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Bioassay method for polyene antibiotics based on the measurement of rubidium efflux from rubidium-loaded yeast cells.

A bioassay method for the polyene antibiotics nystatin and amphotericin B is proposed based on the measurement of the efflux of rubidium ions from a rubidium-loaded yeast culture challenged with the antibiotics. For this purpose a major proportion of the intracellular K(+) ions in a Saccharomyces cerevisiae culture has been substituted by Rb(+) ions. The rubidium leakage is measured by atomic absorption spectrophotometry, and a straight-line, dose-response correlation has been obtained for both antibiotics.

Amphotericin B

[New vistas on rubidium].

Rubidium salts have been used in human therapy since the end of the last century. The fact that an actual use in psychiatry has been considered is mainly due to Meltzer and al's works in 1969. The pharmacological studies do not reveal any psychopharmacological "provile" presently known; they point out that Rubidium has stimulant properties, whick, in certain conditions, can increase activity, central excitability and sometimes aggressiveness, among the animals used for the experiments. The hypothesis of a contingent antidepressant action of Rubidium is essentially based on the existence of properties opposite to those of Lithium. This is especially true in the biochemical field: Rubidium may enhance the release of Norepinephrine whereas Lithium has an opposite effect. The toxicological studies show that, because of some properties common to Rubidium and Potassium, it is necessary to control Potassium intake and to avoid that Rubidium replace too high a percentage of this ion: the extended half-life (about forty days in man) makes necessary the working up of chronical studies to evaluate the toxicity which is linked to its long-term accumulation. For Rubidium blood concentrations superior to 1 mEq/1., the first clinical studies seem to point out that an antidepressant action would exist. Nevertheless a certain delay of action is necessary to the onset of a therapeutic effect; no noticeable adverse effect has been detected.

Animals

Effects of rubidium on behavioral responses to methamphetamine and tetrabenazine.

Different groups of mice were injected subcutaneously every other day with rubidium chloride at three doses (0.41(50), 1.23(150) and 3.69(450) meq/kg (mg/kg)) or with saline as a control for a period of 2-3 weeks. Rubidium administered acutely did not affect spontaneous locomotor activities, while it tended to increase the activities when administered repeatedly though the increase was not statistically significant. The methamphetamine-induced hyperlocomotor activities were potentiated in the rubidium groups as compared with those in the saline group, this effect of ribidium being increased with prolongation of repeated administrations. Monotonic decreases in ambulation after tetrabenazine were not significantly affected in the rubidium-treated animals though the decreases were sometimes preceded by slight increases and recovery from the decrement tended to be more rapid. After tetrabenazine in the rubidium-treated groups, incidences of catalepsy were increased and jumping behavior and Straub tail responses occurred in a few cases. The results suggest that rubidium potentiates the excitatory action of methamphetamine on spontaneous locomotor activities, as contrasted with inhibitory influence of lithium.

Animals

[Influence of lithium and rubidium on exploratory behaviour and locomotor activity in isolated male mice (author's transl)].

The effect of lithium and rubidium on curiosity rearing, and locomotor activity in isolated and group-housed male mice (DBA/2 Han) was investigated. LiCl and RbCl were given with the drinking water (30 mol/l) during 3 weeks. The behaviour of the animals was tested before and after the chronical application of either lithium or rubidium. The results show that lithium was able to enhance the decreased curiosity of the isolated mice which now was no longer different from that of normal group-housed animals. The locomotor activity of the isolated animals was also increased by LiCl, while rearing was not altered. Rubidium even more decreased the already diminished curiosity of the 'fighting mice' but had no influence on rearing and locomotor activity. Three weeks after cessation of treatment the behaviour of the isolated mice returned to predrug levels. Neither lithium nor rubidium had any influence of the behaviour of animals kept in groups.

Animals

Effect of rubidium, lithium and cesium on brain ATPase and protein kinases.

The authors studied the effect of rubidium, lithium and cesium on the ATPase system and c-AMP protein kinase in brain. They demonstrated that rubidium could replace potassium in the Na+K-ATPase system, whereas lithium and cesium had no effect on this enzyme activity in the absence of potassium. K+-dependent ATPase was activated by even low rubidium concentrations; lithium and cesium inhibited it. None of three (rubidium, lithium and cesium) affected c-AMP protein kinase.

Adenosine Triphosphatases

Rubidium shows effects different from lithium on phosphatidylinositol metabolism in a cell line of human neuroblastoma.

In a SK-N-BE human neuroblastoma cell line the incubation of rubidium (1 and 10 mM) for 24 h significantly increased IP2 formation, whereas it apparently did not affect other inositol phosphates. In comparison to lithium (10 mM), which significantly enhanced inositolmonophosphate and IP2 accumulation following carbamoylcholine (1 mM) stimulation, rubidium at the same concentration, was unable to affect inositol phosphate accumulation. In conclusion, the present experiments show that rubidium, compared with lithium, shows a different profile on phosphoinositide metabolism since its main action is an increase in phosphatidylinositol turnover. These results may have some relevance to the use of rubidium as antidepressant in man.

