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Prediction and experimental determination of solute output from a Collison nebulizer.

The total output from a nebulizer is made up of aqueous droplets containing solute and a significant component of water vapor. The solvent loss is reflected in an increase in the nebulizer solution concentration over time and this has been described mathematically. This theory, originally described by Mercer et al., was modified to describe the solute output from a three-jet Collison nebulizer. The influence of concentration, air flow (air pressure), volume, and temperature on the output parameters were then studied. Inlet air pressures were 10 (4.1), 20 (6.4), and 40 (10.0) psig (L/min), starting concentrations were 0.1, 2, and 5% (w/w) and initial solution volumes were 10 and 20 mL. To study temperature effects, solutions were nebulized at fixed temperatures ranging from 4 to 50 degrees C. This was achieved by water-jacketing the nebulizer flask. Test solutions consisted of mannitol and a fixed concentration of 11.1 micrograms/mL carboxyfluorescein. Nebulization was carried out for up to 30 min using dry, filtered air at ambient temperature. Total output was determined gravimetrically while solute output was determined by fluorimetry (495-nm excitation, 515-nm emission). Solution concentration changes were also monitored over time by fluorimetry. The results show that the solution and solvent output parameters are independent of concentration, volume, and air flow within the ranges studied but that solvent output, in particular, is highly dependent upon temperature.

Chemistry, Pharmaceutical↗

Cytochrome P-450 content and activity after cold storage of rat hepatocytes in university of wisconsin and sodium-lactobionate-sucrose solutions.

We compared the capacity of University of Wisconsin (UW) and of sodium-lactobionate-sucrose (SLS) hypothermic preservation solutions to maintain the integrity of the hepatic cytochrome P-450-dependent mono-oxygenase system. Isolated rat hepatocytes were stored for 0, 10, 24, and 48 hours in UW or SLS solution and were subsequently cultured shortly at 37 degrees C. Cell viability declined slightly but significantly in a time-dependent manner during cold preservation in either UW or SLS solution, and warm culture exacerbated this effect. Total cytochrome P-450 declined gradually after cold preservation and warm culture to reach values of 70% and 52% of unstored controls in cells preserved for 24 and 48 hours in cold UW solution, respectively. Storage in cold SLS solution yielded a similar decrease to 79% and 59% of unstored controls for the equivalent preservation times. Cytochrome P-450 activity was assessed by the metabolism of theophylline after various cold preservation times in UW or SLS solutions. Production of the major metabolite 1,3-dimethyluric acid was not significantly affected by extended cold preservation periods in either UW or SLS solutions. Similarly, the amount of residual theophylline remained stable in all groups, suggesting that alternative metabolic routes were not modified. These studies show that cold preservation in SLS solution is as effective as that in UW solution in terms of cell viability, cytochrome P-450 content, and activity toward theophylline. In addition, the significant reduction in cytochrome P-450 in conjunction with unaffected theophylline disposition suggests that certain cytochrome P-450 isoforms are specifically damaged by cold preservation and rewarming.

Adenosine↗

Vitrification of pronuclear-stage mouse embryos on solid surface (SSV) versus in cryotube: comparison of the effect of equilibration time and different sugars in the vitrification solution.

The cryopreservation of pronuclear-stage embryos has particular importance in transgenic technology and human assisted reproductive technology (ART). The objective of this study was to improve the efficiency of cryopreservation of pronuclear-stage mouse embryos. Two vitrification methods (solid surface vitrification (SSV) vs. vitrification in cryotube) have been compared with special emphasis on the effect of the exposure of the embryos to the solutions for various times and the sugar content (trehalose, sucrose, or raffinose) of the vitrification solutions. Pronuclear-stage embryos were either exposed to 1 M dimethyl sulfoxide (DMSO) + 1 M propylene-glycol (PG) solution for 2, 5, 10, or 15 min or not exposed to this "equilibration" solution. The vitrification solutions consisted of 2.75 M DMSO and 2.75 M PG in M2 medium supplemented with 1 M trehalose (DPT), 1 M sucrose (DPS), or 1 M raffinose (DPR). In the cryotube method, groups of 15-25 embryos were transferred into a 1.8 ml cryotube containing 30 microl of DPT, DPS, or DPR. After 30 sec, the cryotubes were directly plunged into liquid nitrogen (LN(2)) and stored for 1 day to 1 month. Vitrified samples were warmed by immersing the cryotubes in a 40 degrees C water bath and then immediately diluted with 300 microl of 0.3 M trehalose, sucrose, or raffinose in M2. In the SSV method, after equilibration 15-20 embryos were exposed to DPT, DPS, or DPR solutions for around 20 sec before being dropped in 2-microl drops onto a pre-cooled (-150 to -180 degrees C) metal surface. Vitrified droplets were stored in cryovials in LN(2). Warming was performed by transferring the vitrified droplets into 0.3 M solutions of trehalose, sucrose, or raffinose at 37 degrees C, respectively. Results showed that both SSV and cryotube vitrification methods can result in high rates of in vitro blastocyst development (up to 58.3 and 68.5% with DPR, respectively), not statistically different from that of the controls (58.3 and 64.4%). Even without the equilibration step prior to vitrification, relatively high-survival rates have been achieved, except for the DPS solution. In conclusion, vitrification of pronuclear-stage mouse embryos can result in high rates of in vitro development to blastocyst, and the use of raffinose in the vitrification solution is advantageous to improve cryosurvival.

