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PubMed1995

Accumulation of glutamate by osmotically stressed Escherichia coli is dependent on pH.

In the present study, we measured the accumulation of glutamate after hyperosmotic shock in Escherichia coli growing in synthetic medium. The accumulation was high in the medium containing sucrose at a pH above 8 and decreased with decreases in the medium pH. The same results were obtained when the hyperosmotic shock was carried out with sodium chloride. The internal level of potassium ions in cells growing at a high pH was higher than that in cells growing in a neutral medium. A mutant deficient in transport systems for potassium ions accumulated glutamate upon hyperosmotic stress at a high pH without a significant increase in the internal level of potassium ions. When the medium osmolarity was moderate at a pH below 8, E. coli accumulated gamma-aminobutyrate and the accumulation of glutamate was low. These data suggest that E. coli uses different osmolytes for hyperosmotic adaptation at different environmental pHs.

Adaptation, Biological↗
PubMed2001

Potassium ion release from a glass ionomer cement matrix.

Glass ionomer was mixed with water alone and with water containing two levels of potassium ion, added as potassium chloride. The dimensions of the specimens varied; discs, cylinders and a rectangular bar were used. Release of potassium was measured until it ceased. A little potassium was released from cement mixed with water but could be accounted for by traces present in the glass. Release took 2 months for the lower level and 1.5 years for the higher level of addition. For some specimen groups not all the potassium added was subsequently released. Potassium release tended towards the pattern found for fluoride release naturally occurring from glass ionomer cements in being proportional to t 1/2 but was less certain at the lower addition. Release was not related to the dimensions of the sample as has been shown for release of intrinsic fluoride.

Cations, Monovalent↗
PubMed2005

Maintenance of eupnea and gasping following alterations in potassium ion concentration of perfusates of in situ rat preparation.

Levels of extracellular potassium ion, above those in vivo, are required for the generation of rhythmic activities of in vitro preparations. Our purpose was to define whether hyperkalemia in the perfusate of an in situ preparation was likewise necessary for the expression of eupnea and gasping. Studies were performed using the in situ preparation of the juvenile rat, in which activity of the phrenic nerve was recorded as an index of the respiratory rhythm. Eupnea and gasping were impervious to modifications of potassium ion of the perfusate. Eupnea was maintained uninterrupted for more than 60 min whether the total concentration of potassium was hypokalemic (2.75 mM), normokalemic (4.0 mM), or hyperkalemic (6.25 and 7.75 mM). Gasping, with identical characteristics, was elicited at any concentration of potassium ion. We conclude that both eupnea and gasping are unaffected by modest changes in the concentration of potassium ion in the perfusate with which the in situ rat preparation is maintained. These results add further support to the conclusion that the in situ preparation represents a model that accurately reproduces mechanisms of rhythm generation of both eupnea and gasping in vivo.

Action Potentials↗
PubMedDate not supplied

[Effect of verapamil and anesthetics on the level of sodium and potassium ions in frog muscles. Role of hydrophobic interactions].

Verapamil, general and local anesthetics influences on ionic contents of frog sartorius and cardiac muscles in respect with these drugs hydrophobicity were studied to clarify the role of verapamil hydrophobic interactions in its physiological activity. It was found that concentration thresholds of the agents toxicities were linearly linked with their hydrophobicity (in logarithmic scale). This relationship provides support for our conclusion that verapamil, general and some local anesthetics are the members of a single drug family. It is a reason to believe that a hydrophobic mechanism of general anesthetic action on cellular structure is the same as a mechanism of verapamil activity in muscles. It is possible that verapamil is accepted as medical drug exactly due to optimum combination of high verapamil hydrophobicity with its structural complementarity to receptor site responsible for use-dependent channel blocking.

Anesthetics↗
PubMed1981

Isolation and partial characterization of active polysomes from calcified and matrix-containing tissues.

