Altered K+ movement in liver mitochondria from alloxan diabetic rats

Summary Potassium movements were monitored in liver mitochondria from control and alloxan diabetic rats with a cationic electrode. There was net accumulation of K⁺ after Ca²⁺ addition to the mitochondria with the diabetic but not with the control.     

The effect of cyclic AMP on Na+ and K+ transport systems in mouse macrophages

Summary Exogenous cyclic AMP (cAMP) inhibits the Na⁺, K⁺-cotransport system and stimulates the Na⁺, K⁺-pump and Na⁺, Ca²⁺ exchange in mouse macrophages. These effects are enhanced by inhibition of phosphodiesterase with methylisobutylxanthine (MIX). MIX alone showed little or no effect. A similar response was observed after stimulation of endogenous production of cAMP by isoproterenol.     

Phospholipid composition of cardiac (Na++K+)-ATPases from various species

Summary There is a difference in phospholipid composition of cardiac (Na⁺+K⁺)-ATPase preparations between species which are sensitive to ouabain and those which are not. Sphingomyelin is higher and phosphatidylcholine is lower in the enzymes from sensitive species than in those from insensitive ones. Lysophosphatidylcholine is detectable only in the latter preparations.     

Further evidence for the dissociation of digoxin-like immunoreactivity from Na+, K+-ATPase inhibitory activity

Summary The effects of adrenalectomy or nephrectomy, carried out one hour previously, on the levels of endogenous digitalis-like factors were determined in rat plasma. Factors were assayed by digoxin-like immunoreactivity and direct Na⁺, K⁺-ATPase inhibitory activity. Digoxin-like immunoreactivity significantly decreased one hour after bilateral ablation of adrenals, while Na⁺, K⁺-ATPase inhibitory activity remained unaltered. There were no changes in either activity one hour after bilateral nephrectomy. These results suggest that digoxin-like immunoreactivity may be derived from the adrenal gland or under adrenal control and the major substances detected by digoxin-like immunoreactivity and direct Na⁺, K⁺-ATPase inhibitory activity may be different.     

Distribution of (Na++K+)-ATPase in the hindgut ofGlossina morsitans morsitans Westwood

Summary (Na⁺+K⁺)-ATPase activity was higher in preparations from the ileum ofGlossina mortisans than in those from the rectum. This result suggests that the ileum as well as the rectum, may play a role in osmoregulation in the tsetse fly.     

Inhibition of the sodium pump does not cause a stoichiometric decrease of ATP-production in energy limited fish hepatocytes

In isolated goldfish hepatocytes underaerobic conditions the energy requirement for the sodium pump (calculated from Rb⁺ flux) is closely matched by the ouabain-sensitive fraction of oxygen consumption, whereas during in vitroanoxia (cyanide inhibition of the electron transport chain) the measured ATP demand of the sodium pump clearly exceeds ouabain-sensitive ATP production by anaerobic glycolysis. We conclude that when the energy status of cells is low, part or all of the ATP spared by the inhibition of a particular function may be used for fuelling other ATP-consuming functions.     

Purification from human plasma of endogenous dodium transport inhibitor(s)

Summary Na⁺, K⁺-ATPase inhibitors extracted from plasma of healthy human subjects displaced³H-ouabain binding to human erythrocytes and inhibited the Na⁺ efflux catalyzed by the Na⁺, K⁺-pump and unexpectedly the Na⁺, K⁺-cotransport system without alteration of the Na⁺, Na⁺-exchange or the Na⁺ passive permeability. This suggests the presence in healthy human plasma of endogenous factors with ouabain-like and furosemide-like activities.Acknowledgments. We are indebted to Dr M. A. Devynck for her advice on chemical measurements and to Dr R. P. Garay for his help with flux measurements     

Action of juvenile hormone on the follicle cells ofRhodnius prolixus: Evidence for a novel regulatory mechanism involving protein kinase C

Summary Juvenile hormone (JH) is known to act on the membranes of the follicle cells ofRhodnius, activating a specific Na⁺, K⁺-ATPase. This leads to a decrease in volume of the cells and the appearance of spaces between them (patency). The addition of an inhibitor of protein kinase C, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), to the medium in vitro inhibits the action of JH on the follicle cells. PDBU (phorbol-12,13-dibutyrate) mimics the action of JH in vitro and the response of the follicle cells to, PDBU is blocked by ouabain. It is concluded that the activation of protein kinase C is a required step in the chain of events leading to activation of the JH-dependent ATPase and set in train by the binding of JH to the membrane.     

