Showing papers in "NMR in Biomedicine in 1999"

Tumour phospholipid metabolism.

Abstract: Following the impetus of early clinical and experimental investigations, in vivo and in vitro MRS studies of tumours pointed in the eighties to the possible significance of signals arising from phospholipid (PL) precursors and catabolites as novel biochemical indicators of in vivo tumour progression and response to therapy. In the present decade, MRS analyses of individual components contributing to the 31P PME (phosphomonoester) and PDE (phosphodiester) resonances, as well as to the 1H 'choline peak', have reinforced some of these expectations. Moreover, the absolute quantification of these signals provided the basis for addressing more specific (although still open) questions on the biochemical mechanisms responsible for the formation of intracellular pools of PL derivatives in tumours, under different conditions of cell proliferative status and/or malignancy level. This article is aimed at providing an overview on: (a) quantitative MRS measurements on the contents of phosphocholine (PCho), phosphoethanolamine (PEtn) and their glycerol derivatives ģlycerol 3-phosphocholine (GPC) and glycerol 3-phosphoethanolamine (GPE)[ in human tumours and cells (with particular attention to breast and brain cancer and lymphomas), as well as in normal mammalian tissues (including developing organs and rapidly proliferating tissues); (b) possible correlations of MRS parameters like PEtn/PCho and PCho/GPC ratios with in vitro cell growth status and/or cell tumorigenicity; and (c) current and new hypotheses on the role and interplay of biosynthetic and catabolic pathways of the choline and ethanolamine cycles in modulating the intracellular sizes of PCho and PEtn pools, either in response to mitogenic stimuli or in relation to malignant transformation.

Multi-component apparent diffusion coefficients in human brain.

Abstract: The signal decay with increasing b-factor at fixed echo time from brain tissue in vivo has been measured using a line scan Stejskal–Tanner spin echo diffusion approach in eight healthy adult volunteers. The use of a 175 ms echo time and maximum gradient strengths of 10 mT/m allowed 64 b-factors to be sampled, ranging from 5 to 6000 s/mm2, a maximum some three times larger than that typically used for diffusion imaging. The signal decay with b-factor over this extended range showed a decidedly non-exponential behavior well-suited to biexponential modeling. Statistical analyses of the fitted biexponential parameters from over 125 brain voxels (15 × 15 × 1 mm3 volume) per volunteer yielded a mean volume fraction of 0.74 which decayed with a typical apparent diffusion coefficient around 1.4 µm2/ms. The remaining fraction had an apparent diffusion coefficient of approximately 0.25 µm2/ms. Simple models which might explain the non-exponential behavior, such as intra- and extracellular water compartmentation with slow exchange, appear inadequate for a complete description. For typical diffusion imaging with b-factors below 2000 s/mm2, the standard model of monoexponential signal decay with b-factor, apparent diffusion coefficient values around 0.7 µm2/ms, and a sensitivity to diffusion gradient direction may appear appropriate. Over a more extended but readily accessible b-factor range, however, the complexity of brain signal decay with b-factor increases, offering a greater parametrization of the water diffusion process for tissue characterization. Copyright © 1999 John Wiley & Sons, Ltd.

Measurement of the extracellular pH of solid tumours in mice by magnetic resonance spectroscopy: a comparison of exogenous 19F and 31P probes

Abstract: Precise measurement of pHein vivo may be of clinical value for both diagnosis and selection of therapy. pHe measurements made by the 31P probe 3-aminopropylphosphonate (3-APP) were compared with those made by the 19F probe, 3-[N-(4-fluor-2-trifluoromethylphenyl)-sulphamoyl]-propionic acid (ZK-150471) in three solid tumour types, human HT29 xenografts, murine RIF-1 fibrosarcomas and Lettre tumours grown subcutaneously in mice. No significant differences were observed when probe measurements of pHe were compared at 20–60 min post-administration, although very low pHe values (ca. 6.0) were recorded in two out of eight Lettre tumours by ZK-150471. The more rapid pHe measurements possible using ZK-150471 showed that during the first 20 min post-administration significant increases occurred in pHe which were greatest in the more necrotic tumours. Since isolated cell experiments showed that ZK-150471 was non-toxic and did not enter the cells, this early increase in pHe may reflect gradual penetration by ZK-150471 of the reportedly alkaline necrotic space in the tumours. The wide chemical shift range, improved signal-to-noise and absence of signal overlap allowed a more rapid and precise measurement of pHe by ZK-150471 compared to 3-APP. These characteristics suggest that ZK-150471 is currently the preferred pHe probe for non-invasive MRS. Copyright © 1999 John Wiley & Sons, Ltd.