Cell Line

Inhibition of activity in rats by rubidium chloride.

Effects of two dosages of rubidium chloride on exploration, locomotion, rearing and immobility were assessed for male and female rats in an exploration box and an open field. In contrast to previous findings, rubidium was found to decrease locomotion and rearing in the exploratory box, and also to decrease locomotion in the open field. Further research in a variety of experimental settings is required before the effects of rubidium on activity can be fully evaluated.

Animals

Effects of rubidium chloride on the course of manic-depressive illness.

Clinical studies of the effects of rubidium ions on the course of manic-depressive illness are reported. It seems that rubidium tends to increase the length of manic phases and possibly reduces the extremes of mood. Rubidium did not seem to produce any severe side effects in the dose administered, but it has a long biological half-life and caution is still required. Some details of the CSF, RBC, saliva and plasma and urine kinetics are also reported.

Adult

Onset of audiogenic seizures in rodents after intake of near-toxic doses of rubidium chloride.

Rats and mice that were not genetically or otherwise predisposed toward audiogenic seizures were rendered susceptible to auditory stimuli by chronic treatment with large doses of rubidium chloride (RbCl). Animals given drinking water, 0.03 N with respect to RbCl, for four weeks responded with convulsive seizures when exposed to signals of 2 to 22 kHz and 74 dbA. The rate of development of audiosusceptibility was dose-related. Less rubidium was needed if the diet was deficient in potassium. No differences were found between tissue rubidium in treated rats which became audiosensitive and their counterparts which remained resistant to auditory stimuli.

Acoustic Stimulation

Simultaneous determination of iron, zinc, selenium, rubidium, and cesium in serum and packed blood cells by neutron activation analysis.

We determined the iron, zinc, selenium, rubidium, and cesium concentrations in serum and packed blood cells by instrumental neutron activation analysis without chemical separations. Lyophilized samples were irradiated for 12 days at a flux of 10(13) neutrons-cm-2-s-1, mineralized by wet digestion, and measured two times with a high-resolution Ge(Li) detector--for 6 h about a month after the irradiation and for 15 h two or three months after the irradiation, The following values were obtained: 163 +/- 0.43 mg/liter (serum iron), 1025 +/- 136 mg/kg wet wt (packed cells iron), 1¿ +/- 0.20 mg/liter (serum zinc), 11.15 +/- 1.83 mg/kg wet wt (packed cells zinc), 0.13 +/- 0.02 mg/liter (serum selenium), 0.16 +/- 0.03 mg/kg wet wt (packed cells selenium), 0.17 +/- 0.04 mg/liter (serum rubidium), 4.28 +/- 0.98 mg/kg wet wt (packed cells rubidium), 0.74 +/- 0.20 microgram/liter (serum cesium), and 4.82 +/- 2.10 microgram/kg wet wt (packed cells cesium).

Blood Cells

The measurement of spleen perfusion in man: a non-invasive method using the ratio between Rubidium-81 and its decay product Krypton-81m.

A non-invasive method has been developed for measuring spleen perfusion in man. This involves recording the gamma-ray energy spectra over the spleen following the localisation of Rubidium-81 within the organ by injecting intravenously labelled heat denatured red cells. The spectra are analysed to provide the ratio of Rubidium-81 to its radioactive decay product Krypton-81 m. This ratio is dependent on the rate of perfusion through the organ. The difficulties encountered in applying this steady state method for monitoring tissue perfusion are illustrated and practical means for their solution presented. The results of applying this method to patients with various splenic disorders are presented and compared with those obtained by other workers using different monitoring techniques.

Humans

Effects of diazepam on photosynthesis, respiration, rubidium uptake, and finestructure of Scenedesmus obliquus in synchronous cultures.

Effects of diazepam (Valium) on photosynthesis, chlorophyll/photosynthesis ratios, respiration, uptake of rubidium ions, and ultrastructure of Scenedesmus obliquus synchronized by a light-dark regimen of 14:10 hrs were determined. 80 and 160 muM diazepam, added to the nutrient medium at the start of the light-dark change (i.e., start of the cell cycle) gradually reduced rates of photosynthesis, below the initial rates from the beginning of the experiment. Contents of chlorophyll, however, remained nearly unaffected. Consequently, the diazepam-treated cells had a higher chlorophyll/photosynthesis ratio--also with regard to respiration in order to calculate the gross photosynthesis. The occurrence of photorespiration cannot be assumed. The net influx of rubidium was slightly reduced by 100 muM diazepam 0.5 and 2.0 hrs after the start of the cell cycle and was strongly inhibited after 5 to 14 hrs. 80 and 160 muM diazepam caused separation of thylakoids, formation of giant mitochondria and enlargement of vacuoles.

Adenosine Triphosphate

Possible mechanism of rubidium-induced hyperactivity in the rat.

This study suggests that replacement of intracellular potassium by rubidium ions might lower the resting membrane potential. Thus rubidium-treated rats were more responsive to depolarizing influences and generated more cyclic AMP in the brainstem and consequently the behavioral changes.

Animals

Investigations on trace elements in normal and senile cataractous lenses. Activation analysis of copper, zinc, manganese, cobalt, rubidium, scandium, and nickel.

