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An explanation for the positive inotropic effect of hypertonic solutions.

Hypertonic solutions of mannitol and sucrose have positive inotropic effects on both cardiac and skeletal muscle in vivo and in vitro. By increasing the tonicity of dog plasma of 50 mOsm in vitro with sucrose or mannitol Ca++ activity. The negative results with calcium proteinate and simple salt solutions suggest that the increased activity increases of 20--50 percent can be measured with a calcium-specific electrode. Increased tonicity had no effect on calcium proteinate complexes. By increasing the tonicity of solutions of small molecular weight complexes, a 5--50 percent increase in Ca++ activity results, depending on the specific complex used. Increasing the tonicity of solutions of inorganic salts such as CaSO4 or CaCl2 did not increase Ca++ activity seen with small molecular weight complexes is caused by dissociation of the Ca++, not by an interaction of the sucrose or mannitol solutions with the electrode. The absolute change inducible in a 10(-3) M solution of a small molecular weight complex is in the range of 0.5 mEq/l, a physiologically significant range. Thus it is suggested that part of the positive inotropic effect of increase osmolarity is a result of increased Ca++ activity from small molecular weight complexes in plasma and interstitial fluid.

Animals

Effect of salt solutions on radiosensitivity of mammalian cells. III. Treatment with hypertonic solutions.

V79 Chinese hamster cells were treated with hypertonic solutions of NaCl or KCl and irradiated rat various times before, during, or after exposure to the solution. In solutions of molarities between 0-2 and 0-5 M, the cellular radiosensitivity increases with the molarity of the bathing solution. At these molarities, the hypertonic solution need not be present during irradiation to sensitize cells. Furthermore, radiosensitivity of cells could be increased by exposing cells for longer times to the hypertonic solution before irradiation. At higher salt concentrations (at 1-5 to 1-8 M), significant radioprotection is observed. Survival curve data showed that this protection was characterized by an increase in DO and a decrease in n, while the survival curves of cells sensitized with 0-465 M NaCl or with lower concentrations exhibited mainly changes in DO. The 1-55 M NaCl solution must be present during radiation to give a protective effect. Prolonged exposure to the salt before irradiation reduced the amount of radioprotection afforded by the salt. The results are discussed in terms of the effects of ions on histones, cellular water structure and the cell-aging cycle.

Cell Survival

Increase in human intestinal permeability following ingestion of hypertonic solutions.

1. A simple oral loading technique involving the ingestion of solutions containing lactulose is described. Timed urinary excretion of lactulose, which is non-metabolizable, is used as an indicator of intestinal permeability, and measured by quantitative paper chromatography. 2. This technique has been used to investigate the intestinal permeability of apparently healthy adults following the ingestion of solutions made hypertonic by the addition of the solutes sucrose, glucose, mannitol, glycerol, urea and sodium chloride. 3. These experiments show that intestinal permeability to lactulose increases as the solute concentration in the ingested solution is increased. Susceptibility to this effect, though consistent for each individual, shows considerable variation between subjects. 4. Factors thought to be pernitent to the enhancement of intestinal permeability by hypertonic solutions, and some possible implications of this, are discussed.

Disaccharides

The sarcoplasmic reticulum and T-system of rat extensor digitorum longus muscles exposed to hypertonic solutions.

Excised rat extensor digitorum longus muscles were soaked in a Krebs solution made hypertonic with extra NaCl and fixed using a combination of acrolein and glutaraldehyde dissolved in the same hypertonic Krebs. Electron microscopic examination showed that the sarcoplasmic reticulum (S.R.) of these muscles occupied the same fraction of fibre volume as it did in control muscles soaked and fixed in normal Krebs solution, contradicting previous studies on frog muscle. It appears unlikely that the S.R. is the rapidly exchanging Na+-compartment of rat muscle which enlarges in hypertonic solutions, or that it is an extracellular compartment. The T-system of hypertonic muscles was swollen, but its volume is insufficient to form the rapidly exchanging Na+-compartment.

Animals

T-tubule swelling in hypertonic solutions: a freeze substitution study.