Animals↗

Water (H2O and D2O) molar absorptivity in the 1000-4000 cm-1 range and quantitative infrared spectroscopy of aqueous solutions.

Water (H2O and D2O) molar absorptivity was measured by Fourier transform infrared transmission spectroscopy in the 1000-4000 cm-1 range at 25 degrees C. A series of assembled cells with path lengths from 1.2 to 120.5 microns was used for these measurements. The optimal path length (the path length of aqueous solution at which the IR spectrum of solute, corrected for water absorbance, has the highest signal-to-noise ratio) was calculated for all water absorbance bands. The results presented here show that the optimal path length does not depend on solute properties and is inversely proportional to the solvent (water) molar absorptivity. The maximal signal-to-noise ratio for measurements of IR spectra of aqueous solution in the 1650 cm-1 spectral region, of primary interest in biological applications, can be obtained at an optimal cell path lengths of 3-4 microns (H2O) and 40-60 microns (D2O). As an example, the signal-to-noise ratio was calculated as a function of the cell path length for the amide I (H2O) and amide I' (D2O) bands of an aqueous lysozyme solution. The molar absorptivities of water bands are several orders of magnitude weaker than those of the strongest bands of biological macromolecules in the same spectral regions. High net water absorbance in aqueous solutions is due simply to the very high molar concentration of water. A method is proposed for the quantitative measuring of the path length of the cell which exploits the molar absorptivity of the strongest water bands (stretching vibrations) or of bands which do not overlap with solute absorbance. A path length in the range from approximately 0.01 micron to approximately 1.0 mm can be determined with high precision using this technique for a samples of known concentration. Problems involved in the proper correction of strong water absorbance in IR spectra of aqueous solutions of biomolecules are discussed, including multiple reflections within the cell, the effects of pH, temperature, and perturbation of water spectral properties by polar solutes, as well as the selection of optimal spectral regions in which one may obtain the most precise absorbance corrections.

Calcium Fluoride↗

Fracture phenomena in an isotonic salt solution during freezing and their elimination using glycerol.

Thermal stress and consequent fracture in frozen organs or cell suspensions have been proposed to be two causes of cell cryoinjury. A specific device was developed to study the thermal stress and the fracture phenomena during a slow cooling process of isotonic NaCl solutions with different concentrations of glycerol (cryoprotectant) in a cylindrical tube. It was shown from the experimental results that glycerol significantly influenced the solidification process of the ternary solutions and reduced the thermal stress. The higher the initial glycerol concentration, the lower the thermal stress in the frozen solutions. Glycerol concentrations over 0.3 M were sufficient to eliminate the fracture of the frozen solutions under the present experimental conditions. To explain the action of glycerol in reducing the thermal stress and preventing the ice fracture, a further study on ice crystal formation and growth of ice in these solutions was undertaken using cryomicroscopy. It is known from previous studies that an increase of initial glycerol concentration reduced frozen fraction of water in the solution at any given low temperature due to colligative properties of solution, which reduced the total ice volume expansion during water solidification. The present cryomicroscopic investigation showed that under a fixed cooling condition the different initial glycerol concentrations induced the different microstructures of the frozen solutions at not only a given low temperature but also a given frozen fraction of water. It has been known that ice volume expansion during solidification is a major factor causing the thermal stress and the interior microstructure is critical for the mechanical strength of a solid. Therefore, functions of glycerol in reducing the total ice volume expansion during water solidification and in influencing interior microstructure of the ice may contribute to reduce the thermal stress and prevent the fracture in the frozen solutions.

