Search PubMed⌕ Search

Biomedical subjects

R Deslauriers

Publications and source records attributed to R Deslauriers.

At least 73 records · Page 4Linked to original sources
PubMed1994

Relationships between cytosolic [ATP], [ATP]/[ADP] and ionic fluxes in the perfused rat heart: A 31P, 23Na and 87Rb NMR study.

In order to assess the relationship between cytosolic [ATP] or [ATP]/[ADP] and the intracellular Na+ concentration ([Na+]i), we have used the phosphate trap 2-deoxy-D-glucose (DG) to alter the high energy phosphate levels in rat cardiomyocytes. Pyruvate-perfused rat hearts were treated with 2 mM DG in the presence of 10IU/l of insulin for 28 min, followed by perfusion with DG without insulin for 60 min. The DG + insulin treatment resulted in dramatic changes in the 31P NMR spectra: phosphocreatine (PCr) and total ATP decreased (to 15 and 35%, respectively) and deoxyglucose-6-phosphate accumulated, with little change in either inorganic phosphate or intracellular pH. These changes corresponded to a decrease in cytoplasmic [ATP] (from 7.6 to 1.8 mM), [ATP]/[ADP] (from 494 to 24) and ATP affinity [A(ATP), by 8.9 kJ/mol] and an increase in [ADP] (five-fold) and free [Mg2+] (two-fold). Subsequent perfusion with DG--insulin resulted in slow recovery of PCr, [ATP]/[ADP] and A(ATP) such that the "low energy" state lasted an additional 16 min during which ATP remained low and constant. There were no detectable changes in the intracellular Na+ content as assessed by shift reagent-aided 23Na NMR at the end of DG + insulin treatment (98 +/- 18%, 28-36 min of the protocol). In addition, there was no change in the Rb+ influx rate as measured by 87Rb NMR at the beginning of insulin washout which was achieved by replacing 20% of the KCl with RbCl ([K+] = 3.76 mM, [Rb+] = 0.94 mM). During DG + insulin treatment the pressure-rate product (PRP) decreased by half and was restored upon insulin washout to 80% of its initial value both in the presence and in the absence of the shift reagent [5 mM Dy (triethylenetetraminehexaacetate)3-]. These data imply that unfavorable thermodynamic [low A(ATP)] and kinetic (low [ATP] and [ATP]/[ADP]) conditions induced by DG treatment do not inhibit Na+, K(+)-ATPase activity. We speculate that during anoxia when changes in [ATP]/[ADP] are comparable to those induced by DG treatment, the observed increase in [Na+]i is not due to inhibition of the Na+ pump by reduced [ATP] or [ATP]/[ADP].

Adenosine Diphosphate↗
PubMed1993

Simultaneous measurement of both lipid and lactate in isolated rat hearts by 1H NMR spectroscopy.

Myocardial lipid and lactate levels are sensitive indicators of biochemical status: lipid levels have been shown to increase in response to high fat diets, disease or metabolic stress and elevated lactate levels are indicative of reduced oxygen supply. Selective measurement of lactate or lipid levels by 1H NMR is not straightforward since both the lactate methyl group and lipid methylene groups resonate at 1.3 ppm. We have overcome this difficulty by employing spectral editing techniques to observe both lipid methylene and lactate methyl resonances, and have measured lipid and lactate levels in perfused rat hearts during control perfusion and in response to metabolic stress. Lactate increased during ischemia and decreased during reperfusion, and the ischemia-induced increase is inhibited by iodoacetate, as expected. In contrast, lipid levels increased during ischemia and remained elevated during reperfusion. Hearts from rats fed high fat diet show elevated lipid levels during control perfusion. Data obtained by 1H NMR are consistent with biochemical data, validating the technique.

Animals↗
PubMed1993

Magnesium enhances function of postischaemic human myocardial tissue.

