MR imaging, proton MR spectroscopy, ultrasonographic, histologic findings in patients with chronic lymphedema.

Lymphedema is a progressive disease with multiple alterations occurring in the dermis. We undertook this study using high-frequency ultrasonography (US), magnetic resonance imaging, proton MR spectroscopy and histology to examine structural changes occurring in the subcutaneous tissue and precisely describe the nature of intralobular changes in chronic lymphedema. Four cutaneous and subcutaneous tissue biopsies from patients with chronic lymphedema during lymphonodal transplantation were studied. We performed US with a 13.5 MHz transducer, TSE T1 and TSE T2 magnetic resonance images with and without fat-suppression, MR Chemical Shift Imaging Spectroscopy and histological evaluation on these biopsies. We found that normal subcutaneous septa are seen as hyperechogenic lines in US and hyposignal lines in MRI and that hyperechogenic subcutis in US can be due to interlobular and intralobular water accumulation and/or to interlobular and intralobular fibrosis. Our study also confirms the usefulness of MR spectroscopy to assess water or fat content of soft tissue. Thus, multiple imaging modalities may be necessary to precisely delineate the nature of tissue alterations in chronic lymphedema.

Removal of local field gradient artefacts in BOLD contrast imaging of head and neck tumours.

Monitoring of oxygenation in tumours is an important issue in predicting the success of anti-cancer treatments such as radiotherapy. Gradient echo (GE) imaging sequences can be used for monitoring changes in tumour blood flow and oxygenation. However, the application of this method in head and neck tumours is hampered by significant artefacts and losses of the MR signal near air-tissue interfaces. We investigated the usefulness of a gradient-echo slice excitation profile (GESEPI) sequence that should keep the oxygen contrast while recovering the signal loss caused by susceptibility artefacts. A tumour model was implanted in the neck and in the leg of mice. MR imaging was performed at 4.7 T. GE and GESEPI sequences were used for monitoring the blood oxygen level dependent (BOLD) contrast after carbogen breathing. The pO2 was also monitored in tumours using an OxyLite probe (Oxford Optronics). Using the tumours implanted in the leg, we found that the variations of signal intensity after carbogen breathing were similarin both sequences. In the tumour implanted in the neck, it was possible, using GESEPI sequences, to recover the signal loss caused by susceptibility artefacts and to monitor the effect of carbogen-induced changes in the tumour.

Non invasive quantification of manganese deposits in the rat brain by local measurement of NMR proton T1 relaxation times.

Up to now, there is no reliable non invasive biomarker for the concentration of manganese (Mn) in the brain after intoxication to this metal. The aim of the present experimental study was to determine the predictive value of the localized measurement of the proton NMR relaxation time T1 as a quantitative estimation of the concentration of Mn in brain. The relationship of the proton relaxation rates (1/T1) was established in rat brain homogenates as a function of the Mn, iron, and copper concentration. Subsequently, an experimental model of Mn neurotoxicity was used: rats were stereotactically injected with increasing amounts of Mn2+ (as MnCl2) in the ventricles. After 3 weeks, local measurements of T1 were carried out in live rats. They were then sacrificed in order to sample the striatum, the cortex, and the cerebellum from the brain and to perform a quantitative determination of the concentration of Mn in these tissues by atomic absorption spectrometry (AAS). The results indicate excellent correlation coefficients between relaxation rates and tissue Mn concentrations (r= 0.84, 0.77 and 0.92 for the striatum, the cortex and the cerebellum, respectively). This methodology offers a unique toolfor monitoring the degree of Mn concentration in different areas of the brain in animal models of Mn intoxication. It will be useful for evaluating the efficacy of treatments aimed at decreasing the metal in the brain. The method could be potentially useful for being transposed in the clinical situation for monitoring Mn-exposed workers.

Changes in tumor oxygenation/perfusion induced by the no donor, isosorbide dinitrate, in comparison with carbogen: monitoring by EPR and MRI.