Serial evaluation of patients with brain tumors using volume MRI and 3D 1H MRSI.

Abstract: Patients with brain tumors are routinely monitored for tumor progression and response to therapy using magnetic resonance imaging (MRI) Although serial changes in gadolinium enhancing lesions provide valuable information for making treatment decisions, they do not address the fate of non-enhancing lesions and are unable to distinguish treatment induced necrosis from residual or recurrent tumor The introduction of a non-invasive methodology, which could identify an active tumor more reliably, would have a major impact upon patient care and evaluation of new therapies There is now compelling evidence that magnetic resonance spectroscopic imaging (MRSI) can provide such information as an add-on to a conventional MRI examination We discuss data acquisition and analysis procedures which are required to perform such serial MRI–MRSI examinations and compare their results with data from histology, contrast enhanced MRI, MR cerebral blood volume imaging and FDG-PET Applications to the serial assessment of response to therapy are illustrated by considering populations of patients being treated with brachytherapy and gamma knife radiosurgery Copyright © 1999 John Wiley & Sons, Ltd

Factors affecting the quantification of short echo in-vivo 1H MR spectra: prior knowledge, peak elimination, and filtering.

Abstract: Short echo 1 H in-vivo brain MR spectra are difficult to quantify for several reasons: low signal to noise ratio, the severe overlap of spectral lines, the presence of macromolecule resonances beneath the resonances of interest, and the effect of resonances adjacent to the spectral region of interest (SRI). This paper outlines several different quantification strategies and the effect of each on the precision of in-vivo metabolite measurements. In-vivo spectra were quantified with no operator interaction using a template of prior knowledge determined by mathematically modeling separate in-vitro metabolite spectra. Metabolite level estimates and associated precision were compared before and after the inclusion of macromolecule resonances as part of the prior knowledge, and following two different methods of handling resonances adjacent to the SRI. The effects of rectangular and exponential filters were also investigated. All methods were tested using repeated in-vivo spectra from one individual acquired at 1.5 T using stimulated echo acquisition mode (STEAM, TE = 20 ms) localization. The results showed that the inclusion of macromolecules in the prior knowledge was necessary to obtain metabolite levels consistent with the literature, while the fitting of resonances adjacent to the SRI concurrent with modeled metabolites optimized the precision of metabolite estimates. Metabolite levels and precision were also affected by rectangular and exponential filtering, suggesting caution must be taken when such filters are used.

Quantitative 13C NMR studies of metabolic compartmentation in the adult mammalian brain

Abstract: Funded by: DGES. Grant Number: PB96-0864-C02 and Community of Madrid. Grant Number: 08/0023/1997.

Structural information in neuronal tissue as revealed by q-space diffusion NMR spectroscopy of metabolites in bovine optic nerve.

Abstract: 1H NMR diffusion experiments performed on the signal of the metabolites in bovine optic nerve showed that the signal decay due to diffusion is bi-exponential with a slow and a fast diffusing component. Diffusion was measured as a function of the diffusion time, and the data were analyzed as a function of b and q values. Bi-exponential fit was used to analyze the data, and the results were compared with the displacement distribution profiles obtained from the q-space analysis of the data. This q-space analysis showed that the fast diffusing component has a broad displacement distribution and appears not to be restricted. On the other hand, the slow diffusing component appears to be highly restricted to milieu in the order of 1–2 µm. The orientation of the sample with respect to the axis for which diffusion was measured affected mainly the relative sizes of the populations of each component, but had only a small effect on the extracted apparent diffusion coefficients. These results from both the b and the q value analyses suggest that the slow diffusing component is related to restricted diffusion of these metabolites in the axonal fibers, while the fast diffusing component represents diffusion of metabolites in cells and along the long axis of the nerve fibers. It is concluded that q-space analysis of metabolite diffusion enables extraction of structural information about the sample, and that the diffusion of the metabolites in optic nerve is dictated mainly by the cellular medium and microstructure of the tissue.

In vivo magnetic resonance methods in pharmaceutical research: current status and perspectives

Abstract: In the last decade, in vivo MR methods have become established tools in the drug discovery and development process. In this review, several successful and potential applications of MRI and MRS in stroke, rheumatoid and osteo-arthritis, oncology and cardiovascular disorders are dealt with in detail. The versatility of the MR approach, allowing the study of various pathophysiological aspects in these disorders, is emphasized. New indication areas, for the characterization of which MR methods have hardly been used up to now, such as respiratory, gastro-intestinal and skin diseases, are outlined in a subsequent section. A strength of MRI, being a non-invasive imaging modality, is the ability to provide functional, i.e. physiological, readouts. Functional MRI examples discussed are the analysis of heart wall motion, perfusion MRI, tracer uptake and clearance studies, and neuronal activation studies. Functional information may also be derived from experiments using target-specific contrast agents, which will become important tools in future MRI applications. Finally the role of MRI and MRS for characterization of transgenic and knock-out animals, which have become a key technology in modern pharmaceutical research, is discussed. The advantages of MRI and MRS are versatility, allowing a comprehensive characterization of a diseased state and of the drug intervention, and non-invasiveness, which is of relevance from a statistical, economical and animal welfare point of view. Successful applications in drug discovery exploit one or several of these aspects. In addition, the link between preclinical and clinical studies makes in vivo MR methods highly attractive methods for pharmaceutical research.

