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R W Briggs

Publications and source records attributed to R W Briggs.

At least 37 records · Page 2Linked to original sources

31P NMR visibility and characterization of rat liver mitochondrial matrix adenine nucleotides.

Compartmentation and NMR visibility of mitochondrial adenine nucleotides were quantitated in isolated rat liver mitochondria respiring on succinate and glutamate in vitro at 8 and 25 degrees C. Intra- and extramitochondrial nucleotides were discriminated by adding the chelator trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA). T1 values of about 0.2-0.3 s for magnesium-bound matrix nucleotides were determined. Adenine nucleotide T1 values were influenced by the ionic environment; only magnesium-free ATP T1's were affected by temperature. Intra- and extramitochondrial adenine nucleotide ratios were varied in ATP-loaded mitochondria with added ATP and phosphate using the mitochondrial inhibitors oligomycin and carboxyatractyloside, and adenine nucleotides were quantitated by using NMR and enzymatic analysis. There was good agreement between matrix ATP concentrations (magnesium-bound ATP) calculated by using NMR and standard biochemical techniques. Although matrix ADP could be detected by NMR, it was difficult to quantitate accurately by NMR. The data indicate that mitochondrial ATP is NMR-visible in isolated mitochondria in vitro.

Adenine Nucleotides↗

Nuclear magnetic resonance study of free and bound water fractions in normal lenses.

Proton NMR relaxation times T1 and T2 were determined for normal lenses excised from sexually mature animals from seven different species. Lenses were immersed in physiological buffer during measurements, and suppression methods were employed to null the buffer signal. This enabled selective analysis of lenticular water. Observed relaxation times were correlated with protein and water content. At 37 degrees C and 1.89 Tesla, single-exponential spin-lattice relaxation was observed, but spin-spin relaxation was found to be double-exponential. It was shown that the short-T2 fraction is proportional to protein concentration; this fraction was attributed to water bound to protein. The long-T2 fraction was attributed to free lenticular water. The amounts of free and bound water thus obtained were used in the spin-lattice relaxation rate equation for rapid exchange in a two-component system to calculate the magnitudes of the two corresponding T1 relaxation components.

Animals↗

In vivo 31P and in vitro 1H nuclear magnetic resonance study of hypoglycemia during neonatal seizure.

To examine the hypothesis that hypoglycemia has an adverse effect on brain energy state during seizure, neonatal dogs were subjected to bicuculline-induced seizure while hyperglycemic, normoglycemic, or hypoglycemic. Cerebral blood flow increased and remained elevated in all animals subjected to seizure, regardless of blood or brain glucose concentration. In vivo 31P nuclear magnetic resonance spectroscopy disclosed a small (10-20%) decrease in adenosine triphosphate levels and a greater (20-40%) decline in phosphocreatine levels in animals experiencing seizure, irrespective of whether they were hyper-, normo-, or hypoglycemic. In vitro analysis of brain extracts with 1H nuclear magnetic resonance spectroscopy disclosed a significant elevation of lactate in all seizing animals. There were differences in brain alanine, glycine, and beta-hydroxybutyrate levels between the hyperglycemia-seizure and hypoglycemia-seizure groups. Alternate substrates such as lactate, fatty acids, or amino acids may be used when neonatal seizure is complicated by hypoglycemia, thereby preventing further deterioration of brain metabolic state.

Animals↗

Brain metabolism after electroshock seizure in the neonatal dog: a [31P]NMR study.

In vivo [31P] nuclear magnetic resonance (NMR) spectroscopy disclosed that a 10 second electroshock seizure in oxygenated neonatal dogs produced prolonged alteration of brain phosphocreatine (PCr), inorganic phosphate (Pi), and lactate. The slow return of these metabolites to baseline may be related to lower endogenous stores of high energy phosphates, or less developed pathways for their regeneration.

Adenosine Triphosphate↗

The effect of hypotension on brain energy state during prolonged neonatal seizure.

Prolonged seizures in the human neonate may be complicated by systemic hypotension. To examine the effect of systemic hypotension on brain metabolic state during seizure, neonatal dogs were made hypotensive (by exsanguination) during bicuculline-induced seizure. Measurement of regional cerebral blood flow showed that moderate hypotension did not impair cerebral perfusion during seizure. Measurement of brain energy state with in vivo 31P nuclear magnetic resonance spectroscopy disclosed a similar pattern of alteration of high energy phosphates in animals subjected to seizure or to the combination of seizure and hypotension. The additional metabolic stress imposed by moderate hypotension during seizure in the neonatal dog appears to be slight.

Animals↗

Relation between work and phosphate metabolite in the in vivo paced mammalian heart.

Nuclear magnetic resonance (NMR) spectroscopy was used to monitor, on a beat-to-beat basis, the concentration of creatine phosphate and adenosine triphosphate during alterations in the work output of canine hearts in vivo. Over a wide range of rate-pressure products (5,000 to 25,000 mmHg/min), the relative amounts of creatine phosphate and adenosine triphosphate within the heart remained constant. The relative concentration of free adenosine diphosphate was calculated under the reasonable assumption that the creatine kinase-catalyzed reaction is near equilibrium in this tissue. The free concentration of adenosine diphosphate also did not change over this range of rate-pressure products. The data demonstrate that the concentration of these compounds is highly regulated in vivo and suggest that factors other than their concentration may be involved in the modulation of steady-state myocardial work output with oxygen consumption.

