Starter sequence for steady-state free precession imaging.

The dynamic equilibrium exploited by balanced steady-state free precession imaging develops slowly because its formation is dependent on both spin-spin and spin-lattice relaxation times. Attempting to image before steady state is established results in artifacts due to transient signal oscillations. Using a starter sequence to precondition the spin system can significantly reduce the delay before imaging. An improved design for a steady-state starter sequence is presented. The new sequence has the advantage of uniformly exciting the steady-state response for all resonance offsets and can be phase cycled to suppress banding artifacts.

Frequency-modulated steady-state free precession imaging.

Exploration of the possibilities of steady-state free precession (SSFP) excitation has led to the discovery that it is tolerant of slow variations in spectral offset frequency. The effect has been used to eliminate banding artifacts from images obtained with the fully balanced SSFP imaging sequence.

FAST sequences optimization for contrast media pharmacokinetic quantification in tissue.

The purpose of this study was to investigate the influence of the fast gradient-recalled echo (GRE) sequence parameters on the contrast dynamic range and signal sensitivity, to optimize the magnetic resonance (MR) sequence for contrast media pharmacokinetic assessment. Effects of the fast low-angle shot (FLASH), Fast acquisition at steady rate (FAST), and radiofrequency-spoiled (RF)-FAST sequence parameters were studied in vitro. The FAST sequence had the highest sensitivity in low gadolinium (Gd) concentration. The FLASH and RF-FAST sequences had a larger contrast dynamic range, but the FLASH images contained side band artifacts. Increasing the flip angle to 90 degrees raised the sensitivity of the FAST sequence and the contrast dynamic range of the RF-FAST sequence. The shortest possible TE was optimal for both contrast dynamics and imaging time. TI had an influence on the sensitivity of the FAST sequence only for small acquisition matrices. This study indicates the optimal parameters for contrast dynamics (RF-FAST, 90 degrees flip angle, shortest possible TE) and sensitivity (FAST, 90 degrees flip angle, long TI(eff)).

Rapid iterative reconstruction for echo planar imaging.

A rapid automated method for reconstructing echo planar imaging (EPI) data has been developed and is shown to improve image quality by suppressing the troublesome ghost artifact. The algorithm can be applied without prior knowledge obtained from either reference scans or operator intervention. It first estimates, then improves iteratively, the parameters for a linear phase correction applied directly to the complex image data derived from odd and even echoes. The theory used to derive the criteria employed in the iteration provides insight into mechanisms that allow the process to work. Magn Reson Med 42:541-547, 1999.

Calibration of the radio frequency field for magnetic resonance imaging.

We have developed and validated the performance of a novel slice selective pulse sequence that allows direct calibration of the RF field using a simple rectangular pulse. The new sequence offers a number of substantial advantages. It operates at steady state and has an accurate calibration response at short repetition times. The slice selection train is insensitive to RF field strength changes caused by patient loading. The issue of patient motion has been addressed in our data collection and analysis routines. The applicability of the method to human scanning has been demonstrated in the automated RF power calibration routine of a commercial imaging system.

On the convergence of the generalized maximum likelihood algorithm for nonuniform image motion estimation.

The generalized maximum likelihood algorithm is a powerful iterative scheme for waveform estimation. This algorithm seeks for the maximum likelihood estimates of the Karhunen-Loeve expansion coefficients of the waveform. The search for the maximum is performed by the steepest ascent routine. The objective of the paper is to obtain conditions that assure the stability in the mean for frame-to-frame image motion estimation. Sufficient conditions are established for the convergence of the algorithm in the absence of noise. Experimental results are presented that illustrate the behavior of the algorithm in the presence of various noise levels.

Sodium-23 and proton nuclear magnetic resonance imaging studies of carbon tetrachloride-induced liver damage in the rat.

Magnetic resonance imaging techniques were used to investigate the response of the liver of the rat in situ to a toxicological challenge by carbon tetrachloride. Proton images were taken as transverse slices through the rat before and after intraperitoneal administration of the hepatotoxin. Two to three hours after carbon tetrachloride was given, a region of high proton signal intensity was observed where the portal vein enters the liver. Sodium-23 images were also taken, and a region of high sodium intensity was observed in the same location within the liver as the increased proton intensity. The results suggest that acute administration of carbon tetrachloride induces localized liver damage in the region where the hepatotoxin first enters the liver. This liver damage is manifest as edema with a buildup of sodium ion and water, which can be readily detected by sodium-23 and proton NMR imaging techniques, respectively.

