Biomedical subjects

D Leibfritz

Publications and source records attributed to D Leibfritz.

At least 37 records · Page 2Linked to original sources

PubMed1999

Detection of homonuclear decoupled in vivo proton NMR spectra using constant time chemical shift encoding: CT-PRESS.

A new pulse sequence, termed CT-PRESS, is presented, which allows the detection of in vivo 1H NMR spectra with effective homonuclear decoupling. A PRESS sequence with a short echo-time TE, used for spatial localization, is supplemented by an additional 180 degrees pulse. The temporal position of this 180 degree pulse is shifted within a series of experiments, while the time interval between signal excitation and detection is kept constant. CT-PRESS is a two-dimensional (2D) spectroscopic experiment as far as data acquisition and processing are concerned, although only diagonal signals are generated in the 2D spectrum. However, since the principle of constant time chemical shift encoding is used in the t1 domain, effective homonuclear decoupling is obtained by projecting the 2D spectrum onto the corresponding f1 axis. Thus, good spectral resolution and high signal-to-noise ratio are obtained. The main advantage, as compared to localized 2D J-resolved MRS, is that optimized experiments can be performed for coupled resonances of interest by choosing the sequence parameters dependent on the type of multiplets, the J-coupling constants and T2. Major fields of application will be parametric studies on coupled resonances, (e.g., T1, diffusion behavior or magnetization transfer) and/or the detection of spatial and temporal changes of metabolites with coupled spin systes.

Animals↗

PubMed1999

Improved proton spectroscopic U-FLARE imaging for the detection of coupled resonances in the rat brain in vivo.

Modifications of the pulse sequence for spectroscopic U-FLARE imaging are discussed to detect not only the predominant singlet signals of N-acetylaspartate, total creatine, and choline containing compounds or the doublet signal of lactate, but also the coupled resonances of glutamate, glutamine, taurine and myo-inositol. Effective homonuclear decoupling is achieved by use of constant time chemical shift encoding. A maximum signal-to-noise ratio (SNR) can be obtained for a certain coupled resonance of interest by optimizing the evolution period t(c) of the J modulated spin echo. Good reproducibility and a high SNR were achieved by combining several methods for water suppression and by using the displaced variant of U-FLARE. Measurements of a 3 mm slice of the rat brain were performed in vivo within 4 min, giving a nominal voxel size of 1.5 x 1.5 x 3.0 mm3 or 1.5 x 0.75 x 3.0 mm3. Thus, optimized spectroscopic U-FLARE is a powerful tool for proton spectroscopic imaging with high spectral, spatial and temporal resolution.

Animals↗

PubMed1999

Homonuclear uncoupled 1H-spectroscopy of the human brain using weighted accumulation schemes.

Homonuclear uncoupled H-spectroscopy based on 2-dimensional (2D) J-resolved spectroscopy is a well suited technique for the assessment of J-coupled metabolite resonances. In comparison with 1-dimensional H-spectroscopy this technique largely reduces problems due to spectral overlap between low molecular weight metabolite resonances and superimposed broad macromolecule resonances, as well as overlap between adjacent multiplets. Usually, 2D J-resolved spectroscopic data were acquired using a constant number of accumulations for each of the various echo times. In contrast to this, a weighted accumulation scheme has been applied in this study, using an echo time dependent number of accumulations. The aims of this modification were to reduce the acquisition time and to improve the signal-to-noise ratio per unit acquisition time (SNRt). Four different sine-bell like accumulation schemes and a reference scheme using a constant number of accumulations have been applied on five normal volunteers. Localized spectra obtained from the parieto-occipital white matter were compared with regard to SNRt and linewidth. A reduction in acquisition time of 45%, related to the reference scheme, and an increase in SNRt of 25-30% were achieved using a sine-bell accumulation scheme in combination with a quarter sine-wave apodization of the short echo time data. Down to an acquisition time of 122 s no significant line broadening has been observed in comparison with the reference scheme.

Brain↗

PubMed1999

Functional MRI of the human motor cortex using single-shot, multiple gradient-echo spiral imaging.