Striated muscles from Rana pipiens have been exposed for variable periods of time to Ringer solutions made hypertonic by addition of either sucrose or sodium chloride. The muscles have been rapid-frozen and then prepared for electron microscopy by either freeze-substitution, freeze-fracture or cryoultramicrotomy. The only compartment greatly affected by hypertonicity is the transverse tubular system, which is visibly swollen. None of the elements of the sarcoplasmic reticulum increase in size.

Animals

Inhibition by hypertonic solutions of Ca-dependent electrogenesis in single crab muscle fibers.

This study describes the effect of hypertonic solutions on isolated muscle fibers of Callinectes danae. Solutions of twice normal tonicity (2.0 T) inhibit both the normal graded membrane responses and the spikes induced by procaine, tetraethylammonium, or barium. The inhibition is maintained throughout exposure to hypertonic solutions prepared by addition of impermeant solutes such as NaCl, sucrose, or Tris-propionate, but is reversible on their withdrawal. In the presence of permeant solutes such as glycerol or acetamide, the inhibition is transient. In both cases the onset of inhibition of the depolarizing Ca electrogenesis is correlated with shrinkage of the fiber. In the case of permeant solutes, the time course of recovery of the graded responses or the spikes follows the recovery of the fiber volume. Changes in the passive electrical characteristics of the fibers due to hypertonic solutions were unrelated to the blockade of membrane Ca activation. The current-voltage relationship in hypertonic sollution revealed no increase in depolarizing K activation. Inhibition of the graded membrane responses and spikes appears to be associated with depression of Ca conductance. Hypertonic solutions might affect the activation of Ca conductance through reduction of the electric field generated by fixed negative surface charges and/or morphological changes in the T tubules. Membrane depolarization elicited little or no tension in 2.0 T solutions while caffeine contracture (10 mM) with an ampliture of 76% of the maximal contractile ability could still be elicited. This indicates that direct effects of hypertonic solutions on the contractile apparatus were not responsible for loss of tension. The latter is attributed to the inhibition of the transmembrane Ca currents.

Acetamides

Enhancement of aorta tonus evoked by hypertonic solutions is endothelium-dependent.

1. The effect of NaCl hypertonic solutions on the contractions induced by noradrenaline or prostaglandin F2 alpha and on the relaxant effect of acetylcholine and sodium nitroprusside was investigated in aortae with or without endothelium isolated from rats. 2. Hypertonic solutions enhanced the contractions elicited by the vasoconstrictor agents in preparations with endothelium and this phenomenon was not altered by previous treatment of the animals with reserpine or the preparations with methylene blue or indomethacin. 3. In hypertonic medium acetylcholine but not sodium nitroprusside vasodilator effect was inhibited. 4. We suggest that NaCl hypertonic solutions modify vascular reactivity probably through: (i) the release of an endothelium-derived contracting factor not sensitive to cyclooxygenase inhibitors; (ii) a diminished release of an endothelium-derived relaxing factor not related to the cGMP system.

Acetylcholine

Antiulcer activity of hypertonic solutions in the rat: possible role of prostaglandins.

The effects of hypertonic solutions on gastric acidity and on experimental gastric and duodenal ulcers as well as on gastric prostaglandins (PGs) were studied in the rat. The oral administration of a 10% NaCl solution resulted in complete absence of free acidity and very significant reductions in total acidity 24 h after pyloric ligation. The antiulcer effect of hypertonic saline was studied in three experimental models. In pyloric-ligated rats, both the incidence and the severity of gastric ulcers were remarkably reduced by hypertonic saline treatment. Indomethacin-induced gastric erosions were significantly reduced by hypertonic NaCl or sorbitol and completely prevented by hypertonic xylitol. Cysteamine-induced duodenal ulcers were also significantly reduced by hypertonic solutions of NaCl, xylitol or sorbitol. In the latter model, indomethacin potentiated the ulcerogenic effect of cysteamine and also reduced the efficacy of the hypertonic NaCl gavage. The possible contribution of PGs to these effects was further investigated by analysing PGE in the gastric mucosa and juice. Rats treated orally with hypertonic NaCl solutions had several-fold higher PGE contents in their gastric mucosa as well as higher PGE levels in the gastric juice. It is concluded that hypertonic solutions stimulate endogenous PGE biosynthesis and also exert profound antiulcer effects in the rat. A causal relationship between the two phenomena is suggested.

