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Biomedical subjects

G Navon

Publications and source records attributed to G Navon.

At least 73 records · Page 4Linked to original sources
PubMed1993

Heat acclimation improves cardiac mechanics and metabolic performance during ischemia and reperfusion.

Cardiac mechanics and metabolic performance were studied in isolated perfused hearts of heat-acclimated (AC) rats (at 34 degrees C for 1 mo) and their age-matched controls (C). Diastolic and systolic pressures, coronary flow, and the appearance of ischemic contracture (IC) were measured during progressive graded ischemia, total ischemia (TI), and reperfusion. ATP, phosphocreatine, and intracellular pH were measured during TI and reperfusion with the use of 31P-nuclear magnetic resonance spectroscopy. Systolic pressure was greater in AC hearts than in C hearts (P < 0.0001). During 50% of perfusion pressure 15 and 46% of AC and C hearts, respectively, showed IC (P < 0.001). During 25% of perfusion pressure 85% of the hearts in both groups developed IC. The onset of IC in AC hearts was delayed compared with in C hearts. On reperfusion 93 and 66% of AC and C hearts, respectively, resumed contraction. Recovery of diastolic pressure was 78 and 36% for the AC and C hearts, respectively (P < 0.05). During TI ATP declined by 0.94 and 1.20 mumol/min in AC and C hearts, respectively, resulting in 21 +/- 2.8% preservation of the ATP pool in AC hearts after 30 min of TI (P < 0.001). The AC group also showed a delayed decline in intracellular pH (P < 0.001). The data suggest beneficial effects of heat acclimation on the heart, which were exhibited by greater pressure generation and by the emergence of protecting features during ischemia and reperfusion, possibly via energy-sparing mechanisms.

Acclimatization↗
PubMed1992

Sodium ion transport in rat hearts during cold ischemic storage: 23Na and 31P NMR study.

The success of heart transplantation is limited by the negative correlation between the length of the cold ischemic storage period and the quality of functional recovery. We use 23Na, 31P NMR spectroscopy, and hemodynamic parameters to describe temperature-dependent changes in sodium influx and the concentration of phosphorus high-energy compounds during different storage periods. Perfusion with Krebs-Henseleit solutions containing Dy(TTHA)3- permitted discrimination of intra- and extracellular sodium during cold ischemic storage. The 23Na NMR visibilities under the acquisition and processing parameters used in our experiments were 40 +/- 4% for the intracellular compartment and 97 +/- 11% for the extracellular compartment. At 4 degrees C, the intracellular Na+ accumulation exceeded that observed at 15 and 22 degrees C. The ATP and PCr depletion rates were much lower at 4 degrees C and the left ventricular contractility was higher after longer periods of storage, as the storage temperature decreased. The intracellular Na+ concentration cannot serve as a marker for the postischemic recovery probability. The relative activity of the Na/K ATPase pumps is not correlated with the preservation success. However, intracellular sodium ion accumulation is a major factor in the time lag of the reperfusion recovery.

Animals↗
PubMed1992

Optimal level of hypothermia for prolonged myocardial protection assessed by 31P nuclear magnetic resonance.

The optimal level of hypothermia during myocardial preservation for cardiac transplantation is not known. Phosphorus 31 nuclear magnetic resonance spectroscopy was used to assess the effect of different preservation temperatures (15 degrees C in group 1, 4 degrees C in group 2) on the myocardial high-energy phosphate profiles during prolonged global ischemia and subsequent reperfusion of isolated rat hearts. Adenosine triphosphate depletion during ischemia was more gradual in group 2, leading to significant differences in myocardial adenosine triphosphate concentrations between the two groups after 3 hours of ischemia. The fall in intracellular pH during ischemia was significantly less pronounced in hearts preserved at 4 degrees C as compared with those at 15 degrees C. The postischemic recovery of both the left ventricular peak systolic pressure and the maximum rate of increase of left ventricular pressure was enhanced in group 2, although the ischemic period was 3 hours longer than in group 1. Hypothermia at 4 degrees C as compared with 15 degrees C appears to prolong myocardial protection with respect to adenosine triphosphate preservation, prevention of the fall in intracellular pH, and the enhancement of postischemic hemodynamic recovery.

