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The effects of varying the cellular and extracellular concentrations of sodium and potassium ions on the uptake of glycine by mouse ascites-tumour cells in the presence and absence of sodium cyanide.

ABSTRACT: 1. Tumour cells were starved to deplete them of ATP and transferred to 0.9mm-glycine in Ringer solutions containing 2mm-sodium cyanide and various Na(+) and K(+) concentrations. The uptake of glycine then usually reached a peak by about 10min. 2. When cellular [Na(+)] and extracellular [Na(+)] were each about 30m-equiv./l., the maximum amount of glycine absorbed increased between 1.2- and 3.0-fold on lowering extracellular [K(+)] from 128 to 10m-equiv./l. 3. When extracellular [Na(+)] was 150m-equiv./l., the ratio, R, of the cellular to extracellular glycine concentrations increased progressively, from near 1 to about 9, when cellular [Na(+)] was lowered from 120 to 40m-equiv./l. 4. When cellular [Na(+)] was almost constant, either at 45 or 70m-equiv./l., R fell about 14-fold when extracellular [Na(+)] varied from 150 to 16m-equiv./l. 5. Values of R near 0.2 were found when cellular [Na(+)] was about four times as large as extracellular [Na(+)]. 6. R fell about threefold when the cells were put with 12mm- instead of 0.9mm-glycine. 7. The results were taken to imply that, under these conditions, the spontaneous movements of both Na(+) and K(+) across the cell membrane, down their respective concentration gradients, served to concentrate the glycine in the tumour cells (Christensen's hypothesis).

SUBMITTER: Eddy AA

PROVIDER: S-EPMC1198835 | biostudies-other | 1968 Jul

REPOSITORIES: biostudies-other

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A net gain of sodium ions and a net loss of potassium ions accompanying the uptake of glycine by mouse ascites-tumour cells in the presence of sodium cyanide.

Project description:1. The tumour cells were starved in a solution lacking Na(+) and then transferred to a Ringer solution containing 2mm-sodium cyanide, 150m-equiv. of Na(+)/l. and 10m-equiv. of K(+)/l. Such cells were depleted of ATP and contained an endogenous pool of various amino acids equivalent to a 26mm solution. 2. At 4min. after the transfer the cellular Na(+) content had increased by about 100% and roughly an equivalent amount of K(+) had left the cells. 3. Under these conditions [(14)C]glycine was absorbed from an 11mm solution and reached the same cellular concentration by about 4min. The pool size increased by approximately the same amount (DeltaGly), so glycine did not simply exchange with the endogenous components. 4. After 4min. with glycine, the cells contained about 20% more Na(+) (DeltaNa(+)) than the control and about 10% less K(+) (DeltaK(+)). The mean values of DeltaNa(+)/DeltaGly and DeltaK(+)/DeltaGly from five experiments were respectively 0.90+/-0.11 and 0.62+/-0.11equiv./mole. 5. A further indication that these two ratios were not equal was that the cells absorbed more water than the movement of glycine itself required. The excess of water was osmotically equivalent to 0.95+/-0.16equiv. of solute/mole of glycine absorbed. 6. The variation of DeltaNa(+)/DeltaGly with the duration of the incubation was consistent with the stimulated uptake of Na(+) being linked to the actual transport of glycine. The same may apply to the movement of K(+), though the time-dependence was not examined in that case. 7. The observations were analysed in terms of a model in which both K(+) and Na(+) moved with a glycine-carrier system without ATP being involved. The analysis supported the idea that the spontaneous movements of the ions through the system might concentrate glycine in the cells significantly by purely physical means (Christensen's hypothesis).

| S-EPMC1198793 | biostudies-other

The effects of sodium ions and potassium ions on glycine uptake by mouse ascites-tumour cells in the presence and absence of selected metabolic inhibitors.

Project description:1. The initial rate, v, of glycine uptake by ascites-tumour cells respiring their endogenous nutrient reserves was studied as a function of the respective extracellular concentrations of glycine, Na(+) and K(+). With the extracellular concentration of Na(+)+K(+) constant at 158m-equiv./l. and that of glycine either 4 or 12mm, v tended to zero as the extracellular concentration of Na(+) approached zero. Glycine appeared to enter the cells as a ternary complex with a carrier and Na(+). K(+) competed with Na(+) for one of the carrier sites, whereas glycine was bound at a second site. The values of the five relevant binding constants showed that the two sites interacted. 2. The glycine uptake rate at various extracellular concentrations of glycine and Na(+) was scarcely affected by starving the cells for 30min. in the presence of 2mm-sodium cyanide provided that cellular Na(+) and K(+) contents were kept at the normal values. When the cells took up Na(+), however, v decreased approximately threefold. 3. When their Na(+) content was relatively small and the extracellular concentration of Na(+) was large, the starved cells accumulated glycine in the presence of cyanide for about 15min. Glycine then tended to leave the cells. An average of about 5mumoles of glycine/ml. of cell water was taken up from a 1mm solution, representing about 20% of the accumulation observed during respiration. Studies with fluoride, 2,4-dinitrophenol and other metabolic inhibitors supported the view that ATP and similar compounds were not implicated. The relation between the transient accumulation of glycine that occurred in these circumstances and the normal mode of active transport was not established.

| S-EPMC1270492 | biostudies-other

Further observations on the inhibitory effect of extracellular potassium ions on glycine uptake by mouse ascites-tumour cells.

