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J J Ackerman

Publications and source records attributed to J J Ackerman.

At least 37 records · Page 2Linked to original sources

Determination of intracellular calcium in vivo via fluorine-19 nuclear magnetic resonance spectroscopy.

Fluorine-19-nuclear magnetic resonance (19F-NMR) spectroscopic detection of the NMR-active Ca2+ indicator 5-fluoro-1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (5F-BAPTA) is one method for measuring cytosolic free Ca2+ concentration ([Ca2+]i) and has been used previously to measure [Ca2+]i in isolated cells and perfused organs. The aim of the present investigation was to demonstrate the feasibility of determining [Ca2+]i in vivo and in situ using 19F-NMR and 5F-BAPTA. Experiments were performed on male Sprague-Dawley rats with a surface-coil antenna employed for NMR interrogation. The Ca2+ indicator, 5F-BAPTA, was infused either intravenously (kidney, spleen) or intraventricularly (brain) as a 100 mg/ml solution of the cell-permeant acetoxymethyl ester (5F-BAPTA-AM) in dimethyl sulfoxide. Rats tolerated intravenous infusion without evident change in mean arterial blood pressure. In all tissues examined, kidney, spleen, and brain, [Ca2+]i was approximately 200 nM. To our knowledge, these results represent the first in vivo and in situ determinations of [Ca2+]i employing 19F-NMR.

Animals↗

An evaluation of the sensitivity of the intravoxel incoherent motion (IVIM) method of blood flow measurement to changes in cerebral blood flow.

To evaluate the sensitivity of the intravoxel incoherent motion (IVIM) technique to changes in cerebral blood flow, we made measurements of IVIM parameters in rat brain under conditions of altered arterial pCO2. The arterial pCO2 was varied over a range which would be expected to change cerebral blood flow from roughly 50 to 500 ml/(100 g.min). The IVIM measurements were made with suppression of extravascular water signal. The parameters f' (the apparent fraction of spins which have "fast" pseudodiffusion), D1* (the "fast-pseudodiffusion") coefficient), and D2* (the "slow-pseudodiffusion" coefficient) all showed statistically significant positive linear correlations with arterial pCO2. These results suggest that the IVIM method, when used with suppression of extravascular water signal, is sensitive to changes in blood flow.

Animals↗

Increased intracellular Ca2+: a critical link in the pathophysiology of sepsis?

Severe bloodstream-borne infection--i.e., sepsis--and the resulting multiorgan failure are now the most common cause of death in many intensive care units. One of the most fundamentally important and controversial issues concerning the pathophysiology of sepsis is the role of intracellular free calcium concentration ([Ca2+]i) in this disorder. Because of the critical role of calcium as an intracellular second messenger and as a potential cellular toxin, resolution of this issue is crucial. Using 19F NMR spectroscopy and the calcium indicator 5,5'-difluoro-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetate we demonstrate in the intact perfused organ, the rat thoracic aorta, that [Ca2+]i in aortic smooth muscle is increased > 2-fold during sepsis. Furthermore, we determined that sodium dantrolene, a drug that decreases release of calcium from the sarcoplasmic reticulum and that is lifesaving in malignant hyperthermia (a disorder due to increased [Ca2+]i), is able to reduce the elevated [Ca2+]i in sepsis to control values when added in vitro or when given in vivo to the animal. These results suggest that an increase in [Ca2+]i is an early event in sepsis and that increased [Ca2+]i may be responsible for, or contribute to, cellular injury. Dantrolene may offer a therapeutic strategy in the treatment of sepsis.

Animals↗

In vivo NMR imaging and spectroscopic investigation of renal pathology in lean and obese rat kidneys.

