Daniel Jirák

MRI of transplanted pancreatic islets

A promising treatment method for type 1 diabetes mellitus is transplantation of pancreatic islets containing β‐cells. The aim of this study was to develop an MR technique to monitor the distribution and fate of transplanted pancreatic islets in an animal model. Twenty‐five hundred purified and magnetically labeled islets were transplanted through the portal vein into the liver of experimental rats. The animals were scanned using a MR 4.7‐T scanner. The labeled pancreatic islets were clearly visualized in the liver in both diabetic and healthy rats as hypointense areas on T2*‐weighted MR images during the entire measurement period. Transmission electron microscopy confirmed the presence of iron‐oxide nanoparticles inside the cells of the pancreatic islets. A significant decrease in blood glucose levels in diabetic rats was observed; normal glycemia was reached 1 week after transplantation. This study, therefore, represents a promising step toward possible clinical application in human medicine. Magn Reson Med 52:1228–1233, 2004.

Texture analysis of human liver.

To classify healthy and diseased livers by texture analysis (TA).

Effects of MRI acquisition parameter variations and protocol heterogeneity on the results of texture analysis and pattern discrimination: an application-oriented study.

MRI texture features are generally considered to be sensitive to variations in signal-to-noise ratio and spatial resolution, which represents an obstacle for the widespread clinical application of texture-based pattern discrimination with MRI. This study investigates the sensitivity of texture features of different categories (co-occurrence matrix, run-length matrix, absolute gradient, autoregressive model, and wavelet transform) to variations in the number of acquisitions (NAs), repetition time (TR), echo time (TE), and sampling bandwidth (SBW) at different spatial resolutions. Special emphasis was placed on the influence of MRI protocol heterogeneity and implications for the results of pattern discrimination. Experiments were performed using two polystyrene spheres and agar gel phantoms with different nodular patterns. T2-weighted multislice multiecho images were obtained using a 3.0 T scanner equipped with a microimaging gradient insert coil. Linear discriminant analysis and k nearest neighbor classification were used for texture-based pattern discrimination. Results show that texture features of all categories are increasingly sensitive to acquisition parameter variations with increasing spatial resolution. Nevertheless, as long as the spatial resolution is sufficiently high, variations in NA, TR, TE, and SBW have little effect on the results of pattern discrimination. Texture features derived from the co-occurrence matrix are superior to features of other categories because they enable discrimination of different patterns close to the resolution limits for the smallest structures of physical texture even for datasets that are heterogeneous with regard to different acquisition parameters, including spatial resolution.

Human Induced Pluripotent Stem Cells Improve Stroke Outcome and Reduce Secondary Degeneration in the Recipient Brain

Human induced pluripotent stem cells (hiPSCs) are a most appealing source for cell replacement therapy in acute brain lesions. We evaluated the potential of hiPSC therapy in stroke by transplanting hiPSC-derived neural progenitor cells (NPCs) into the postischemic striatum. Grafts received host tyrosine hydroxylase-positive afferents and contained developing interneurons and homotopic GABAergic medium spiny neurons that, with time, sent axons to the host substantia nigra. Grafting reversed stroke-induced somatosensory and motor deficits. Grafting also protected the host substantia nigra from the atrophy that follows disruption of reciprocal striatonigral connections. Graft innervation by tyrosine hydoxylase fibers, substantia nigra protection, and somatosensory functional recovery were early events, temporally dissociated from the slow maturation of GABAergic neurons in the grafts and innervation of substantia nigra. This suggests that grafted hiPSC-NPCs initially exert trophic effects on host brain structures, which precede integration and potential pathway reconstruction. We believe that transplantation of NPCs derived from hiPSCs can provide useful interventions to limit the functional consequences of stroke through both neuroprotective effects and reconstruction of impaired pathways.

Cyclodextrin‐Based Bimodal Fluorescence/MRI Contrast Agents: An Efficient Approach to Cellular Imaging

A novel bimodal fluorescence/MRI probe based on a cyclodextrin scaffold has been synthesized and characterized. The final agent employs the fluorescein (F) functionality as a fluorescence marker and the Gd(III) complex of a macrocyclic DOTA-based ligand (GdL) having one aminobenzyl-phosphinic acid pendant arm as an MRI probe, and has a statistical composition of (GdL)(6.9)-F(0.1)-beta-CD. Slow rotational dynamics (governed by a very rigid cyclodextrin scaffold) combined with fast water exchange (ensured by the chosen macrocyclic ligand) resulted in a high relaxivity of approximately 22 s(-1) mM(-1) per Gd(III) or approximately 150 s(-1) mM(-1) per molecule of the final conjugate (20 MHz, 25 degrees C). In vitro labelling of pancreatic islets (PIs) and rat mesenchymal stem cells has been successfully performed. The agent is not cytotoxic and is easily internalized into cells. The labelled cells can be visualized by MRI, as proved by the detection of individual labelled PIs. A fluorescence study performed on mesenchymal stem cells showed that the agent stays in the intracellular space for a long time.