OBJECTIVES: The effect of Mg2+ on the developed force and concentrations of high energy phosphate metabolites in isolated human atrial trabeculae has been investigated. METHODS: Human atrial trabeculae, obtained from right atrial appendages of patients undergoing cardiac surgery requiring cardiopulmonary bypass, were dissected at room temperature in modified Krebs-Henseleit buffer containing 1.2 or 16 mM Mg2+, mounted on muscle stands, and rewarmed to 34 degrees C in the same buffer. After 30 minutes, their mechanical function was assessed. At the end of the protocol, trabeculae were fast frozen for measurement of concentrations of metabolites of high energy phosphates. RESULTS: Trabeculae collected and rewarmed in 16 mM Mg2+ Krebs-Henseleit buffer showed significantly higher mean developed force (0.59(SEM 0.10) g, p < 0.01) than those rewarmed in 1.2 mM Mg2+ Krebs-Henseleit buffer (0.32(0.03) g). Trabeculae that had a developed force > or = 0.8 g, a resting force < or = 0.7 g, and a cross sectional area < or = 1 mm2 ("functional" trabeculae) were selected for further comparison. New reverse phase high performance liquid chromatography techniques developed for the analysis of small samples (0.5-5 mg dry weight) were used to measure nucleotide, nucleoside, and creatine compounds. Total adenylate (ATP+ADP+AMP) concentrations in trabeculae revived in the presence of 16 mM Mg2+ (15.4(1.1) mumol.g-1 dry weight) were significantly higher (p < 0.01) than in those revived with 1.2 mM Mg2+ (11.8(1.0) mumol.g-1), but lower (p < 0.01) than in trabeculae fast frozen immediately after removal from the patient (22.6(1.0) mumol.g-1). There were no significant differences in NAD and total creatine (phosphocreatine+creatine) concentrations between the three groups. CONCLUSIONS: The presence of high Mg2+ during the rewarming of human atrial trabecular preparations maintains a significantly higher developed force and a significantly higher total adenylate pool than does collection and rewarming with normal concentrations of Mg2+.

Aged↗
PubMed1993

Perspectives on myocardial protection: warm heart surgery.

Continuous normothermic blood cardioplegia (CNBC), was recently introduced into clinical cardiac surgery and has generated great interest. CNBC represents the evolution of concepts which were initially described in the 1950s but which were forgotten as hypothermia became the main ingredient in all cardioplegic techniques and in heart surgery in general. The historical background to the development of CNBC is presented, as well as a review of the current state of knowledge about normothermic heart surgery. The limitations and pitfalls of the method are highlighted, along with future developments and perspectives.

Forecasting↗
PubMed1992

Phasing spin-echo-acquired 31P spectroscopic images using complex conjugate data reversal.

A simple method has been developed for phasing 31P spectroscopic images acquired with short echo-time (1-2 ms) spin-echo sequences. The technique is based on reconstructing complete echoes in the time domain by the reversal of the complex conjugate of the data. After Fourier analysis, a magnitude reconstruction is used, which no longer broadens the lines. Advantages of the method compared to other phasing procedures are discussed.

Animals↗
PubMed1991

An NMR probe to study function and metabolism simultaneously in isolated human cardiac tissue.

Trabeculae isolated from human atrial appendages have been used to study preservation of donor hearts for cardiac transplantation. We have developed a perifusion system equipped with a fiber optic strain gauge to study mechanical performance of human atrial trabeculae (10-20 mg) while simultaneously observing the energetic compounds by 31P NMR spectroscopy. The NMR probe consists of an eight-turn solenoid coil (2.3 mm i.d. x 5 mm length, 24-gauge wire) double tuned to allow observation of 1H and 31P nuclei. The probe and the perifusion system are temperature regulated and permit preservation studies using a variety of small muscles at low temperatures (down to 4 degrees C) as well as at physiological temperature (37 degrees C). 31P NMR spectra suitable for quantification can be obtained from approximately 10 mg of human atrial trabeculae in 15 min. Spectra of 8- to 12-mg mouse extensor digitalis longus or soleus muscle can be obtained in less than 10 min.

Animals↗
PubMed1991

23Na and flame photometric studies of the NMR visibility of sodium in rat muscle.