PURPOSE: In an effort to improve radiotherapy treatments, methods aimed at increasing the quantity of oxygen delivered to tumors were investigated. The aim of this study was to evaluate the effect of one nitric oxide (NO) donor (isosorbide dinitrate) on pO(2) and blood flow in a murine tumor model. The effect was compared to carbogen, used as a reference treatment. METHODS AND MATERIALS: Thirty-six liver tumors implanted in mouse thighs were imaged using magnetic resonance imaging (MRI) at 4.7 Tesla with dynamic Gd-DTPA and blood oxygen level-dependent (BOLD) contrast-enhanced imaging after administration of isosorbide dinitrate or carbogen. The effect on the pO(2) was also tested by EPR oximetry (1.1 GHz) on 52 mice. RESULTS: A significant increase in MRI intensity was observed for both treatments in comparison with the control group. EPR oximetry showed a dose-dependant increase in tumor pO(2) for isosorbide dinitrate (by 5.9 mmHg at 0.2 mg/kg) and a substantially greater change for carbogen breathing (by 23 mmHg). CONCLUSION: Both tumor blood flow and pO(2) were increased by isosorbide dinitrate and carbogen. Carbogen is more efficient than isosorbide dinitrate in increasing the BOLD image intensity, as well as the tumor pO(2), but as efficient as isosorbide dinitrate in the Gd-DTPA contrast-enhanced imaging. We conclude that the effects of carbogen on improving tumor pO(2) involve both improved blood flow and improved hemoglobin oxygenation, whereas the effects of isosorbide dinitrate are predominantly mediated by improved blood flow alone.

Effect of exogenous creatine supplementation on muscle PCr metabolism.

31P NMR was used to assess the influence of two weeks creatine supplementation (21g x d(-1)) on resting muscle PCr concentration, on the rate of PCr repletion (R(depl)), and on the half-time of PCr repletion (t 1/2). Body mass (BM) and volume of body water compartments were also estimated by impedance spectroscopy. Fourteen healthy male subjects (20.8+/-1.9 y) participated in this double-blind study. PCr was measured using a surface coil placed under the calf muscle, at rest and during two exercise bout the duration of which was 1 min. They were interspaced by a recovery of 10 min. The exercises comprised of 50 plantar flexions-extensions against weights corresponding to 40% and 70% of maximal voluntary contraction (MVC), respectively. Creatine supplementation increased resting muscle PCr content by approximately 20% (P= 0.002). R(depl) was also increased by approximately 15% (P< 0.001) and approximately 10% (P = 0.026) during 40% and 70% MVC exercises, respectively. No change was observed in R(repl) and t1/2. BM and body water compartments were not influenced. These results indicate that during a standardized exercise more ATP is synthesized by the CK reaction when the pre-exercise level in PCr is higher, giving some support to the positive effects recorded on muscle performance.

Evidence for a neuroprotective effect of pyrid-3-yl-sulphonyl-urea in photochemically induced focal ischaemia in rats: magnetic resonance imaging evaluation.

A neuroprotective effect can be obtained with N-[(4-cycloheptylaminopyrid-3-yl)sulphonyl]N'-cycloheptyl urea (BM27), a pyrid-3-yl-sulphonylurea structurally related to torasemide, a loop diuretic. We have investigated the neuroprotective effect of BM27 by magnetic resonance imaging and use of the photothrombotic model of cerebral infarction in the rat. This method enables non-invasive quantification of the extent of the cerebral oedema from T2-weighted spin-echo images. This article reports the evolution of the extent of oedema with time (0.5, 1, 2, 4, 6, 24 and 48 h, 7 and 15 days and 1 month after induction of the lesion) in rats pretreated with 5 mg kg(-1) BM27 or an appropriate control. At all times, the rats treated with BM27 had, on average, smaller lesions than control rats (30% decrease between 2 h and 6 h). These results strongly suggest a significant (P < 0.01) but modest neuroprotective effect of BM27 in ischaemic cerebral stroke. Further investigations should be performed to determine if BM27 or its analogues are of clinical interest.

Multiple echo frequency-domain image contrast: improved signal-to-noise ratio and T2 (T2*) weighting.

