Valproic acid intoxication identified by1H and1H-13C correlated NMR spectroscopy of urine samples

Analysis of biological fluids by proton and carbon nuclear magnetic resonance spectroscopy (¹H and¹³C NMR) is a promising tool in clinical biology. We used this method for rapid toxicological screening in the case of two suicide attempts. For each case, a urine sample was analysed at 300 MHz by 1D and 2D sequences (TOCSY and HMBC) in a short experimental time. Quantification was performed by peak integration on the 1D¹H NMR spectrum. For the two patients, results showed the same resonances of the major metabolite, valproyl-O-glucuronide at concentrations of 121 and 44 mmol/l.     

Validation of glutathione quantitation from STEAM spectra against edited 1H NMR spectroscopy at 4T: application to schizophrenia

Objective: Quantitation of glutathione (GSH) in the human brain in vivo using short echo time ¹ H NMR spectroscopy is challenging because GSH resonances are not easily resolved. The main objective of this study was to validate such quantitation in a clinically relevant population using the resolved GSH resonances provided by edited spectroscopy. A secondary objective was to compare several of the neurochemical concentrations quantified along with GSH using LCModel analysis of short echo time spectra in schizophrenia versus control. Materials and Methods: GSH was quantified at 4T from short echo STEAM spectra and MEGA-PRESS edited spectra from identical volumes of interest (anterior cingulate) in ten volunteers. Neurochemical profiles were quantified in nine controls and 13 medicated schizophrenic patients. Results: GSH concentrations as quantified using STEAM, 1.6 ± 0.4 μmol/g (mean ± SD, n=10), were within error of those quantified using edited spectra, 1.4 ± 0.4 μmol/g, and were not different (p=0.4). None of the neurochemical measurements reached sufficient statistical power to detect differences smaller than 10% in schizophrenia versus control. As such, no differences were observed. Conclusions: Human brain GSH concentrations can be quantified in a clinical setting using short-echo time STEAM spectra at 4T.     

Accurate work-rate measurements during in vivo MRS studies of exercising human quadriceps

INTRODUCTION: Given that we have reached a point in the field of muscle energetics where absolute measurements are warranted to take the area forward, we designed an ergometer, including two force and two displacement transducers, allowing dynamic and isometric knee extension within a MR system and accurate measurements of power output. METHODS: On the basis of repeated measurements, the force and displacement transducers accuracy was 1% for values ranging from 0 to 394 N and 4% for values ranging from 0 to 20 cm. In addition, measurements were not affected by magnetic field. MRS experiments in exercising muscle were conducted in eight subjects. They performed two standardized dynamic alternate leg extension exercises (25 and 35% of MVC) while the corresponding metabolic changes were measured using (31)P-MRS. RESULTS: The mean power output produced during both exercises were 63 +/- 16 and 81 +/- 15 W while the eccentric work was reduced i.e. 12 +/- 14 and 21 +/- 6 W for the moderate and heavy exercise respectively. The corresponding metabolic changes were significant with a 20-40% PCr depletion and an end of exercise pH ranging from 0.02 to 0.70 pH units. CONCLUSION: Overall, the present ergometer allows quadriceps exercise in a MR system and should be useful for future metabolic studies for which reliable and absolute quantification of power output is warranted.     

In vivo quantification of response to treatment in patients with multiple myeloma by <Superscript>1</Superscript>H magnetic resonance spectroscopy of bone marrow

Object Magnetic resonance imaging (MRI) is the gold standard non-invasive technique to detect malignant disease in the bone marrow. Proton magnetic resonance spectroscopy (MRS) can be performed as a quick adjunct to routine spinal MRI. We performed proton MRS to patients with multiple myeloma (MM) at diagnosis and after treatment to investigate the possible correlation of MRS data with response to therapy. Patients and methods Twenty-one patients with newly diagnosed MM underwent combined MRI/MRS explorations of a transverse center section in the fifth lumbar vertebral body. MRS was acquired with STEAM and 40 ms TE. Areas of unsuppressed water and lipid resonances were used to calculate the lipid-to-water ratio (LWR). Results No association was detected between initial LWRs and the clinical characteristics of patients. Post treatment MRS was available in 16 patients of whom 11 (69%) presented an LWR increase, this included all complete responders (8/8, 100%, P = 0.012). A post-treatment LWR value equal to or larger than one is proposed as a non-invasive marker of complete response to treatment. Conclusion Only patients responding to treatment presented a significant increase in bone marrow LWR after therapy. MRS may provide an adequate quantification of response to chemotherapy in patients with MM.     