Diffusional anisotropy is induced by subcellular barriers in skeletal muscle.

Abstract: The time- and orientational-dependence of phosphocreatine (PCr) diffusion was measured using pulsed-field gradient nuclear magnetic resonance (PFG-NMR) as a means of non-invasively probing the intracellular diffusive barriers of skeletal muscle. Red and white skeletal muscle from fish was used because fish muscle cells are very large, which facilitates the examination of diffusional barriers in the intracellular environment, and because they have regions of very homogeneous fiber type. Fish were cold-acclimated (5 degrees C) to amplify the contrast between red and white fibers. Apparent diffusion coefficients, D, were measured axially, D(axially) and radially, D(radially), in small muscle strips over a time course ranging from 12 to 700 ms. Radial diffusion was strongly time dependent in both fiber types, and D decreased with time until a steady-state value was reached at a diffusion time approximately 100 ms. Diffusion was also highly anisotropic, with D(axially) being higher than D(radially) for all time points. The time scale over which changes in D(radially) occurred indicated that the observed anisotropy was not a result of interactions with the thick and thin filament lattice of actin and myosin or restriction within the cylindrical sarcolemma, as has been previously suggested. Rather, the sarcoplasmic reticulum (SR) and mitochondria appear to be the principal intracellular structures that inhibit mobility in an orientation-dependent manner. This work is the first example of diffusional anisotropy induced by readily identifiable intracellular structures.

Tumour response to hypercapnia and hyperoxia monitored by FLOOD magnetic resonance imaging.

Abstract: Flow and oxygenation dependent (FLOOD) MR images of GH3 prolactinomas display large intensity increases in response to carbogen (5% CO 2 /95% O 2 ) breathing. To assess the relative contributions of carbon dioxide and oxygen to this response and the tumour oxygenation state, the response of GH3 prolactinomas to 5% CO 2 /95% air, carbogen and 100% O 2 was monitored by FLOOD MRI and pO 2 histography. A 10-30% image intensity increase was observed during 5% CO 2 /95% air breathing, consistent with an increase in tumour blood flow, as a result of CO 2 -induced vasodilation, reducing the concentration of deoxyhaemoglobin in the blood. Carbogen caused a further 40-50% signal enhancement, suggesting an additional improvement due to increase blood oxygenation. A small 5-10% increase was observed in response to 100% O 2 , highlighting the dominance of CO 2 -induced vasodilation in the carbogen response. Despite the large FLOOD response, non-significant increases in tumour pO 2 were observed in response to the three gases. Tissue pO 2 is determined by the balance of oxygen supply and demand, hence increased blood flow/oxygenation may not necessarily produce a large increase in tissue pO 2 . The FLOOD response is determined by the level of deoxygenation of blood, the size of this response relating to vascular density and the potential of high-oxygen content gases to improve the oxygen supply to tumour tissue.

Magnetic resonance angiography of the rat cerebrovascular system without the use of contrast agents.

Abstract: We describe and discuss the application of three-dimensional (3D) time-of-flight (TOF) magnetic resonance angiography (MRA) to visualize non-invasively the cerebral vasculature of the rat. MR angiograms of healthy spontaneously hypertensive rats were obtained without the use of contrast agents. Total imaging time ranged from 1 to 50 min for a 3D data set. The influences of the data matrix and the inflow delay on the image quality and the total imaging time are assessed and discussed. Varying the inflow delay yielded in addition semiquantitative information on hemodynamics. The method was applied to obtain angiograms in rat models of permanent and temporal middle cerebral artery occlusion. Occlusion and reopening of the vessel could easily be verified by MRA. However, after reperfusion a slight reduction in blood flow was observed.

In vivo magnetic resonance spectroscopy and dynamic contrast enhanced imaging of the prostate gland.

Abstract: The quantitation of in vivo H MR spectroscopy and dynamic contrast enhanced MR imaging is described for patients with histologically confirmed prostate adenocarcinoma and benign prostatic hypertrophy (BPH). Results are presented which suggest that combined use of these techniques may be helpful in improving the characterization of prostate pathologies and ultimately increase the staging accuracy of magnetic resonance. Copyright © 1999 John Wiley & Sons, Ltd.