Diabetic nephropathy is a major cause of end-stage renal failure. While our understanding of the pathogenesis of nephropathy is incomplete, progressive glomerular injury appears to play a significant role in the decline of renal function. Proton NMR spectroscopy and imaging techniques were used to address changes in renal pathology associated with glomerular mesangial expansion in vivo in kidneys from spontaneously obese and lean (control) littermate Zucker rats. Fully functioning rat kidneys were surgically exposed and externalized for direct NMR signal detection via a coil placed around the organ. High-resolution (78 microns in plane) proton images were obtained at 4.7 T magnetic field strength revealing fine structure within the well-defined cortical and medullary regions. The obese rat kidney images were distinct in appearance from the lean kidney images and exhibited marked cortical expansion as well as increased overall kidney size. Enlargement of mean glomerular diameter was verified histologically in the obese kidneys as compared with the lean kidneys. Proton T1 and T2 relaxation times were determined from the entire kidney using standard spectroscopic techniques, and from specific regions within the kidney from multiple T1- and T-2 weighted images. Additionally, image contrast enhancement resulting from saturation transfer between protons in restricted-mobility environments and mobile water protons within the kidney was investigated in the lean and obese rat kidneys using magnetization-transfer imaging techniques. At the early stage of renal injury examined in this study, diseased and healthy kidneys could not be differentiated on the basis of relaxation times alone. The magnitude of saturation transfer obtained in cortical tissue in the lean and obese kidneys was also not statistically significantly different. However, the magnitude of saturation transfer achieved in the medullary tissue of obese kidneys was statistically significantly less than that achieved in lean kidneys.

Animals↗

Effect of sepsis on brain energy metabolism in normoxic and hypoxic rats.

Neurological abnormalities including agitation, confusion, disorientation, lethargy, and obtundation are early characteristic findings in patients with sepsis. The etiology of the changes in mental status that occur during severe infection is unknown. We investigated the effects of sepsis on intermediary metabolism and bioenergetics in the brain during normoxia and moderate hypoxia (8% inspired O2 concentration) in rats 36-42 hr following cecal ligation and perforation. The rats were anesthetized with halothane, and brains frozen using the funnel-freezing technique. Perchloric acid extracts of brains were analyzed with fluorometric enzymatic methods and 31P nuclear magnetic resonance spectroscopy. There was no impairment in bioenergetics or intermediary metabolism in septic brain, and sepsis did not compromise the ability of the brain to maintain high-energy phosphates during hypoxia. Hypoxia did cause the brain lactate-to-pyruvate ratio to increase equivalently in both septic and control rats from approximately 9:1 to 20:1 (P < 0.001). We conclude that the neurologic changes which are characteristic of sepsis are unlikely to be due to alterations in cellular energy stores or intermediately metabolism. In addition, there is no evidence that sepsis results in brain cellular hypoxia.

Animals↗

Quantification of the accumulation and degradation of beta-very-low-density lipoproteins in vivo using a 19F-containing residualizing label and n.m.r. spectroscopy.

beta-Very-low-density lipoproteins (beta-VLDL)O were conjugated to the 19F-containing residualizing label, NN-dilactitol-3,5-bis(trifluoromethyl)benzylamine (DLBA), to determine whether the metabolism of this lipoprotein fraction could be characterized in vivo with n.m.r. spectroscopy. Solution state 19F high-resolution n.m.r. spectroscopy of DLBA-beta-VLDL, containing either intact apoproteins or selectively enzymically digested products, demonstrated that the extent of degradation could be distinguished by differences in spin-spin relaxation times (T2 times). DLBA-beta-VLDL was injected intravenously into rabbits, and accumulation of 19F in hepatic tissue was quantified non-invasively by n.m.r. spectroscopy 5 and 30 h after injection. In addition to quantifying the accumulation of DLBA-beta-VLDL in hepatic tissue, a marked decrease (approx. 100 Hz) in the linewidth of 19F resonance from labelled lipoproteins was observed at 30 h compared with the 5 h interval in continuously monitored animals. The change in linewidth was consistent with a decrease in molecular size that occurred during protein degradation, resulting in increased T2 times. To demonstrate that T2 times can be used as an index to quantify apoprotein degradation in vivo, relaxation measurements were performed on livers excised 20 h after injection of DLBA-beta-VLDL into rabbits. Two molecular motional fractions were revealed by relaxation profiles representing either an intact or an extensively degraded form of apoprotein. The amplitudes of each component were compared with results from trichloroacetic acid precipitation of liver homogenates acquired from rabbits 20 h after injection of beta-VLDL labelled with the radioiodinated analogue of DLBA, dilactitol-125I-tyramine. The amount of degraded apoprotein determined by n.m.r. spectroscopy and acid precipitation was 68.6 +/- 7.0% and 58.7 +/- 7.5% (n = 4) respectively. The results of this study demonstrate that 19F n.m.r. spectroscopy can be used to define the temporal characteristics of the hepatic metabolism of lipoproteins in vivo by quantifying both the tissue-specific accumulation and extent of apoprotein degradation. The methodology developed offers promise for the non-invasive, sequential and longitudinal evaluation of lipoprotein metabolism in vivo.