Animals

Effects of hypertonic solutions on conjunctival epithelium and mucinlike glycoprotein discharge.

A simple, semiquantitative, histochemical method using periodic acid-Schiff (PAS) staining was developed to measure the mucinlike glycoprotein content of bathing solutions exposed to rabbit conjunctiva. Rabbit eyes were bathed in vivo in solutions of various tonicity (160, 260, 300, 330, and 363 mOsm/L). Eyes bathed in hypotonic or isotonic solutions (160, 260, and 300 mOsm/L) exhibited a steady-state glycoprotein secretion (50-100 micrograms/h). In hypertonic solutions (330 and 363 mOsm/L), glycoprotein discharge increased to 150-360 micrograms/h when the solutions were exchanged hourly for 6 h, but glycoprotein discharge increased to a much lower rate (100-167 micrograms/h) after prolonged bathing for 6 h without solution exchange. Periodic acid-Schiff--positive glycoprotein aggregates were noted more frequently on the filters of hypertonic solutions than on those of other tonicity. After eyes were bathed in hypertonic solutions, increased disruption of the surface epithelium, increased abnormal discharge of mucin granules, and decreased goblet cell density were noted in the biopsied conjunctiva. These data suggest that mucinlike glycoprotein secretion from the conjunctival goblet cells can be enhanced by the tonicity of the bathing solution, and that nonphysiological hypertonic solutions can also affect the conjunctival epithelium.

Animals

Some effects of hypertonic solution on the properties of spontaneous transmitter release in the hypogastric ganglia of guinea-pigs.

1. The effects of hypertonic solutions on spontaneous transmitter release at synapses in the hypogastric ganglia of guinea-pigs were studied using intracellular recording techniques. 2. Hypertonic solutions were found to cause a marked hyperpolarization of the ganglion cells and this was accompanied by an increase in both the threshold and input resistance of these cells. 3. The frequency of spontaneous quantal release was found to depend exponentially on the tonicity of the bathing solution with a 50% increase in tonicity producing a 7-9 fold increase in the frequency of release. 4. Hypertonic solutions were also found to increase the degree of bursting within the spontaneous process so that the proportion of multiquantal spontaneous potentials was found to be increased in such solutions. 5. Two different models, the 'charge-screening' model (Van der Kloot & Kita, 1973) and the 'Ca-increase' model (Shimoni, Alnaes & Rahamimoff, 1977), for the mechanism underlying the effects of hypertonic solution were considered.

Action Potentials

Mechanism of inhibition of gastric acid secretion by hypertonic solutions and vasopressin.

An experiment was performed in female rats in order to assess the influence and mechanism underlying the effects of hyperglycemia, hypertonic saline and vasopressin upon the gastric acid secretion and mucosal blood flow (MBF). Infusion of isotonic saline did not alter acid output and gastric clearance of plasma aminopyrine whereas hypertonic solutions (20% glucose or 3% NaCl) significantly increased plasma osmolality and decreased the acid secretion within 30 min and recovered to normal levels after 2 h. Vasopressin also effectively inhibited acid secretion. Both hypertonic solutions and vasopressin decreased the mucosal blood flow. However, the ratio (R) of MBF to gastric secretory rate which is a helpful guide to the mechanism of secretory inhibition did not significantly change in either case. We concluded that all three agents probably had a direct action on secretion rather than decreasing MBF. The mechanism of inhabition of acid secretion and its relationship to MBF was suggested and discussed.

Animals

Excitation of squid giant axons in hypotonic and hypertonic solutions.

