Magnetic resonance imaging studies with cluster algorithm for characterization of brain edema after controlled cortical impact injury (CCII).

Objective of this study was the characterization of traumatic brain injury induced by a "Controlled Cortical Impact" with magnetic resonance imaging techniques. The impact was applied to the intact dura of the left hemisphere in Sprague-Dawley rats. The pneumatic impactor was accelerated to a velocity of 7 m/s contusing the left temporo-parietal hemisphere to a depth of 2 mm. Posttraumatic hemispheric swelling and water content were determined gravimetrically, Evans Blue extravasation photometrically, and volume of ischemia by TTC-staining and planimetry. Magnetic resonance imaging was performed by a Bruker biospec 24/40, 90 min, 24 and 72 h post trauma using a T2w RARE sequence, a T1w sequence, before and after application of contrast agent, and a set of diffusion weighted images for calculation of ADC-maps. Data analysis was performed using a cluster algorithm enabling to interpret corresponding image pairs simultaneously. T2w imaging indicates the maximum edema about 24 h post trauma. Blood-brain barrier damage, detected by T1w imaging, is more predominant in the early posttraumatic phase. The cluster algorithm detects different edema components: from the necrotic core to the perifocal vasogenic rim. MRI in combination with the cluster algorithm will hopefully be a valuable tool in testing neuroprotective agents.

Liposome-encapsulated superparamagnetic iron oxide particles as markers in an MRI-guided search for tumor-specific drug carriers.

Three different types of liposomes containing superparamagnetic iron oxide particles (SPIOs) as the contrast agent were tested for intratumoral enrichment by magnetic resonance imaging (MRI). The liposome-encapsulated SPIOs were investigated in comparison with AMI-227, dextran-coated SPIOs in two different dosages using the CC531 adenocarcinoma in the liver of WAG/RIJ rats as a model. Reduction of the relative signal intensity (SI) in the tumor in T2-weighted MR images was assumed as a measure of the liposome enrichment in the tumor or adjacent tissue. The samples were tested at dosages with isomolar iron content and at dosages producing the same MR relaxivity R2. For reverse phase evaporation vesicles and small unilamellar vesicles behavior differed remarkably according to the applied dosage. A steady, strong reduction in SI, starting immediately after injection and extending up to 48 h, was observed for small unilamellar vesicles sterically stabilized with polyethylene glycol.

Characterisation of brain edema following "controlled cortical impact injury" in rats.

Significance, origin and nature of posttraumatic brain edema are still being debated. Recently, a "controlled cortical impact injury" (CCII) was introduced to model traumatic brain injury. Purpose of this study was to investigate the development and nature of brain edema following CCII. Traumatic brain injury was applied to the intact dura of the left hemisphere in Sprague-Dawley rats (n = 52, 250-350 g b.w.). Ketamine/xylazine-anesthesia or inhalation-anesthesia were used. A pneumatic impactor with a diameter of 5 mm contused the temporo-parietal cortex with a velocity of 7 m/s and an impact depth of 2 mm. 24 hours post injury the brains were removed. Posttraumatic hemispheric swelling and water content were determined gravimetrically, Evans blue extravasation spectrophotometrically, area and volume of ischemia by staining with TTC. MRI studies were performed with T1-,T2- and diffusion-weighted sequences. Posttraumatic swelling following CCII was 14.3 +/- 3.1%. Brain water content increased to 82.5 +/- 0.5% in lesioned hemisphere compared to 79.9 +/- 0.2% in control hemisphere. Following TTC staining, the average ischemic tissue volume was 56.7 +/- 19.2 mm3. There was a moderate uptake of Evans blue into the lesioned hemisphere. MRI studies demonstrated edema in 35.4 +/- 9.5 mm3 of the lesioned hemisphere. Gd-DTPA was taken up early after trauma only. A significantly decreased ADC (apparent diffusion coefficient) indicates the cytotoxic (ischemic) component of edema in this model. In conclusion, CCII produces significant posttraumatic brain swelling and edema which is both, of vasogenic and cytotoxic nature. Thus, the CCII models the human cortical contusion more appropriately and opens new avenues for therapeutical studies focussing on cortical contusions.

Simultaneous determination of 5-fluorouracil and its active metabolites in serum and tissue by high-performance liquid chromatography.