Metabolite and diffusion changes in the rat brain after Leksell Gamma Knife irradiation

Our study describes the time course of necrotic damage to the rat brain resulting from Leksell Gamma Knife (LGK) irradiation at a dose that was previously considered to be subnecrotic. A lesion induced in the rat hippocampus by 35 Gy irradiation was monitored by MRI, MRS, and DW‐MRI for 16 months. T2‐weighted images revealed a large hyperintense area with an increased apparent diffusion coefficient of water (ADCw), which occurred 8 months after irradiation, accompanied by metabolic changes (increase of lactate (Lac) and choline (Cho), and decrease of creatine (Cr) and N‐acetyl aspartate (NAA), as determined by MRS) that indicated an edema. In two animals, the hyperintensity persisted and a postnecrotic cavity connected to enlarged lateral ventricles developed. In the rest of the animals, the hyperintensity started to decrease 9 months post‐irradiation (PI), revealing hypointense areas with a decreased ADCw. Histology confirmed the MRI data, showing either scar formation or the development of a postnecrotic cavity. Magn Reson Med 52:397–402, 2004.

Selective in vitro anticancer effect of superparamagnetic iron oxide nanoparticles loaded in hyaluronan polymeric micelles.

Due to its native origin, excellent biocompatibility and biodegradability, hyaluronan (HA) represents an attractive polymer for superparamagnetic iron oxide nanoparticles (SPION) coating. Herein, we report HA polymeric micelles encapsulating oleic acid coated SPIONs, having a hydrodynamic size of about 100 nm and SPION loading capacity of 1-2 wt %. The HA-SPION polymeric micelles were found to be selectively cytotoxic toward a number of human cancer cell lines, mainly those of colon adenocarcinoma (HT-29). The selective inhibition of cell growth was even observed when the SPION loaded HA polymeric micelles were incubated with a mixture of control and cancer cells. The selective in vitro inhibition could not be connected with an enhanced CD44 uptake or radical oxygen species formation and was rather connected with a different way of SPION intracellular release. While aggregated iron particles were visualized in control cells, nonaggregated solubilized iron oxide particles were detected in cancer cells. In vivo SPION accumulation in intramuscular tumor following an intravenous micelle administration was confirmed by magnetic resonance (MR) imaging and histological analysis. Having a suitable hydrodynamic size, high magnetic relaxivity, and being cancer specific and able to accumulate in vivo in tumors, SPION-loaded HA micelles represent a promising platform for theranostic applications.

Phosphonate-titanium dioxide assemblies: platform for multimodal diagnostic-therapeutic nanoprobes.

Multimodal imaging-therapeutic nanoprobe TiO(2)@RhdGd was prepared and successfully used for in vitro and in vivo cell tracking as well as for killing of cancer cells in vitro. TiO(2) nanoparticles were used as a core for phosphonic acid modified functionalities, responsible for contrast in MRI and optical imaging. The probe shows high (1)H relaxivity and relaxivity density values. Presence of fluorescent dye allows for visualization by means of fluorescence microscopy. The applicability of the probe was studied, using mesenchymal stem cells, cancer HeLa cells, and T-lymphocytes. The probe did not exhibit toxicity in any of these systems. Labeled cells were successfully visualized in vitro by means of fluorescence microscopy and MRI. Furthermore, it was shown that the probe TiO(2)@RhdGd can be changed into a cancer cell killer upon UV light irradiation. The above stated results represent a valuable proof of a principle showing applicability of the probe design for diagnosis and therapy.

Dual imaging probes for magnetic resonance imaging and fluorescence microscopy based on perovskite manganite nanoparticles

The present study reveals the potential of magnetic nanoparticles based on the La0.75Sr0.25MnO3 perovskite manganite for magnetic resonance imaging (MRI). Moreover, it describes the development of the dual imaging probe where the magnetic cores are combined with a fluorescent moiety while the improved colloidal stability is achieved by a two-ply silica shell. At first, the magnetic cores of La0.75Sr0.25MnO3 are coated with a hybrid silica layer, comprising a covalently attached fluorescein moiety that is subsequently covered by a pure silica layer providing the enhanced stability. The detailed characterization of the intermediate and the final product reveals the importance of the complex two-ply shell. Viability tests show that the complete particles are suitable for biological studies. Internalization of the particles and their presence in intracellular vesicles are observed by fluorescence microscopy in different cell types. Further experiments prove no fatal interference with the vitality and insulin releasing ability of labeled pancreatic islets. Relaxometric measurements confirm high spin–spin relaxivities at magnetic fields of B0 = 0.5–3 T, while visualisation of in vitro labeled pancreatic islets by MRI is successfully tested.

Two distinct tumor phenotypes isolated from glioblastomas show different MRS characteristics

We have developed a human brain tumor model in immunodeficient rats that gradually changes its phenotype by serial passages in vivo, from a highly infiltrative, non‐angiogenic one with numerous stem cell markers [low‐generation (LG) tumor] to a more typical glioblastoma one with extensive angiogenesis and necrosis [high‐generation (HG) tumor]. In this study we determined the metabolic properties of these two phenotypes, using 1H MRS. The LG tumors showed an intact blood–brain barrier and normal vascular morphology, as shown by MRI and Hoechst staining. In contrast, the HG tumors exhibited vascular leakage and necrosis. The animals with HG tumor had raised concentrations of choline and myo‐inositol, and decreased concentrations of glutamate and N‐acetylaspartate. In the LG tumor group, similar changes in metabolic concentrations were detected, although the alterations were more pronounced. The LG tumors also had higher concentrations of choline, taurine, and lactate. Subdividing the LG and HG tumors into large and small tumors revealed a significant increase in choline and decrease in glutamate as the LG tumors increased in size. Our results show that metabolic profiles produced by 1H MRS can be used to distinguish between two distinct glioblastoma phenotypes. More pronounced anaerobic metabolism was present in the LG stem‐cell‐like tumors, suggesting a more malignant phenotype. Copyright