98, Mo-99, Mo-100, Tc-99, Ru-96, Ru-98, Ru-99, Ru-100, Ru-101, Ru-102, Ru-103, Ru-104, Ru-106, Rh