Spin-lattice relaxation time in water/graphene-oxide dispersion.

We present the results of the calculations of the spin-lattice relaxation time of water in contact with graphene oxide by means of all-atom molecular dynamics simulations. We fully characterized the water-graphene oxide interaction through the calculation of the relaxation properties of bulk water and of the contact angle as a function of graphene oxide oxidation state and comparing them with the available experimental data. We then extended the calculation to investigate how graphene oxide alters the dynamical and relaxation properties of water in different conditions and concentrations. We show that, despite the diamagnetic nature of the graphene oxide, the confining effects of the bilayers strongly affect the longitudinal relaxation properties of interfacial water, which presents a reduced dynamics due to hydrogen bonds with oxygen groups on graphene oxide. This property makes graphene oxide an interesting platform to investigate water dynamics in confined geometries and an alternative contrast-agent for magnetic resonance imaging applications, especially in view of the possibility to functionalize graphene oxide from theranostic perspectives.

Hypoxia induces peroxisome proliferator-activated receptor α (PPARα) and lipid metabolism peroxisomal enzymes in human glioblastoma cells.

Glioblastoma multiforme (GBM) represents the most severe type of glioma, the most common brain tumor. Their malignancy shows a relationship with an increased proliferation and a poorly organized tumor vascularization, an event that leads to inadequate blood supply, hypoxic areas and at last to the formation of necrotic areas, a feature of glioblastoma. Hypoxic/necrotic tumors are more resistant to chemotherapy and radiation therapies, thus it is crucial to formulate new therapeutic approaches that can render these tumors more sensitive to the action of conventional therapies. It has been demonstrated that under hypoxia, gliomas accumulate lipid droplets and that this event is positively correlated with the degree of malignancy, glioblastoma being the most endowed with lipid droplets. We have previously demonstrated in ex vivo glioma specimens a grade-dependent lipid metabolism perturbation. Here we studied the lipid pathways and the presence of stemness markers in glioma primary cultures, obtained from surgical specimens of patients affected by glioma at different grade of malignancy, GBM primary cultures cultured under both hypoxic and normoxic conditions, as well as normal human astrocytes. The results obtained demonstrate that hypoxia plays a crucial role in regulating the expression of lipid metabolism peroxisomal enzymes, the lipid droplets accumulation as well as the transcription factor PPARα.

Lipid metabolism impairment in human gliomas: expression of peroxisomal proteins in human gliomas at different grades of malignancy.

Gliomas are histologically graded by cellularity, cytological atypia, necrosis, mitotic figures, and vascular proliferation, features associated with biologically aggressive behaviour. However, abundant evidence suggests the presence of unrecognized, clinically relevant subclasses of the diffuse gliomas, both in respect to their underlying molecular phenotype and their clinical response to therapy. It is well-known that patient prognosis and therapeutic decisions rely on accurate pathological grading. Recently, it was reported that human gliomas accumulate lipid droplets during progression, suggesting a lipid metabolism impairment. Considering the crucial role of peroxisomes in lipid metabolism, in the present work we studied the expression profiles of proteins either exclusively localized to peroxisomes, such as peroxin14 (PEX14), peroxisomal membrane protein 70Kda (PMP70), acyl-CoA oxidase, thiolase, or partially associated to peroxisomes such as Hydroxymethylglutaryl-CoA reductase (HMGCoA-red) and peroxisomal-related proteins, namely PPARalpha, in human glioma specimens at different grades of malignancy. Moreover, Nile red staining of lipid droplets, thin layer chromatography (TLC) and proton nuclear magnetic resonance spectroscopy (NMR) were carried out in order to correlate the biochemical results with the lipid content of tumor tissues. The results obtained indicate that correlating the malignancy grade with the expression of peroxisomal genes and proteins, may constitute a sensitive tool to highlight possible subtypes not recognized by the classical histological techniques.

Practical design of a 4 Tesla double-tuned RF surface coil for interleaved 1H and 23Na MRI of rat brain.