Animals↗

Nonglycolytic acidification of murine radiation-induced fibrosarcoma 1 tumor via 3-O-methyl-D-glucose monitored by 1H, 2H, 13C, and 31P nuclear magnetic resonance spectroscopy.

The effects of 3-O-methyl-D-glucose (3-OMG) on subcutaneously implanted murine radiation-induced fibrosarcoma 1 tumor were examined with 2H, 13C, and 31P nuclear magnetic resonance (NMR) in situ. Using 31P NMR, changes in tumor high-energy phosphate metabolism were monitored for 2.5 h after i.p. administration of 3-OMG (8.1 g/kg body weight); tumor pH decreased by a mean maximum of 0.52 +/- 0.05 (SE) (n = 10), [PCr] decreased by 54%, [NTP] decreased by 35%, and [Pi] increased by 36%. Tumor blood flow, as measured by 2H NMR monitoring of D2O washout kinetics, decreased by 40% at 1 h and by 47% at 2 h after 3-OMG injection (n = 4). This substantial tumor acidification (pH decrease much greater than 0.1), expected to require a glycolytic substrate (Hwang et al., Cancer Res., 51: 3108-3118, 1991), is surprising in light of the previously documented metabolically inert nature of 3-OMG. In situ 13C NMR spectroscopy, following [6-13C]3-OMG i.p. injection, examined the possibility of the glycolytic metabolism of 3-OMG. However, only the C-6 resonance of 3-OMG was detected (n = 6); no resonances from [6-13C]3-OMG-6-phosphate or [3-13C]lactate were observed. These results confirmed that 3-OMG was not metabolized in radiation-induced fibrosarcoma 1 tumor. At the completion of the in situ 13C NMR experiments, tumors were freeze clamped, and perchloric acid extraction was performed. High-resolution 1H NMR measurement of lactate concentrations showed no statistically significant difference in control tumor extracts (from mice not receiving i.p. injection; n = 5) and in tumor extracts from mice administered i.p. [6-13C]3-OMG (n = 5), indicating that there was no significant increase in lactate level in the tumor extracts from mice administered i.p. 3-OMG due to increased plasma glucose concentration. The results of these 1H and 13C NMR studies indicated that the radiation-induced fibrosarcoma 1 tumor acidification caused by i.p. administration of 3-OMG was not due to a direct (3-OMG----lactate) or an indirect (systemic glucose----lactate) increase in tumor lactic acid levels.

3-O-Methylglucose↗

Concurrent quantification of tissue metabolism and blood flow via 2H/31P NMR in vivo. I. Assessment of absolute metabolite quantification.

In a series of three papers, we demonstrate and validate an approach for concurrent absolute quantification in situ of blood flow and energy metabolism with a modification of the NMR method for absolute concentration determination put forth by Thulborn and Ackerman [J. Magn. Reson. 55, 357 (1983)] and later expanded upon by Tofts and Wray. In this first paper of the series, we briefly review the theoretical basis for the concentration measurement and present, for the first time, a successful paired validation of metabolite quantification via 31P surface-coil NMR through corroborative in vitro enzymatic assays. The paired radiolabeled microsphere validation of blood flow measurement via 2H surface-coil NMR employing D2O as a freely diffusible tracer and the concurrent determination of blood flow and energy metabolism in a septic rat model are presented in the accompanying second and third paper to complete the series. In this article a classical RF tank circuit is employed to describe the effect of conductive sample loading on the NMR receiver by considering its apparent series resistance. It is shown in an easily visualized generalizable manner that the effect of sample loading on the observed NMR signal intensity can be accounted for quantitatively by monitoring changes in 90 degrees pulse width at constant power at a fixed reference point, i.e., Ssample = Sphantom (PW90phantom/PW90sample). In a series of paired experiments the absolute concentrations of high energy phosphates obtained from resting rat leg muscle (n = 4) in situ (NMR) and in vitro (enzymatic) were determined as follows: [PCr]NMR = 17.2 +/- 0.8 SD, [PCr]enzymatic = 17.3 +/- 2 SD, [ATP]NMR = 5.1 +/- 0.8 SD, [ATP]enzymatic = 5.0 +/- 0.2 SD mmol/kg tissue wet wt. Results of these two independent methods of concentration determination were not statistically different (P = 0.94 and P = 0.74 respectively) and serve to rigorously validate the Thulborn approach for absolute quantification of phosphorous metabolites in situ via NMR. Furthermore, these results strongly suggest that ATP and PCr in resting rat leg muscle under normal physiologic conditions are 100% NMR visible. The free cytosolic [ADP]NMR was estimated from the creatine kinase reaction equilibrium expression to be 0.022 +/- 0.003 SD mmol/kg tissue wet wt.