Adenosine Triphosphate↗
PubMed1992

The effectiveness of University of Wisconsin solution on prolonged myocardial protection as assessed by phosphorus 31-nuclear magnetic resonance spectroscopy and functional recovery.

The effectiveness of the University of Wisconsin solution on extended myocardial preservation was examined in this study using phosphorus 31-nuclear magnetic resonance spectroscopy. Isolated perfused rat hearts were arrested and stored in four preservation solutions: group 1, modified Krebs-Henseleit solution; group 2, modified St. Thomas' Hospital solution; group 3, oxygenated modified St. Thomas' Hospital solution containing 11 mmol/L glucose; and group 4, University of Wisconsin solution. The changes in myocardial high energy phosphate profiles and the intracellular pH values were measured during 12 hours of cold (4 degrees C) global ischemia and 90 minutes of normothermic reperfusion. Following ischemia, the hearts were assessed for hemodynamic recovery and myocardial water content. During ischemia, adenosine triphosphate depletion was observed in all groups; however, after 5 hours of ischemia, the adenosine triphosphate levels were significantly higher in group 3 compared with the other groups (adenosine triphosphate levels at 6 hours in mumol/gm dry weight: group 3, 7.6; group 4, 3.2; group 2, < 1; p < 0.025). The tissue water content at the end of ischemia was lower with the University of Wisconsin solution compared with the modified St. Thomas' Hospital solution or the oxygenated modified St. Thomas' Hospital solution (in ml/gm dry weight: group 4, 3.0; group 2, 4.4; group 3, 3.9; p < 0.05). The adenosine triphosphate repletion during reperfusion was greater with the University of Wisconsin solution compared with the modified St. Thomas' Hospital solution or the oxygenated modified St. Thomas' Hospital solution (12 mumol/gm dry weight in group 4; 8.1 in group 2; 9.0 in group 3; p < 0.05). Similar findings were obtained for the recovery of left ventricular pressure (in percent of preischemic control: group 4, 70%; group 2, 42%; group 3, 52%; p < 0.01) and coronary flow (group 4, 61%; group 2, 49%; group 3, 49%; p < 0.05). These data suggest that preservation with the University of Wisconsin solution affords improved hemodynamic recovery, enhanced adenosine triphosphate repletion, and reduced tissue edema upon reperfusion; however, oxygenated St. Thomas' Hospital solution with glucose is associated with the preservation of higher myocardial adenosine triphosphate levels during prolonged cold global ischemia. In conclusion, these data indicate that the University of Wisconsin solution might improve graft tolerance of ischemia in clinical heart transplantation.

Adenosine↗
PubMed1992

[31-phosphor-NMR spectroscopy for determining optical preservation temperature during long-term myocardial ischemia].

The ideal level of hypothermia during myocardial preservation for cardiac transplantation remains unknown. Therefore 31P-NMR spectroscopy was applied to assess the effect of different preservation temperatures (15 degrees C in group 1, 4 degrees C in group 2) on time dependent changes of myocardial high energy phosphorous compounds during prolonged global ischemia (5 hours in group 1, 8 hours in group 2) and subsequent reperfusion of isolated rat hearts preserved with modified St. Thomas Hospital solution. ATP-depletion during ischemia was slower in group 2 leading to a significant difference in myocardial ATP-concentrations between both groups after 3 hours of ischemia. The drop of intracellular pH during ischemia was significantly less pronounced in hearts preserved at 4 degrees C compared to 15 degrees C. Postischemic recovery of both left ventricular (LV) peak systolic pressure and its +dP/dt max. was superior in group 2, although the ischemic time was 3 hours longer than in group 1. Hypothermia at 4 degrees C appears favourable for prolonged myocardial protection compared to 15 degrees C with regard to preservation of ATP, prevention of intracellular acidosis and postischemic hemodynamic recovery.

