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.

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.

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.

Comparison of gadolinium-DTPA and polylysine-gadolinium-DTPA--enhanced magnetic resonance imaging of hepatocarcinoma in the rat.

RATIONALE AND OBJECTIVES: To compare the magnetic resonance (MR) imaging characteristics of gadolinium-DTPA (Gd-DTPA), a low-molecular-weight contrast agent, and polylysine-Gd-DTPA, a macromolecular contrast agent, in two types of hepatocarcinomas (HCC) in the rat. METHODS: T1-weighted spin-echo images were obtained in 13 rats with chemically induced HCC and 26 rats with Novikoff HCC before and 3 minutes to 60 hours after administration of either Gd-DTPA or polylysine-Gd-DTPA. RESULTS: Three minutes after polylysine-Gd-DTPA administration, the tumor-to-liver contrast of the two types of HCC increased significantly (positive contrast for chemically induced HCC and negative contrast for Novikoff HCC). At 30 minutes and 60 hours, the tumor-to-liver contrast remained above baseline values in chemically induced HCC and returned progressively to baseline values in Novikoff HCC. No significant increase in tumor-to-liver contrast was observed after Gd-DTPA administration. CONCLUSIONS: These results suggest that polylysine-Gd-DTPA provides a higher and more prolonged increase in tumor-to-liver contrast than Gd-DTPA.

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.

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.

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.

Evaluation of nonionic nitroxyl lipids as potential organ-specific contrast agents for magnetic resonance imaging.

Considering their intrinsic properties of accumulation in the hepatic tissue, we have synthesized nitroxyl-containing lipids as potential organ-specific contrast agents for magnetic resonance imaging (MRI). Their resistance to reduction by ascorbate and in liver homogenates, and their relaxivity in different media were investigated and compared to those of free carboxyl-Proxyl (3-carboxy-2,2,5,5-tetramethylpyrrolidine-1-oxyl) and Tempamaine (4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl). With respect to the reduction rates by ascorbate, the lipid derivatives show the same well-known order of reactivity as carboxy-Proxyl and Tempamine, the five-membered nitroxyls being more stable than the six-membered compounds. However the binding of the piperidinoxyl compounds to the fatty acids confers to those lipid derivatives a markedly increased stability. Similarly, in liver homogenates, the nitroxyl lipids remained unchanged more than 20 min, contrarily to carboxy-Proxyl and Tempamine. The measurements of spin-lattice relaxation time (T1) in biological media have demonstrated a higher relaxivity of nitroxyl lipids, which can be related to their interaction with proteins. Tested in vivo, one of the synthesized compounds (0.75 mmol/kg) produced an enhancement of 44 +/- 12% of the hepatic signal 5 min after intraportal injection in T1-weighted images. The potential applicability of the other nitroxyl lipids as contrast agents for MRI was limited in the in vivo studies by an unexpected toxicity. Work is currently in progress to improve the therapeutic index of the present class of nitroxyl lipids.

Spin labelled arabinogalactan as MRI contrast agent.

In this study, we report the synthesis and the evaluation as MRI contrast agent of arabinogalactan/pyrrolidinoxyl radicals (PCA) covalent adduct (SLAG:Spin Labelled ArabinoGalactan). Arabinogalactan was used as targeting device, as it is recognized by the asialoglycoprotein receptor specific to the hepatocytes. The higher relaxivity R1 in water of SLAG, compared with small hydrophilic nitroxyl radicals, was explained by the molecular dynamics study using EPR spectroscopy that showed some immobilization of the radical into the polysaccharide. A binding study on isolated hepatocytes revealed that SLAG still recognizes the asialoglycoprotein receptor. MR imaging was performed using spin-echo T1 weighted images on mice to compare the contrast effect obtained with SLAG and PCA after IV injection (1 mmol/kg free radical). The percent signal enhancement observed in the liver 5 min after IV injection was 40 +/- 3% and 13 +/- 5% for SLAG and PCA, respectively. The signal was also dramatically increased in the renal cortex. This latter effect as well as the prolonged duration of the contrast (+/- 3 h), indicates at least a partial nonselective biodistribution; the high concentration needed to obtain a contrast effect could account for the saturation of the asialoglycoprotein receptor and hence for the apparent nonselective biodistribution.

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.

Magnetic resonance imaging of lower abdominal and pelvic lesions: assessment of oral magnetic particles as an intestinal contrast agent.