MRI is proving to be a very useful tool for sodium quantification in animal models of stroke, ischemia, and cancer. In this work, we present the practical design of a dual-frequency RF surface coil that provides (1)H and (23)Na images of the rat head at 4 T. The dual-frequency RF surface coil comprised of a large loop tuned to the (1)H frequency and a smaller co-planar loop tuned to the (23)Na frequency. The mutual coupling between the two loops was eliminated by the use of a trap circuit inserted in the smaller coil. This independent-loop design was versatile since it enabled a separate optimisation of the sensitivity and RF field distributions of the two coils. To allow for an easy extension of this simple double-tuned coil design to other frequencies (nuclei) and dimensions, we describe in detail the practical aspects of the workbench design and MRI testing using a phantom that mimics in vivo conditions. A comparison between our independent-loop, double-tuned coil and a single-tuned (23)Na coil of equal size obtained with a phantom matching in vivo conditions, showed a reduction of the (23)Na sensitivity (about 28 %) because of signal losses in the trap inductance. Typical congruent (1)H and (23)Na rat brain images showing good SNR ((23)Na: brain 7, ventricular cerebrospinal fluid 11) and spatial resolution ((23)Na: 1.25 x 1.25 x 5mm(3)) are also reported. The in vivo SNR values obtained with this coil were comparable to, if not better than, other contemporary designs in the literature.

Versatile coil design and positioning of transverse-field RF surface coils for clinical 1.5-T MRI applications.

Clinical MRI/MRS applications require radio frequency (RF) surface coils positioned at an arbitrary angle alpha with respect to B(0). In these experimental conditions the standard circular loop (CL) coil, producing an axial RF field, shows a large signal loss in the central region of interest (ROI). We demonstrate that transverse-field figure-of-eight (FO8) RF surface coils design are not subject to the same amount of signal loss in the central ROI as loop coils when their orientations are changed. The 1.5-T CL and FO8 prototypes (diameter = 10 cm) were built on Plexiglas using copper strips (width = 4 mm, thickness = 100 mum). The two linear elements of the FO8 coil were 1 cm apart. Axial spoiled gradient echo (SPGR) images of a phantom containing doped water were acquired with the coil plane at alpha=0 degrees , 45 degrees , and 90 degrees . As alpha increases, the CL images show, in the central ROI, a signal that decreases from a maximum value to zero. Whereas the FO8 images show, in the same ROI, a signal that varies little from the maximum value (20%). Optimized FO8 coils can be oriented with the coil plane positioned along any direction with respect to B(0) without significant signal loss. Transverse RF coil design should be useful for clinical MRS studies and also for parallel imaging techniques where versatile RF coils disposed along arbitrary directions are required.

Thalamic neurodegeneration in relapsing-remitting multiple sclerosis.

OBJECTIVE: To define the extent of neuronal injury and loss in thalamic gray matter in patients with relapsing-remitting (RR) MS and to characterize how these neuronal pathologic changes are related to disease duration. METHODS: The authors studied 14 patients with RRMS (Expanded Disability Status Scale score, mean 3.25, range 2.0 to 6.0) and 14 (8 men, 6 women) age-matched healthy controls. Structural MR and MRS studies were performed in a single scanning session using a 3T MR system. RESULTS: N-acetylaspartate (NAA) concentrations (a measure of the apparent neuronal density) were decreased approximately 11% in the thalami of the patients with RRMS relative to controls (p < 0.05). The patients with RRMS also had an almost 25% lower mean normalized thalamic volume than controls (p < 0.005). Decreases in thalamic NAA concentration correlated strongly with thalamic volume loss for individual patients (r = 0.85, p < 0.01). Both the NAA concentration (r = -0.48, p = 0.044) and normalized thalamic volume (r = -0.60, p = 0.01) were correlated inversely with disease duration. There was a trend for a correlation between the thalamic NAA/creatine (Cr) ratio and the NAA/Cr in the frontal normal-appearing white matter (r = 0.56, p < 0.08). CONCLUSIONS: The reduction of both NAA concentration and thalamic volume suggests that a neurodegenerative component may contribute to the pathology of MS even in the earlier RR stage. The trend toward a relationship between thalamic NAA/Cr and distant normal-appearing white matter changes implies that there may be a common mechanism for the white matter axonal loss and thalamic neuronal injury.

Radio frequency magnetic field mapping of a 3 Tesla birdcage coil: experimental and theoretical dependence on sample properties.