Adenosine Triphosphate↗

Concurrent quantification of tissue metabolism and blood flow via 2H/31P NMR in vivo. II. Validation of the deuterium NMR washout method for measuring organ perfusion.

The deuterium washout technique of measuring tissue blood flow is based upon NMR detection of HOD (administered as D2O saline, but typically detected as HOD because of rapid proton-deuteron exchange) as a freely diffusible tracer. Though this method is coming into more general use, it has not yet been rigorously validated in vivo against an accepted, independent measure of tissue blood flow. To this end, simultaneous radiolabeled microsphere and HOD washout blood flow measurements were made in rat gastrocnemius muscle. D2O saline was administered either intramuscularly or intraarterially (near the aortic bifurcation), and the sciatic nerve was electrically stimulated to increase the muscle blood flow rate. Over a range of flows of 2 to 80 ml/(100 g.min), comparison of microsphere and HOD washout measurements showed good agreement, with r = 0.92 (n = 16) for intramuscular administration and r = 0.91 (n = 12) for intraarterial administration. These data strongly suggest that the HOD washout technique provides accurate blood flow measurements in skeletal muscle.

Animals↗

Concurrent quantification of tissue metabolism and blood flow via 2H/31P NMR in vivo. III. Alterations of muscle blood flow and metabolism during sepsis.

In the conclusion of this series of reports, the application of 31P/2H NMR to investigate the pathophysiology of sepsis in rat hindlimb muscle is demonstrated. Sepsis decreased muscle [PCr] by 18%, 18 +/- 4 SD vs 22 +/- 4 SD mmol/kg tissue wet wt (P = 0.01) in control rats but [ATP] was unchanged, 6 mmol/kg tissue wet wt (P = 0.2). The derived free cytosolic [ADP] in the two groups was similar, [ADP]septic = 0.023 +/- 0.004 SD and [ADP]control = 0.021 +/- 0.003 SD mmol/kg tissue wet wt, and not statistically different (P = 0.14). Likewise [Pi] in the septic and control groups was not statistically different, [Pi]septic = 1.1 +/- 0.5 SD and [Pi]control = 1.2 +/- 0.4 SD mmol/kg tissue wet wt (P = 0.2). Septic rats presented the symptom of respiratory alkalosis evidenced by elevated blood pH. Sepsis decreased muscle blood flow by 33%, P = 0.003, but examination of individual subjects did not demonstrate a correlation with the reduction in [PCr]. Thus, a metabolic energy deficit caused by cellular ischemia/hypoxia is not a likely cause of cellular abnormality in rat hindlimb muscle during sepsis.

Adenosine Diphosphate↗

Effect of methylation inhibitors on gene expression in HL-60 cells.

The methylation inhibitors Neplanocin A (Nep A), 3'-deazaadenosine (dzAdo), and 3-deaza(+/-)aristeromycin (Dari) were tested for their effect on the expression of histone H2B, actin, and the protooncogenes c-myc, and v-fos. Nep A and Dari bind to the S-adenosylhomocysteine hydrolase resulting in the accumulation of S-adenosylhomocysteine, while dzAdo served as a substrate for the enzyme. With dzAdo, inordinant amounts of 3-deazaadenosylhomocysteine (dzAdoHcy) accumulated in the cell, provided L-homocysteine (Hcy) was present. When added at sublethal concentrations, the methylation inhibitors had little or no effect on c-myc, v-fos, histone H2B, or actin expression, nor did any significant number of the drug-treated cells demonstrate myeloid characteristics. However, growth and gene expression were markedly inhibited upon the addition of Hcy and dzAdo. One of the earliest effects of dzAdoHcy on HL-60 cells was the disappearance of c-myc mRNA. Within 1 h of the addition of dzAdo and Hcy, only trace amounts of c-myc mRNA were detectable. After 4-5 h v-fos, histone H2B, and actin mRNAs also decreased to about 40% of control levels. Differences in the stability of preexisting mRNAs would appear to account for these results. Within 1 h following the addition of dzAdo and Hcy, the synthesis of rRNA and mRNA were completely blocked as measured by the incorporation of [3H]uridine.