In this study, we combined the advantages of a fast multi-slice spiral imaging approach with a multiple gradient-echo sampling scheme at high magnetic field strength to improve quantification of BOLD and inflow effects and to estimate T2* relaxation times in functional brain imaging. Eight echoes are collected with echo time (TE) ranging from 5 to 180 ms. Acquisition time per slice and echo time is 25 ms for a nominal resolution of 4 x 4 x 4 mm3. Evaluation of parameter images during rest and stimulation yields no significant activation on the inflow sensitive spin-density images (rho or I0-maps) whereas clear activation patterns in primary human motor cortex (M1) and supplementary motor area (SMA) are detected on BOLD sensitive T2*-maps. The calculation of relaxation times and rates of the activated areas over all subjects yields an average T2* +/- standard deviation (SD) of 46.1+/-4.5 ms (R2* of 21.8+/-2.2 s(-1)) and an average increase (deltaT2* +/- SD) of 0.93+/-0.47 ms (deltaR2* of -0.4+/-0.14 s(-1)). Our findings demonstrate the usefulness of a multiple gradient echo data acquisition approach in separating various vascular contributions to brain activation in fMRI.

Adult↗

PubMed1999

Fast spectroscopic imaging for non-invasive thermometry using the Pr[MOE-DO3A] complex.

The praseodymium complex of 10-(2-methoxyethyl)-1,4,7,10-tetraaza-cyclododecane-1,4,7-tr iacetate) was evaluated as a temperature-sensitive contrast agent using the temperature dependence (approximately 0.12 ppm degrees C(-1)) of the chemical shift of its methoxy side group signal. Pr[MOE-DO3A] was employed in combination with spectroscopic imaging (SI) methods for the determination of spatially resolved 2D and 3D temperature distributions in phantoms. Conventional SI and fast echo planar SI sequences (EPSI) were implemented on a 4.7 T MR imaging system fulfilling the demands for non-invasive thermometry (NIT) with respect to thermal and temporal resolution, being <1 degree C and <20 s total measuring time, respectively. The sequences are based on a fast spin echo SI method taking into account the very short relaxation times of the Pr complex methoxy group (T1 = 28 ms, T2 = 13 ms) and its chemical shift difference (-24 ppm) from water. Calibration curves were measured in a uniformly heated water phantom and 2D SI methods were applied to dynamic heating experiments. The average differences between the temperatures measured via fibreoptic thermometer and those derived from the spectroscopic methods were < or =0.2 degrees C. Furthermore, 3D EPSI experiments with a 16 x 16 x 16 matrix size yielded temperature measurements within 17 s from voxels of size 3 x 3 x 3 mm3.

Calibration↗

PubMed1999

Evaluation of individual and combined neurotoxicity of the immunosuppressants cyclosporine and sirolimus by in vitro multinuclear NMR spectroscopy.

Neurotoxicity, a crucial side effect of immunosuppressive therapy with cyclosporine, also has been demonstrated in vitro for sirolimus, a novel macrolide immunosuppressant, which is under clinical investigation in combination with cyclosporine. NMR spectroscopy was used to study the separate and combined effects of cyclosporine and sirolimus on cerebral metabolism, both in brain cells and in perfused rat brain slices. The high-energy phosphate metabolism was already affected significantly at cyclosporine concentrations as low as 100 micrograms/liter: phosphocreatine was reduced by 10 +/- 2% [half-maximal inhibition concentration (IC50) = 1850 +/- 600 micrograms/liter], and nucleoside triphosphate was reduced by 11 +/- 5% (IC50 = 1110 +/- 420 micrograms/liter; n = 4, P <.05). At 500 micrograms/liter cyclosporine, N-acetylaspartate and glutamate were decreased by 13 +/- 7% (IC50 = 1100 +/- 330 micrograms/liter) and 22 +/- 9% (IC50 = 360 +/- 220 micrograms/liter; n = 4, P <.05), respectively. As evaluated using an algorithm based on Loewe isobolograms, combination of cyclosporine and sirolimus resulted in a synergetic reduction of high-energy phosphate metabolites. Addition of sirolimus to the perfusion medium increased brain slice concentrations of cyclosporine. It is concluded that cyclosporine significantly reduced high-energy phosphate metabolism in brain tissue at in vivo relevant concentrations. Combination with sirolimus resulted in synergism, which, in part, is explained by a greater distribution of cyclosporine into the brain tissue in the presence of sirolimus.