Animals↗

Oxygen transport by erythrocyte/hemoglobin solution mixtures in an in vitro capillary as a model of hemoglobin-based oxygen carrier performance.

Oxygen transport behavior of erythrocyte/extracellular hemoglobin mixtures flowing in microvessels was studied as a model of hemoglobin-based oxygen carrier (HBOC) performance. An experimental in vitro 25-microm-diameter capillary model was used to provide detailed oxygen flux measurements for hemoglobin solutions, erythrocyte suspensions, and erythrocyte/hemoglobin solution mixtures. The experimental apparatus includes computerized data acquisition and control coupled to a dual wavelength microspectrophotomer. This apparatus had been previously validated by good agreement of experimental measurements with predictive mathematical models of oxygen transport for either erythrocyte suspensions or hemoglobin solutions. The experimental methodology was extended to measurement of oxygen transport in erythrocyte/hemoglobin solutions. The hemoglobin solutions consisted of either purified or gluteraldehyde polymerized bovine hemoglobin. Dose-response plots were generated by varying the extracellular to intracellular hemoglobin ratio while holding the overall hemoglobin concentration constant. Measurements were also made on unmixed erythrocyte suspensions and hemoglobin solutions to generate limiting cases for comparison. Direct comparison of experimental results showed that both types of hemoglobin solutions were substantially more efficient than erythrocyte suspension in uptake and release of oxygen. Increased extracellular hemoglobin concentration increased oxygen transport efficiency for both uptake and release, even when total hemoglobin concentration was held constant. When only 10% of the total hemoglobin was extracellular, approximately half of the increased efficiency of pure hemoglobin solutions was reached. When 50% of the total hemoglobin was extracellular, the increased efficiency was virtually equal to that of pure hemoglobin solutions.

Animals↗

Standardization of nitric oxide aqueous solutions by modified Saltzman method.

Nitric oxide (NO) aqueous solutions were prepared by saturating pure NO gas and hydrolyzing 1 mM 1-hydroxy-2-oxo-3-(N-methyl-3-aminoethyl)-3-methyl-1-triazene (NOC-7), a NO donor, under anerobic conditions. The modified Saltzman method was employed for standardization of the NO aqueous solutions. NO and NO(2) in the solutions were driven with nitrogen gas stream into the first Saltzman solution to measure NO(2) and the leaked NO was driven with air stream through an oxidizing solution into the second Saltzman solution to measure NO, and NO(-)(2) and NO(-)(3) in the residual solutions were determined directly and after reduction with nitrate reductase, respectively. The concentrations of nitrogen oxide species in the NO solutions were about 1.8 mM NO/0.01 mM NO(2)/0.1 mM NO(-)(2)/0.1 mM NO(-)(3), and unchanged during keeping at 20 degrees C for 1 h under anerobic conditions but became 0.05 mM NO/0.01 mM NO(2)/1.7 mM NO(-)(2)/0.1 mM NO(-)(3) by keeping at 20 degrees C for 10 min under aerobic conditions. Instability of NO under aerobic conditions was supported by consumption of 1/4 equivalent amount of dissolved oxygen, and by loss of ability to convert 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO) to carboxy-PTI. Simultaneous quantification of nitrogen oxide species by the modified Saltzman method was found to be useful for practical standardization of NO aqueous solutions.

Electron Spin Resonance Spectroscopy↗

Chemical analysis of freshly prepared and stored capsaicin solutions: implications for tussigenic challenges.

The purpose of this study was to assess the stability of stored capsaicin solutions and the actual concentrations of prepared solutions. Capsaicin solutions ranging in concentration from 0.5 to 128 microM were mixed and analyzed using high performance liquid chromatography. Samples of varying concentrations were then stored under 4 environmental conditions: 4 degrees C and protected from light, room temperature (RT) exposed to light, RT protected from light, and -20 degrees C and protected from light. The concentrations were measured every other month for 1 year. Actual concentrations of freshly prepared solutions were on average 88.3% of predicted. For solutions stored at 4 degrees C, there was a decrease only in the lower concentrations (0.5, 1, and 2 microM) after 2 months (P=0.003). Solutions stored at RT exposed to light decreased in concentration after 6 months (P=0.020), and solutions stored at RT protected from light decreased in concentration after 4 months (P=0.026). The group stored at -20 degrees C decreased in concentration after 1 year (P=0.033). We conclude that the actual concentration of capsaicin solution is less than predicted, and solutions of 4 microM or higher concentration are stable for 1 year if stored at 4 degrees C protected from light.