The RF B(1) distribution was studied, theoretically and experimentally, in phantoms and in the head of volunteers using a 3 T MRI system equipped with a birdcage coil. Agreement between numerical simulation and experiment demonstrates that B(1) distortion at high field can be explained with 3D full-Maxwell calculations. It was found that the B(1) distribution in the transverse plane is strongly dependent on the dielectric properties of the sample. We show that this is a consequence of RF penetration effects combined with RF standing wave effects. In contrast, along the birdcage coil z-axis the B(1) distribution is determined mainly by the coil geometry. In the transverse plane, the region of B(1) uniformity (within 10% of the maximum) was 15 cm with oil, 6 cm with distilled water, 11 cm with saline, and 10 cm in the head. Along z the B(1) uniformity was 9 cm with phantoms and 7 cm in the head.

Compensating for B(1) inhomogeneity using active transmit power modulation.

The effect of poor B(1) homogeneity on MRI images not only affects the appearance of the images, but produces difficulty in automated segmentation and in certain quantification methods. While improved RF coil design is the first line in reducing such artifact, compensation methods can significantly improve the quality of images. Existing methods of compensation typically apply a filter during the image reconstruction. Here a method is presented that compensates for part of the inhomogeneity by actively modulating the RF transmit power as a function of slice position. The method is demonstrated both quantitatively on a phantom and qualitatively on a human brain.

Temporal profile of serum anti-inflammatory and pro-inflammatory interleukins in acute ischemic stroke patients.

The presence of an inflammatory response in the pathophysiology of acute brain ischemia is relatively well established, but less is known about the anti-inflammatory mechanisms. The aim of the present study was to evaluate part of the immune response in acute stroke patients and to analyze a possible correlation with other hematological parameters, clinical outcome, size of infarct and subtypes of strokes. We prospectively studied 42 stroke patients, without signs of infections or inflammatory diseases, at days 0, 1, 3, 7 and 14, and 39 healthy control subjects. We measured serum levels of the anti-inflammatory cytokine interleukin-10 (IL-10) and the pro-inflammatory cytokine interleukin-6 (IL-6) by ELISA method. We observed a highly inverse correlation between these two molecules in control subjects (r=-0.78, p=0.0000001), and this correlation was lost in stroke patients. Patients had significantly lowered IL-10 serum levels soon after the acute event (p=0.00005), with a slight increase at the seventh day. On the other hand, patients had increased IL-6 serum levels compared with controls after day one until day 14 (p<0.04), with a maximum increase at day 3. Interleukin-6 correlated with clinical outcome whereas interleukin-10 did not. Low levels of interleukin-10 indicate that the antiinflammatory response is down-regulated in acute stroke patients. The pro-inflammatory response begins 24 hours after the onset of acute cerebral ischemia, as indicated by the increased serum levels of interleukin-6. The physiological balance between these two molecules is altered in acute stroke patients.

Omega-space adaptive acquisition technique for magnetic resonance imaging from projections.

An omega-space adaptive acquisition technique for MRI from projections is presented. It is based on the evaluation of the information content of a set composed of four initial projections, measured at angles 0 degrees, 45 degrees, 90 degrees, and 135 degrees, followed by the selection of new angles where the information content is maximum. An entropy function is defined on the power spectrum of the projections that is useful for evaluating the information content of each projection. The method makes it possible to reduce the total acquisition time with little degradation of the reconstructed image and it adapts to the arbitrary shape of the sample. For this reason, it can be particularly useful in those applications where acquisition from projections is strongly recommended to save acquisition time, such as functional MRI, imaging of species having very short T(2), or angiography. The method has been tested both on simulated data and on experimental data collected by a commercial MRI apparatus. The method has also been compared to the regular acquisition method, that is, the standard acquisition method in MRI from projections.

Pulsed EPR imaging: image reconstruction using selective acquisition sequences.

Pulsed nuclear magnetic resonance imaging (NMRI) apparatus has developed very quickly. On the other hand, the development of apparatus for pulsed electron paramagnetic resonance imaging (EPRI) has been very slow. This fact is due to the extremely reduced relaxation times of the paramagnetic probes. EPR linewidths are larger than typical NMR linewidths. These large linewidths are also responsible for a substantial worsening of spatial resolution. Due to the brevity of the electronic relaxation times, not all the acquisition/reconstruction techniques currently used in NMRI (such as spin-echo, gradient-echo, etc) can be applied in pulsed EPRI. In fact, the usable sequences in pulsed EPRI are only acquisitions from projections, where it is possible to use stationary magnetic field gradients. Moreover, the use of high fixed magnetic field gradients induces a short decay time constant T2*. The low T2* value can make it impossible for the analogue to digital conversion system (ADC) to reproduce signal variations during the whole acquisition interval and the resolution can worsen. A new pulsed EPRI acquisition sequence from projections, based on selective reception, is presented that is particularly useful in solving the problems of worsening of spatial resolution associated with the use of an ADC. In order to demonstrate the capabilities of our acquisition method, simulated numerical tests will also be reported.