In vivo proton nuclear magnetic resonance imaging and spectroscopy studies of halocarbon-induced liver damage.

Proton magnetic resonance imaging and localized NMR spectroscopy were used to study the rat liver in situ. Respiratory gating was used in both the imaging and the localized spectroscopy studies to control for the movement of the upper abdomen of the rat during breathing. After administration of carbon tetrachloride, bromotrichloromethane, or halothane, localized regions of high proton signal intensity were observed in the NMR images of the liver. Localized (VOSY) proton NMR spectra from within these regions indicated that the increase in a signal intensity was due to a longer T2 relaxation time for the water resonance, indicating acute edema in the region of tissue damage.

Nuclear magnetic resonance studies of carbohydrate metabolism and substrate cycling in Fasciola hepatica.

We have been interested in clarifying unique features of glycolytic metabolism in parasitic trematodes and in developing improved methods for monitoring the effects of pharmacologic agents that may alter functions of the pathway. In the present study metabolism of [1-13C]glucose by the common liver fluke, Fasciola hepatica, was studied both directly with 13C NMR and indirectly by observation of 13C-induced multiplet splitting of the 1H NMR resonances from the glycolytic end-products propionate and acetate. The extent of 13C enrichment of the end-products demonstrated that exogenous glucose was the predominant source of glycolytic substrate under the incubation conditions used. Specific enrichments of propionate and acetate in 13C were similar and enrichments at the acetate C-1 carboxyl and C-2 methyl were identical, demonstrating that acetate is generated preferentially from pyruvate formed by the malic enzyme reaction. End-product synthesized in substrate-free medium following incorporation of a small fraction of [1-13C]glucose into endogenous glycogen demonstrates that glucose equivalents from the most recently synthesized polymeric chains, which have a specific activity in 13C equal to that of the exogenous glucose, are preferentially used for glycogenolysis. Stimulation of flukes with 0.1 mM serotonin results in a reduction of the propionate/acetate 13C enrichment ratio consistent with functional "compartmentation" of glycogen pools having different structures and/or specific enrichment in 13C. Glucose equivalents were incorporated into glycogen in intact flukes with label at both the C-1 and C-6 positions during perfusion with [1-13C]glucose as a consequence of "substrate cycling" at the phosphofructokinase/fructosebisphosphatase enzyme couple. The observed glycogen C-6/C-1 labeling ratio of 0.42 and the net glycolytic flux of 11 mumol/g wet weight/hr imply a total forward flux of about 29 mumol/g wet weight/hr through phosphofructokinase with a reverse flux of about 17 mumol/g wet weight/hr through fructosebisphosphatase. Net glycolytic flux is therefore a poor estimate of the true flux through phosphofructokinase in this preparation.

31P-NMR studies of metabolite compartmentation in Fasciola hepatica.

Fasciola hepatica, the common liver fluke, is an anaerobic parasitic worm. Possible compartmentation of metabolites between different cell types, metabolic compartments, and free and macromolecule-bound species was investigated using 31P-NMR. A spectrum of the intact worm shows unusual metabolic features, among which are large amounts of glycerolphosphorylcholine, phospholipids mobile on the NMR time-scale, and free cytosolic ADP. Spectra from cells as different as those in oral sucker tissue and eggs showed similar features. Acidosis after serotonin administration was associated with parallel changes in chemical shifts of intracellular Pi and glucose 6-phosphate, suggesting that they are in the same metabolic compartment. Although 13.4 +/- 1.1 mumol/g wet wt. (n = 3) Mg2+ is present in fluke tissue, a considerable fraction is sequestered out of the cytosol. The intracellular free [Mg2+] was independently estimated from the chemical shifts of ATP and ADP as 1.6 +/- 0.5 mM and 2.9 +/- 0.7 mM, respectively. Quantitation of observable phosphate-containing metabolites in whole tissue and in perchlorate extracts demonstrated that 60% of the total ADP and 50% of the total Pi are 'NMR-invisible' in the intact fluke and therefore probably bound to macromolecules in the cells. The apparent ATP/ADP X Pi free concentration ratio is much lower in this anaerobic tissue than in mammalian oxidative tissues.