Administration, Inhalation↗

A superfusion bath for single-cell recording with high-precision optical depth control, temperature regulation, and rapid solution switching.

A superfusion system for isolated cell recording, which uses a novel optical technique for feedback control of solution depth (+/- 5 microns), is described. A retroreflective optosensor senses the distance between itself and a white hydrophobic plastic float, supported by surface tension on the surface of the solution. The combination of optical depth sensing with stepper-motor-driven pumps, pneumatic ("noise-free") solution switching and Peltier-effect solution heating/cooling provides a high degree of control over the cell superfusion environment. The solution flow rate to the bath is selected over the range 0.08-5.00 (+/- 0.01) ml min-1 with provision for solution recirculation. The temperature control range of the Peltier element is 4.0 degrees-70.0 degrees C. At temperatures selected over the range 16.0 degrees-36.0 degrees C, and at flow 3.00 ml min-1 the temperature gradient through the central working position of the bath is not more than 0.2 degrees C. At this flow, bath solution exchange on valve actuation is 90% complete within 0.7 s. This superfusion system is particularly suited to conventional and fluorescence imaging techniques where accurate control of the solution level ensures constant optical conditions. In electrophysiological experiments this apparatus will also provide stabilization of electrode capacitance with adjustment of solution depth. The robust and compact design of the system allows it to be carried between different applications.

Cell Physiological Phenomena↗

Neurohumoral mechanism involved in augmentation of canine jejunal absorption following oral rehydration solutions.

Throughout the world diarrheal diseases kill over 5 million children annually. Oral rehydration therapy, initially using glucose-based solutions and more recently cereal-based solutions, prevents complications and death from dehydration. These experiments compared the effect of these two rehydration solutions and a mixed meal on jejunal water and ionic transport. Five dogs had 25-cm proximal jejunal Thiry-Vella fistulae constructed. Following recovery, jejunal absorption studies (N = 40) were performed using an isotonic electrolyte solution containing [14C]PEG to calculate net fluxes of water, sodium, and chloride. Each study consisted of a 1-hr basal period, followed by a 3-hr experimental period. Each animal was randomly studied in each of four study groups: control, mixed meal, glucose-based and cereal-based rehydration solution. In the mixed meal, glucose-based, and cereal-based solution groups there were significant increases (P < 0.0001) in jejunal Thiry-Vella fistula water and ion absorption following the stimuli, in the absence of direct luminal nutrient contact with the Thiry-Vella fistula. There were no differences between the observed responses to the glucose-based or cereal-based rehydration solutions. Glucose-based and cereal-based rehydration solutions were equally effective in stimulating jejunal absorption of water and electrolytes, but less effective than a mixed meal. Both food and oral rehydration solutions appear to increase jejunal absorption partially via a neurohumoral mechanism that is independent of luminal nutrient contact with the Thiry-Vella fistula.

Animals↗

The effect of fructose-1, 6-diphosphate and HTK solution on protecting primary cardiac muscle cells of rat with cold preservation.

In this study we tried to investigate the effect of fructose-1,6-diphosphate and HTK solution on protecting primary cardiac muscle cells of rat with cold preservation. The primary cardiac muscle cells of rat were cultured in vitro with four preservation solutions respectively: 0.9% sodium chloride solution (group A), FDP (group B), HTK solution (group C) and a mixture of FDP and HTK solution (group D). The cells were preserved for 6, 8 and 10 h at 0-4 degrees C. The values of AST and LDH-L and the Na+-K+ ATPase activity in cardiac muscle cells were detected, and the survival rate of cardiac muscle cells was detected with trypan blue staining. The values of AST and LDH-L in group C and group D were remarkable lower those in group A and group B (P<0.001), while the Na+-K+ ATPase activity and the survival rate of cells in group C and group D were much higher than those in group A and group B (P<0.001). The values of AST and LDH-L after 6 hours in group D decreased much more than those in group C (P<0.01), while the Na+-K4 ATPase activity and the survival rate of cells in group D improved more than those in group C (P<0. 01). Both of the HTK solution and the mixture of HTK and FDP solution have an evident effect on protecting the primary cardiac muscle cells of rat in vitro with cold preservation, Compared with the HTK solution, the mixture solution has a better short-term protective effect.