Metabolic changes in the rat brain after a photochemical lesion treated by stem cell transplantation assessed by 1H MRS

Object  Metabolite changes in an experimental lesion in the rat cortex and in the contralateral hemisphere after the intravenous administration of mesenchymal stem cells (MSCs) were assessed by proton MR spectroscopy to verify the impact of the cell treatment on the brain tissue. Materials and methods  Wistar rats with a photochemical cortical lesion and transplanted MSCs or sham transplanted rats were examined. Proton spectra were obtained from the lesion and from the contralateral cortex. Results  Magnetic resonance spectroscopy revealed a gradual recovery of the damaged tissue; however, we found no significant differences in metabolite concentrations in the lesioned hemisphere between treated and untreated animals. Higher concentrations of glutamate and N-acetyl aspartate were found in the contralateral hemisphere in cell-treated animals compared to untreated ones. Lesioned animals showed neurogenesis in the contralateral hemisphere; the number of newly generated cells in stem cell-treated animals was 50% higher than those observed in untreated animals. Conclusion  No direct impact of cell transplantation was observed in the lesion. However, changes in the contralateral hemisphere suggest that the transplanted MSCs might stimulate repair processes and plasticity resulting in the generation of newborn cells, which might enable the faster adoption of the damaged tissue’s function by healthy tissue.     

Glycogen resynthesis in liver and muscle after exercise: measurement of the rate of resynthesis by13C magnetic resonance spectroscopy

We have used natural abundance¹³C magnetic resonance spectroscopy (MRS) to measure glycogen content of muscle and liver before and after heavy exercise, and after consumption of different carbohydrate-based drinks. After an overnight fast, five healthy men (mean±SEM age 23±1 years) exercised to exhaustion at 75% of VO_2max on two occasions (mean work rate 165±8 W for 78±14 min) and then, in a single blind random order, consumed either of two drinks containing the same carbohydrate load (177 g). Spectra were recorded over Vastus Lateralis muscle and the liver before and after exercise, and hourly for 5 h after the carbohydrate load. In muscle, glycogen content after exercise was 37% and 31% of basal (preexercise) concentration before consuming the drinks. After carbohydrate loading, glycogen concentration had increased significantly (p<0.05) to 70% and 64% of basal concentration respectively after 5 h. Hepatic glycogen concentration did not change significantly throughout. The study demonstrates the feasibility of sequential MRS measurement of muscle and liver glycogen before and after exercise and after carbohydrate loading.     

Multi-spin-echo J-resolved spectroscopic imaging without water suppression: application to a rat glioma at 7 T

Two-dimensional J-resolved spectroscopy may be used to separate resonances which overlap in 1D NMR spectra. Coupled with spectroscopic imaging (SI), it would give unequivocal information on the distribution of such resonances. Multi-echo acquisition decreases the minimum experimental time of such 4D experiments. The water peak may be used for phase and chemical-shift reference. This study aimed to demonstrate the feasibility of J-resolved SI based on a multi-echo sequence and without water suppression, and its ability to separate the peaks for lactate and mobile lipid in a rat glioma. Experiments were performed on rat brain, without water suppression, at 7 T. The water signal was used for correcting the phase of the echoes. A FOCSY-like acquisition was used to collect the first part of the echoes at short echo times. Two different data processing methods were tested to overcome the problem of contaminations of metabolite signals by the intense water signal. Maps of N-acetylaspartate, choline, creatine, lactate and mobile lipids were obtained in vivo on a rat glioma in 70 min. The in-plane resolution was 2 mm². The 2D spatially resolved, 2D J-resolved spectra enabled the separate mapping of lactate and mobile lipids.     

Proton magnetic resonance spectroscopy of the central,transition and peripheral zones of the prostate: assignments and correlation with histopathology

Proton magnetic resonance spectroscopy (MRS) is used to compare the chemistry of the transition, central and peripheral zones of the prostate. The assignments are made using two-dimensional correlated spectroscopy and the results compared with histopathology. The chemistry associated with benign prostatic hyperplasia (BPH), prostate intraepithelial neoplasia (PIN) and malignant biopsy tissues are described. There are distinct MR spectral patterns for glandular and stromal BPH, PIN and adenocarcinoma. Importantly, there are also different spectral patterns from BPH in the transitional and central zones when compared to BPH in the peripheral zone. A pattern recognition method is used to analyze the MR spectra from the biopsy specimens. The resultant mathematical classifiers generated a high level of accuracy (sensitivity and specificity of 100 and 97%). It was found that for this accuracy to be achieved, the classifiers need to be developed by comparing the spectra with specialist serial sectioned histopathology. With serial sectioned pathology the pattern recognition method was capable of identifying less than 5% of adenocarcinoma in a given piece of tissue. Many of the chemicals identified in the biopsy specimens are available for inspection from the prostate, in vivo, at 3 T.     