Project description:1. The initial rate of uptake of glycine by the tumour cells was measured as a function of the Na(+) and K(+) concentrations in the solution in which the cells were suspended. When [Gly] was 1mm or 12mm, the rate in the absence of Na(+) was independent of [K(+)] and about 3% or 10% respectively of the rate when [Na(+)] was 150m-equiv./l. 2. The Na(+)-dependent glycine entry rate, v, at a given value of [Na(+)] was successively lowered when [K(+)] was increased from 8 to 47 to 96m-equiv./l. A kinetic analysis indicated that K(+) competitively inhibited the action of Na(+). The results were in fair agreement with previous determinations of the kinetic parameters. 3. The presence of 2mm-sodium cyanide and 10mm-2-deoxyglucose lowered the cellular ATP content to less than 3% of the value in the respiring cells. Although v was then about 50% smaller, the relative effects of K(+) and Na(+) on the system were similar to those observed during respiration. 4. A theoretical analysis indicated that the variation of v with [K(+)] is not a reliable guide to the extent to which the K(+) gradient between the cells and their environment may contribute to the net transport of glycine.

| S-EPMC1184968 | biostudies-other

Diagnostic usefulness of the spot urine sodium/potassium ratio in cirrhotic patients with ascites.

Project description:Background and aimsThe low-salt diet is considered important for control of ascites in cirrhotic patients. To validate whether the spot urine sodium (Na)/potassium (K) ratio could replace 24-h urine Na (uNa) excretion in assessing low-salt diet compliance.MethodsWe prospectively studied 175 patients. 24-h urine collection and spot urine collection were performed. Subsequently, 24-h uNa, urine creatinine (uCr), and spot urine Na and K were assessed. A complete urine collection was confirmed based on 24-h uCr excretion levels of 15mg/kg/day for men and 10mg/kg/day for women. The area under the receiver operating characteristic (AUROC) curve analysis was performed to evaluate the feasibility of spot urine Na/K ratio in predicting 24-h uNa greater than 78mmol/day.ResultsOut of 175 patients, 24-h urine samples were completely collected in 57 patients only. Moreover, urine samples were not completely collected in 118 patients because their 24-h uCr excretion level was less than the established criteria. In complete urine collection group, AUROC curve for spot urine Na/K ratio in predicting 24-h uNa greater than 78mmol/day was 0.874±0.051 (P<0.001). In the incomplete urine collection group, the AUROC was 0.832±0.039 (P<0.001). In complete urine collection group, the classical cutoff value greater than 1.0 of spot urine Na/K ratio showed 90.9% sensitivity and 56.0% specificity.ConclusionsThe spot urine Na/K ratio reflects 24-h uNa, but the AUROC value obtained in this study is lower than that of a previous study. Considered the large number of patients with incomplete urine collection, validating 24-h complete urine collection criteria is necessary.

| S-EPMC8224901 | biostudies-literature

Rates of effux and intracellular concentrations of potassium, sodium and chloride ions in isolated fat-cells from the rat.

Project description:1. The metabolism of K(+), Na(+) and Cl(-) has been investigated in isolated fat-cells prepared from the epididymal adipose tissue of rats. 2. Methods are described for measuring the intracellular water space, the rates of loss of intracellular (42)K(+), (22)Na(+) and (36)Cl(-) and the intracellular concentrations of K(+), Na(+) and Cl(-) in isolated fat-cells. 3. The intracellular water space, measured as the [(3)H]water space minus the [carboxylic acid-(14)C]inulin space, was 3.93+/-0.38mul./100mg. cell dry wt. 4. The first-order rate constants for radioisotope effluxes from isolated fat-cells were 0.029min.(-1) for (42)K(+), 0.245min.(-1) for (22)Na(+) and 0.158min.(-1) for (36)Cl(-). 5. The intracellular concentrations of K(+), Na(+) and Cl(-) were 146m-equiv./l., 18.6+/-2.9m-equiv./l. and 43+/-2.4m-equiv./l. respectively. 6. The total intracellular K(+) content of isolated fat-cells was determined by atomic-absorption spectrophotometry to confirm the value obtained from the radioisotope-efflux data. 7. The ion effluxes from isolated fat-cells were: K(+), 1.5pmoles/cm.(2)/sec., Na(+), 1.6pmoles/cm.(2)/sec., and Cl(-), 2.4pmoles/cm.(2)/sec. 8. The membrane potential of isolated fat-cells calculated from the Cl(-) distribution ratio was -28.7mv.

| S-EPMC1185227 | biostudies-other

Interctions between potassium ions and glycine transport in the yeast Saccharomyces carlsbergensis.