Adenosine Diphosphate↗

Pulse techniques for the suppression of individual components in multiexponential relaxation curves.

NMR pulse sequences have been investigated which selectively suppress undesired signal components from multiple-exponential spin-spin and spin-lattice relaxation curves. Suppression is achieved by exploiting differences in the relaxation times of sample components. Special attention is devoted to discussion of practical details necessary for implementation of these methods, as well as their relative advantages and disadvantages. The techniques are illustrated using multiple-compartment aqueous phantoms and are then employed in measurements of proton T1 and T2 values for freshly isolated guinea pig lenses immersed in aqueous buffer solutions to prevent dehydration and maintain viability. These methods obviate many of the difficulties inherent in obtaining accurate relaxation times for minor components of multiple-exponential curves and should be useful in many applications of NMR to the study of intact tissue, both in vivo and in vitro.

Animals↗

Appearance of Pantopaque on MR images.

Pantopaque was shown to be both a radiographic and magnetic resonance (MR) contrast agent in two patients with suspected cord abnormalities who underwent MR examination following myelography. The Pantopaque appeared as a linear band of high signal intensity along the dependent portion of the spinal canal on MR images obtained with a repetition time of 500 msec and an echo time of 30 msec. MR images of two dogs--one injected with Pantopaque in the cervical subarachnoid space and the other with Amipaque--confirmed that the high signal arose from the Pantopaque. This high signal intensity is due to Pantopaque's short T1 relaxation time. Although Pantopaque will probably not be used as an MR contrast agent, it is important to recognize this property so that its high signal is not mistaken for fat or hemorrhage in the subarachnoid space on MR images.

Aged↗

Gated in vivo examination of cardiac metabolites with 31P nuclear magnetic resonance.

Phosphorus-31 nuclear magnetic resonance (31P NMR) spectroscopy was used to study the temporal aspects of metabolism of canine heart in vivo. An NMR catheter coil was passed through the jugular vein of a dog into the apex of the right ventricle, and spectra were recorded at four points in the cardiac cycle by triggering from the blood pressure trace of the animal. The 31P spin-lattice relaxation times of phosphocreatine (PC) and the gamma-, alpha-, and beta-phosphates of ATP at 1.89 Tesla are 4.4, 1.8, 1.7, and 1.6 s, respectively. The ratio of PC to ATP is 2.0. No changes in PC/ATP were noted in any of the four portions of the cardiac cycle examined, and difference spectra exhibited no observable signals, in contrast to previously reported results for glucose-perfused rat hearts. On the assumption that intracellular pH and the total creatine pool were constant, the equilibrium expression for the creatine kinase reaction was used to deduce that free ADP concentrations were invariant throughout the cardiac cycle. This is in apparent disagreement with the proposed regulatory role for ADP in heart oxidative phosphorylation.

2,3-Diphosphoglycerate↗

31P nuclear magnetic resonance study of the effect of hypoxemia on neonatal status epilepticus.

Prolonged neonatal seizures are often accompanied or exacerbated by hypoxemia. To determine the effects of hypoxemia on neonatal status epilepticus, we determined cerebral blood flow and cerebral metabolic state in groups of neonatal dogs subjected to hypoxia, to seizures during normoxia, or to seizures during hypoxia. The compensatory increase in cerebral blood flow was greatest in animals subjected to seizures during normoxia and somewhat less pronounced in animals made hypoxic. However, blood flow failed to increase in forebrain structures when animals were subjected to the combination of seizures and hypoxia. Accordingly, levels of adenosine triphosphate in forebrain (measured both by in vitro enzymatic analysis and by in vivo phosphorus-31 nuclear magnetic resonance spectroscopy) were depleted to the greatest degree in animals who were seizing while hypoxic. In addition, brain glucose was significantly reduced only in the seizure-hypoxia group. Systemic factors such as hypoxemia may play a critical role in the disruption of cerebral energy balance during neonatal status epilepticus.

Animals↗

A comparison of in vivo catalysis by creatine kinase in avian skeletal muscles with different fibre composition.

The maximum activity of creatine kinase in vitro is similar in the pectoralis major muscle of the chicken and the duck. However, the flux (phosphocreatine to ATP) as measured by 31P saturation transfer NMR in vivo is almost 2-fold higher in the duck. This apparent discrepancy can be accounted for by the differences in the cytosolic free ADP concentrations in resting muscle.

Adenosine Diphosphate↗

31P NMR study of cerebral metabolism during prolonged seizures in the neonatal dog.