23Na nuclear magnetic resonance spectroscopy (NMR) is increasingly being used to study Na+ gradients and fluxes in biological tissues. However, the quantitative aspects of 23Na NMR applied to living systems remain controversial. This paper compares sodium concentrations determined by 23Na NMR in intact rat hindlimb (n = 8) and excised rat gastrocnemius muscle (n = 4) with those obtained by flame photometric methods. In both types of samples, 90% of the sodium measured by flame photometry was found to be NMR-visible. This is much higher than previously reported values. The NMR measurements for intact hindlimb correlated linearly with the flame photometric measurements, implying that one pool of sodium, predominantly extracellular, is 100% visible. From measurements on excised muscle, in which extracellular space is more clearly defined, the NMR visibility of intracellular Na+ was calculated to be 70%, assuming an extracellular space of 12% of the total tissue water volume and an extracellular NMR visibility of 100%. 23Na transverse relaxation measurements were carried out using a Hahn spin echo on both intact hindlimb (n = 1) and excised muscle (n = 2) samples. These showed relaxation curves that could each be described adequately using two relaxation times. The rapidly relaxing component showed a T2 value of 3-4 ms and the slowly relaxing component a T2 of 21-37 ms. A spin lattice relaxation (T1) measurement on intact hindlimb yielded a value of 51 ms. These relatively long relaxation times show that the quadrupolar relaxation effect of Na+ complexing to large macromolecules or being otherwise motionally restricted is relatively weak. This is consistent with the high NMR visibilities reported here.

Animals↗
PubMed1991

Cardiac hypothermia: 31P and 1H NMR spectroscopic studies of the effect of buffer on preservation of human heart atrial appendages.

31P and 1H nuclear magnetic resonance spectroscopy has been used to follow noninvasively the time course of energetic metabolite levels in human heart atrial appendages preserved under various temperatures and buffer conditions. From sample harvest up to the normal 5-h time limit for heart preservation, ATP levels in human atrial appendages are much better maintained in 0.9% saline and PIPES-buffered preservation solutions at 12 degrees C than at 4 degrees C. Furthermore, preservation at 12 degrees C can be improved considerably by using high extracellular buffer concentrations. The increased buffer concentration allows better maintenance of the intracellular pH and leads to a faster glycolytic rate as measured by lactate production. At 4 degrees C, ATP levels decline rapidly during the first 5 h but reached a stable plateau, which is well maintained over 15-20 h. At this temperature, the rate of lactate production is similar at all buffer concentrations (20, 60, and 100 mM PIPES). As a consequence of these observations, we postulate that the mechanisms of ATP production and utilization at 4 degrees C and at 12 degrees C are different. At 4 degrees C, the rate of glycolysis is temperature limited whereas at 12 degrees C, low intracellular pH inhibits glycolysis.

Adenosine Triphosphate↗
PubMed1991

The effect of high buffer cardioplegia and secondary cardioplegia on cardiac preservation and postischemic functional recovery: a 31P NMR and functional study in Langendorff perfused pig hearts.

High buffer cardioplegia may provide protection against ischemic damage by reducing the extent of intracellular acidosis. Secondary cardioplegia may improve postischemic recovery by restoration of high energy phosphates, ionic gradients, and intracellular pH. To test these hypotheses, pig hearts were arrested with high buffer (150 mM MOPS) cardioplegia or modified St. Thomas' solution II and then kept ischemic at 12 degrees C for 8 h. High energy phosphates and intracellular pH were followed during the period of ischemia, using 31P nuclear magnetic resonance spectroscopy, and functional recovery was followed during reperfusion. The hearts arrested by high buffer cardioplegia showed significantly higher intracellular pH than hearts preserved with St. Thomas' solution, but there were no significant differences in high energy phosphates. There were no significant differences in functional recovery. We found, however, that secondary cardioplegia abolished ventricular fibrillation, and resulted in improved functional recovery after 8 h of ischemic preservation compared with the hearts reperfused with Krebs-Henseleit solution alone. Our results suggest that despite attenuating the decreases in intracellular pH, high buffer cardioplegia does not improve recovery following 8 h of preservation at 12 degrees C. Secondary cardioplegia reduces the incidence of ventricular fibrillation and improves postischemic functional recovery of the myocardium.

Adenosine Triphosphate↗
PubMed1990

Optimization of magnetization transfer measurements: statistical analysis by stochastic simulation. Application to creatine kinase kinetics.