Diffusion- and perfusion-weighted MR imaging of transient focal cerebral ischaemia in mice

Abstract: Temporary focal ischaemia was induced in wild-type C57Black/6 mice by thread occlusion of the middle cerebral artery (MCA). Recirculation was started after 60 min and maintained for 24 h, after which the mouse brain was frozen in situ. Development of the cerebral infarct was monitored by diffusion-, perfusion- and T2-weighted magnetic resonance imaging (MRI) during ischaemia, during the early reperfusion period of 90 min, and at 24 h after reperfusion. Ischaemia caused a marked reduction of the perfusion signal intensity and of the apparent diffusion coefficient (ADC) of tissue water in the ipsilateral MCA territory. In sham-operated control animals ADC remained unchanged. Hemispheric lesion volume after 1 h MCA occlusion was 53 ± 6% (n = 6), as defined by an ADC decrease of more than 20%. Recirculation reduced hemispheric lesion volume to only 27 ± 13%, while there was a trend towards secondary lesion growth at 24 h. Post-ischaemic recovery of perfusion was slow, heterogeneous and incomplete. A region-of-interest analysis showed only partial and transient recovery of the ADC, particularly in the dorsolateral cortex and lateral caudate putamen, which may be explained by inadequate reperfusion in these regions. Detailed MRI studies of cerebral ischaemia and reperfusion may now also be performed in the transgenic mice. Copyright © 1999 John Wiley & Sons, Ltd.

In vivo magnetic resonance spectroscopy of human fetal neural transplants.

Abstract: To better define the survival and cellular composition of human fetal neurotransplants in vivo, we performed quantitative 1H MRS to determine the concentration of the neuronal amino acid [N-acetylaspartate] within MRI-visible grafts. In all, 71 grafts in 38 patients [24 Parkinson's disease (PD), 14 Huntington's disease (HD)] were examined, as well as 24 untreated PD and HD patients and 13 age-matched normal controls. MRI appearances of edema were present in three out of 71 grafts, the remainder being consistent with histologically identified viable neural transplant tissue. N-acetylaspartate (NAA), creatine, choline, myoinositol and glutamine plus glutamate (Glx) were identified in all post-transplant putamens, with abnormal metabolites, lactate and/or lipid detectable in only three patients. Of 71 grafts, 19 occupied more than 60% of the MRS-examined volume (VOI) (mean 84.2 ± 3%; range 61–100%). In those, [NAA] was 8.50 ± 0.99 mM in eight PD spectra and 6.59 ± 0.81 mM in 11 HD spectra, and was not significantly different from controls. In contrast, transplanted fetal neurones contain less than 0.4 mM of the neuronal amino acid NAA. This suggests that established fetal neurotransplants in the human putamen of both PD and HD patients are populated by adult neurones, axons and dendrites. Copyright © 1999 John Wiley & Sons, Ltd.

Regional and directional anisotropy of apparent diffusion coefficient in rat brain

Abstract: Quantitative diffusion maps were recorded in normal rat brain. In multi-slice sections covering the whole brain, strong variation of the apparent diffusion coefficient (ADC) was observed depending on slice position at constant gradient direction. Furthermore, a varying difference between apparent diffusion coefficients depending on gradient direction was found, reaching 32% in the cortex of the ventral-most horizontal sections while showing equal ADC on the dorsal cortex side. The regional variation and directional anisotropy of the ADC was not restricted to white matter but was described for both cortical and subcortical brain tissue. From diffusion coefficients along the three major field gradient directions (ADCx, ADCy, ADCz), the average ADC (ADCaverage) was determined from the trace of the diffusion tensor (D) as 653+/-28 microm2/s for parietal cortex and 671+/-32 microm2/s for lateral cortex, independent of position along the sagittal direction. From these observations about the regional diffusion anisotropy, a more stringent protocol for the description of ischemic ADC changes is proposed.