Polysomes were isolated from calcified and matrix-containing tissues, such as rat calvaria, rat chondrosarcoma and chick embryos. The method of isolation involves preliminary swelling of the tissues in hypotonic buffer containing heparin and cycloheximide. After homogenization, differential centrifugation is used to separate membrane-bound and non-bound polysomes. Each fraction is rehomogenized in the presence of detergent (Triton X-100) and potassium ions (0.25M). Polysomes are harvested by centrifugation through sucrose cushions in the continued presence of high levels of potassium ions and heparin. Total, non-bound, and membrane-bound polysomes prepared in this manner are equally active in protein synthesizing activity in an heterologous cell-free system. The distribution between non-bound and membrane-bound polysomes in the 12 day old chick embryo is 43 and 57 per cent respectively. Sucrose gradient profiles of polypeptide chains on polysomes labeled in organ culture correlate well with the protein synthetic activity of the isolated polysomes. Much of the protein synthetic activity is devoted to collagen. Polysomal fractions obtained from sucrose gradients show preferential incorporation of 3H-proline and nearly 60 per cent of trichloroacetic acid precipitable material is susceptible to collagenase digestion. Products of synthesis are also substrates for collagen specific enzyme, prolyl hydroxylase. The method described in this communication overcomes the inherent difficulties in obtaining active polysomes from calcified and matrix-containing tissues.

Animals↗
PubMed2004

Acid-base balance parameters and ionic composition of arterial, venous and capillary blood in goats.

The aim of the present study was to compare the acid-base balance parameters of arterial, venous and capillary blood in clinically healthy goats, and to determine the electrolyte content of venous and arterial blood. The experiment was performed on ten adult goats. It was found that the acid-base balance parameters of venous blood differed from those of arterial blood, whereas the parameters of capillary and arterial blood were similar. The levels of sodium and chloride ions in arterial and venous blood were similar, whereas the level of potassium ions was higher in venous blood.

Animals↗
PubMed2005

The effect of propofol on acid-base balance and ionic composition of venous and arterial blood in goats.

The aim of the present study was to determine the effect of propofol on acid-base balance and ionic composition of arterial and venous blood in clinically healthy goats. The experiment was performed on ten adult goats. Propofol was administered intravenously at bolus dose of 6 mg/kg bw. The heart and breath rate, acid-base balance (pH, pCO2, pO2, HCO3-, BE, O2SAT, ctCO2) and ionic composition (Na+, K+, Cl-) of arterial and venous blood were measured before injection and 3, 6 and 15 min. after. The propofol infusion induced increase of heart rate, decrease of breath rate and compensated respiratory acidosis in venous and arterial blood. It was found that changes of acid-base balance parameters in arterial blood arose faster than in venous blood. The levels of sodium and chloride ions in both types of blood were similar, whereas the level of potassium ions was higher in venous blood during entire experiment.

Acid-Base Equilibrium↗
PubMed1994

Na(+)-Ca2+ exchanger-like immunoreactivity and regulation of intracellular Ca2+ levels in fish retinal ganglion cells.

1. We have used two experimental approaches to examine regulation of intracellular calcium ion levels in fish retinal ganglion cells. In the first set of experiments, we ratio-imaged fura-2 emission intensity to estimate the concentration of free intracellular calcium ions ([Ca2+]i) in isolated goldfish retinal ganglion cells depolarized by increases in extracellular levels of potassium ions ([K+]o), in the presence and absence of extracellular sodium ions (Na+). Stepwise increases in [K+]o from 5 mM to as high as 60 mM produced stepwise increases in [Ca2+]i. These increases were sustained in the absence of external Na+, but transient and smaller in the presence of external Na+. The decline of [Ca2+]i in high-K, Na(+)-containing saline could be reversed by application of the ionophore monensin, or by replacement of external Na+ with either N-methyl-D-glucamine or lithium. In Na(+)-containing saline, [Ca2+]i fell to control levels after [K+]o was restored to control levels. 2. In the second set of experiments, we assessed Na(+)-Ca2+ exchanger-like immunoreactivity in goldfish retinal ganglion cells with the use of a polyclonal antiserum directed against Na(+)-Ca2+,K+ exchanger purified from bovine rod outer segments. This antiserum specifically stained the somata, neurites, and growth cones of isolated ganglion cells, the outer segments of rod photoreceptors, and (on Western blots prepared from mechanically isolated rods) protein displaying an apparent molecular mass of 210 kDa. 3. These results provide measurements of changes in [Ca2+]i of retinal ganglion cells depolarized in Na(+)-containing saline, and the distribution and apparent molecular weight of Na(+)-Ca2+ exchanger-like immunoreactivity in teleost retina.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2005

Genetic requirements for potassium ion-dependent colony spreading in Bacillus subtilis.