Optimization of axial RF field distribution in low-frequency EPR loop-gap resonators.

A novel coupling method that optimizes the axial RF distribution of low-frequency EPR loop-gap resonators is presented. It consists of a resonant coupling loop positioned at the centre of a two-section loop-gap resonator. This arrangement ensures a symmetrical distribution of the radio frequency field along the axis of the resonator. The design of a central coupling system suitable for EPR resonators operating at about 220 MHz is described. Experimental results show that with the central coupling system the RF field is symmetrical and has a very good axial homogeneity (100% of the resonator length).

Low field (10 mT) pulsed dynamic nuclear polarization.

EPR irradiation by a train of inverting pulses has potential advantages over continuous-wave EPR irradiation in DNP applications; however, it has previously been used only at high field (5 T). This paper presents the design and testing of an apparatus for performing pulsed DNP experiments at 10 mT with large samples (17 ml). Experimental results using pulsed DNP with an aqueous solution of a narrow-linewidth paramagnetic probe are presented. A maximum DNP enhancement of about -36 with a train of inverting pulses (width 500 ns, repetition time 4 micros) was measured. A preliminary comparison showed that, when the same enhancement value is considered, the pulsed DNP technique requires an average power that is about three times higher than that required with the CW irradiation. However, for in vivo DNP applications it is very important to minimize the average power deposited in the sample. From the experimental results reported in this work, when considering the maximum enhancement, the pulsed technique requires only 2% of the average power necessary with the CW DNP technique. We believe that this reduction in the average power can be important for future DNP studies with large biological samples.

Fourier reconstruction as a valid alternative to filtered back projection in iterative applications: implementation of Fourier spectral spatial EPR imaging.

The qualitative equivalence between the Fourier reconstruction (FR) algorithm and the filtered back projection (FBP) algorithm is demonstrated when all the different phase errors that can occur in FR are eliminated. The causes of phase errors are underlined and methods to eliminate them are presented. The practical comparison between FR and FBP has been evaluated on a numerical test image and the results are reported, demonstrating the qualitative equivalence. FR has the advantage of being very computationally efficient. In fact, the time spent to obtain the FR image was 1/20 of that used to obtain the FBP image. Because of the computational efficiency of FR and the good quality of the results obtained, an iterative version of FR has been used to implement the spectral-spatial imaging (SSI) algorithm in the field of electron paramagnetic resonance imaging (EPRI). An experimental example, demonstrating its good performance, is reported.

Two-dimensional 220 MHz Fourier transform EPR imaging.

In the last decade radiofrequency continuous-wave EPR spectrometers have been developed to detect and localize free radicals in vivo. Only recently, pulsed radiofrequency EPR spectrometers have been described for imaging applications with small samples. In the present work, we show the first two-dimensional image obtained at 220 MHz on a large phantom (40 ml) that simulates typical conditions of in vivo EPR imaging. This pulsed EPR apparatus has the potential to make the time required for three-dimensional imaging compatible with the biological half-life of normally used paramagnetic probes.

Nitroxide free radical clearance in the live rat monitored by radio-frequency CW-EPR and PEDRI.

The use of RF (100 to 300 MHz) PEDRI and CW-EPR techniques allows the in vivo study of large animals such as whole rats and rabbits. Recently a PEDRI instrument was modified to also allow CW-EPR spectroscopy with samples of similar size and under the same experimental conditions. In the present study, this CW-EPR and PEDRI apparatus was used to assess the feasibility of the detection of a pyrrolidine nitroxide free radical (2,2,5,5,-tetramethylpyrrolidine-1-oxyl-3-carboxylic acid, PCA) in the abdomen of rats. In particular, we have shown that after the PCA administration (4 mmol kg(-1) b.w.): (i) the PCA EPR linewidth does not show line broadening due to concentration effects; (ii) a similar PCA up-take phase is observed by EPR and PEDRI; and (iii) the PCA half-lives in the whole abdomen of rats measured with the CW-EPR (T1/2=26+/-4 min, mean+/-sd, n=10) and PEDRI (T1/2=29+/-4 min, mean+/-sd, n=4) techniques were not significantly different (p > 0.05). These results show, for the first time, that information about PCA pharmacokinetics obtained by CW-EPR is the same as that from PEDRI under the same experimental conditions.

pH-sensitive imaging by low-frequency EPR: a model study for biological applications.