Excitability of intracellularly perfused squid giant axons was maintained in hypotonic solutions (down to 300 mOSM) and in hypertonic solutions (up to about 10 OSM), when osmolalities of internal and external solutions were adjusted to be equal with glycerol, glucose, or sucrose. Molar concentrations of ions were kept constant during one series of experiments. The resting potential and the amplitude of the action potential did not change in both hypotonic and hypertonic solutions. With reduction of osmolality, the duration of action potential decreased and the maximum rate of rise and conduction velocity increased. By raising osmolality, the duration was prolonged and the maximum rate of rise and the conduction velocity decreased. Effects of osmolality change were almost reversible. However, these effects were not directly related to the osmolality change but seemed to be related to the viscosity change of the solutions. When the osmolality of external solution was raised with NaCl (up to 2.6 M NaCl), the overshoot increased in porportion to the logarithm of the NaCl concentration. The slope of increase was about 50 mV/decade. However, the resting potential showed little change. With increase of the NaCl concentration, the duration of the action potential increased.

Animals

[The effect of hypertonic solutions on contracture of a tonic muscle fiber].

Effect of glycerol, sucrose and NaC1 hypertonic solutions exerted on potassium contracture and resting membrane potential in isolated frog tonic muscle fibres were studied. Sucrose and NaC1 hypertonic solutions induce a stable depolarization and depress potassium contracture, while glycerol results in a transient depolarization followed by recovery of resting membrane potential and contracture reactions. The removal of 400 mM glycerol induces an insignificant depolarization and disappearance of contracture reactions, however, after the removal of sucrose of NaC1 hypertonicity both the parameters tend to restore. The isolated phasic and tonic fibres lose irreversibly potassium contracture reactions after the removal of 100 mM and 400 mM glycerol, respectively, the latter being more resistant to this procedure. Nevertheless it is concluded that the T-system is equally important for excitation--contraction coupling in tonic and in phasic fibres.

Animals

Correlation between non-repairable DNA lesions and fixation of cell damage by hypertonic solutions in Chinese hamster cells.

In Chinese hamster HA-1 cells, killing induced by gamma-rays was enhanced by post-irradiation treatment with hypertonic solution (0.5 mol/l NaCl in phosphate buffered saline, pH 7.2) for 20 min. The initial DNA double-strand breaks (dsb) induced by gamma-rays were repaired during post-irradiation treatment with hypertonic solution. However, hypertonic treatment following gamma-irradiation enhanced the frequency of non-repairable dsb, as compared with the frequency after incubation at 37 degrees C following gamma-irradiation. Hypertonic treatment did not affect the initial half-time for rejoining of dsb. Hypertonic treatment did not enhance cell killing, nor did it enhance the non-repairable dsb when the irradiated cells were incubated at 37 degrees C for 2 h. These results suggest that fixation of gamma-ray-induced potentially lethal damage by hypertonic treatment results from inhibition of the rejoining of dsb.

Animals

Hypertonic solutions in the treatment of hypovolemic shock: a prospective, randomized study in patients admitted to the emergency room.

BACKGROUND: The infusion of small volumes of hypertonic saline solution or hypertonic saline plus dextran 70 is remarkably effective in restoring adequate hemodynamic conditions after hypovolemic shock. This prospective double-blind study compares the immediate hemodynamic effects of a bolus infusion of 7.5% NaCl or 7.5% NaCl plus 6% dextran 70 (both 2400 mOsm/L) in severe hypovolemia. METHODS: One hundred five adult patients admitted in hypovolemic shock (systolic blood pressure less than 80 mm Hg) were revived on arrival to the emergency room and administration of a 250 ml intravenous bolus of hypertonic saline solution (n = 35), hypertonic saline plus dextran (n = 35), or isotonic saline solution (n = 35). This infusion was immediately followed by standard crystalloid and blood replacement until systolic pressure reached 100 mm Hg. Mean arterial pressure (MAP) was measured every 5 minutes, and all intravenous infusions were registered. Plasma volume expansion was calculated from plasma protein concentration measurements. Patients were followed up throughout their hospital course, and results of treatment were recorded. RESULTS: At the end of the infusion period, and 5 and 10 minutes after infusion, MAP was significantly higher in patients receiving either hypertonic solution, compared with the group receiving isotonic solution. All groups showed similar trends toward restoration of hemodynamic parameters thereafter. The calculated plasma volume expansion, immediately after the bolus infusion, was significantly higher (24.1% +/- 1.8% and 24.9% +/- 1.1%) in the hypertonic groups, compared with isotonic groups (7.9% +/- 1.3%). Significantly greater volumes of fluids were required to restore systolic pressure in the patients receiving isotonic saline solution than in the groups receiving hypertonic solution. There were no significant differences between the groups receiving hypertonic solutions. The incidence of complications was low, and the mortality rate was similar in all groups. CONCLUSIONS: Infusion of 250 ml hypertonic saline solution in patients with severe hypovolemia was not related to any complications, nor did it affect mortality rates; it improved MAP significantly, acutely expanded plasma volume by 24%, and reduced significantly the volumes of crystalloids and blood required in their resuscitation.