Conventional T2- and T2*-weighted image contrasts are produced by waiting a TE period for the transverse magnetic resonance (MR) signals to decay to differentiate tissue types with distinct relaxation rates. Significant image signal-to-noise ratio (SNR) is compromised by this contrast-producing process. In this report, a multiple echo frequency-domain image contrast (MEFIC) method is presented. During the conventional TE period, a multiple echo train modulated by T2 or T2* decay is acquired. A third Fourier transform along the echo direction produces an image set with pixel signal intensity modulated by the spectrum of the decay curve. This method simultaneously enhances image contrast with a large increase in SNR. Experimental studies of cerebral vasogenic edema in immature rats and functional MR imaging studies of the human motor cortex have demonstrated that the MEFIC method produces superior image quality over conventional methods for generating T2- and T2* weighted images.

Expression of truncated utrophin improves pH recovery in exercising muscles of dystrophic mdx mice: a 31P NMR study.

31P NMR spectroscopy was used to study the energy metabolism of dystrophin-deficient skeletal muscle of mdx mice, an animal model of Duchenne muscular dystrophy, in which expression of a truncated form of utrophin has been obtained through transgenesis technology. Measurements of ATP, phosphocreatine (PCr), inorganic phosphates (Pi) and intracellular pH (pHi) were made at rest, during a fatigue protocol and during the subsequent recovery. Mechanical fatigue of transgenic muscles was similar to normal muscle, while mdx muscle showed larger force loss. At rest, muscles of all groups had similar values for [ATP], [PCr], [Pi] and pHi. During fatigue, [PCr] decreases mirrored [Pi] increases and were similar in all groups. The major difference between mdx muscles and the group of normal and trc-utrophin muscles concerned the values and evolution of pHi. The mdx muscles showed a more severe intracellular acidosis during exercise and a slower and incomplete post-exercise recovery of normal pHi. In contrast, in trc-utrophin muscles, the kinetics and amplitude of pHi changes were remarkably close to normal behaviour. We conclude that the impaired proton washout which is present in mdx muscles, is corrected to a great extent by the expression of trc-utrophin.

Small particles of fusinite and carbohydrate chars coated with aqueous soluble polymers: preparation and applications for in vivo EPR oximetry.

The development of oxygen-sensitive paramagnetic materials is being pursued actively because of their potential applications in in vivo EPR oximetry. Among these materials, several charcoals and carbohydrate chars are of special interest because of their desirable EPR properties: high sensitivity of the EPR linewidth to the partial pressure of oxygen, simple EPR spectra, and high spin density. Their potential use in humans, however, is limited by the need to demonstrate that they will not lead to deleterious effects. A strategy was used to optimize the biocompatibility of the oxygen-sensitive materials by decreasing the size of the particles and coating them with suspending or surfactive agents such as arabic gum, poloxamer (Pluriol 6800), and polyvinylpyrrolidone. The coated particles of a carbohydrate char and fusinite were characterized in vitro for their size, stability, and pO2 sensitivity. The feasibility of performing pO2 measurement was examined in vivo by inducing ischemia in the gastrocnemius muscle of mice. The use of arabic gum for coating the fusinite particles preserved the pO2 sensitivity in vivo, whereas the other surfactive agents led to a loss of the pO2 sensitivity in vivo. Small particles of fusinite coated by arabic gum and intravenously administered to mice accumulated in the liver, whereas the uncoated fusinite was toxic when injected intravenously due to the large size and aggregation of the particles. Histological studies performed up to 6 months after the injection in muscles of mice did not indicate any toxicity from the materials used in the present study.

Comparison of unenhanced and gadoxetate disodium-enhanced spin-echo magnetic resonance imaging for the detection of experimental hepatocellular carcinoma in the rat.

RATIONALE AND OBJECTIVES: The authors compare the potential value of unenhanced and gadoxetate disodium-enhanced spin-echo images for the detection of hepatocellular carcinoma in a rat model. METHODS: Eleven rats with chemically induced hepatocellular carcinoma underwent unenhanced T2-weighted fast spin-echo imaging followed by T1-weighted spin-echo imaging before and at 5 minutes, 30 minutes, 3 hours, 1 day, and 3 days after intravenous administration of 60 micromol/kg gadoxetate disodium at 4.7 tesla. Tumor and liver enhancement, and tumor-to-liver contrast-to-noise (C/N) ratio were calculated. RESULTS: After gadoxetate disodium administration, the tumors showed less enhancement than the liver. Tumor-to-liver C/N ratio increased from 5.5 +/- 0.8% on unenhanced T1-weighted images to 12.9 +/- 2.4% on gadoxetate-enhanced T1-weighted images (P = 0.02). However, the C/N ratio on unenhanced T2-weighted images (23.5 +/- 3.6%) remained higher than that on gadoxetate-enhanced T1-weighted images, a difference that is statistically significant (P < 0.01). CONCLUSIONS: In the experimental setting of our study, the higher tumor-to-liver C/N ratio on unenhanced T2-weighted spin-echo images suggests that unenhanced T2-weighted spin-echo images are superior to gadoxetate disodium-enhanced T1-weighted spin-echo images for the detection of hepatocellular carcinoma.