Adenosine↗

Modulation of murine radiation-induced fibrosarcoma-1 tumor metabolism and blood flow in situ via glucose and mannitol administration monitored by 31P and 2H nuclear magnetic resonance spectroscopy.

The hyperglycemia-induced in situ metabolism and blood flow changes produced in s.c. implanted murine radiation-induced fibrosarcoma-1 tumors, grown on the flanks of female C3H/HeJ mice, were examined with 31P and 2H nuclear magnetic resonance. Initial experiments verified a hyperglycemic tumor acidification similar to that reported earlier with a different substrain of mice, C3H/AnF (J.L. Evelhoch et al., Proc. Natl. Acad. Sci. USA, 81: 6496-6500, 1984). Changes in the tumor pH, phosphorus metabolites, and blood flow were then compared after administration of saline, glucose, or mannitol (a nonmetabolizable glucose analogue) using a mole-equivalent dose of the sugars (i.e., 0.8 mmol/20g mouse). Neither saline (n = 8) nor mannitol (n = 6) administration had any marked effect upon tumor pH, whereas glucose administration produced a mean maximum tumor pH reduction of 0.74 +/- 0.09 (SE; n = 9) during the 2.5 h post-glucose injection. No significant changes in high energy phosphate concentrations were observed during the same period after saline injection. After glucose injection, the [phosphocreatine] gradually decreased by 64% (P = 0.0001). After the initial 1 h post-glucose injection, the [inorganic phosphate] increased by 58% (P = 0.0001), and the [nucleoside triphosphates] decreased by 29% (P = 0.0001) during the following 1.5 h. After mannitol injection, while there was no change in [inorganic phosphate] over time (P = 0.37), the [phosphocreatine] decreased by 33% (P = 0.0001) and the [nucleoside triphosphates] decreased by 21% (P = 0.0015) within 20 min, then both the [phosphocreatine] and [nucleoside triphosphates] remained at constant levels during the following 2 h. In parallel experiments, the volumetric rate of tumor blood flow and perfusion was measured by 2H nuclear magnetic resonance monitoring of 2H2O washout kinetics (S-G. Kim and J. J. H. Ackerman, Cancer Res., 48: 3449-3453, 1988); tumor blood flow decreased by 80% (P = 0.0001, n = 11), 60% (P = 0.0031, n = 4), and 20% (P = 0.058, n = 10) at 2 h after glucose, mannitol, or saline injections, respectively. These results suggest that anaerobic glycolysis is a requirement for hyperglycemic tumor acidification. However, the decrease in tumor blood flow accompanying hyperglycemic acidification suggests that flow reduction also may be a contributing or a required cofactor for acidification via inhibition of lactic acid egress.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

1H and 2H NMR studies of water in work-free wheat flour doughs.

Proton and deuterium NMR relaxation methods were used to characterize water compartmentalization and hydration in work-free wheat flour doughs. Transverse (spin-spin) relaxation measurements define three motionally unique water compartments in the work-free dough preparations. The apparent occupancy fraction and relative mobility of each water domain are found to be functions of moisture content, temperature, and flour type. Additionally, the number of relaxation-resolved water compartments and their characteristic relaxation rate constants are found to depend critically on both moisture content and the interpulse-delay employed for the multi-pulse relaxation experiments. Under controlled experimental conditions, dynamics between the three water compartments can be observed to be consistent with the onset of flour hydration. The most notable observation during the initial period of hydration is a loss of "free" or "loosely bound" water to environments characterized by less mobility. Freezing studies show that hard wheat doughs have slightly less amorphous, non-freezable water than do soft wheat flour doughs prepared under similar conditions.

Chemical Phenomena↗

Sepsis does not impair tricarboxylic acid cycle in the heart.

