William M. Brooks

Proton polarization transfer enhancement for a nucleus with arbitrary spin quantum number from n scalar coupled protons for arbitrary preparation times

Abstract Polarization transfer enhancement of the NMR resonance from a nucleus with spin quantum number l (l = 1 2 , 1, 3 2 , …) from n scalar coupled protons using the pulse sequence (90 x H)-τ-(180 x H)(180 x I)-τ-(90 y H)(90 x I)-Δ-(acquire with or without H decoupling) is studied theoretically. It is shown that the enhancement of the components of the I multiplet varies with τ as Σ m I m I sin (4 πJm I τ ) where J is the scalar coupling constant and m I takes the values l, l - l, …, -l . Values of τ opt which result in the maximum enhancement are calculated, as well as the size of these maximum enhancements. Values of Δ opt which yield the maximum decoupled signal are also computed, along with the maximum decoupled enhancements. The theory is confirmed experimentally for l = 3 2 and l = 3 by studying proton polarization transfer spectra of the 10 B and 11 B resonances in NaBH 4 .

Polarization transfer via an intermediate multiple-quantum state of maximum order

Description dun type de sequences dimpulsion POMMIE qui peut devenir une technique de remplacement de la methode DEPT des linstant ou lamelioration de linstrumentation permettra lutilisation des deplacements de phase

31P nuclear magnetic resonance study of the recovery characteristics of high energy phosphate compounds and intracellular pH after global ischaemia in the perfused guinea-pig heart.

The recovery of high energy phosphate compounds in perfused guinea-pig heart at 20 degrees C after a 12 min period of global ischaemia was examined using 31P-NMR with a time resolution of 12 s. This time resolution was achieved by overlaying the data acquired from five successive ischaemic periods by arresting and restoring the flow of perfusion fluid to the heart in synchrony with the data acquisition sequence. The rate of creatine phosphate resynthesis after the ischaemic period proceeded 14 times faster than its rate of loss during the ischaemic period. ATP levels did not decrease during ischaemia and ADP was undetectable at any time. Estimates of intracellular pH from the chemical shift of the inorganic phosphate peak were impossible in normal guinea-pigs since the inorganic phosphate peak was not clearly defined. During the ischaemic period the inorganic phosphate peak increased in size and shifted upfield. On restoration of flow, the inorganic phosphate peak collapsed in a complex way following a different path to its formation during ischaemia.

Application of the reverse dept polarization-transfer pulse sequence to monitor in vitro and in vivo metabolism of 13C-ethanol by 1H-NMR spectroscopy

Using the reverse 13C----1H DEPT polarization-transfer pulse sequence the metabolism of 13C ethanol in vitro and in vivo has been monitored by 1H-NMR spectroscopy. Using yeast alcohol dehydrogenase, acetaldehyde, the hydrated form of acetaldehyde and acetate were identified as metabolites of [2-13C]-ethanol. The ratio of hydrated to free acetaldehyde was dependent upon the protein concentration of the reaction mixture. Binding of acetaldehyde in an irreversible Schiffs base resulted in optimal enzyme activity. Hepatocytes from rats fasted for 20 h, metabolised [1-13C] and [2-13C]ethanol in a linear fashion, but no [13C]acetaldehyde was detected. Metabolic integrity of the hepatocytes was confirmed with [2-13C]acetate. The addition of disulfiram (50 micron) to hepatocyte suspensions which had been incubated with [1-13C]ethanol, resulted in the resynthesis of [13C]ethanol. The amount of [13C]ethanol resynthesized under these conditions represents intracellular acetaldehyde whose concentration was in the range of 400-800 mumol/g wet weight of hepatocytes when 50 mM ethanol had been originally incubated with the hepatocyte suspension. These studies show how NMR-polarization transfer pulse sequences can be used to monitor the metabolism of 13C-ethanol in vivo, and provide a unique tool to measure in vivo concentrations of acetaldehyde. The studies also suggest that cytoplasmic aldehyde dehydrogenase may play a major role in hepatic ethanol metabolism.

Relation between the phosphocreatine to ATP ratio determined by 31P nuclear magnetic resonance spectroscopy and left ventricular function in underperfused guinea-pig heart.

The relation between the PC/ATP ratio and left ventricular function was examined in the Langendorff-perfused guinea-pig heart over a range of perfusion flow rates. PC/ATP ratios were determined from the 31P-nuclear magnetic resonance spectra of hearts obtained at 80.98 MHz and ventricular function estimated by measuring pressure in the left ventricle. When flow rates were increased over the range 0.6 to 6.0 ml/min, the PC/ATP ratio increased from 0.64 +/- 0.05 at 0.6 ml/min to 1.82 +/- 0.12 at 3.8 ml/min with no further increase up to a flow rate of 6.0 ml/min. Developed pressure (DP) increased with the flow rate up to 6.0 ml/min but the end diastolic pressure (EDP) also increased. The DP/EDP ratio was found to correlate closely with the PC/ATP ratio over the range of flow rates examined. The PC/ATP ratio may be a practical index of myocardial function available to the clinician when the topical magnetic resonance technique is fully developed.