1H-NMR analysis of microbial-derived organic acids in primary root carious lesions and saliva

Abstract: In addition to lowered pH values, the molecular profile and concentrations of microbial-derived organicacids in carious dentin are important demineralization parameters involved in the induction, development and progression of dental caries. High-resolution proton ( 1H) NMR spectroscopy was employed to examine the organic acid status of primary root carious lesions. 1H-NMR analysis of post-neutralized perchloric acid extracts of active carious lesions revealed that at an operating frequency of 600 MHz, the 1H-NMR-detectable organic acid composition of carious dentin samples (mean molecular percentage content ± standard error; the mean molecular percentage content is defined here as the mean of the concentration of each 1H-NMR-visible organic acid/anion expressed as a percentage of total 1H-NMR-detectable organic acid/anion level in each sample) was acetate 51 ± 2%, formate 37 ± 2%, lactate 5 ± 1%, propionate 3 ± 0.8%, pyruvate 2.4 ± 0.3%, n-butyrate 1.2 ± 0.2%; succinate 0.1 ± 0.1%; iso-butyrate, n- and iso-valerate, and n- and iso-caproate (total) <0.2%. Further components detectable included alanine, glycine, choline, phosphorylcholine, trimethylamine oxide, methanol, glycolate and assorted saccharides. In view of their high dissociation constants (Ka), our results demonstrate that formic and pyruvic acids (Ka = 1.77 x 10-4 and 3.20 x 10-3 mol/dm3, respectively) contribute substantially to the decreased pH values associated with active caries lesions (cf. lactate Ka = 1.40 x 10-4 mol/dm3).

Myocardial manganese elevation and proton relaxivity enhancement with manganese dipyridoxyl diphosphate. Ex vivo assessments in normally perfused and ischemic guinea pig hearts.

Abstract: Manganese (Mn) dipyridoxyl diphosphate (MnDPDP) is the active component of a contrast medium for liver MRI. By being metabolized, MnDPDP releases Mn(2+), which is taken up and retained in hepatocytes. The study examined whether MnDPDP elevates Mn content and enhances proton relaxivity in normal myocardium, but not in ischemic myocardium with reduced coronary flow and impaired metabolism. Isolated guinea pig hearts were perfused at normal flow or low flow, inducing global subtotal ischemia. Ventricular ATP and Mn contents, T(1) and T(2) were measured. At normal flow tissue Mn content increased from the control level of 4.1 to 70.4 micromol/100g dry wt with MnDPDP (3000 microM), while low-flow perfusion with MnDPDP (3000 microM) resulted in a Mn content of 16.6 micromol/100 g dry wt. Prolonged ischemia (35 and 90 min) reduced tissue Mn down to the control level. T(1) shortening closely paralleled myocardial Mn elevations during both normal and low-flow perfusion. The use of a Mn(2+)-releasing contrast agent like MnDPDP may be a promising principle in MRI assessments of myocardial function and viability in coronary heart disease by revealing a differential pattern of changes in T(1) relative to coronary flow, cell Mn uptake and retention, ion channel function and metabolism.

Proton MRS of oral creatine supplementation in rats. Cerebral metabolite concentrations and ischemic challenge

Abstract: Proton magnetic resonance spectroscopy (MRS) was employed to determine the concentrations of N-acetylaspartate (NAA), total creatine (tCr), choline-containing compounds (Cho), myo-inositol (Ins), glucose (Glc), and lactate (Lac) in rat brain before and after 10 days of oral supplementation of 2.6 g Cr-monohydrate per kg body weight per day. Measurements were performed both in vitro (n = 16) and in vivo (n = 6). The neuroprotective potential of oral Cr was assessed by dynamically monitoring brain Glc and Lac in response to transient global ischemia (12 min). In comparison to controls the in vitro concentrations of Cr (13.1 ± 9.3%) and Ins (12.7 ± 14.0%) were significantly increased in Cr-fed rats. Under in vivo conditions, the data revealed trends for elevated tCr (4.7%) and Ins (10.6%) which were enhanced in the concentration ratios of tCr:Cho (10.2%) and Ins:Cho (17.8%). Together with an increased Glc level (27.3%), the observation of a statistically significant decrease of brain Lac (−38.5 ± 19.3%) in Cr-fed rats may reflect a shift of the energy metabolism from non-oxidative toward oxidative glycolysis. One hour after global ischemia most of the metabolic differences between Cr-fed rats and controls were retained. The increased Glc level (44.4 ± 33.3%) reached statistical significance, but the accumulation of Lac and its time course during ischemia and early reperfusion showed no differences between Cr-fed rats and controls. Copyright © 1999 John Wiley & Sons, Ltd.

Brain metabolic alterations in Cushing's syndrome as monitored by proton magnetic resonance spectroscopy.

Abstract: Proton magnetic resonance spectroscopy ( 1 H MRS) was used to evaluate changes in cerebral metabolites in 13 patients with Cushing's syndrome (including seven with pituitary corticotroph adenomas and six with primary adrenal disease) as compared to 40 normal subjects. Data were recorded in the frontal, thalamic and temporal areas; quantification of the MRS signals demonstrated a statistically significant decrease of the Cho/Cr ratio in the frontal and thalamic areas but not in the temporal area for patients with Cushing's syndrome. The largest decrease in Cho/Cr was measured in the thalamic area of patients with a Cushing's syndrome secondary to an adrenal disease. No statistically significant changes in the NAA/Cr ratio were measured in any of the areas studied. These results suggest that the quantification of choline levels could be helpful for monitoring the cerebral metabolite alterations in patients with hypercortisolism.