Undomesticated strains of Bacillus subtilis exhibit extensive colony spreading on certain soft agarose media: first the formation of dendritic clusters of cells, followed by spreading (pellicle-like) growth to cover the entire surface. These phases of colonization are dependent on the level of potassium ion (K(+)) but independent of flagella, as verified with a mutant with a hag gene replacement; this latter finding highlights the importance of sliding motility in colony spreading. Exploring the K(+) requirement, directed mutagenesis of the higher-affinity K(+) transporter KtrAB, but not the lower-affinity transporter KtrCD, was found to inhibit surface colonization unless sufficient KCl was added. To identify other genes involved in K(+)-dependent colony spreading, transposon insertion mutants in wild-type strain 3610 were screened. Disruption of genes for pyrimidine (pyrB) or purine (purD, purF, purH, purL, purM) biosynthetic pathways abolished the K(+)-dependent spreading phase. Consistent with a requirement for functional nucleic acid biosynthesis, disruption of purine synthesis with the folic acid antagonist sulfamethoxazole also inhibited spreading. Other transposon insertions disrupted acetoin biosynthesis (the alsS gene), acidifying the growth medium, glutamine synthetase (the glnA gene), and two surfactin biosynthetic genes (srfAA, srfAB). This work identified four classes of surface colonization mutants with defective (i) potassium transport, (ii) surfactin formation, (iii) growth rate or yield, or (iv) pH control. Overall, the ability of B. subtilis to colonize surfaces by spreading is highly dependent on balanced nucleotide biosynthesis and nutrient assimilation, which require sufficient K(+) ions, as well as growth conditions that promote sliding motility.

Acetoin↗
PubMed1979

Potassium fluxes and ouabain binding in growing, density-inhibited and Rous sarcoma virus-transformed chicken embryo cells.

Potassium fluxes, ouabain binding, and Na+ and K+ intracellular concentrations were determined for cultures of growing normal, density-inhibited and Rous sarcoma virus-transformed chicken embryo fibroblasts. No significant differences in K+ influx or ouabain binding were detected between growing normal cells and Rous sarcoma virus-transformed cells; however, ouabain binding and ouabain-sensitive K+ influx were 1.5- to 1.8-fold lower in density-inhibited cells. Thus, potassium influx in this system can be classified as a growth-related, but not transformation-specific change. As determined by both flame photometry and radioisotopic (42K) equilibration, growing normal and density-inhibited cells had similar potassium contents, whereas transformed cells exhibited 1.4-fold higher potassium levels. Sodium ion levels, as measured by flame photometry, were also 2- to 4.5-fold higher in transformed than normal or density-inhibited cells. Complementary studies of potassium efflux showed a 1.3- to 1.5-fold higher rate (based on the percentage of pool exiting the cell) in growing normal versus density-inhibited or transformed fibroblasts. Because of the larger potassium pool in transformed cells, efflux based on absolute number of potassium ions is similar in normal and transformed chicken embryo fibroblasts.

Animals↗
PubMed1992

Osmotic adaptation of Escherichia coli with a negligible proton motive force in the presence of carbonyl cyanide m-chlorophenylhydrazone.

It has been reported that Escherichia coli is able to grow in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP) when ATP is produced by glycolysis (N. Kinoshita et al., J. Bacteriol. 160:1074-1077, 1984). We investigated the effect of CCCP on the osmotic adaptation of E. coli growing with glucose. When E. coli growing in rich medium containing CCCP was transferred to medium containing sucrose, its growth stopped for a while and then started again. This lag time was negligible in the absence of CCCP. The same results were obtained when the osmolarity was increased by N-methylglucamine-maleic acid. In addition to adapting itself to the hyperosmotic rich medium, E. coli adapted itself to hyperosmolarity in a minimal medium containing CCCP, again with a lag time. Hyperosmotic shock decreased the internal level of potassium ion rather than causing the accumulation of external potassium ion in the presence of CCCP. The internal amount of glutamic acid increased in cells growing in hyperosmotic medium in the presence and absence of CCCP. Large elevations in levels of other amino acids were not observed in the cells adapted to hyperosmolarity. Trehalose was detected only in hyperosmosis-stressed cells in the presence and absence of CCCP. These results suggest that E. coli can adapt to changes in the environmental osmolarity with a negligible accumulation of osmolytes from the external milieu but that the accumulation may promote the adaptation.