Removal of local field gradient artifacts in T2*-weighted images at high fields by gradient-echo slice excitation profile imaging.

Development of high magnetic field MRI techniques is hampered by the significant artifacts produced by B0 field inhomogeneities in the excited slices. A technique, gradient-echo slice excitation profile imaging (GESEPI), is presented for recovering the signal lost caused by intravoxel phase dispersion in T2*-weighted images. This technique superimposes an incremental gradient offset on the slice refocusing gradient to sample k-space over the full range of spatial frequencies of the excitation profile. A third Fourier transform of the initial two-dimensional image set generates an image set in which the artifacts produced by the low-order B0 inhomogeneity field gradients in the sample are separated and removed from the high-order microscopic field gradients responsible for T2* contrast. Application to high field brain imaging, at 3.0 T for human and at 9.4 T for immature rat imaging demonstrates the significant improvement in quality of the T2*-weighted contrast images.

Effects of riluzole on the evolution of focal cerebral ischemia: a magnetic resonance imaging study.

The aim of this study was to investigate the effects of riluzole on the lesion induced by a permanent middle cerebral artery occlusion (MCAO) in rats. Riluzole at 4 or 8 mg/kg i.v. significantly reduced the cortical ischemic brain damage. With the most effective dose of 8 mg/kg, the time evolution of the lesion was assessed by T2-weighted magnetic resonance imaging (MRI) repeated on the same animals after MCAO. MRI obtained at 24, 48, and 72 hours after MCAO showed a progressive increase of the ischemic lesion, except in the cortex of the riluzole-treated rats (8 mg/kg i.v.). Furthermore, there was no difference between lesion volumes as measured by MRI or by histology. This study indicates that MRI may be a valuable method to quantify in vivo the neuroprotective profile of a drug.

Free magnesium concentration in isolated rabbit hearts subjected to high dose isoproterenol infusion: a 31P NMR study.

The hypothesis of magnesium deficiency in isoproterenol (ISO) induced myocardial injury has been investigated by 31P nuclear magnetic resonance spectroscopy. High energy phosphate concentrations, pHi, and intracellular free magnesium concentration ([Mg2+]i) were measured in isolated rabbit hearts perfused at constant flow and subjected to 10(-6)M isoproterenol during 30 min. Recent calibrations were used for [Mg2+]i measurements, and uncertainties on [Mg2+]i estimated values were calculated. During isoproterenol infusion, pHi, [PCr], and [ATP] decreased, while [P(i)] increased. When it was stopped, [PCr] completely repleted, whereas only a partial restoration was observed for pHi and [P(i)]. A rise of end-diastolic pressure and perfusion pressure expressed a contracture, concomitant with a lack of [ATP] recovery, which remained at 59 +/- 13% of the rest value. These results establish that 10(-6) M isoproterenol caused severe myocardial injury. [Mg2+]i increased from 0.70 mM at rest to 0.88 mM at the end of the isoproterenol period. Considering the estimated uncertainties on the [Mg2+]i values, this increase was not significant. After isoproterenol infusion, [Mg2+]i progressively decreased to reach 0.72 mM at 45 min recovery. It is concluded that isoproterenol myocardial toxicity may not be related to [Mg2+]i deficiency.

31P NMR saturation transfer study of the creatine kinase reaction in human skeletal muscle at rest and during exercise.