Inflammatory granulomas: evaluation with proton MRS

Abstract: Intracranial inflammatory lesions consisting mainly of tuberculomas (n = 28), neurocysticercosis (NCC) (n = 10), and non-specific inflammatory granulomas (IG) (n = 22) were evaluated with proton MRS. Water-suppressed proton spectra were acquired from 60 patients using the STEAM sequence with an echo time of 135 ms and metabolite ratios determined from the spectra. Student's paired t-test and chi2-test were used to analyse the data. Statistically significant differences were observed for the following ratios between the three patient categories: NAA/ Cr (p < 0.0001), NAA/Cho (p = 0.001) and Cr/Cho (p = 0.02) for the non-specific IG and NCC, NAA/Cho (p = 0.03) for non-specific IG and tuberculoma, and NAA/Cr (p < 0.0001) for NCC and tuberculoma. While lipids were seen in 86% of the tuberculomas, they were observed in only 20% of the NCCs (chi2 = 6.81, p = 0.009), and 21% of the non-specific IGs (chi2 = 10.75, p = 0.0001). While the presence of lipid can be used for differentiating tuberculomas from both non-specific IG and NCC, the extremely low levels of metabolites together with a poor signal/noise ratio could itself act as a marker for NCC.

fMRI assessment of hemispheric language dominance using a simple inner speech paradigm

Abstract: Hemispheric language dominance (HLD) has been determined by means of functional MRI (fMRI) using a simple, inner speech, word fluency paradigm. During the task periods, subjects perform mental imagery of visual scenes and generate silently the nouns of all objects visualized. During the control periods, subjects attend to the scanner noise. Activated areas have been identified by means of cross-correlation analysis. HLD indices have been determined by comparing the number of activated pixels detected in both hemispheres within predefined cortical areas (Brodmann areas 6, 9, 10, 39, 40 and 44-47). The paradigm has been assessed on 10 healthy, right-handed volunteers. A volume 35 mm thick, centered on the inferior frontal gyrus, was imaged. A conventional GRE MR sequence was used on a 1.5 T clinical MR scanner. HLD indices were compared with those determined for overt speech. Robust fMRI responses were obtained. HLD indices indicated left hemispheric language dominance for all subjects examined. They correlated well with those obtained for overt speech (R(2) = 0.93, regression coefficient = 0.998, with p < 10(-4)). Thus, an inner speech paradigm based on visual imagery is well adapted for assessment of HLD by means of fMRI.

Effects of ischemia on skeletal muscle energy metabolism in mice lacking creatine kinase monitored by in vivo 31P nuclear magnetic resonance spectroscopy.

Abstract: The aim of this study was to provide in vivo experimental evidence for the proposed biological significance of the creatine kinase (CK)/phosphocreatine (PCr) system in the energy metabolism of skeletal muscle. As a test system we compared hindlimb muscle of knockout mice lacking the cytosolic M-type (M-CK(-)/(-)), the mitochondrial ScMit-type (ScCKmit(-)/(-)), or both creatine kinase isoenzymes (CK(-)/(-)), and in vivo 31P-NMR was used to monitor metabolic responses during and after an ischemic period. Although single mutants show some subtle specific abnormalities, in general their metabolic responses appear similar to wild type, in contrast to CK(-)/(-) double mutants. This implies that presence of one CK isoform is both necessary and sufficient for the system to be functional in meeting ischemic stress conditions. The global ATP buffering role of the CK/PCr system became apparent in a 30% decline of ATP in the CK(-)/(-) mice during ischemia. Both M-CK(-)/(-) and CK(-)/(-) showed increased phosphomonoester levels during ischemia, most likely reflecting adaptation to a more efficient utilization of glycogenolysis. While in M-CK(-)/(-) muscle PCr can still be hydrolyzed to provide Pi for this process, in CK(-)/(-) muscle only Pi from ATP breakdown is available and Pi levels increase much more slowly. The experiments also revealed that the system plays a role in maintaining pH levels; the CK(-)/(-) mice showed a faster and more pronounced acidification (pH = 6.6) than muscles of wild type and single knockout mutants (pH = 6.9).