Adenosine Triphosphate↗
PubMed1991

Acetylcholine interactions with tryptophan-184 of the alpha-subunit of the nicotinic acetylcholine receptor revealed by transferred nuclear Overhauser effect.

Acetylcholine interactions with three genetically engineered fusion proteins containing peptides from the nicotinic acetylcholine receptor were studied by 1D and 2D nuclear magnetic resonance methods. The three proteins were Torpedo alpha 184-200, Torpedo alpha 186-198, and human alpha 183-204 of the acetylcholine receptor fused to the first 323 residues of the E. coli protein trpE. Nuclear Overhauser effect studies revealed interactions of bound acetylcholine with tryptophan-184 present in the Torpedo alpha 184-200, and the human alpha 183-204 sequences. These interactions are between the N(CH3)3+ and CH3 groups of acetylcholine with the aromatic protons of tryptophan. The appearance of these cross-peaks indicates a distance of less than 5 A between tryptophan and the bound ligand; however, direct contact has yet to be proven.

Acetylcholine↗
PubMed1991

Molecular dissection of cholinergic binding sites: how do snakes escape the effect of their own toxins?

Snakes have evolved a novel binding site demonstrating selective biorecognition. The snake nicotinic acetylcholine receptor is sensitive to acetylcholine while resistant to the effect of the lethal neurotoxins secreted in their own venom. By subjecting recombinant binding sites to point mutagenesis, biochemical analyses and NMR spectroscopy the binding characteristics of three cholinergic ligands have been measured. The amino acid residue at position 189 has been found to be of particular importance to toxin binding.

Amino Acid Sequence↗
PubMed1991

23Na, 59Co and 2H NMR studies of experimental acute pancreatitis.

Multinuclear MR was applied to the study of experimental acute pancreatitis. A continuous increase, up to 700%, of total, NMR visible, sodium content was detected during the development of the disease through the edematous stage to necrosis. The effect of the elevation of sodium content is amplified by an increased 23Na NMR visibility during the disease. Interstitial and total water volume of pancreases were estimated using the distribution of Co(CN)3-(6) and D2O, by 59Co and 2H NMR, respectively. In healthy pancreases the interstitial compartment was found to comprise 35% of the tissue volume. In the diseased pancreases the penetration volume of Co(CN)3-(6) reached 90% of the total tissue volume, indicating extensive membrane injury. The time dependence of the 23Na NMR in the presence of the shift reagent Na7Dy(tripolyphosphate)2.3NaCl provided additional evidence of the increased permeability of the membranes in the diseased organs.

Animals↗
PubMed1991

Sodium-23 NMR relaxation times in nucleated red blood cells and suspensions of nuclei.

The relaxation behavior of intracellular 23Na in suspensions of chicken erythrocytes and of their nuclei was investigated. The transverse magnetization was found to decay biexponentially. The average relaxation rates for the nucleated chicken erythrocytes are considerably shorter than the average relaxation rates obtained for dog and human nonnucleated red blood cells. Of particular significance is the twofold decrease in the short component of T2. Calculations based on the measured 23Na NMR relaxation rates in suspensions of nuclei indicate that most of the difference between the relaxation rates in the mammalian as compared to the chicken erythrocytes, can be accounted for by the contribution of the nuclei in the latter.

Animals↗
PubMed1990

Direct measurement of agonist binding to genetically engineered peptides of the acetylcholine receptor by selective T1 NMR relaxation.