The creatine kinase reaction has been studied by 31P NMR in exercising human calf muscle. Quantitative analysis of high energy phosphates and saturation transfer study of the creatine kinase flux in the direction of ATP synthesis (Vfor) were performed at rest and during exercise. As expected, exercise induced a [PCr] decrease (from 28.5 +/- 0.9 to 21.9 +/- 1.5 mM, P < 0.01) matched by a Pi increase (from 4.5 +/- 0.2 to 8.9 +/- 1.8 mM, P = 0.06). pHi and [ATP] remained unchanged. Vfor did not change from rest (12.4 +/- 0.9 mM s(-1)) to moderate exercise and decreased at the highest exercise level (8.4 +/- 1.4 mM s(-1), P = 0.006). This observation differs from the prediction of the creatine kinase rate equation, showing an increase in the flux with exercise intensity. Computations suggest that this discrepancy arises from metabolite compartmentalization and/or from the reaction kinetics of a dead end complex stabilized by planar anions.

Detection and segmental location of malignant hepatic tumors: comparison of ferumoxides-enhanced gradient-echo and T2-weighted spin-echo MR imaging.

OBJECTIVE: The aim of our prospective study was to compare the values of ferumoxides-enhanced gradient-echo and T2-weighted spin-echo MR imaging for the detection and segmental location of malignant hepatic tumors. SUBJECTS AND METHODS: Eighteen patients underwent ferumoxides-enhanced T2 weighted spin-echo and steady-state gradient-echo imaging before surgery. Intraoperative sonography was used as the reference examination and showed 28 malignant tumors involving 39 of the 144 hepatic segments. The MR images were reviewed independently by two observers who recorded the number of tumors and their segmental location. The results of the segmental location were subjected to receiver-operating-characteristic analysis. Tumor-liver, vessel-liver, and tumor-vessel contrast-to-noise ratios were calculated. RESULTS: Detection on the gradient-echo images did not differ significantly from that on the T2-weighted spin-echo MR images (observers A and B detected 23 and 24 tumors, respectively, on the gradient-echo images; the two observers each detected 22 tumors on the T2-weighted spin-echo images). Segmental localization was significantly better on the gradient-echo images than on the T2-weighted spin-echo images (mean area under the receiver-operating-characteristic curve: 0.974 versus 0.895 for observer A and 0.962 versus 0.906 for observer B, p = .024). The highest contrast-to-noise ratios for vessel to liver and tumor to vessel were obtained on the gradient-echo images and the highest tumor-liver contrast-to-noise ratio was obtained on the T2-weighted spin-echo images. CONCLUSION: In our study, ferumoxides-enhanced gradient-echo imaging was as accurate as T2-weighted spin-echo imaging for revealing malignant hepatic tumors and was superior for showing their segmental location.

Ferumoxides and Tc-99m sulfur colloid: comparison of the tumor-to-liver uptake in focal nodular hyperplasia.

The tumor-to-liver uptake of two reticuloendothelial agents, namely ferumoxides and technetium-99m (Tc-99m) sulfur colloid, was compared in focal nodular hyperplasia (FNH). Twelve patients with FNH who had undergone ferumoxides-enhanced MR imaging and planar Tc-99m sulfur colloid scintigraphy within 1 year were included from the study. Fourteen patients with FNH with a diameter larger than 3 cm were selected for the comparison. The tumor-to-liver ferumoxides uptake was calculated and the Tc-99m sulfur colloid uptake was assessed visually. Fermuoxides uptake was observed in all but one patient with FNH (mean tumor-to-liver ratio = .36). The six tumors showing normal (n = 5) or increased (n = 1) radiocolloid uptake when compared to the liver accumulated more ferumoxides than the eight tumors showing decreased radiocolloid uptake (P < .01). However, in some tumors, no direct relation was observed between ferumoxides and Tc-99m sulfur colloid uptake. Our observations suggest that ferumoxides uptake might not exactly mimic Tc-99m sulfur colloid uptake in FNH.

Detection of reperfused ischemia of the rat intestine: value of magnetic resonance imaging with small-molecular-weight dysprosium and gadolinium chelates.