31P and 1H NMR spectroscopic studies of liver extracts of carbon tetrachloride‐treated rats

Abstract: NMR spectroscopy was used to examine hepatic metabolism in cirrhosis with a particular focus on markers of functional cellular hypoxia. P and H NMR spectra were obtained from liver extracts from control rats and from rats with carbon tetrachloride-induced cirrhosis. A decrease of 34% in total phosphorus content was observed in cirrhotic rats, parallelling a reduction of 40% in hepatocyte mass as determined by morphometric analysis. Hypoxia appeared to be present in cirrhotic rats, as evidenced by increased inorganic phosphate levels, decreased ATP levels, decreased ATP:ADP ratios (1.72 ± 0.40 vs 2.48 ± 0.50, p < 0.01), and increased inorganic phosphate:ATP ratios (2.77 ± 0.48 vs 1.62 ± 0.24, p < 0.00001). When expressed as a percentage of the total phosphorus content, higher levels of phosphoethanolamine and lower levels of NAD and glycerophosphoethanolamine were detected in cirrhotic rats. Cirrhotic rats also had increased phosphomonoester:phosphodiester ratios (5.73 ± 2.88 vs 2.53 ± 0.52, p < 0.01). These findings are indicative of extensive changes in cellular metabolism in the cirrhotic liver, with many findings attributable to the presence of intracellular hypoxia.

Quantifying 1H decoupled in vivo 31P brain spectra.

Abstract: Our objective was to develop a precise method for quantification of in vivo proton decoupled 31 P spectra from the human brain. This objective required that an appropriate spectral model be created and that the quantification was performed using a non-subjective fitting technique. The precision of the quantification was assessed using Cramer-Rao standard deviations and compared using two different spectral models: one containing a pair of peaks representing 2,3-diphosphoglycerate the other excluding this metabolite. The data was quantified using a Marquardt-Levenberg (ML) algorithm incorporating prior knowledge with a Hankel singular value decomposition (HSVD) performed initially to provide parameter estimates for the ML algorithm. Quantification was performed on two different in vivo 2-D CSI 31 P data sets: the first examined 11 normal controls, the second examined a single individual six times. Spectra from a region in the parieto-occipital cortex were analyzed. The Cramer-Rao standard deviations were significantly lower for some metabolites with 2,3-diphosphoglycerate in the model: in the repeat study mobile phospholipids (p = 0.045) and phosphocholine (p = 0.034), and in the 11 controls mobile phospholipids (p = 0.003) and P i (p = 0.002). Copyright © 1999 John Wiley & Sons, Ltd.

Non-invasive observation of acetyl-group buffering by 1H-MR spectroscopy in exercising human muscle.

Abstract: The observation of a previously unidentified peak in localized 1H magnetic resonance (MR) spectra of human muscle during and after a work load is reported. Basic NMR properties of this resonance, as well as physiologic circumstances of its observation, suggest that it is due to the acetyl group of acetylcarnitine. The relatively large pool of muscular carnitine acts as a buffering system stabilizing the ratio of acetylated to free coenzyme A. Free carnitine can be acetylated to a large extent whenever a mismatch occurs between the fluxes through pyruvate dehydrogenase and the TCA cycle. Results of initial applications of 1H MR spectroscopy in several muscles and under different exercise regimens are in agreement with earlier invasive measurements of acetylcarnitine. It is demonstrated that the detailed dynamics of acetyl group formation are now likely to be observable non-invasively in humans by localized 1H magnetic resonance spectroscopy on standard MR imaging systems, and that acetylcarnitine buffering as a function of exercise type, oxygenation states, diet and pathology could thus be studied repeatedly and in various muscle groups with much improved temporal resolution.

Frontal lobe in vivo (31)P-MRS reveals gender differences in healthy controls, not in schizophrenics.

Abstract: Phosphorus-31 magnetic resonance spectroscopy ((31)P-MRS) has gained much interest in schizophrenia research in recent years since it allows the non-invasive measurement of high-energy phosphates and phospholipids in vivo. However, until now only differences in metabolite concentrations between certain brain areas of schizophrenic patients and healthy controls have been examined. We investigated the influence of gender on the concentrations of different phosphorus compounds. For this purpose, well-defined volumes in the frontal lobe of 32 healthy controls and 51 schizophrenic in-patients were examined with an image selected in vivo spectroscopy (ISIS) sequence on a whole-body scanner at 1.5 T. Healthy females exhibited increased values of inorganic phosphate (P(i)) and decreased values of phosphocreatine (PCr) in comparison to their male counterparts. In schizophrenic patients such gender differences were not present. Thus, the results can be interpreted in the sense that frontal energy demanding processes are enhanced in female compared to male healthy volunteers; schizophrenia seems to reduce these gender differences.

Metabolite changes in neonatal rat brain during and after cerebral hypoxia-ischemia: a magnetic resonance spectroscopic imaging study.