Animals↗

PubMed1998

Expression of aquaporins in Xenopus laevis oocytes and glial cells as detected by diffusion-weighted 1H NMR spectroscopy and photometric swelling assay.

Expression of aquaporins (AQP) and water permeability were studied in Xenopus laevis oocytes and immobilized glial cells by a pulsed-field gradient spin echo NMR technique and a photometric swelling assay. Oocytes injected with poly(A) RNA from C6-BU-1 cells showed increased swelling behavior under hypoosmotic stress due to expressed water channels as compared to control oocytes. The swelling could be reversibly inhibited by HgCl2. Furthermore, the intracellular relaxation time and the apparent intracellular diffusion coefficient of water in oocytes were determined by diffusion-weighted 1H NMR experiments to be T2=36 ms and Dapp, intra=0.18x10-3 mm2/s. In immobilized C6 and F98 cells the mean exchange time of intracellular water was found to be 51 ms which increased to 75 ms upon chronic treatment (4 days) in hypertonic medium. Additional hybrid depletion experiments with antisense oligonucleotides directed against AQP1 were performed on oocytes and C6 cells. Moreover, different water channel subtypes of glial cells were assessed by a reverse transcriptase polymerase chain reaction assay. With this, the mRNA encoding AQP1 could be detected in primary cultures and glial cell lines, whereas AQP4 mRNA was found in astroglia-rich primary cultures, but not in F98 and C6 cells. Our results show that water permeability in glial cells is mainly mediated by water channels which play an important role in the regulation of water flow in brain under normal and pathological conditions.

Animals↗

PubMed1998

Direct administration and utilization of [1-13C]glucose by fetal brain and liver tissues under normal and ischemic conditions: 1H, 31P, and 13C NMR studies.

Three distinct, maternal-independent routes (e.g. intraamniotic, intraperitoneal and intracerebral), for [1-13C]glucose utilization by fetal brain and liver tissues, were examined by multinuclear magnetic resonance (NMR) spectroscopy before and after vascular occlusion of the maternal-fetal blood flow. Labeled lactate was the major glycolytic product by all routes, but in addition labeled TCA cycle products were also generated. Fractional 13C enrichment in both glucose and lactate were always higher in the ischemic state compared to controls using either one of the three routes studied. After intraperitoneal injection total glucose in the fetal brain was decreased by 85% after 20 min reperfusion following 20 min ischemia, but was elevated up to 170% after 60 min. [1-13C]glucose increased continuously by up to 370% after 60 min. Total glucose in the fetal liver remained unchanged while [1-13C]glucose increased up to 380%. Total lactate level in brain was 50-80% above the control apart from a transient increase (140%) notable after 40 min reperfusion. The kinetics of [3-13C]lactate followed a similar time course. At the same time when lactate was transiently increased in fetal brain, total lactate as well as 13C-labeled lactate showed a transient decrease in liver after 40 min. While the ways of mobilization of energy substrates for maintaining adequate metabolic activity in the fetal brain remain still unclear, the present 13C NMR studies suggest that both liver glucose and lactate can contribute to brain metabolism particularly under ischemic stress.

Adenosine Triphosphate↗

PubMed1998

Monitoring of cell volume and water exchange time in perfused cells by diffusion-weighted 1H NMR spectroscopy.

Diffusion of intracellular water was measured in perfused cells embedded in basement membrane gel threads. F98 glioma cells, primary astrocytes, and epithelial KB cells were used and were exposed to osmotic stress, immunosuppressiva, the water channel blocker p-chloromercuriobenzenesulfonate (pCMBS), and apoptotic conditions. With diffusion-weighted 1H NMR spectroscopy changes in the intracellular signal could be monitored and quantified with single signal (ss), constant diffusion time (ct), and constant gradient strength (cg) experiments. The temporal resolution of the ss monitoring was 3.5 s with a standard deviation of 0.5% of the signal intensity and 32 s (3%) with ct monitoring, respectively. A mean intracellular residence time of water was determined with the cg experiment to about 50 ms. Changes of this exchange time from (51.9 +/- 1.0) to (59.0 +/- 1.1) ms were observed during treatment with pCMBS. The changes in the diffusion attenuated signal could be simulated analytically varying the intracellular volume fraction and exchange time by combination of restricted diffusion (Tanner model) and water exchange (Kärger model). This sensitive and noninvasive NMR method on perfused cells allows to determine changes in the intracellular volume and residence time in a simple and accurate manner. Many further applications as anoxia, volume regulation, ischemia and treatment with various pharmaceuticals are conceiveable to follow up their effect on the cell volume and the exchange time of intracellular water.