Project description:A study has been made of the effects of both varying the pH and extracellular [K(+)] on the initial rate of uptake of glycine (v) by a strain of Saccharomyces carlsbergensis that concentrated the amino acid, with respect to the extracellular phase, by up to 1400 times. When no other substrate than glycine was provided and [glycine] was relatively small (</=0.2mm) (1) v increased fivefold when the pH was lowered from 7 to 4.5; (2) v fell by up to about 80% as [K(+)] rose, K(+) behaving as a non-competitive inhibitor of the system, with K(i) 0.33mequiv./l at pH7; (3) the absorption of glycine caused up to about 2 or 3 equiv. of K(+) to leave the yeast cells. These three phenomena were each less evident when glucose was present. An analogy is drawn between the respective interactions of H(+) and K(+) with the yeast system and the well recognized effects of Na(+) and K(+) on amino acid transport in certain mammalian systems.

| S-EPMC1179679 | biostudies-other

Free concentrations of sodium, potassium and calcium in chromaffin granules.

Project description:We have measured the contents of Na+ and K+ in isolated chromaffin granules. Total contents varied between 227 and 283 nmol/mg of protein, equivalent to matrix concentrations of 53-66 mM. The value found depended on the isolation buffer used, and the ratio of the two ions reflected the composition of the buffer. We then measured the free concentration of each of these ions, and of Ca2+, in the matrix, by using a null-point method with acridine-fluorescence quenching. This monitored H+ fluxes induced by an ionophore in the presence of known concentrations of the ion in the supporting medium. In contrast with organic constituents of the matrix, which have low activity coefficients, Na+ and K+ were found to have activity coefficients around 0.8 Ca2+, however, was strongly bound: its free concentration was only 0.03% of the total.

| S-EPMC1138534 | biostudies-other

Anomeric and Enantiomeric 2'-Deoxycytidines: Base Pair Stability in the Absence and Presence of Silver Ions.

Project description:Dodecamer duplex DNA containing anomeric (α/β-d) and enantiomeric (β-l/β-d) 2'-deoxycytidine mismatches was studied with respect to base pair stability in the absence and presence of silver ions. Stable duplexes with silver-mediated cytosine-cytosine pairs were formed by all anomeric and enantiomeric combinations. Stability changes were observed depending on the composition of the mismatches. Most strikingly, the new silver-mediated base pair of anomeric α-d-dC with enantiomeric β-l-dC is superior to the well-noted β-d/β-d-dC pair in terms of stability. CD spectra were used to follow global helical changes of DNA structure.

| S-EPMC8362019 | biostudies-literature

A sodium ion concentration gradient formed during the absorption of glycine by mouse ascites-tumour cells.

Project description:1. To deplete them of ATP the tumour cells were starved at 37 degrees in a Ringer solution containing 33m-equiv. of Na(+)/l., 131m-equiv. of Li(+)/l., 2mM-sodium cyanide and 0.1mm-ouabain. The cellular content of K(+) was largely replaced by Li(+), but cellular [Na(+)] remained near 33m-equiv./l. 2. The addition of 12mm-glycine to the system caused cellular [Na(+)] to increase, during the next 4min., by about 4m-equiv./l., so that it slightly exceeded extracellular [Na(+)]. This occurred in parallel with the absorption of glycine. 3. The cellular K(+) content fell by an amount representing about 10% of the amount of Na(+) absorbed. 4. The results provide a clear demonstration that the flow of glycine into the cells is linked to a parallel movement of Na(+); K(+) appears to play a facultative role in the carrier system, whereas Li(+) is almost inert. 5. The effects produced by glycine were not reproduced by l-arabinose.

| S-EPMC1185130 | biostudies-other

Electrochemical and Fluorescent Properties of Crown Ether Functionalized Graphene Quantum Dots for Potassium and Sodium Ions Detection.

Project description:The concentration of sodium and potassium ions in biological fluids, such as blood, urine and sweat, is indicative of several basic body function conditions. Therefore, the development of simple methods able to detect these alkaline ions is of outmost importance. In this study, we explored the electrochemical and optical properties of graphene quantum dots (GQDs) combined with the selective chelating ability of the crown ethers 15-crown-5 and 18-crown-6, with the final aim to propose novel composites for the effective detection of these ions. The results obtained comparing the performances of the single GQDs and crown ethers with those of the GQDs-15-crown-5 and GQDs-18-crown-6 composites, have demonstrated the superior properties of these latter. Electrochemical investigation showed that the GQDs based composites can be exploited for the potentiometric detection of Na+ and K+ ions, but selectivity still remains a concern. The nanocomposites showed the characteristic fluorescence emissions of GQDs and crown ethers. The GQDs-18-crown-6 composite exhibited ratiometric fluorescence emission behavior with the variation of K+ concentration, demonstrating its promising properties for the development of a selective fluorescent method for potassium determination.

| S-EPMC8625964 | biostudies-literature

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