In order to observe the biodistribution of 5-fluorouracil (5-FU) and its main metabolites in different kinds of tissue (tumor, liver, kidney, spleen, mucosa, lungs, heart, peritoneum, pancreas) and serum according to various novel application forms, a simple and rapid method for the simultaneous determination of 5-FU and its active metabolites 5-fluorouridine (5-FUrd) and 5-fluoro-2'-deoxyuridine (5-FdUrd) has been established. Proteins in serum and tissue samples were precipitated by perchloric acid after addition of the internal standard 5-bromouracil. The compounds were separated using an ODS Hypersil (5 microm) column and detected by UV absorbance (254 nm). Specificity, linearity, reproducibility, intermediate precision and accuracy of the method were established. The lower limit of quantitation (LOQ) for the compounds in serum and various tissue samples was determined. Data on the recovery of the compounds and the internal standard are provided.

Evaluation of efficient chemoembolization mixtures by magnetic resonance imaging therapy monitoring: an experimental study on the VX2 tumor in the rabbit liver.

To find effective chemoembolization mixtures, we tested combinations of carboplatin with the embolizates Spherex and Gelfoam in comparison to a therapy with NaCl-solution, a treatment with the cytostatic drug only, and a therapy with each of the embolizates alone. The experiments were carried out using as a model the VX2 tumor in the liver of male chinchilla rabbits (five for each group). Carboplatin was revealed by the 3-(4,5-dimethylthiazole-2)-yl-2,5-diphenyltetrazolium bromide test to be a potent cytostatic drug for VX2 rabbit tumor cells. We used magnetic resonance imaging to examine the tumor volume and signal intensity enhancement up to 15 min after Gd-DTPA administration within the tumor and liver before and after the different therapies. These parameters allowed us to evaluate tumor growth and vitality as well as liver injury for the different therapy types. The results found by magnetic resonance imaging corresponded very well to those obtained by histological analysis of the tumors. The chemoembolization therapies were significantly more efficient than the other therapies, as indicated by the reduction of signal intensity enhancement after contrast agent administration within the tumor and by the histologically determined necrotic fraction after therapy. In addition, we found a significant decrease of the tumor volume and no significant live injury for a therapy with Carboplat and Gelfoam.

Interaction between paramagnetic metal complexes and intracellular water of single cells.

Oocytes from Xenopus laevis were used as well-established model to investigate spatially resolved changes in relaxation time (T1) within a cell during exposure to copper complexes by means of 1H-NMR-microscopy. T1 relaxation of intracellular water was shortened in dependence on the complex concentration, the water content, and the water mobility in various cell compartments. A relatively constant T1 decrease was observed in the cytoplasm of the vegetal pole, irrespective of the type and concentration of the permeable complexes used. Since the lowest content of free mobile water molecules was detected at the vegetal pole, we concluded that this quantity was as important for the relaxation time decrease as that of the chelated paramagnetic ions. Experiments using 14C-labeled inulin demonstrated that the paramagnetic metal complexes entered the oocytes without gross injury to their plasma membranes.

Estimation of water content and water mobility in the nucleus and cytoplasm of Xenopus laevis oocytes by NMR microscopy.

NMR microscopy is a noninvasive approach for studying cell structure and properties. Spatially resolved measurements of the relaxation times T1 and T2 provided information on the water proton spin density and water mobility in different parts of Xenopus laevis oocytes. The spin-lattice relaxation time T1 was determined using a saturation-recovery sequence and the common spin-echo sequence with increasing repetition times, while the transverse relaxation time T2 was measured by means of the spin-echo sequence with varying echo times. From the relaxation times, the mole fractions of possible reorientational correlation times tau c for different types of intracellular water were calculated according to a simple two-phase model. The values for T1, T2, and proton spin density (i.e., water content) are: nucleus >> animal cytoplasm > vegetal cytoplasm. Based on the estimation of tau c, nearly 90% of the nuclear water and 74.4% of the water of the animal pole was considered as free mobile water, whereas 55.5% of the water of the vegetal pole appeared as bound water.

Study of the membrane permeability of a paramagnetic metal complex on single cells by NMR microscopy.

A new procedure has been developed for investigating the ability of paramagnetic metal complexes to penetrate the plasma membrane of eukaryotic cells without decomposition. Defolliculated Xenopus laevis oocytes formed the biological system to test N,N-ethylenebis-(1,5,5-trimethyltetramic-acid-3-acetiminato) copper (II). An increase of the signal intensities in spin-echo (SE) images of oocytes treated with the tested substance indicated that the complex was able to penetrate biological membranes due to the arrangement of hydrophobic and hydrophilic groups within the ligand. In contrast, the treatment with the commonly used contrast agent gadolinium-DTPA/dimeglumine did not enhance the signal intensity in NMR images of oocytes after time periods of exposure comparable to those used for the copper complex. After microinjection into Xenopus oocytes the copper complex was released into the extracellular medium without degradation, as shown by HPLC measurements.