The effects of prolonged bicuculline-induced seizures on cerebral blood flow and metabolism were determined in paralyzed, mechanically ventilated neonatal dogs. Transient changes occurring early in the course of status epilepticus included significant arterial hypertension, hypocarbia, elevation of plasma norepinephrine levels, and decline in brain glucose concentration. Cerebral blood flow remained elevated throughout the 45 minutes of seizure. Determination of cerebral metabolite values by in vivo phosphorus 31 nuclear magnetic resonance spectroscopy and by in vitro enzymatic analysis of frozen brain samples showed significant decreases in the level of phosphocreatine and relatively less change in ATP values. Progressive intracellular acidosis occurred, coincident with elevation of brain lactate concentrations. We conclude that the physiological and metabolic alterations that occur during prolonged seizures are not uniform, but change with time. Any hypothesis advanced to explain the mechanism of neuronal injury during prolonged seizures must take into account these temporally related changes.

Adenosine Triphosphate↗

31P spin-lattice relaxation times and resonance linewidths of rat tissue in vivo: dependence upon the static magnetic field strength.

Phosphorus-31 NMR spin-lattice relaxation times and resonance linewidths of rat leg muscle, brain, and liver metabolites in vivo have been examined at 1.9-, 4.7-, and 8.5-T static magnetic field strengths. The resonance linewidths expressed in ppm that have been measured are independent of field strength. The spin-lattice relaxation times of muscle and brain phosphorus metabolites decrease linearly with increasing field strength while those of liver are constant over the range of static fields examined.

Animals↗

Phosphorus-31 nuclear magnetic resonance studies of pig adrenal glands.

We have used phosphorus-31 nuclear magnetic resonance to study the secretion of adenosine 5'-triphosphate (ATP) from the medulla of perfused pig adrenal glands. The resonances of the nucleotide pools in the chromaffin granules and cytoplasm are clearly resolved and therefore the intragranular and cytoplasmic processes involving ATP can be monitored simultaneously in the gland during secretion. Secretion of nucleotide during a 3 h continuous stimulation by infusion of acetylcholine chloride was monitored by the decrease in intensity of the intragranular ATP resonances. Up to 40% of the total intragranular nucleotide was released under these conditions. The rate of secretion decreased with duration of stimulation. No significant changes in the steady-state levels of cytoplasmic ATP or in oxygen consumption were observed. The intragranular pH in ischaemic glands was 5.52 + 0.15, while in glands which had been perfused until their cytoplasmic nucleotide levels had recovered, the intragranular pH was 5.76 +/- 0.16. These results provide the first estimate of the internal pH of the chromaffin granules in intact perfused adrenal glands and show that no net acidification occurs in the presence of cytoplasmic ATP. However, the isolated chromaffin granule possesses a proton-pumping adenosine 5'-triphosphatase which, in the presence of a permeant counter-ion, such as chloride, acidifies the granule interior. It is, therefore, suggested that in the intact cell, the cytoplasmic concentration of permeant counter-ions is too low to allow electrically neutral proton accumulation in the granules.

Acetylcholine↗

In vivo 31P nuclear magnetic resonance measurements in canine heart using a catheter-coil.

We have developed a new method of obtaining high resolution 31P nuclear magnetic resonance spectra from canine heart without the need for major surgery. This is done by passing an elliptical nuclear magnetic resonance receiver coil through a peripheral blood vessel into the right or left ventricle. This technique enables spectra to be obtained from a defined region of myocardium in less than 7 minutes (for the right ventricle) with a signal:noise ratio of greater than 9:1. With this catheter-coil, useful cardiac metabolic information will be obtainable, not only from specified regions of the heart, but also from various layers of myocardium.

Animals↗

Evidence for site-selective metal binding in calf liver metallothionein.

Two isoproteins of calf liver metallothionein (MT) have been isolated, purified, and characterized by atomic absorption, ultraviolet absorption, electron spin resonance, and 113Cd nuclear magnetic resonance spectroscopy. Native calf liver MT was found to contain both Cu+ and Zn2+ in a mole ratio of approximately 0.75. Selective replacement of the native Zn2+ with 113Cd2+ can be accomplished in vitro by adding 113CdCl2 to the homogenate before chromatography. Both isoproteins of metallothionein thus prepared contain approximately 3.9 g atoms of Cd2+ and 2.6 g atoms of Cu+/mol of protein. No ESR signal was found, indicating that either Cu+ or antiferromagnetically coupled Cu2+ is the form of copper present. Arguments in support of the former state are presented. Unlike the native 113Cd,Zn MT from rabbit liver, calf liver 113Cd,Cu MT exhibits a remarkably simple 113Cd NMR spectrum. Four major resonances were found for each isoprotein, in the same positions as the resonances assigned to the metals in the four-metal cluster A of rabbit liver metallothionein. This conclusion was confirmed by homonuclear decoupling experiments. This result in conjunction with the stoichiometry of bound metal ions found in the native protein suggests that Cu+ is bound selectively to the three-metal cluster B sites, and that one homogeneous protein fraction predominates. Three minor resonances to higher field are observed in the 113Cd NMR spectrum of calf liver MT-1 and one in calf liver MT-2, which may be attributed to a small fraction of cluster B with one Cu+ replaced by 113Cd2+. The possible biological significance of the different metal ion specificities of cluster A versus cluster B is discussed.

Animals↗