Abstract: Cerebral metabolite concentrations were measured in infant rats using proton magnetic resonance spectroscopic imaging. Measurements were made prior to, during and after exposure of rats (6- and 7-day-old) to unilateral cerebral hypoxia-ischemia (right carotid artery occlusion +2h 8% oxygen). Data clustered according to age and outcome-6-day-old animals with no infarct and 7-day-old animals with infarct. In 6-day-old animals, cerebral lactate concentration increased during hypoxia-ischemia, particularly ipsilateral to the occlusion, and returned to normal soon after the end of hypoxia. There were no major changes in N-acetyl-aspartate levels (NAA) in this group and no regions of hyperintensity on T2 or DW weighted images at 24 h. In the 7-day-old animals, lactate increased during hypoxia-ischemia and remained elevated in the first hour after reperfusion. Furthermore, lactate remained at 258+/-117% and 233+/-56% of pre-hypoxic levels, 24 and 48 h post-hypoxia, respectively. NAA concentrations ipsilateral to the occlusion decreased to 55+/-14% during hypoxia, recovered early post-hypoxia and again decreased to 61+/-25% and 41+/-28% at 24 and 48 h post-hypoxia-ischemia, respectively. The infarct volumes measured by diffusion weighted and T2 weighted MRI at 48 h post-hypoxia were 152+/-40 mm3 and 172+/-35 mm3, respectively. Thus, irreversible damage correlated well with measured in vivo lactate and NAA changes. Those animals in which NAA was unaltered and lactate recovered soon after hypoxia did not show long-term damage (6-day-old animals), whereas those animals in which NAA decreased and lactate remained elevated went on to infarction (7-day-old animals).

Effects of sedation, stimulation, and placebo on cerebral blood oxygenation: a magnetic resonance neuroimaging study of psychotropic drug action.

Abstract: The effects of pharmacologic depression and stimulation of cerebral activity were investigated in seven healthy young volunteers using blood oxygenation-sensitive MRI at 2.0 T. Dynamic gradient-echo imaging (7 min) was performed before, during and after the intravenous application of 10 mg diazepam and 15 mg metamphetamine as well as of the corresponding drug placebos (isotonic saline) in a brain section covering frontotemporal gray matter, subcortical gray matter structures, and cerebellum. The MRI responses were significantly different for the two drugs applied (p = 0.01). Relative to signal strength during injection, metamphetamine elicited a signal increase of 0.97 ± 0.03% (mean ± SD, p = 0.02) within the whole section 4–5 min after injection. Similarly, both placebo conditions led to a small signal increase, i.e. 0.50 ± 0.03% (n.s.) for the metamphetamine placebo and 0.40 ± 0.07% (p = 0.03) for the diazepam placebo. Diazepam abolished this signal increase. A topographic analysis revealed the metamphetamine-induced signal increase to be more pronounced in subcortical gray matter structures (p = 0.01) and cerebellum (p = 0.02) than in frontotemporal cortical gray matter (p = 0.04). This finding is in agreement with the hypothesis that pertinent responses not only reflect global cerebral hemodynamic adjustments, but also localized perfusion changes coupled to alterations in synaptic activity. The occurrence of a placebo response is best explained by expectancy and may provide a confounding factor in the design of functional activation experiments. Copyright © 1999 John Wiley & Sons, Ltd.

Magnetization transfer attenuation of creatine resonances in localized proton MRS of human brain in vivo.

Abstract: To assess putative magnetization transfer effects on the proton resonances of cerebral metabolites in human brain, we performed quantitative proton magnetic resonance spectroscopy (2.0 T, STEAM, TR/TE/ TM = 6000/40/10 ms, LCModel data evaluation) of white matter (7.68 mL, 10 healthy young subjects) in the absence and presence of fast repetitive off-resonance irradiation (2.1 kHz from the water resonance) using a train of 100 Gaussian-shaped RF pulses (12.8 ms duration, 120 Hz nominal bandwidth, 40 ms repetition period, 1080° nominal flip angle). A comparison of pertinent metabolite concentrations revealed a magnetization transfer attenuation factor of the methyl and methylene resonances of creatine and phosphocreatine of 0.87 0.05 (p< 0.01). No attenuation was observed for the resonances of N-acetylaspartate and N-acetylaspartylglutamate, glutamate and glutamine, choline-containing compounds, and myo-inositol. The finding for total creatine is in excellent agreement with data reported for rat brain. The results are consistent with the hypothesis of a chemical exchange of mobile creatine or phosphocreatine molecules with a small immobilized or 'bound' pool. Copyright © 1999 John Wiley & Sons, Ltd.