Potential hazards of NMR imaging. No evidence of the possible effects of static and changing magnetic fields on cardiac function of the rat and guinea pig

Clinical proton NMR imaging uses magnetic field strengths in the range 0.1 to 0.5 T. In addition to the large static magnetic field, patients are exposed to magnetic field gradients during imaging and under extreme conditions, such as power failure or quenching, the field may collapse precipitously. A potential source of hazard to patients under these conditions is the induction of thoracic currents which may trigger ventricular fibrillation. In the present experiments, a 0.16 T resistive magnet with a time constant of 60 ms, powered by a programmable power supply, was used to examine any possible effects of static and changing magnetic field on the ECG and arterial blood pressure of anesthetized rats and guinea pigs. Animals were exposed to the following field conditions: static fields of 0.16 T; sine, triangular, and square wave modulated fields from 0.1 to 2 Hz; rapid field switches in excess of 2.0 T/s for 25 ms timed to occur at different stages of the cardiac cycle, including the vulnerable period during ventricular repolarization; and AC fields of 50 Hz. No change was observed in the blood pressure, heart rate, or ECG under any of the field conditions examined.

Reverse polarization transfer through maximum order multiple-quantum coherence: A reverse pommie sequence

A reverse POMMIE sequence [(randomize 1 H)-½π[C, x ] -(2 J ) −1 -½π[H, j ]½π [H, ψ] π[C]-(2 J ) −1 -½π[C, y ] π[H, k ]-(2 J ) −1 -½π[H, ±ψ], acquire 1 [H; (decouple 13 C)] is proven both theoretically and experimentally. This sequence, which utilises a selective multiple-quantum filter, will be useful for editing 1 H spectra based on 13 C- 1 H scalar coupling constants. Because 1 H H 2 O NMR signals can be effectively eliminated by application of an appropriate 1 H randomization process prior to the commencement of the pulse train, the major application of the method will be in metabolic studies of 13 C-labelled material. Suppression of H 2 O 1 H signals using this technique can exceed 10 5 : 1, a factor which should be adequate even for demanding applications.

In vivo determination of 31P spin relaxation times (T1, T2, T1ϱ) in rat leg muscle. Use of an off-axis solenoid coil

A probe using a solenoid coil tilted 45 degrees off-axis has been used to study the 31P NMR relaxation characteristics of the resonances arising from phosphorus metabolites in rats in vivo. T1, T1 rho and T2 values have been determined for phosphocreatine and ATP in leg muscle. The ratio of 31P T1(1700ms) to T2(12ms) for ATP was in excess of 200:1 compared with a ratio of 5:1 for 1H T1:T2. Of major significance was the observation that T2 values for phosphocreatine (230ms) were markedly longer than T2 values for ATP (12ms). Thus by use of appropriate delay times in spin echo sequences ATP signals can be nulled, and discrete 31P imaging of phosphocreatine in muscle may be possible provided the overall signal-to-noise is satisfactory.

In Vivo 13C nuclear magnetic resonance spectroscopy using a solenoid transmit/receiver coil

Topical nuclear magnetic resonance spectroscopy (NMR) using a solenoid transmit/receiver coil has been used to monitor the kinetics of intragastric utilization of [1-13C] glucose in mice in vivo. Using a double-tuned solenoid coil, signal to noise response was excellent and the natural abundance in vivo 13C spectra obtained with acquisition times of 10-15 minutes compares more than favourably with comparable studies using surface or saddle coils. This study clearly shows the potential of solenoid coils to monitor the kinetics of 13C-labelled metabolites in vivo.

Determination of intracellular pH in the Langendorff-perfused Guinea-pig heart by 31P nuclear magnetic resonance spectroscopy

The 31P NMR spectrum of freshly prepared guinea-pig heart perfused in the Langendorff mode at 37 degrees C with medium containing either glucose or pyruvate as an energy source exhibited no inorganic phosphate peak making the determination of intracellular pH from the chemical shift of this peak impossible. By incorporating 2-deoxyglucose-6-phosphate into the heart to act as an alternative pH indicator, intracellular pH was determined to be 6.98 +/- 0.02 (mean +/- S.E.M. for eight animals). 2-Deoxyglucose was not taken up and phosphorylated by guinea-pig heart in the presence of glucose. Uptake and phosphorylation did proceed if glucose was omitted from the perfusion medium, but a large inorganic phosphate peak was present in the initial spectrum prior to 2-deoxyglucose addition. With pyruvate as sole energy source, an inorganic phosphate peak was absent initially and 2-deoxyglucose was taken up and phosphorylated. During a 12 min period of global ischaemia at 20 degrees C, when inorganic phosphate was visible together with 2-deoxyglucose-6-phosphate, a close correlation was found between the pH estimated from the chemical shift of inorganic phosphate and 2-deoxyglucose-6-phosphate over the pH range 6.3 to 6.9.