Animals↗

PubMed1998

Restricted diffusion and exchange of intracellular water: theoretical modelling and diffusion time dependence of 1H NMR measurements on perfused glial cells.

Intracellular diffusion properties of water in F98 glioma cells immobilized in basement membrane gel threads, are investigated with a pulsed-field-gradient spin-echo NMR technique at diffusion times from 6 to 2000 ms and at different temperatures. In extended model calculations the concept of 'restricted intracellular diffusion at permeable boundaries' is described by a combined Tanner-Kärger formula. Signal components in a series of ct experiments (constant diffusion time) are separated due to different diffusion properties (Gaussian and restricted diffusion), and physiological as well as morphological cell parameters are extracted from the experimental data. The intracellular apparent diffusion coefficients strongly depend on the diffusion time and are up to two orders of magnitude smaller than the self diffusion constant of water. Propagation lengths are found to be in the range of 4-7 microns. Hereby intracellular signals of compartments with a characteristic diameter could be selected by an appropriate gradient strength. With cg experiments (constant gradient) a mean intracellular residence time for water is determined to be about 50 ms, and the intrinsic intracellular diffusion constant is estimated to 1 x 10(-3)mm2/s. Studying the water diffusion in glial cells provides basic understanding of the intracellular situation in brain tissue and may elucidate possible influences on the changes in the diffusion contrast during ischemic conditions.

Animals↗

PubMed1998

A 1H/13C inverse 2D method for the analysis of the polyamines putrescine, spermidine and spermine in cell extracts and biofluids.

The polyamines putrescine, spermidine and spermine are involved in the regulation of various metabolic processes. It is therefore desirable to detect and quantify the polyamines with NMR. We present the proton and carbon assignments for all polyamine signals obtained from PCA extracts of F98 glioma cells with high resolution using a semi-selective HSQC 2D-experiment. The biosynthesis of the polyamines in cell culture was examined using the labeled substrates [U-13C]glucose and [U-13C]glutamate. In such studies the high resolution of the semi-selective HSQC experiment at very high magnetic fields (14-19 T) allows the analysis of carbon-carbon couplings, and isotopomer patterns. The different effects of osmotic stress on the concentrations of polyamines and amino acids are also reported.

Animals↗

PubMed1998

Magnetization transfer attenuates metabolite signals in tumorous and contralateral animal brain: in vivo observations by proton NMR spectroscopy.

Tumorous and contralateral rat brain was examined by in vivo single voxel proton NMR spectroscopy. Magnetization transfer (MT) experiments cause attenuation of various metabolite signals. Selective saturation of immobile metabolites was achieved by pulsed RF preirradiation. The method is compared with continuous wave MT generation. In contralateral tissue, MT attenuation is detected for both the CH3 and the CH2 protons of (phospho-)creatine (Cr + PCr) and for a signal at 3.44 ppm ascribed to taurine. Significant attenuation is also observed for a signal at 3.78 ppm that is commonly ascribed to the alphaCH proton of glutamate and glutamine (Glx); however, no effect is observed for the gammaCH2 protons of Glx. Within implanted F98 glioma tumors, only the CH3 signal of Cr + PCr shows significant MT attenuation. Although the MT effect detected for lactate in the tumors fails to reach significance, a significant effect is observed for the lactate signal acquired during 3 to 9 min postmortem.

Animals↗

PubMed1998

Temporal and regional changes during focal ischemia in rat brain studied by proton spectroscopic imaging and quantitative diffusion NMR imaging.