ESMRMB 2009 Congress,Antalya, Turkey, 1–3 October: Abstracts,Thursday

Object  To determine whether glycine can be measured at 7 T in human brain with ¹H magnetic resonance spectroscopy (MRS). Materials and methods  The glycine singlet is overlapped by the larger signal of myo-inositol. Density matrix simulations were performed to determine the TE at which the myo-inositol signal was reduced the most, following a single spin-echo excitation. ¹H MRS was performed on an actively shielded 7 T scanner, in five healthy volunteers. Results  At the TE of 30 ms, the myo-inositol signal intensity was substantially reduced. Quantification using LCModel yielded a glycine-to-creatine ratio of 0.14 ± 0.01, with a Cramér–Rao lower bound (CRLB) of 7 ± 1%. Furthermore, quantification of metabolites other than glycine was possible as well, with a CRLB mostly below 10%. Conclusion  It is possible to detect glycine at 7 T in human brain, at the short TE of 30 ms with a single spin-echo excitation scheme.     

Monitoring the development of hepatocellular carcinoma in woodchucks using 31P-MRS

The woodchuck is one of the only lab animal models of chronic viral hepatitis infection and the development of hepatocellular carcinoma. Using this model, changes in tissue energetics in the liver due to the development of hepatocellular carcinoma can be monitored by repeated magnetic resonance imaging and localized phosphorus spectroscopy. Age- and sex-matched control (n=5) and chronically infected (n=5) adult woodchucks were imaged four times in a six-month period in a 7-T horizontal-bore magnet. Using a custom-built doubly tunable quadrature volume coil, sagittal and axial FLASH images (128×128, slice thickness = 5 mm, TR/TE=1000/4.1, 8 averages) were acquired to locate the largest portion of the liver with the least amount of signal contamination from surrounding abdominal muscle. Two-dimensional 31P chemical-shift imaging (2D-CSI) was acquired (16×16 data matrix, 24×24×2 cm³, 1024 data points, 16 averages) for all animals. The extent of liver injury was determined using serum gamma glutamyltransferase (GGT). The livers of infected woodchucks showed a significant increase (p=0.01) in phosphomonoesters (PME):β-adenosine triphosphate (NTP). Chronically infected woodchucks had higher levels of serum GGT compared to uninfected woodchucks (p=0.002). An increase in the PME:β-NTP ratio indicates cellular proliferation within the malignant tumor.     

Proton MR spectroscopy of prostatic tissue focused on the etection of spermine, a possible biomarker of malignant behavior in prostate cancer

To investigate whether polyamines may be valuable diagnostic and prognostic markers in prostate cancer, the presence of alyamines was studied in various human prostatic tissues using both proton magnetic resonance (MR) spectroscopy and righ-pressure liquid chromatography (HPLC). The HPLC results showed that normal and benign hyperplastic prostatic tissues re characterized by a high content of spermine. Spermine levels were reduced in tumor tissue, especially in prostatic carcinoma h metastases, and in xenografts of human prostatic carcinoma cells. These preliminary results indicate that spermine may be d as a biomarker for malignant behavior. The MR spectroscopy study showed that it is possible to detect spermine resonances prostatic biopsy material by one-dimensional and two-dimensional J-resolved MR spectroscopy at high field (600 MHz). ocalized one-dimensional in vitro MR spectra obtained at the clinical field strength of 1.5 T showed spermine signals in the region between 3.0 and 3.3 ppm. In in vivo MR spectra of the human prostate, however, these signals were obscured by esonances of choline (3.2 ppm) and creatine (3.0 ppm).     

Fast three-dimensional sodium imaging of human brain

A three-dimensional sodium imaging technique with a minimum echo time of 0.9 ms is described in a 2.0 Tesla whole-body system. The relaxation behaviour in vivo of sodium was analysed: a lastT ₂ ^* relaxation component between 1.2 and 1.6 ms and a slowT ₂ ^* relaxation component between 7.1 ms and 8.4 ms were quantified in brain tissue of three volunteers. Three-dimensional sodium images of the human brain were acquired in 8.5 min with a resolution of 4.7 × 4.7 × 10 mm (0.2 cc voxel size) and a signal-to-noise ratio of 20 in brain tissue and 30 in cerebrospinal fluid.     