Adult

The effect of hypertonic solutions on the rate of relaxation of contracture tension in Mytilus smooth muscle.

1. The effect of the bathing solution tonicity on the mechanical properties of the anterior byssal retractor muscle (ABRM) of Mytilus edulis was examined. 2. The rate of relaxation of contracture tension produced by acetylcholine (ACh) was greatly reduced when the bathing solution tonicity was increased by adding NaCl, KCl or LiCl after the removal of ACh, whereas a decreased tonicity increased the rate of relaxation. 3. The contracted ABRM in hypertonic solutions showed no active shortening after an isotonic release and barely redeveloped active tension after a quick release. 4. The resistance to stretch increased with increasing tonicity of the bathing solution. 5. The wet weight of the ABRM decreased much more markedly in sucrose-hypertonic solution than in Na-, K- or Li-hypertonic solutions, but the decrease in the rate of relaxation was less marked in the former, indicating that there may be little relation between the rate of relaxation and the degree of osmotic deformation of the ABRM fibres. 6. It is suggested that the elevated ionic strength in the myoplasm may be related to a reduction in the rate of detachment of the cross-linkages between the thick and thin filaments.

Animals

Coagulation changes during second-trimester abortion induced by intra-amniotic prostaglandin E2 and hypertonic solutions.

The coagulation system was studied in twenty-seven patients undergoing second-trimester abortion induced by intra-amniotic prostaglandin (P.G.) E2 alone and in combination with a hypertonic solution of urea or glucose. Changes consistent with intravascular coagulation, namely a rise in fibrin degradation products and a fall in plasma-fibrinogen and platelet-count, were observed in those patients treated using P.G.E2 with hypertonic urea. Similar but less pronounced changes were found in the group treated using P.G.E2 with hypertonic glucose. In patients treated with P.G.E2 alone no changes suggestive of intravascular coagulation were detected. One patient treated using P.G.E2 with hypertonic urea who did not abort for 26 hours demonstrated changes indicative of a pronounced degree of disseminated intravascular coagulation. These findings show that when abortion is induced using P.G.E2 and a hypertonic solution, particularly hypertonic urea, disseminated intravascular coagulation can occur as a result of a hypertonic agent being used.

Abortion, Induced

Volume changes in sarcoplasmic reticulum of rat hearts perfused with hypertonic solutions.

To explore whether morphometry of intracellular membrane-limited subcompartments can be used to follow physiological volume changes in such subcompartments in hearts rapidly fixed by perfusion fixation, we have measured osmotically induced volume changes in electron micrographs of longitudinally oriented sarcoplasmic reticulum (LSR) and terminal cisterns (TC) of rat left ventricular myocardial cells. Vascular perfusion with solutions whose osmolality varied from 0.67 to 1.88 isomolal showed that in the hyperosmolal range LSR volume decreased linearly. Approximately 79% of LSR luminal volume participated in the osmotic rey unresponsive. By contrast, we found that the TC responded by dilation when hearts were perfused with hypersomolal NaCl, NaI, LiCl, or sucrose. Furthermore, with hyperosmolal NaCl the dilation developed within 1 minute; its rate and extent of development were concentration-dependent; it manifested an obligate association with prior or concomitant T-tubular dilation and was not readily reversible. We conclude that (1) the technique sensitively measures in situ changes of LSR volume; (2) most of LSR luminal water is osmotically responsive, but a significant fraction may not be; (3) exposure to hyperosomolal solutions may bring about (perhaps irreversible) structural changes in the diadic membrane complex, leading to changes in its solute permeability.

Animals