Interactions of four ligands of the nicotinic acetylcholine receptor with genetically engineered peptides have been studied by NMR. A recombinant cholinergic binding site was prepared as a fusion protein between a truncated form of the bacterial protein trpE and a peptide corresponding to the sequence alpha 184-200 from the Torpedo californica receptor. This construct binds alpha-bungarotoxin while the trpE protein alone does not, and thus serves as a negative control [Aronheim, A., Eshel, Y., Mosckovitz, R., & Gershoni, J. M. (1988) J. Biol. Chem. 263, 9933-9937]. In this study agonist binding to alpha 184-200 is demonstrated by monitoring the T1 relaxation of the ligand's protons in the presence and absence of the recombinant binding site. This binding is specific as it can be competed with alpha-bungarotoxin. Quantitative analyses of such competitions yielded the concentration of binding sites, which corresponded to 3.3% and 16.5% of the total protein, for partially purified and affinity-purified alpha 184-200 constructs, respectively. The KD values for the binding of acetylcholine, nicotine, d-tubocurarine, and gallamine to the affinity-purified construct were 1.4, 1.4, 0.20, and 0.21 mM, respectively, while KD's with the nontoxin binding protein were all above 10 mM. Thus, this is a direct demonstration that the toxin binding domain alpha 184-200 may comprise a major component of the cholinergic agonist site.

Acetylcholine↗
PubMed1990

Effects of 2-deoxyglucose on drug-sensitive and drug-resistant human breast cancer cells: toxicity and magnetic resonance spectroscopy studies of metabolism.

The glycolytic inhibitor 2-deoxyglucose (2-DG) was tested as a potential chemotherapeutic agent for drug-resistant cancer cells. Previously it was found that Adriamycin-resistant human MCF-7 breast cancer cells (ADR) exhibit an enhanced rate of glycolysis compared to their parent wild-type (WT) cell line (R. C. Lyon et al., Cancer Res., 48: 870-877, 1987). We now describe a specific toxic effect of 2-DG on the ADR cells, which is more than 15-fold greater than for WT cells. Using 31P magnetic resonance spectroscopy of perfused MCF7 cells we continuously monitored the accumulation of 2-deoxyglucose 6-phosphate together with concomitant changes in other phosphate-containing metabolites. Kinetic measurements demonstrated that ADR cells accumulated 2-deoxyglucose 6-phosphate faster and to a greater extent than WT cells, while their depletion of high energy compounds (ATP, phosphocreatine) was more pronounced and became irreversible earlier. The phosphorylation of 2-DG could be followed more effectively by the use of 13C magnetic resonance spectroscopy of 2-DG enriched with 13C at C-6, since the signals of 2-DG and 2-deoxyglucose 6-phosphate are clearly resolved and, unlike 31P magnetic resonance spectroscopy, there are no other interfering signals. With the use of this technique with ADR and WT cells the rate of phosphorylation of 2-DG was found to be 11.2 x 10(-4) and 6.5 x 10(-4) mmol/min/mg protein, respectively. The results of these studies indicate that differences in the biochemistry of energy metabolism of resistant cells may make them targets for energy antimetabolites.

Antineoplastic Agents↗
PubMed1990

An apparatus for applying a mechanical massage to rat hearts inside a wide-bore NMR spectrometer.

A device for applying mechanical massage to the isolated perfused rat heart inside a wide-bore NMR spectrometer was developed. This device exerts a squeezing pressure on a fibrillating heart placed inside a NMR spectrometer without interfering with the NMR measurements. It appears that the apparatus is important for obtaining reliable results in the postischemic reperfusion period.

Animals↗
PubMed1990

Differences in metabolite content between intact pancreases and their perchloric acid extracts. A 2D 1H/31P correlation NMR study.

Perchloric and hydrochloric acid extracts of intact pancreases from healthy rats and from rats with experimental acute pancreatitis were analysed using 2D 1H/31P correlation NMR spectroscopy. Major differences were found in the 2D maps between the extracts and the intact tissues. In the case of the intact diseased pancreas the prominent 31P signal at the phosphodiester region has been assigned in our previous work as lecithin/taurocholate complex. However, the signal found in the same chemical shift region in extracts of diseased pancreases as well as healthy ones, is identified here as the phosphodiester residue of oligoribonucleotides. In these extracts additional 31P signals were found and assigned as phosphomonoester and phosphodiester hydrolysis products of RNA. The amounts of these compounds, as a function of the acid concentrations were determined, and conditions for their minimization were defined.