Preliminary characterization of an experimental breast cancer cells brain metastasis mouse model by MRI/MRS

PURPOSE: Chemotherapy increases survival in breast cancer patients. Consequently, cerebral metastases have recently become a significant clinical problem, with an incidence of 30-40% among breast carcinoma patients. As this phenomenon cannot be studied longitudinally in humans, models which mimic brain metastasis are needed to investigate its pathogenesis. Such models may later be used in experimental therapeutic approaches. MATERIAL AND METHODS/RESULTS: We report a model in which 69% of the animals (9/13 BALB/c nude mice) developed MR-detectable abnormal masses in the brain parenchyma within a 20 to 62-day time window post intra-carotid injection of 435-Br1 human cells. The masses detected in vivo were either single (7 animals) or multiple (2 animals). Longitudinal MR (MRI/MRS) studies and post-mortem histological data were correlated, revealing a total incidence of experimental brain metastases of 85% in the cases studied (11/13 animals). ADC maps perfectly differentiated edema and/or CSF areas from metastasis. Preliminary MRS data also revealed additional features: decrease in N-acetyl aspartate (NAA) was the first MRS-based marker of metastasis growth in the brain (micrometastasis); choline-containing compounds (Cho) rose and creatine (Cr) levels decreased as these lesions evolved, with mobile lipids and lactate also becoming visible. Furthermore, MRS pattern recognition-based analysis suggested that this approach may help to discriminate different growth stages. CONCLUSIONS: This study paves the way for further in vivo studies oriented towards detection of different tumor progression states and for improving treatment efficiency.     

Regional metabolite levels of the normal posterior fossa studied by proton chemical shift imaging

MR spectroscopy of the posterior fossa is pitted with numerous technical difficulties. It is, however, of great clinical interest in the study of the degenerative diseases and tumors of this area. We have developed a method to perform 2D CSI of this area, by using a sagittal slice and a careful positioning of outer volume saturation. We performed this acquisition in 30 healthy volunteers to determine the normal metabolic ratios in five voxels of this area (mesencephalon. pons. medulla oblongata, vermis, cerebellar white matter). The main technical difficulty was magnetic field inhomogeneity in the lower brainstem generated by dental alloys. However, 88% of the voxels were of sufficient quality to be analyzed. The statistically significant regional variations were a higher NAA/Cr ratio in the pons than in the medulla oblongata, higher Cho/Cr in the pons than in the mesencephalon and higher Cho/ Cr in the cerebellar white matter than in the vermis. We conclude that 2D CSI of the brainstem, although technically delicate can be performed in most patients.     

Mri and mrs of human tendon xanthoma at 1.5 t: Anin vivo study

We studied a xanthomatous Achilles tendon and a normal Achilles tendon with the proton magnetic resonance imaging (MRI) and spectroscopy (MRS) at 1.5 T in a standard head coil. TheT ₂ maps and the localized proton spectra of the Achilles tendon were reconstructed. The normal tendon revealed no MR signal, whereas the xanthomatous tendon image consisted of variable signal intensities, for which the value ofT ₂ was significantly shorter (p=0.0002) than that of adipose tissue. The proton spectrum of this tendon xanthoma showed an increased water peak and unsaturated olefinic group intensity compared with the spectrum of the normal Achilles tendon area. The complex cholesterol molecule itself cannot be proven directly in a xanthomatous tendon, but its presence can be revealed with the help of the increased methyl and methylene groups of the fatty acids of the cholesteryl esters. This and other typical features describedin vitro for atheromatous tissue can be detectedin vivo in xanthomas.Additional reprints of this chapter may be obtained from the Reprints Department, Chapman & Hall, One Penn Plaza, New York, NY 10119.     