RATIONALE AND OBJECTIVES: The authors assessed whether the small-molecular-weight magnetic resonance (MR) imaging contrast agents dysprosium diethylenetriamepentaacetic acid bismethylamide (sprodiamide injection), which enhances T2*, and gadolinium diethylenetriamepentaacetic acid bismethylamide (gadodiamide injection), which enhances T1, could improve the detection of reperfused ischemia of the rat intestine. METHODS: Eighteen rats were subjected to vascular occlusion of the distal ileum for 30 minutes, followed by reperfusion. Ten minutes after reperfusion, T1- and T2-weighted spin-echo (SE) images were obtained before and after administration of sprodiamide, gadodiamide, or both. The same imaging protocol was applied in another group of 18 rats subjected to 10 minutes of occlusion and reperfusion. Histologic examination of the intestine was performed after MR imaging. RESULTS: Villous injury (ie, denudation) was observed in most cases after 30 minutes of occlusion, but not after 10 minutes of occlusion. After 30 minutes of occlusion, the superficial part of the ischemic intestine was hyperintense to the normal intestine on unenhanced T2-weighted images. Administration of sprodiamide improved the contrast between the normal and ischemic intestine on T2-weighted images, and administration of both gadodiamide and sprodiamide improved the contrast on T1- and T2-weighted images. After 10 minutes of occlusion, no contrast was discernible before or after contrast material administration. CONCLUSION: These results suggest that the detection of reperfused intestinal ischemia of sufficient duration to cause villous injury can be improved by using sprodiamide injection alone or in combination with gadodiamide.

Gene therapy of rat C6 glioma using adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene: long-term follow-up by magnetic resonance imaging.

The herpes simplex virus thymidine kinase gene was transferred into C6 glioma cells by infection with a recombinant adenovirus. In vitro, a 10 microM ganciclovir concentration was able to kill 100% of the infected cells. For in vivo experiments, brain tumors were established by stereotactic injection of C6 glioma cells in the caudate nucleus of rats. Five days later, the recombinant adenovirus was inoculated into the tumors and the animals were treated by intraperitoneal injections of ganciclovir for 14 days. At the end of ganciclovir therapy, histological examination revealed a 28-fold decrease in tumor volumes in the treated animals, as compared with control animals. In long-term studies, the mean survival time of the treated animals were four-fold longer than that of control ones. Magnetic resonance imaging demonstrated an apparent complete tumor regression in 62% of the animals. However, late tumor recurrence was observed in the treated animals. Repeated inoculation of C6 glioma cells in the contralateral hemisphere of long-term surviving animals resulted in either tumor rejection or slowly growing tumors. These findings demonstrate the potential efficacy of adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene and ganciclovir administration in the treatment of rat gliomas.

Development of biocompatible implants of fusinite for in vivo EPR oximetry.

The development of oxygen-sensitive paramagnetic materials is being actively pursued because of their potential applications for in vivo electron paramagnetic resonance (EPR) oximetry. Among these materials, fusinite is of particular interest because of the high sensitivity of the EPR linewidth to the partial pressure pO2. Although this material has led to a number of very useful results in experimental systems, its potential use in humans is limited by the need to prove that it will not cause deleterious effects. The strategy used in this study to optimize the biocompatibility of the oxygen-sensitive materials was to prepare small silicon implants containing the fusinite. The use of silicon permits the diffusion of oxygen inside the implant while the material does not have contact with the biological environment. Radiosterilization did not affect the pO2 sensitivity of the material. The feasibility of performing pO2 measurement was verified in vivo by periodically inducing ischemia in the gastrocnemius muscle of mice over a period of 6 weeks.

Reperfused ischemia of the rat intestine: detection by MR imaging with polylysine-Gd-DTPA enhancement.

To detect reperfused ischemia of the rat intestine, T2-weighted spin-echo images were acquired, followed by T1-weighted images before and after administration of polylysine-Gd-DTPA or Gd-DTPA. Before administration of the contrast agent, the reperfused intestine was hyperintense on T2-weighted images, and to a lesser extent on T1-weighted images. After administration of polylysine-Gd-DTPA, the reperfused intestine enhanced more than the normal one, giving a significantly better contrast-to-noise (CNR) ratio than on unenhanced images. Gd-DTPA induced the same enhancement of the reperfused and the normal intestine and the CNR was lower than on unenhanced T2-weighted images. Reperfused intestinal ischemia could thus be better detected on polylysine-Gd-DTPA-enhanced MR images than on unenhanced images or on Gd-DTPA-enhanced images.