Sepsis has been reported to cause mitochondrial dysfunction and inhibition of key enzymes that regulate the tricarboxylic acid (TCA) cycle. We investigated the effect of sepsis on high-energy phosphates, glycolytic and TCA cycle intermediates, and specific amino acids that are involved in regulating the size of the TCA cycle pool during changes in metabolic state of the heart. Sepsis was induced in 12 female rats by the cecal ligation and perforation technique under halothane anesthesia; seven control rats underwent cecal manipulation without ligation. At 36-42 h postsurgery, the rats were reanesthetized, the chest was opened, and the hearts were freeze-clamped. Perchloric acid extracts of the hearts were analyzed with fluorometric enzymatic methods and 31P nuclear magnetic resonance spectroscopy. There were no significant differences in the levels of the TCA cycle intermediates or high-energy phosphates between the septic and control rats. The major metabolic changes were the 28% decrease in alanine and the 31% decrease in glutamate in the septic hearts compared with control (P less than 0.05 and P less than 0.005, respectively). Phosphocholine, a component of membrane phospholipids, was increased by 91% in the septic hearts (P less than 0.01). We conclude that sepsis does not impair the TCA cycle or induce significant cellular ischemia in the heart. The increase in phosphocholine may represent significant cellular membrane disruption during sepsis.

Adenosine Triphosphate↗

Quantification of regional blood flow by monitoring of exogenous tracer via nuclear magnetic resonance spectroscopy.

Deuterium and fluorine nuclear magnetic resonance spectroscopy have been employed for quantification of regional blood flow in concert with nonradiative, exogenous, freely diffusible tracers such as D2O and freon gas. Typically, the tracer residue washout was monitored by NMR over time following tracer administration by bolus injection or inhalation. The theory, including compartmental analysis, required to quantitatively derive volumetric tissue blood flow and perfusion is reviewed herein. Applications of NMR tissue blood flow measurement techniques to tumor, muscle, liver, and brain are presented with discussion of the advantages and disadvantages of NMR methods.

Animals↗

Spatially-localized NMR spectroscopy employing an inhomogeneous surface-spoiling magnetic field gradient. 1. Phase coherence spoiling theory and gradient coil design.

The use of a current-generated, inhomogeneous, surface-spoiling magnetic field gradient for enhancing magnetic resonance spatial localization, by rapidly inducing spin phase incoherence in surface lying regions of a sample is examined theoretically. A geometrically simple surface-spoiling magnetic field gradient coil design is presented and its gradient field characterized via computer simulations. Mathematical expressions describing the time dependence of net sample spin phase coherence under influence of the spoiling gradient are developed for application with homogeneous (B1) volume coils. The dependence of spoiling efficiency on the magnetogyric ratio of the nuclide under investigation and on the current driving the gradient field is described. Spoiling periods of ca 1-2 ms with driving currents of ca 0.5-1.0 A are predicted to be adequate for surface-spoiling experiments with rat, e.g., for noninvasive monitoring of liver. A companion article (W. Chen and J.J.H. Ackerman, NMR Biomed., 3, 158-165 (1990)) describes implementation of the surface-spoiling technique with multicompartment models (phantoms) and with rat in vivo.

Equipment Design↗

Spatially-localized NMR spectroscopy employing an inhomogeneous surface-spoiling magnetic field gradient. 2. Surface coil experiments with multicompartment phantom and rat in vivo.

The use of inhomogeneous surface-spoiling magnetic field gradients for elimination of signal from surface lying regions of a sample was theoretically examined in the companion article (W. Chen and J.J.H. Ackerman, NMR Biomed. 3, 147-157 (1990)). Using the spoiling gradient coil design described therein, this article presents experimental verification of the feasibility of such an approach to enhanced spatial localization. Single coil mode 31P NMR surface coil interrogation of both a two compartment phantom and rat in vivo are shown to provide excellent suppression of surface lying regions with minimal degradation of signal from the deep lying region of interest. Both pulse-and-collect and spin echo sequences were highly efficient in concert with spoiling gradient periods of 0.5-2 ms and driving currents of 0.5-2 A. The use of a current-generated surface spoiling gradient offers a robust means to remove surface tissue signal contributions and can be implemented with a wide range of localizing pulse sequences and imaging protocols.

Animals↗