To determine the value of oral magnetic particles (OMP) as a superparamagnetic MR contrast agent for the gastrointestinal tract in lower abdominal and pelvic lesions, 30 patients underwent spin-echo imaging before and after ingestion of OMP at a dose of approximately 80 mg of iron in 800 ml water. The preparation was divided into four portions and taken by the patient over a 2-h period. Two readers independently reviewed the MR images. The contrast material was well tolerated and the distribution of the contrast material was good to excellent in the proximal and pelvic small bowel, but was not sufficient in the colon with the dose and timing used in the study. Postcontrast images showed a significantly better delineation of the lesions, the small bowel, and the paraaortic region, but no significant improvement in the delineation of the colon, the iliac vessels area, the bladder or genital tract. Compared with precontrast images, confidence in defining or excluding disease on postcontrast images was better, equal or worse in 40, 60 and 0% of cases, respectively (P less than 0.001) with a substantial agreement between readers (kappa = 0.71). OMP produced susceptibility artefacts of significant intensity in only one case. These results indicate that OMP may be useful in the delineation of lower abdominal and pelvic lesions at MR imaging. Marking of the colon by a contrast agent might improve the results.

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.

Cervical myelopathy: MRI evaluation of cord compression.

In order to assess the consequences of cervical spinal cord compression in cervical myelopathy, MRI measurements of the sagittal diameter of the cervical spinal cord were obtained in 50 normal volunteers and 50 patients suffering from cervical myelopathy. Whatever the degree of stenosis, the values obtained in the latter group are significantly inferior to those of the normal control group (average mean diameter: normal group: 8.2 mm; cervical myelopathy: 6.2 mm).

Validation of automated brain contour determination in normal and abnormal cerebral single-photon emission tomography.

A contour detection algorithm for cerebral studies, using the method of Tomitani, has been implemented on a single-photon emission tomographic (SPET) system. It is based on the detetion by threshold of the brain edge in the sinogram and does not depend on the reconstruction algorithm. Thirteen normal subjects underwent an examination on both computed tomography (CT) and SPET using a head holder to ensure the reproducibility of the positioning. The CT scan contour of the brain was drawn manually according to the brain parenchyma limits. The SPET brain contour was obtained by use of the Tomitani algorithm after the threshold had been determined on an active cylindrical phantom. Using a threshold of 37% of the maximum uptake, the length of the contour as well as the area obtained with SPET and CT were not found to be statistically different. The method of Tomitani, which is simpler and faster then previous methods, provides contours which superimpose very well with CT scan images. Application to patients with unilateral pathological defects is possible by requiring that the contour is symmetrical.

A gating and triggering system dedicated to nuclear magnetic resonance studies of isolated perfused heart.

This work reports a low-cost and versatile electronic device designed to trigger NMR acquisitions from the cardiac cycle of an isolated perfused heart, or to perform electrical stimulation of the heart. The triggering is synchronised with the pressure curve of the perfused heart. The cardiac pacing is achieved from pulses of the NMR system, or by an internal pulse generator, in order to be operated separately from the NMR instrument.

Ganciclovir mediated regression of rat brain tumors expressing the herpes simplex virus thymidine kinase imaged by magnetic resonance.

Using a rat C6 brain tumor model, we studied the antitumor effects of Herpes simplex virus type 1 thymidine kinase (HSV-tk) gene transfer followed by ganciclovir treatment. C6 glioma cells were transfected in vitro with the HSV-tk gene, and tested for their sensitivity to ganciclovir. Although there was no surviving cell at a 30 microM ganciclovir concentration, unmodified C6 cells were not affected by the drug. For in vivo experiments, intracerebral tumors were induced in rats by stereotactic injection of 10(4) HSV-tk-modified C6 cells. Ten days later, the animals were treated with intraperitoneal injections of ganciclovir for 21 days. The tumors evolution was evaluated by high resolution magnetic resonance imaging. In 33% of the rats, the signal intensity of the tumors became heterogeneous, with development of highly hyperintense areas, and a complete tumor regression was subsequently noted. Histological examination of successfully treated tumors revealed progressive necrosis with formation of cysts. The survival time of the HSV-tk/ganciclovir treated animals was consistently increased, all rats surviving more than 30 days and 33% of them being still alive after 80 days.

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.

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.