Animals↗

Effect of IGL-1, a new preservation solution, on kidney grafts (a pre-clinical study).

Ischemia-reperfusion injury conditions short-term and long-term graft function. The effects of the inversion of K+ and Na+ concentrations and substitution with polyethylene glycol for hydroxyethyl starch in University of Wisconsin (K-UW) solution were evaluated in isolated perfused rat kidneys and in autotransplanted pig kidneys. In the rat model kidneys were cold-stored for 24 h in K-UW or Na-UW or Na-PEG UW solutions (IGL-1 solution). Fractional sodium reabsorption and glomerular filtration rate was better in kidneys preserved in Na-UW and IGL-1 solution than those preserved in K-UW solution. In the pig model the left kidney was harvested and preserved in K-UW or IGL-1 solution for 24 h and then transplanted. In the autotransplanted pig model, kidneys preserved in IGL-1 solution showed a better function and a significant reduction of MHC class II expression, cellular apoptosis and interstitial fibrosis. In conclusion, kidneys preserved in IGL-1 solution tolerated ischemia/reperfusion injury better than those preserved in K-UW solution.

Animals↗

Scientific basis for the use of hypotonic solutions with ultrasonic liposuction.

BACKGROUND: A number of plastic surgeons have advocated using hypotonic solution in ultrasound lipoplasty, theorizing that induced adipocyte swelling increases membrane susceptibility to ultrasonic disruption. Additionally, it has been theorized that potassium increases membrane permeability. This study aimed to determine the effect of solution osmolality on adipocyte diameter, the time course of hypotonic solution action, and the effect of potassium addition on adipocyte diameter. METHODS: Base solutions with three different osmolalities were prepared: normal saline (NS) (154 mOsm/l), 1/2NS (77 mOsm/l), and 1/4NS (38.5 mOsm/l). Each solution was modified to contain 0, 5, and 10 mEq/l of potassium and adjusted to starting osmolality. Adipocytes of six patients were suspended in the nine solutions, and diameters were determined at 0, 15, 30, and 45 min. Diameters were measured using imaging software (Kodak ID 3.6). RESULTS: At time 0, the average adipocyte diameter was 79 +/- 8 microm, and no difference was seen in any of the solutions. Cells in the NS group showed no significant increase in diameter over 45 min. The 1/2NS group achieved an 8% +/- 1.9% increase in diameter at 45 min (p < 0.05). The 1/4NS group showed an increase by 14% +/- 2.4% (p < 0.01) at 15 min, and 15% +/- 2.3% (p < 0.01) at 45 min. Potassium had no independent effect on cell diameter. CONCLUSIONS: Hypotonic solution can significantly increase human adipocyte diameter. The findings showed that 1/2NS had a significant effect within 15 min. Tumescent solutions with an osmolality of 1/4NS may be useful in facilitating ultrasonic lipoplasty.

Adipocytes↗

Neutral-pH peritoneal dialysis solution improves peritoneal function and decreases matrix metalloproteinase-2 (MMP-2) in patients undergoing continuous ambulatory peritoneal dialysis (CAPD).