Shortening velocity of single muscle cells isolated from a molluscan smooth muscle

Summary The maximal unloaded shortening velocity (V_max) of smooth muscle cells isolated from the pedal retractor muscle ofMytilus was more than twice as large as that of the whole muscle, suggesting the presence of extracellular components which resist the contraction of the whole muscle. The V_max of the isolated cells was almost constant at cell lengths ranging between 0.5 and 0.831₀ (1₀, optimal length for tension generation) indicating that the intracellular resistance to contraction is negligible within this range of lengths.     

Effect of various stressors on the levels of lipid peroxide,antioxidants and Na+, K+-ATPase actrivity in rat brain

The level of malondialdehyde (MDA), an index of lipid peroxidation, and the antioxidants superoxide dismutase (SOD) and glutathione (GSH), as well as the activity of Na⁺, K⁺-ATPase, were assessed in whole rat brain after immobilization, anemic hypoxia (NaNO₂) and 72 h starvation. The effect of these stressors on plasma glucose and corticosterone levels was also observed. Hypoxia and starvation stimulated the lipidj peroxide formation in braini as indicated by an increase in the level of MDA, being higher after starvation than hypoxia. Brain SOD activity was also increased in response to hypoxia and starvation while GSH content was only diminished ini hypoxia. However, neither MDA nor antioxidants were affected by immobilization. On the other hand, the activity of brain Na⁺, K⁺-ATPase was significantly increased by immobilization and hypoxia but decreased in starvation. A similar pattern of change was also observed in plasma glucose and corticosterone levels in response to these stressors. These results elucidate differences in the biochemical response of animals towards various types of stress, with increased lipid peroxide formation in hypoxia and starvation.     

The effect of a phorbol ester upon the cholinergic regulation of potassium permeability in the rat submandibular gland

Acetylcholine releases calcium from cytoplasmic stores and permits an influx of calcium in salivary acinar cells. The resultant rise in [Ca²⁺]_i causes an increase in potassium permeability which is an important part of the secretory response. We have investigated the effects of 12-0-tetradecanoyl phorbol-13-acetate, a potent activator of protein kinase C, upon this regulation of potassium permeability in superfused pieces of rat submandibular salivary gland. This compound inhibited the initial [Ca²⁺]_o-independent component of the response of acetylcholine but had no effect upon the subsequent [Ca²⁺]_o-dependent phase. This compound does not, therefore, appear to inhibit receptor-regulated calcium influx.     

Enzymology of NAD+ homeostasis in man

This review describes the enzymes involved in human pyridine nucleotide metabolism starting with a detailed consideration of their major kinetic, molecular and structural properties. The presentation encompasses all the reactions starting from the de novo pyridine ring formation and leading to nicotinamide adenine dinucleotide (NAD⁺) synthesis and utilization. The regulation of NAD⁺ homeostasis with respect to the physiological role played by the enzymes both utilizing NAD⁺ through the nonredox NAD⁺-dependent reactions and catalyzing the recycling of the common product, nicotinamide, is discussed. The salient features of other enzymes such as NAD⁺ pyrophosphatase, nicotinamide mononucleotide 5-nucleotidase, nicotinamide riboside kinase and nicotinamide riboside phosphorylase, described under miscellaneous, are likewise presented.Received 30 April 2003; received after revision 9 June 2003; accepted 20 June 2003     

Selective blockade of components of potassium activation inMyxicola axons

Summary The K⁺ conductance inMyxicola giant axons activates in two phases which are pharmacologically separable. The fast phase of K⁺ activation is specifically inhibited by 4-aminopyridine and by the substitution of D₂O for H₂O. We suggestMyxicola giant axons, like the amphibian node of Ranvier, may possess more than one variety of K⁺ channel.     