Role of magnetic resonance methods in the evaluation of prostate cancer: an Indian perspective

The challenges in detection, localization, and staging of prostate cancer have prompted the investigation of the role of various magnetic resonance (MR) methodologies in a large cohort of men prior to biopsy. The identification of suspicious areas of malignancy was carried out using magnetic resonance imaging (MRI), magnetic resonance spectroscopic imaging (MRSI) and diffusion-weighted imaging (DWI). Our data shows that apparent diffusion coefficient (ADC) may be a reliable marker to differentiate normal, benign, and malignant prostate tissues similar to the metabolite ratio. Also, the combined use of MRSI and DWI improves the diagnosis of prostate cancer. In this review, we present our experience on the use of MRI, MRSI and DWI methods in the assessment of prostate cancer in Indian men. Further, analysis of the comparison of the ADC and the metabolite ratio values reported in the literature across various patient populations are presented. An erratum to this article can be found at     

Muscle glycogen recovery after exercise measured by13C-magnetic resonance spectroscopy in humans: effect of nutritional solutions

The rate of glycogen resynthesis in human skeletal muscle after glycogen-depleting exercise is known to depend on carbohydrate intake and is reported to reach a platean after an adequate amount of carbohydrate (CHO) consumption. Efforts to maximize the rate of glycogen storage by changing the type and form of CHO, as well as by adding proteins or lipids have yielded inconsistent results. The objective of this study was to assess whether isocaloric addition of proteins and arginine to a CHO diet in the first 4 h after an endurance exercise would increase the rate of glycogen synthesis. The CHO solution, given twice at a 2 h interval according to earlier optimized protocols, contained 1.7 g CHO kg^{body weight}. The effects of this solution were compared to those of an isocaloric solution containing 1.2 g CHO/kg^{body weight} plus 0.5 g protein/kg^{body weight} (including 5 g arginine). Glycogen was measured in quadriceps muscle in vivo with natural abundance¹³C-magnetic resonance spectroscopy before exercise and twice after exercise, before and at the end of a 4-h period following the intake of one of the solutions. Eight subjects took part in a randomized cross-over trial separated by at least 1 week. Glycogen synthesis was found to be significantly increased with both regimes compared to a zero-caloric placebo diet, but no significant difference in glycogen resynthesis was found between the CHO-only diet and the one supplemented by proteins and arginine. It is estimated that significance would have been reached for an increase of 34%, while the effectively measured synthesis rates only differed by 5%.     

Cerebral blood flow response to a hypoxic-ischemic insult differs in neonatal and juvenile rats

To compare the cerebral blood flow (CBF) response to a transient episode of hypoxia–ischemia producing damage in neonatal and juvenile rats. One- and four-week-old rats were subjected to unilateral carotid artery occlusion plus hypoxia (8% oxygen). Perfusion MR images were acquired either in sham controls or in hypoxic–ischemic rats before, during, 1 h and 24 h after hypoxia–ischemia. At 24 h post hypoxia–ischemia, T₂ maps and histology were used to assess damage. In sham controls, CBF increased twofold between the age of one and four weeks. Reductions in CBF ipsilateral to the occlusion occurred during hypoxia–ischemia followed by a substantial recovery at 1 h post in both age groups. However, contralaterally, hyperemia occurred during hypoxia–ischemia in four-week but not one-week-old rats. Similarly, hyperemia occurred ipsilaterally at 24 h post hypoxia–ischemia in four-week but not one-week-olds, corresponding to the distribution of elevations in T_2. Despite CBF differences, extensive cell death occurred ipsilaterally in both age groups. The CBF responses to hypoxia–ischemia and reperfusion differ depending on postnatal age, with hyperemia occurring in juvenile but not neonatal rats. The results suggest a greater CBF responsiveness and differential relationship between post-ischemic vascular perfusion and tissue injury in older compared with immature animals.     

Time-domain quantitation of <Superscript>1</Superscript>H short echo-time signals: background accommodation

Quantitation of ¹H short echo-time signals is often hampered by a background signal originating mainly from macromolecules and lipids. While the model function of the metabolite signal is known, that of the macromolecules is only partially known. We present time-domain semi-parametric estimation approaches based on the QUEST quantitation algorithm (QUantitation based on QUantum ESTimation) and encompassing Cramér–Rao bounds that handle the influence of nuisance parameters related to the background. Three novel methods for background accommodation are presented. They are based on the fast decay of the background signal in the time domain. After automatic estimation, the background signal can be automatically (1) subtracted from the raw data, (2) included in the basis set as multiple components, or (3) included in the basis set as a single entity. The performances of these methods combined with QUEST are evaluated through extensive Monte Carlo studies. They are compared in terms of bias–variance trade-off. Because error bars on the amplitudes are of paramount importance for diagnostic reliability, Cramér–Rao bounds accounting for the uncertainty caused by the background are proposed. Quantitation with QUEST of in vivo short echo-time ¹H human brain with estimation of the background is demonstrated.