A total of 125 human lenses (97 senile cataractous and 28 clear) were analyzed by means of neutron activation analysis for trace element concentrations of copper, zinc, manganese, cobalt, rubidium, scandium, and nickel. Tt was found that the rubidium concentration remarkably decreased and the copper concentration moderately increased with the progression of cataract. The concentrations of manganese, zinc, and cobalt showed some fluctuations without a definite increase or decrease, and the scandium and nickel content was found to be very low.

Age Factors

Effect of rubidium on responses of rabbit vas deferens to transmural stimulation and to noradrenaline.

The effects of 2 mM RbCl on adrenergic transmission in rabbit vas deferens were investigated. Rubidium, but not potassium, markedly potentiated the sustained, secondary contractile response to transmural stimulation at 2--16 Hz. This effect was not abolished by indomethacin. Significant spontaneous desensitization to (--)-noradrenaline occurred, but this was prevented by the addition of 2 mM RbCl or KCl. Addition of 2 mM RbCl, but not of 2 mM KCl, significantly increased the amount of [3H] (+/-)-metaraminol released on transmural stimulation at 5 Hz, without affecting the uptake of [3H] (+/-)-metaraminol over a 30 min period. It is suggested that rubidium mainly potentiated responses of rabbit vas deferens to transmural stimulation by increasing transmitter release, possibly as a result of prolongation of the action potential.

Animals

Alcohol consumption in rats treated with lithium carbonate or rubidium chloride.

Wistar-NTRU rats, offered a free choice between tap water and a 10% ethanol solution (v/v) in the absence of reinforcement, were injected for five days with Li2 CO3, RbCl or placebo. Lithium-treated group consumed 25% more liquid per day but chose to take 14.5% less alcohol than controls (p less than 0.05). By contrast, rubidium-treated animals consumed 15% less liquid but 70% more alcohol than control animals (p less than 0.005). Rubidium-treated rats were strikingly more active than the other two groups: their motility index was 60.0 as compared to 33.6 for lithium-treated and 29.4 for control rats. Serum glucose and urea nitrogen concentration were not significantly affected by the treatment but serum alcohol content was low in lithium-treated and high in rubidum-treated animals.

Alcohol Drinking

Effects of replacing medium sodium by choline, caesium, or rubidium, on water and ion contents of renal cortical slices.

1. Renal cortical slices from rat, rabbit, and guinea-pig were incubated in media in which choline, caesium or rubidium replaced sodium.2. Slices of rabbit and guinea-pig renal cortex incubated in oxygenated choline Ringer decreased in volume initially and did not swell over 3 hr at 25 degrees C. There was a steady loss of potassium. Inhibition of metabolism (N(2) + 1 mM iodoacetamide) caused some swelling. Ouabain, 10 mM, in choline Ringer affected neither loss of potassium nor tissue water content.3. Slices of rat renal cortex similarly incubated in choline Ringer swelled over 3 hr at 25 degrees C whether or not metabolism was inhibited; ouabain (15 mM) affected neither tissue potassium loss nor tissue water content.4. Incubation in choline Ringer containing either 0.2 mMp-chloromercuribenzoic acid, or 1 mM ethacrynic acid increased the tissue water content of guinea-pig renal cortical slices.5. Depletion of cellular potassium (by preliminary incubation in oxygenated potassium-free sodium Ringer with 10 mM ouabain at 30 degrees C) resulted in increased tissue water content when rabbit renal cortical slices were subsequently incubated in oxygenated choline Ringer at 25 degrees C for 3 hr.6. There was no evidence of energy-dependent extrusion of water or ions from either equilibrated rat or rabbit renal cortical slices leached at 0.5 degrees C and then reincubated at 25 degrees C in choline Ringer.7. Rat and guinea-pig renal cortical slices leached at 0.5 degrees C and reincubated at 25 degrees C swelled in rubidium Ringer and in caesium Ringer. There was no evidence of energy-dependent water or ion extrusion when metabolism was restored after leaching in either of these media. Metabolizing rat slices but not guinea-pig slices swelled faster than slices whose metabolism was inhibited.8. These results lend no support to the mechano-chemical hypothesis which ascribes cellular volume regulation to a contractile mechanism squeezing isotonic extracellular fluid from the cells. Instead it is suggested that cellular water content in these experiments reflects the balance between the rate of loss of potassium (and chloride) from the cells and the rate of uptake of extracellular cation (and chloride) into the cells - these rates reflecting both the electrochemical potential gradients of the ions and membrane permeability to them. The implications in relation to the hypothesis of ouabain-insensitive cellular volume regulation are discussed.

Animals

Nerve fibre refractory period in patients treated with rubidium and lithium.

Nerve fibre refractory period distributions have been measured on the median nerves of six manic-depressive patients controlled with lithium carbonate, three chronic patients (two manic-depressives and one catatonic schizophrenic) controlled with rubidium chloride, and eight normal volunteers. Rubidium prolonged the refractory periods of all nerve fibres while lithium increased only the longer refractory periods.

Adult