Selectivity of action between pyrethroids and combined DDT-pyrethroid insecticides on Na+ influx into mammalian neuroblastoma

Summary Several of the most active synthetic pyrethroid insecticides in the presence of sea anemone toxin II, induced a dose related influx of sodium ion into the C9 mouse neuroblastoma. The influx of sodium ion into this mammalian cell did not take place with a DDT analogue, EDO and several new combined DDT-pyrethroid insecticides, although these have been reported to cause excess sodium influx into arthropod axons, related to their insecticidal activity. This difference between species in the action of the new insecticides at the nerve sodium channel explains their low mammalian toxicity.     

Intracellular injection of cAMP and cGMP into snail neurones induces an increase in Na+-conductance

Summary Injection of cAMP and more rarely cGMP into the neurones of the snailHelix pomatia induces an increase in membrane conductance, membrane depolarization and excitation. The effect is theophylline-dependent and has a reversal potential near — 10 mV.     

Effects of methylene blue on electrical behavior of myenteric neurons

Summary Intracellular recording methods were used to investigate the action of methylene blue on electrical behavior of myenteric neurons in guinea pig small intestine. The neurophysiological studies were done in parallel with studies on contractile activity of the intestinal musculature. Methylene blue depolarized the membranes, increased the input resistance, augmented excitability and reduced postspike hyperpolarizing potentials in AH/Type 2 myenteric neurons. These effects, with the exception of suppression of postspike hyperpolarization, were reversed by exposure to elevated calcium. The mechanism of action of methylene blue appeared to be suppression of calcium-dependent potassium conductance in the neuronal membranes. The neuronal action of methylene blue was manifest as a release of excitatory neurontransmitter substances with evoked contraction of the small intestinal longitudinal muscle.     

Protective effect of Shigyaku-to,a traditional Chinese herbal medicine,on the infection of herpes simplex virus type 1 (HSV-1) in mice

The antiviral activity of Shigyaku-to (TJS-109), a traditional Chinese herbal medicine, was investigated in mice infected with herpes simplex virus type 1 (HSV-1). TJS-109 is a combination of the medicinal plant extracts fromZingiberis siccatum rhizoma,Aconiti tuber andGlycyrrhizae radix in a specific proportion. Mice infected with a 10 LD₅₀ dose of HSV-1 were treated with TJS-109 orally at doses of 1.25 to 20 mg/kg 2 days before, and 1 and 4 days after the infection. The treated groups had 80% (1.25 mg/kg), 40% (5 mg/kg) and 23% (20 mg/kg) mortality rates 25 days after the infection as compared with a 100% mortality rate in control mice treated with saline. When HSV-1 infected mice (recipients) received CD8⁺T cell fractions derived from spleens of mice treated with TJS-109 (donors), 70% of recipients survived, as compared with 0% survivors in the groups of mice treated with saline, B cell fractions, CD4⁺ T cell fractions or macrophage-enriched fractions prepared from the same donors. TJS-109 did not show any virucidal activities against HSV-1 or any virostatic activities on the growth of HSV-1 in Vero cells. These results suggest that TJS-109 protected mice exposed to lethal amounts of HSV-1 through the activation of CD8⁺ T cells.     

Calcium ionophore plus excision induce a large conductance chloride channel in membrane patches of human colon carcinoma cells HT-29cl.19A

In excised inside-out membrane patches of the human colon carcinoma HT-29cl.19A cells a large conductance (373±10 pS) chloride channel was found. Channel activity could only be observed after excision of patches from cells incubated with calcium ionophore. The channel was never observed in cell-attached patches. The channel was strongly voltage dependent, being open only between +30 and –30 mV clamp potentials. The selectivity sequence among anions, deduced from reversal potentials, was I>Br>Cl>F>gluconate. The P_Na/P_Cl was 0.09. Although a similar type of channel, has been described earlier, this is the first report stating its appearance in patches of intestinal epithelial cells requiring the combined action of Ca²⁺ ionophore and excision, suggesting its control by an intracellular compound.