Roger Demeure

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.

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.

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.

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.

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.

Multiple echo frequency-domain image contrast : Improved signal-to-noise ratio and T2 (T*2) weighting

Conventional T2‐ and T  *2 ‐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 T  *2 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 T  *2 ‐weighted images. Magn Reson Med 41:423–428, 1999.

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.

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.

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 quantifyin vivo the neuroprotective profile of a drug.

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 tumorto-liver contrast than Gd-DTPA.