The early development of focal ischemia after permanent occlusion of the right middle cerebral artery (MCA) was studied in six rats using interleaved measurements by diffusion-weighted NMR imaging (DWI) of water and two variants of proton spectroscopic imaging (SI), multiecho SI (TE: 136, 272, 408 ms) and short TE SI (TE: 20 ms). Measurements on a 4.7-T NMR imaging system were performed between the control phase and approximately 6 h postocclusion. In the center of the ischemic lesion of all rats, the apparent diffusion coefficient (ADC) decreased rapidly to 84.4 +/- 4.2% (mean +/- SD) of the control values approximately 2 min postocclusion. Approximately 6 h postocclusion, the ADC was reduced to 67.1 +/- 5.9%. In contrast, large differences between the animals were observed for the temporal increase of lactate (Lac) in the ipsilateral hemisphere. The maximum Lac signal was reached in four rats after 0.5-1.5 h, and in two rats was not reached even after 6 h postocclusion. Six h postocclusion, SI spectra measured at a TE of 136 ms revealed a decrease in the CH3 signal of N-acetylaspartate (NAA) to 67 +/- 13% of the control values. Differences were observed between the spatial regions of decreased NAA and increased Lac. In the lesions, a T2 relaxation time of Lac of 292 +/- 40 ms, considering a J-coupling constant of 6.9 Hz, was measured. Furthermore, a prolongation of the T2 of the CH3 signal of creatine/phosphocreatine (Cr/PCr) was observed in the lesion, from 163 +/- 22 ms during control to 211 +/- 41 ms approximately 6 h postocclusion. The experiments proved that DWI and proton SI are valuable tools to provide complementary information on processes associated with brain infarcts.

Animals↗

PubMed1998

Fast 1H spectroscopic imaging using a multi-element head-coil array.

Fast proton magnetic resonance spectroscopic imaging (MRSI) using a multi-element head-coil array is examined with respect to three aspects: the coil design, the use of an appropriate signal combination method, and the design of the MRSI pulse sequence itself. An eight-element head-coil array has been developed to increase the signal-to-noise ratio (SNR) of MRSI in the human brain. The flexible wraparound design optimally fits different head sizes and thus provides high sensitivity. The signal combination of the individual coil elements is based on the approach proposed by Roemer et al. (Magn. Reson. Med. 16, 192 (1990)). An additional short prescan is performed to provide a good estimate of the complex coil sensitivity profiles, which are used in the signal combination procedure to correct the spectroscopic imaging data for the spatially varying intensity. The use of coil arrays in MRSI has some effect on the requirements for both water and lipid suppression. These techniques and a MRSI pulse sequence that provides a high spectroscopic resolution are described and discussed. Experimental results at 1.5 T show that metabolite maps of N-acetylaspartate (NAA), choline (Cho), phosphocreatine (PCr)/creatine (Cr) can be obtained within a 5-min acquisition time.

Aspartic Acid↗

PubMed1998

Water signal attenuation in diffusion-weighted 1H NMR experiments during cerebral ischemia: influence of intracellular restrictions, extracellular tortuosity, and exchange.

The "concept of restricted intracellular water diffusion at permeable boundaries", which was recently used to model diffusion-weighted 1H NMR experiments on glioma cells, was applied to measurements on the rat brain in vivo. Combined with the "concept of extracellular tortuosity", various physiological states of the brain were simulated. Hereby, a variable intracellular volume fraction, intracellular exchange time, and extracellular tortuosity factor were considered for young, adult, and ischemic rat brains. The model simulated the cytotoxic shift of extracellular water, changes in membrane permeability and tissue morphology, and was able to explain the diffusion time dependence as well as the non-monoexponentiality of the diffusion attenuation curves. Preliminary diffusion time dependent experiments on the healthy rat brain (1H NMR imaging) agreed well with the theoretical concept. Hereby, the intracellular water signal was separated from extracellular signal contributions by large diffusion weighting. It showed the characteristic of restricted diffusion as well as a signal decay due to the exchange of intracellular water across the plasma membrane. A map of the mean intracellular exchange time for water in living animal brain was determined, and the upper limit in rat brain was evaluated to 15 ms. The presented methods can be applied to correlate local differences in a map of exchange times with tissue morphology and to detect pathological deviations of the exchange time, e.g., during ischemia.

Animals↗

PubMed1998

Absolute metabolite quantification by in vivo NMR spectroscopy: I. Introduction, objectives and activities of a concerted action in biomedical research.