BACKGROUND: Conventional lactate-buffered peritoneal dialysis (PD) solutions have several bioincompatible characteristics, including acidic pH, lactate buffer, and the presence of glucose degradation products (GDPs), and these characteristics contribute to membrane dysfunction in PD patients. The formation of GDPs can be reduced by separating the glucose component of the solution from the lactate component during sterilization. This study was carried out to evaluate the clinical effect of dual-chambered neutral-pH PD solution in patients on continuous ambulatory peritoneal dialysis (CAPD). METHODS: Thirteen CAPD patients using conventional PD solution were enrolled in this study. The fast peritoneal equilibration test (fast PET) was performed periodically before and after treatment with neutral PD solution. The concentration of matrix metalloproteinase-2 (MMP-2) in dialysate effluent was measured using 4-h dwelling 2.5% glucose dialysis solution. The patients were categorized into two groups, according to the value of the initial dialysate/plasma (D/P) creatinine ratio: i.e., lower transporters (group L, D/PCr < 0.65) and higher transporters (group H, D/PCr >/= 0.65). RESULTS: The mean D/P creatinine ratio measured by fast PET, was significantly decreased (0.72 +/- 0.09 to 0.60 +/- 0.06; P < 0.03) after treatment with neutral PD solution in group H. The mean glucose level in 4-h dwelling dialysate effluent was elevated (824.6 +/- 195.9 mg/dl to 942.6 +/- 147.8 mg/dl; P < 0.022) in all subjects. In group H, a significant decrease of MMP-2 in the dialysate effluent was recognized from 15 months after the beginning of treatment with the neutral PD solution (141.4 +/- 52.5 ng/ml to 91.3 +/- 15.1 ng/ml; P < 0.05), with the lowest value being shown at 21 months (80.0 +/- 31.8 ng/ml; P < 0.03). CONCLUSIONS: Neutral-pH peritoneal dialysis solution decreased the MMP-2 level in dialysate and improved peritoneal function in high-transporter patients with CAPD treatment.

Adult↗

Membrane potentials in Rana temporaria muscle fibres in strongly hypertonic solutions.

Conventional microelectrode methods were used to measure variations in resting membrane potentials, E(m), of intact amphibian skeletal muscle fibres over a wide range of increased extracellular tonicities produced by inclusion of varying extracellular concentrations of sucrose. Moderate increases in extracellular tonicity to up to 2.6x normal (2.6tau) under Cl(-) free conditions produced negative shifts in E(m) that followed expectations for the K(+) Nernst equation (E(K)) applied to a perfect osmometer containing a conserved intracellular K(+) content despite any accompanying cell volume change. In contrast, E(m) remained stable in fibres studied in otherwise similar Cl(-) containing solutions, consistent with E(m) stabilization despite negative shifts in E(K) through inward cation-Cl(-) co-transport activity. Short exposures to higher tonicities (>3tau) similarly produced negative shifts in E(m) in Cl(-) free but not Cl(-) containing solutions. However, prolonged exposures to solutions of >3tau caused gradual net positive changes in E (m) in both Cl(-) containing and Cl(-) free solutions suggesting that these changes were independent of cation-Cl(-) transport. Indeed, there was no evidence of cation-Cl(-) co-transport activity in strongly hypertonic solutions despite its predicted energetic favourability, suggesting its possible regulation by E (m) in muscle. Additional findings implicated a failure to maintain greatly increased transmembrane [K(+)] gradients in these E(m) changes. Thus: (1) halving or doubling [K(+)](e) produced negative or positive shifts in E(m), respectively in isotonic or moderately hypertonic (<2.7tau), but not strongly hypertonic (>3tau) solutions; (2) subsequent restoration of isotonic extracellular conditions produced further positive changes in E(m) consistent with a dilution of the depleted [K(+)](i) by fibres regaining their original resting volumes; (3) quantitative modelling similarly predicted a gradual net efflux of K(+) as the balance between active and passive [K(+)] fluxes altered due to increased transmembrane [K(+)] gradients in hypertonic and low [K(+)](e) solutions. However, the observed positive changes in E(m) in the most strongly hypertonic solutions eventually exceeded these predictions suggesting additional limitations on Na(+)/K(+)-ATPase activity in strongly hypertonic solutions.

Animals↗

Myocardial preservation related to magnesium content of hyperkalemic cardioplegic solutions at 8 degrees C.