Continuous perfusion of mammalian cells embedded in agarose gel threads.

A method for perfusing cells by embedding them in fine agarose gel threads is described and characterized. The rate of diffusion of a metabolite into the gel threads is determined by 31P-NMR spectroscopy. This perfusion method is shown to enable Chinese hamster lung fibroblasts (CHLF) to remain in a metabolically active state with high levels of intracellular ATP for many hours.

The inhibition of erythrocyte glyceraldehyde-3-phosphate dehydrogenase. In situ PMR studies.

The kinetics of inhibition of human erythrocyte glyceraldehyde-3-phosphate dehydrogenase by iodoacetate were studied in the intact cell and in vitro. The kinetics were determined using 1H-NMR to follow solvent exchange of 1H and 2H at the C-2 position of lactate. The exchange occurs via a series of enzyme-catalysed reactions, including that catalysed by glyceraldehyde-3-phosphate dehydrogenase. A direct assay with quenching of the inhibition was also used to check the results. Iodoacetate was shown to act as an active site-directed inhibitor of the dehydrogenase. The enzyme inhibition patterns, which are characterised by a binding step and a kinetic step, are similar in situ and in vitro. Membrane binding, however, was found to alter the inhibition pattern for the enzyme in vitro.

Studies of lactate dehydrogenase in the purified state and in intact erythrocytes.

Lactate dehydrogenase in intact erythrocytes was studied by observing isotope exchange between lactate and pyruvate by p.m.r. The inhibition of the enzyme in intact cells by both oxalate and pyruvate was found to be similar to that of the purified enzyme. The activity of the enzyme in intact cells indicates that the free solution NAD+ + NADH concentration in erythrocytes is about 10 microM whereas the total extractable NAD+ + NADH is about 80 nmol/ml of cell water.

A p.m.r. isotope-exchange method for studying the kinetic properties of dehydrogenases in intact cells.

A method to determine the activity of dehydrogenases in an intact-cell system is described. The method involves the use of n.m.r. to monitor bulk isotope exchange. The approach is illustrated by application to the isotope equilibration of pyruvate and lactate as catalyzed by lactate dehydrogenase in intact erythrocytes. Particular problems peculiar to bulk isotope exchange and its observation by n.m.r. are considered.

A 1H n.m.r. study of isotope exchange catalysed by glycolytic enzymes in the human erythrocyte.

The exchange of hydrogen and deuterium atoms between the C-2 position of lactate and solvent was monitored in suspensions of human erythrocytes by using a non-invasive spin-echo p.m.r. method that permits continuous assessment of the rate and the extent of exchange. Exchange rates were measured in cells suspended in buffers made in 2H2O and 1H2O after the addition of L-[2-1H]lactate and L-[2-2H]lactate respectively. The rate of exchange is dependent on the activities of four glycolytic enzymes (fructose bisphosphate aldolase, triose phosphate isomerase, glyceraldehyde phosphate dehydrogenase and lactate dehydrogenase) and on the concentrations of their substrates. The dependence of the exchange on the following substrates was studied: (1) lactate, (2) the triose phosphates and fructose 1,6-bisphosphate and (3) pyruvate. Observation of the exchange in vitro, in a system produced by mixing the isolated enzymes, permits determination of the individual isotope-exchange equilibrium velocities of the enzymes. The dependence of the equilibrium velocity of human erythrocyte lactate dehydrogenase on NAD+ + NADH concentration was measured. Possible applications of these methods are discussed.

Studies of pyruvate-water isotope exchange catalysed by erythrocytes and proteins.

Erythrocyte suspensions in buffer made with 2H2O catalyse the exchange of pyruvate protons. This process can be easly observed by spin-echo proton magnetic resonance. The dominant exchange process is shown to be due to the formation of Schiff-base links between pyruvate and amino groups of haemoglobin. Other proteins with free alpha-amino groups also catalyse the exchange. The pH*-dependence of the exchange rate due to hen-egg-white-lysozyme reflects the dissociation of the alpha-amino group.