Adaptation, Physiological↗
PubMed1985

Influence of environmental factors on the metabolism of glucose by preimplantation mouse embryos in vitro.

The metabolism of glucose by late preimplantation mouse embryos was studied in a variety of media whose composition had been changed to reflect the environmental conditions in the uterus more closely than do standard culture media. The effects of combinations of energy substrates, the presence or absence of amino acids and the level of potassium in the medium were investigated. The use of energy substrates for in vitro culture at levels present in the uterine environment resulted in rates of synthesis and degradation of glycogen pools similar to those obtained using standard in vitro culture conditions but elevated incorporation into non-glycogen macromolecules. Amino acids influenced the metabolism of glucose by limiting the entry of glucose carbon into the non-glycogen macromolecular pool and directing more glucose into the synthesis of acid-soluble glycogen. Increasing the K+ concentration to 60 mM in the culture medium caused a small but significant increase in the number of eight-cell embryos degenerating during culture for 24 h but the metabolism of glucose was unaffected over this time. At the time of morula transformation to the blastocyst this level of potassium ions suppressed glycogen synthesis by 50% over 5 h but did not affect its turnover during chase culture. It is concluded that factors other than those studied here contribute to the maintenance of the low glycogen levels found in uterine embryos.

Amino Acids↗
PubMed2005

Imaging TOF-SIMS of rat kidney prepared by high-pressure freezing.

Phosphocholine, potassium ions, and sodium ions were localized in rat kidney with imaging TOF-SIMS. Tissue preparation was performed with high-pressure freezing, freeze-fracturing and freeze-drying. The distribution of sodium ions was visualized by imaging the signal at m/z 23 of positively charged secondary ions, and the distribution of potassium ions was visualized by imaging the signal at m/z 39. Potassium was found localized within cells of the proximal tubulus epithelium and within cells of the glomeruli. High signals of sodium ions were seen in the interstitial tissue and also in epithelial cells of the collecting ducts and in glomeruli. The overlay image showed that the distribution of sodium ions and potassium ions were largely complementary with color mixing in glomeruli and in the interstitium surrounding proximal tubules. The ion distribution was further analyzed by correlation analysis. Phosphocholine-containing phospholipids were visualized by imaging the phosphocholine head group at m/z 184 of positively charged ions. The m/z 184 signal shows a ubiquitous distribution with a high intensity of phosphocholine in epithelial cells. Overlay image of m/z 184, m/z 39, and m/z 23 and multivariate analysis showed that the localization of high levels of phosphocholine colocalizes with high levels of potassium ions, as expected for an ion with intracellular localization.

Animals↗
PubMed1990

Induction of fine structural alteration in cardiac Purkinje cells by ischaemic metabolites depends on oxygen tension.

The effects of major ischaemic metabolites on the fine structure of false tendon Purkinje cells from the dog heart were investigated in vitro under oxygenated and oxygen-deficient conditions. High levels of potassium ion induced potentially reversible alteration consisting of moderate cell swelling, swelling of sarcoplasmic reticulum, and nuclear swelling with moderate aggregation of chromatin irrespective of gas tension. Other metabolites induced alteration only in oxygen-deficient solutions. Ischaemic levels of lactate and inorganic phosphate both caused mitochondrial swelling, which was most severe in the presence of lactate. At acidic pH intramitochondrial dense inclusions, the hallmark of irreversible injury, also developed. Severe aggregation of nuclear chromatin and gross dilatations of the sarcoplasmic reticulum were focal and much less prominent than in contractile myocardium exposed to excess hydrogen ion. These observations indicate that the efflux of ischaemic metabolites from a developing infarct may be arrhythmogenic by impairing the function of the subjacent Purkinje network without damaging its cell structure, since luminal blood flow ensures it is always adequately oxygenated.

Animals↗