By utilizing achievements and results of two previous concerted research projects on magnetic resonance imaging and spectroscopy (MRS), the EU BIOMED 1 Concerted Action on "Cancer and brain disease characterization and therapy assessment by quantitative MRS" was specifically aimed at: 1) developing at a multicentre level harmonized methodologies and protocols for quantitative and reproducible MRS measurements, as a basis for validating these procedures in well controlled clinical and experimental conditions; and 2) providing multicentre critical reviews on the present understanding of the significance of MRS parameters as possible new markers of diagnosis, prognosis and response to therapy. The programme comprised the following main areas of collaborative research and multicentre evaluation: a) development of methods and protocols for quality assessment, calibration and absolute metabolite quantification in in vivo localized, volume-selective MRS; b) design and validation of a new method for assessing localization performance in spectroscopic imaging (MRSI); c) interlaboratory comparison of different methods of signal processing and data analysis, for improving signal quantification in vivo and in vitro MRS spectra; d) quality assessment of high resolution MRS analyses of biological fluids; e) protocol for assembling a pilot data base of MR spectra of tumour extracts for pattern recognition analysis; f) multicentre review on evaluation of the significance of MRS parameters in monitoring lipid metabolism and function in cancer; and g) multicentre review on evaluation of drug pharmacokinetics and metabolism using MRS. The main results and conclusions of four multi-centre trials on items (a), (b) and (c), which involved 24 teams, are reported in the accompanying papers of this series.

Brain↗

PubMed1998

Rat brain primary neurons immobilized in basement membrane gel threads: an improved method for on-line 13C NMR spectroscopy of live cells.

In vivo MRS studies on intact brain reflect the metabolism of all cells present, but do not distinguish between different cell types. NMR studies of immobilized cultured primary cells, such as neurons and astrocytes, are a useful model to monitor the specific differences in metabolism of the various cell types in the brain. The present study shows that primary rat neuronal cells can be cultured in basement membrane gel threads. After 4 days of incubation the threads are filled with viable cells, and represent a population of morphologically differentiated neuronal cells with less than 5% of non-neuronal cells, i.e., astrocytes. These threads were placed into a NMR tube and used for on-line monitoring of neuronal metabolism. Under these conditions cells remained viable and metabolically active for several days. The energy status was monitored by using 31P NMR spectroscopy. To study neuronal glucose metabolism [1-13C]glucose was added to the perfusion medium and 30 min later 13C-labeled metabolites were detectable by 13C NMR spectroscopy. Immobilized neurons synthesized glycolytic products such as [3-13C]lactate and [3-13C]alanine, as well as several tricarboxylic acid cycle products, i.e., [2-13C]glutamate, [3-13C]glutamate, [4-13C]glutamate, [2-13C]aspartate, and [3-13C]aspartate. In summary, 31P and 13C NMR spectra can be recorded from live neuronal cells for up to 24 h using the newly designed procedure described in the present communication.

Animals↗

PubMed1998

Alterations in glial cell metabolism during recovery from chronic osmotic stress.

NMR spectroscopy of F98 glioma cell extracts showed that chronic hypertonic conditions largely increased the intracellular content of small, osmotically active molecules. Moreover, hypertonic stress decreased the incorporation of 13C-labeled amino acids into the cellular proteins albeit their cytosolic concentrations were increased, which reflects an inhibition of protein synthesis under these conditions. Reincubation with isotonic medium restored almost completely the control values for the cytosolic metabolites but not for amino acid incorporation into the protein. An increased amount of 13C label was found in the phospholipids, which indicates stimulation of membrane synthesis processes due to the recovery-induced cell swelling. On the other hand, chronic hypotonic conditions largely decreased the steady state concentration and synthesis of small, cytosolic molecules, whereas the effect on the incorporation of 13C-labeled amino acids into the cellular proteins was variable. Reincubation with isotonic medium partially restored the depressed cytosolic metabolite content and also the incorporation of labeled amino acids into cellular protein, but induced an inhibition of phospholipid synthesis. The results verify that 'readaptation' of glial cell metabolism during recovery from chronic osmotic stress is impaired or at least seriously retarded.

Adaptation, Physiological↗