The use of pH-sensitive nitroxides, in conjunction with low-frequency EPR, offers a unique opportunity for non-invasive assessment of pH values (in the range 0 to 14) in living animals. In the present study, we have investigated the potential use of pH-sensitive nitroxide free radicals in conjunction with EPR imaging techniques at low and very low frequencies (280 MHz-2.1 GHz). In particular, we have measured the hyperfine splitting (hfs) of a pH-sensitive probe at three different EPR frequencies: 280 MHz, 1.1 GHz and 2.1 GHz. We have also developed EPR imaging experiments with phantoms simulating in vivo conditions, using pH-sensitive probes at 280 MHz (spatial-spatial) and 1.1 GHz (spectral-spatial). Finally, we discuss the actual sensitivity/resolution limits of the EPR imaging techniques at low frequencies. Practical applications of this method in the biomedical field are suggested for the continuous and non-invasive localization of pH in vivo.

Plasma and platelet amino acid concentrations in patients affected by major depression and under fluvoxamine treatment.

Plasma and platelet levels of 18 amino acids were measured in 29 outpatients (mean age +/- SD 47.41 +/- 10.85 years; 14 F, 15 M) affected by major depression (DSM IV) and in 28 healthy volunteers (mean age 42.46 +/- 14.19 years; 12 F, 16 M). Plasma and platelet levels of amino acids tended to be higher in depressed patients than in healthy controls. In particular, glutamate, taurine and lysine plasma levels and aspartate, serine and lysine platelet levels were significantly higher. Tryptophan/large neutral amino acids ratio (trp/LNAAs) was significantly lower in depressed patients. Fluvoxamine treatment did not influence plasma and platelet levels of amino acids or trp/LNAAs ratio.

Serotonin metabolism in cluster headache.

Despite some evidence of the involvement of the serotonergic system in cluster headache (CH) pathophysiology, the serotonin (5HT) metabolism has so far been poorly studied. The aim of this study was to investigate plasma and platelet levels of 5HT and 5-hydroxyindoleacetic acid (5HIAA) in CH patients in the active period of the disease. Nineteen CH sufferers and 17 sex- and age-matched healthy controls were studied. CH patients showed significantly higher plasma levels of 5HT and 5HIAA compared to controls (5HT: 5.7+/-6.1 ng/ml vs 0.2+/-0.2 ng/ml; p=0.02; 5HIAA: 34.7+/-46.1 ng/ml vs 0.6+/-0.7 ng/ml; p=0.004). In platelet 5HT levels were slightly reduced in CH patients in comparison with those of control subjects (662.4+/-522.3 ng/10(-8) platelets vs 832.4+/-587.9 ng/10(-8) platelets; n.s.) and 5HIAA levels resulted significantly lower in CH sufferers than in control subjects (3.2+/-2.6 ng/10(-8) platelets vs 6.7+/-4.8 ng/10(-8) platelets; p=0.04). Our data suggest that CH is characterized by an increase of plasma serotonergic metabolism that could reflect an involvement of the central serotonergic system in the pathogenesis of CH.

A Radiofrequency (220-MHz) Fourier Transform EPR Spectrometer

Radiofrequency continuous wave EPR spectrometers for detecting and localizing free radicals in vivo in samples of 50-100 g have been developed. The main limitation of these EPR instruments is the slow acquisition time, and a sensible improvement is expected by the adoption of pulsed EPR techniques. We present here a Fourier transform EPR spectrometer operating at 220 MHz suitable for large volume samples (up to 50 ml). A detailed description of the transmitter and receiver sections, including the EPR resonator, is given. Representative free induction decay data obtained from a sample with a relaxation time of about 900 ns are reported. Copyright 1998 Academic Press.