Animals↗
PubMed1989

Identification of a characteristic 31P NMR signal in acute experimental pancreatitis with the aid of 1H-31P correlated 2D measurements of intact pancreas.

Two-dimensional 1H-31P correlation spectroscopy and relaxation time measurements of intact diseased pancreas were performed in order to identify an unknown peak appearing at -0.18 +/- 0.04 ppm in a 31P NMR spectrum of taurocholate-induced experimental acute pancreatitis in rats. The signal was identified as a solubilized lecithin taurocholate complex.

Animals↗
PubMed1989

Mechanical, biochemical, and structural effects of vitamin D deficiency on the chick heart.

The effects of vitamin D deprivation on the chick heart were investigated from three aspects: cardiac contractility (+/- dP/dT), intracellular high-energy phosphorus compounds, and structural differences. Four-week-old vitamin D-deficient chicks were divided into four groups: Group A served as the normal group and received subcutaneous injections of cholecalciferol; Groups B and C were vitamin D-deficient hearts but perfused differently; Group D received daily subcutaneous injections of 5 micrograms of 1,25(OH)2D3. When the isolated spontaneously beating hearts (modified Langendorff preparation) were perfused with Krebs-Henseleit (KH) solution containing a calcium concentration of 2.5mM, the myocardial contractility of the vitamin D-deficient hearts was significantly increased when compared with group A. After the isolated heart had beaten for one hour, the myocardial contractility in the vitamin D-deficient hearts was found to decline to significantly lower values. Presacrifice administration of 1,25(OH)2D3 improved cardiac performance. Vitamin D deficiency resulted in an enhanced rate of decline of the intracellular high-energy phosphorus compounds. No differences were found in the microscopic study. These observations suggest that vitamin D has a role in cardiac function.

Adenosine Triphosphate↗
PubMed1988

Measuring relative acetylcholine receptor agonist binding by selective proton nuclear magnetic resonance relaxation experiments.

A method is presented that uses selective proton Nuclear Magnetic Resonance (NMR) relaxation measurements of nicotine in the presence of the acetylcholine receptor to obtain relative binding constants for acetylcholine, carbamylcholine, and muscarine. For receptors from Torpedo californica the results show that (a) the binding constants are in the order acetylcholine greater than nicotine greater than carbamylcholine greater than muscarine; (b) selective NMR measurements provide a rapid and direct method for monitoring both the specific and nonspecific binding of agonists to these receptors and to the lipid; (c) alpha-bungarotoxin can be used to distinguish between specific and nonspecific binding to the receptor; (d) the receptor--substrate interaction causes a large change in the selective relaxation time of the agonists even at concentrations 100x greater than that of the receptor. This last observation means that these measurements provide a rapid method to monitor drug binding when only small amounts of receptor are available. Furthermore, the binding strategies presented here may be useful for the NMR determination of the conformation of the ligand in its bound state.

Acetylcholine↗
PubMed1988

Conformation of acetylcholine bound to the nicotinic acetylcholine receptor.

We report here the biologically active conformation of acetylcholine when bound to the high-affinity state of the receptor from Torpedo californica. The acetylcholine conformation was determined in the free and bound states by proton NMR two-dimensional nuclear Overhauser effects. In agreement with x-ray crystallographic data, acetylcholine in solution has an extended conformation with an average distance between the acetyl methyl and choline methyl protons of approximately equal to 5 A. When bound to the acetylcholine receptor, acetylcholine adopts a conformation where the acetyl methyl group is close (3.3 A) to the methyl groups of the choline moiety. This bent conformation places the oxygens adjacent to one another and allows the methyl groups to form an uninterrupted hydrophobic surface over the rest of the acetylcholine molecule. The significant difference between the free- and bound-state conformations implies that structure-activity studies based solely on molecular modeling strategies must be approached with caution.

Acetylcholine↗