A systematic study was performed to optimize the accuracy of kinetic parameters derived from magnetization transfer measurements. Three techniques were investigated: time-dependent saturation transfer (TDST), saturation recovery (SRS), and inversion recovery (IRS). In the last two methods, one of the resonances undergoing exchange is saturated throughout the experiment. The three techniques were compared with respect to the accuracy of the kinetic parameters derived from experiments performed in a given, fixed, amount of time. Stochastic simulation of magnetization transfer experiments was performed to optimize experimental design. General formulas for the relative accuracies of the unidirectional rate constant (k) were derived for each of the three experimental methods. It was calculated that for k values between 0.1 and 1.0 s-1, T1 values between 1 and 10 s, and relaxation delays appropriate for the creatine kinase reaction, the SRS method yields more accurate values of k than does the IRS method. The TDST method is more accurate than the SRS method for reactions where T1 is long and k is large, within the range of k and T1 values examined. Experimental verification of the method was carried out on a solution in which the forward (PCr----ATP) rate constant (kf) of the creatine kinase reaction was measured.

Adenosine Diphosphate↗
PubMed1990

A high-sensitivity, high-B1 homogeneity probe for quantitation of metabolites.

Accurate quantitation of metabolites in biological samples of irregular shape and inhomogeneous composition is generally acknowledged to be difficult. The difficulties are less pronounced with a probe having excellent B1 field homogeneity, high sensitivity, and a resonant frequency independent of sample size and composition. A prototype probe that aims to fulfill these requirements in wide-bore horizontal magnets is described. It comprises four separate tuned rings on a spherical surface which give a B1 field that is flat to +/- 1% over the design volume. Inter-ring coupling, and to a fifth ring used for matching, is by induction, and the mathematics of the tuning of the system are derived. It is shown that resonant frequency variation with sample size is negligible, and that the sensitivity closely approaches the theoretical limit.

Animals↗
PubMed1990

Elimination of signal strength dependency upon coil loading--an aid to metabolite quantitation when the sample volume changes.

The phase, height, and width of NMR spectral lines from a constant number of nuclei are frequently dependent upon changes in probe Q factor and tuning, caused by motion of the sample and/or changes in its electrical conductivity and size. Thus quantitation of metabolites in biological samples can be difficult. When probe tuning and matching are rendered independent by the use of a tuned coupling coil for matching, use of a very low input impedance preamplifier can virtually eliminate such dependencies in the received signal. Likewise, by reciprocity, the use of a low output impedance transmitter greatly reduces the dependence of pulse flip angle upon sample characteristics. Experimental results verifying these statements are presented, and the time course of signals from a swelling, perfused pig heart and an external reference are plotted.

Animals↗
PubMed1989

1H magnetic resonance of human tumours. Analysis of the transverse relaxation of the methylene protons using continuous distributions of relaxation times.

In tumours, the decay of 1H transverse magnetization (relaxation profile) of the methylene resonances is usually not a single exponential. The conventional sum-of-exponentials approximation often leads to several solutions which fit the experimental data; choosing one solution over another is highly subjective. We have therefore analyzed transverse relaxation data for the methylene resonance of lipids in human tumours, assuming a continuous distribution of relaxation times. We have studied 89 colon tumours (27 with metastasis), 12 'normal' colon tissues, 40 breast tumours (24 with metastasis) and 13 malignant lymph nodes. All were primary tumours and the patients had received no previous treatment. Two continuous distribution approximations were tested. The two-parameter lognormal distribution provides a simple representation of the relaxation profile. The constrained regularization method (Contin) complements and extends the lognormal approach. The average T2 values, mean value of T2, whether derived from the lognormal or the Contin approximations, are consistent and comparable. Tumour and 'normal' colon tissue taken from the same patient show important differences in relaxation behaviour. A considerable broadening of the lognormal distribution, with mean value of T2 shifted to shorter values, is observed for the colon tumours. This is verified by the constrained regularization method. The 'normal' colon tissues are usually characterized by a single, relatively narrow distribution, while tumours show one or more broad peaks. A substantial broadening of the distribution of relaxation times is observed in colon tumours, whether metastasized or not. A similar broadening is noted for breast tumours and malignant lymph nodes, although the range is slightly less than for colon tumours.(ABSTRACT TRUNCATED AT 250 WORDS)

Breast Neoplasms↗
PubMed1989

31P NMR studies of the metabolic status of pig hearts preserved for transplantation.