This study investigates whether the addition of magnesium to a hyperkalemic cardioplegic solution containing 0.1 mM ionized calcium improves myocardial preservation, and whether there is an optimal magnesium concentration in this solution. Isolated perfused rat hearts were arrested for two hours by this cardioplegic solution, which was fully oxygenated and infused at 8 degrees C every 15 minutes to simulate clinical conditions. The cardioplegic solution contained either 0, 2, 4, 8, 16, or 32 mM magnesium. At end-arrest, the myocardial creatine phosphate concentration (nanomoles per milligram of dry weight) was 20.7 +/- 2.1, 22.9 +/- 1.7, 24.8 +/- 2.0, 31.3 +/- 1.4, 33.1 +/- 1.8, and 31.6 +/- 0.8, respectively, in hearts given cardioplegic solution containing these magnesium concentrations. Thus, the concentration of creatine phosphate was significantly higher at end-arrest when the cardioplegic solution contained 8, 16, or 32 mM than 0 or 2 mM magnesium (p less than 0.002) or 4 mM magnesium (p less than 0.02), and highest with 16 mM magnesium. Also, creatine phosphate was more sensitive to the magnesium concentration of the cardioplegic solution than was end-arrest adenosine triphosphate levels, which did not differ among the experimental groups. Aortic flow, expressed as a percentage of prearrest aortic flow, was 60.3 +/- 5.0, 70.2 +/- 5.5, 71.6 +/- 4.4, 71.8 +/- 4.8, 81.0 +/- 5.0, and 71.8 +/- 5.3, respectively. The addition of magnesium to the cardioplegic solution improved recovery of aortic flow (p less than 0.05, 16 mM versus 0 mM magnesium). We conclude from these data that with deep myocardial hypothermia and at an ionized calcium concentration of 0.1 mM, the addition of magnesium, over a broad concentration range, improved preservation of myocardial creatine phosphate and, at a concentration of 16 mM, improved aortic flow. The optimal magnesium concentration in the cardioplegic solution was 16 mM.

Adenine Nucleotides↗

Role of potassium concentration in cardioplegic solutions in mediating endothelial damage.

We studied the effect of potassium concentration in cardioplegic solutions on endothelial function by examining its influence on 5-hydroxytryptamine- (5-HT) and nitroglycerin-induced vasodilation in the isolated rat heart. Forty-eight rat hearts were perfused on a modified Langendorff preparation. After a baseline record of increase in coronary flow induced by 10(-7) M 5-HT and 10 micrograms/mL nitroglycerin, the hearts were perfused for 30 or 60 minutes with either St. Thomas' solution or Bretschneider solution containing 20 mmol/L of potassium or for 30 minutes with either solution containing 30 mmol/L of potassium (n = 8 in each). Initially, 5-HT and nitroglycerin caused a 39.0% +/- 3.3% and 39.7% +/- 2.8% increase in coronary flow, respectively. After 30 or 60 minutes' perfusion with St. Thomas' solution containing 20 mmol/L of potassium, there was little change in the response to 5-HT or nitroglycerin (5-HT, 43.1% +/- 4.1%; nitroglycerin, 38% +/- 3.2%). Similarly, perfusion with Bretschneider solution (20 mmol/L K+) for 30 or 60 minutes did not alter the degree of vasodilation (5-HT, 39.2% +/- 2.9%; nitroglycerin, 38.0% +/- 3.3%). However, perfusion with St. Thomas' solution containing 30 mmol/L of potassium for 30 minutes abolished the endothelial-dependent 5-HT-induced vasodilation (5-HT, -1.6% +/- 1.4%; nitroglycerin, 36.9% +/- 2.2%). Perfusion with Bretschneider solution (30 mmol/L K+) gave similar results (5-HT, -2.1% +/- 1.2%; nitroglycerin, 36.4% +/- 1.7%). We conclude that the concentration of potassium in cardioplegic solutions plays a critical role in causing functional endothelial damage.

Animals↗

Influence of pH of preservation solution on lung viability.

Recently, various solutions for organ preservation have been introduced. Despite much debate regarding the composition of preservation solutions, the ideal pH of a preservation solution has not been determined. We investigated the effects of the pH of a lung preservation solution on viability using an isolated, ventilated lung perfusion model. Initially, we performed flush perfusion with solutions of five different pH groups (6.60, 7.00, 7.40, 7.80 and 8.20, measured at 10 degrees C). Lungs perfused with pH 6.60 solution had a significantly higher pulmonary vascular resistance than those perfused with pH 7.40, 7.80 and 8.20 solutions. Blood was washed out completely and uniformly with solutions of pH 7.40 and 7.80. After preservation the arterial oxygen tension was significantly greater in pH 7.40 and 7.80 groups than in pH 6.60, 7.00, or 8.20 groups. Moreover, pulmonary artery pressure, airway pressure, and wet/dry weight ratios in the pH 7.40 and 7.80 groups were lower than in the others. We conclude that for both pulmonary flush and storage preservation, solutions of pH 7.40 to pH 7.80 are optimal for lung preservation.

Animals↗