31P NMR spectroscopy has been used to evaluate the metabolic status of cardioplegically arrested pig hearts. Hearts were stored with Plegisol for up to 12 hours at either 5 degrees C or 12 degrees C. Results indicated that the ATP content of hearts could be maintained (greater than 70% of initial values) for up to 5 hours in the ischemic storage state. The ATP loss was greater at 12 degrees C. PCr was lost exponentially under the same conditions. Functional testing by reperfusing the stored hearts in vitro indicated a good correlation between the ATP content and survivability of the preparations. Twenty-four hour preservation of pig hearts using slow perfusion with a modified cardioplegic solution (Wicomb) allowed for preservation of both PCr and ATP, in all cases, reperfusion of hearts revealed a loss of NMR- visible ATP and PCr.

Adenosine Triphosphate↗
PubMed1989

The effects of temperature and buffer concentration on the metabolism of human atrial appendages measured by 31P and 1H NMR.

In this study we measured changes in intracellular ATP and pH together with lactate production in isolated ischemic human atrial tissue. The measurements were made using 31P and 1H NMR. ATP preservation is improved as temperature is reduced from 20 degrees C to 1 degree C because of a progressive decrease in energy demand. At a constant temperature (12 degrees C), ATP preservation is improved by increasing the extracellular buffer capacity with PIPES buffer at concentrations up to 100 mM. Under these conditions, energy demand appears to increase but the ATP level is kept relatively constant for periods of 10 hours or longer. This appears to be due to a tighter regulation between supply and demand in which glycolysis is driven faster at relatively lower ADP and Pi levels. This tight regulation may be attributed to the better maintenance of intracellular pH.

Adenosine Triphosphate↗
PubMed1989

Preservation of high-energy phosphates in human myocardium. A phosphorus 31-nuclear magnetic resonance study of the effect of temperature on atrial appendages.

After prolonged exposure to low temperatures (1 degree and 4 degrees C), human atrial trabeculae show poor recovery of contraction. At somewhat higher temperatures (12 degrees and 20 degrees C), recovery is much better (Keon and associates. Ann Thorac Surg 1988;46:337-41). Although better preservation of adenosine triphosphate and therefore improved contractile recovery might be expected after exposure to lower temperatures, it remained possible that, below a certain temperature, adenosine triphosphate-generating mechanisms could be slowed more than adenosine triphosphate utilization. To investigate this phenomenon further, we followed the time course of metabolic changes in human atrial appendages, harvested during cardiac bypass operations, at 1 degree, 4 degrees, 12 degrees, and 20 degrees C using high-resolution 31P and 1H nuclear magnetic resonance spectroscopy. The results are quantitated by correlation with data obtained from biochemical assays on quick-frozen tissues. Initial adenosine triphosphate levels in myocytes of human atrial appendages are 3.3 to 4.3 mumol.gm-1 tissue wet weight. At 20 degrees C, adenosine triphosphate disappears after 6 hours; at 12 degrees C, about half the initial adenosine triphosphate is still observable at this time; at 4 degrees C or 1 degree C, the decline is still slower. Only a small contribution toward adenosine triphosphate maintenance comes from creatine phosphate, since creatine phosphate, inorganic phosphate, and total creatine levels in the appendage are low (less than 2 mumol.gm-1 tissue wet weight). Glycolysis is active at all temperatures; the rate of glycolysis correlates positively with increasing temperature. Adenosine triphosphate generated by glycolysis falls just short of demand at all temperatures, but the difference is small at 1 degree and 4 degrees C. These studies lead us to conclude that the relatively poor recovery of contractile response of human atrial trabeculae, together with contracture reported previously at lower temperatures (1 degree and 4 degrees C), is not due to a failure to maintain adenosine triphosphate levels.

Adenosine Triphosphate↗