Parkinson's disease: From bench to bedside.

Parkinson's disease is a very complex public health problem with different etiologies and a wide range of clinical features including motor and non-motor symptoms. The aim of the current report is to review the latest advances in the understanding of PD etiologies, diagnosis, epidemiology and an overview of the earlier and latest advances in the management of PD, from L-DOPA to adult astroglia reprogramming into dopaminergic neurons in vivo.

An electrical characterisation methodology for identifying the switching mechanism in TiO2 memristive stacks.

Resistive random access memories (RRAMs) can be programmed to discrete resistive levels on demand via voltage pulses with appropriate amplitude and widths. This tuneability enables the design of various emerging concepts, to name a few: neuromorphic applications and reconfigurable circuits. Despite the wide interest in RRAM technologies there is still room for improvement and the key lies with understanding better the underpinning mechanism responsible for resistive switching. This work presents a methodology that aids such efforts, by revealing the nature of the resistive switching through assessing the transport properties in the non-switching operation regimes, before and after switching occurs. Variation in the transport properties obtained by analysing the current-voltage characteristics at distinct temperatures provides experimental evidence for understanding the nature of the responsible mechanism. This study is performed on prototyped device stacks that possess common Au bottom electrodes, identical TiO2 active layers while employing three different top electrodes, Au, Ni and Pt. Our results support in all cases an interface controlled transport due to Schottky emission and suggest that the acquired gradual switching originates by the bias induced modification of the interfacial barrier. Throughout this study, the top electrode material was found to play a role in determining the electroforming requirements and thus indirectly the devices' memristive characteristics whilst both the top and bottom metal/oxide interfaces are found to be modified as result of this process.

X-ray spectromicroscopy investigation of soft and hard breakdown in RRAM devices.

Resistive random access memory (RRAM) is considered an attractive candidate for next generation memory devices due to its competitive scalability, low-power operation and high switching speed. The technology however, still faces several challenges that overall prohibit its industrial translation, such as low yields, large switching variability and ultimately hard breakdown due to long-term operation or high-voltage biasing. The latter issue is of particular interest, because it ultimately leads to device failure. In this work, we have investigated the physicochemical changes that occur within RRAM devices as a consequence of soft and hard breakdown by combining full-field transmission x-ray microscopy with soft x-ray spectroscopic analysis performed on lamella samples. The high lateral resolution of this technique (down to 25 nm) allows the investigation of localized nanometric areas underneath permanent damage of the metal top electrode. Results show that devices after hard breakdown present discontinuity in the active layer, Pt inclusions and the formation of crystalline phases such as rutile, which indicates that the temperature increased locally up to 1000 K.

Spatially resolved TiOx phases in switched RRAM devices using soft X-ray spectromicroscopy.

Reduction in metal-oxide thin films has been suggested as the key mechanism responsible for forming conductive phases within solid-state memory devices, enabling their resistive switching capacity. The quantitative spatial identification of such conductive regions is a daunting task, particularly for metal-oxides capable of exhibiting multiple phases as in the case of TiOx. Here, we spatially resolve and chemically characterize distinct TiOx phases in localized regions of a TiOx-based memristive device by combining full-field transmission X-ray microscopy with soft X-ray spectroscopic analysis that is performed on lamella samples. We particularly show that electrically pre-switched devices in low-resistive states comprise reduced disordered phases with O/Ti ratios around 1.37 that aggregate in a ~100 nm highly localized region electrically conducting the top and bottom electrodes of the devices. We have also identified crystalline rutile and orthorhombic-like TiO2 phases in the region adjacent to the main reduced area, suggesting that the temperature increases locally up to 1000 K, validating the role of Joule heating in resistive switching. Contrary to previous studies, our approach enables to simultaneously investigate morphological and chemical changes in a quantitative manner without incurring difficulties imposed by interpretation of electron diffraction patterns acquired via conventional electron microscopy techniques.

Reduced brain choline in homocystinuria due to remethylation defects.

OBJECTIVE: To investigate whether secondary impairment of the transmethylation pathway is a mechanism underlying the neurologic involvement in homocystinuria due to remethylation defects. METHODS: Twelve patients with neurologic disease due to remethylation defects were examined by brain magnetic resonance spectroscopic imaging ((1)H MRSI). Brain N-acetylaspartate, choline-containing compounds (Cho), and creatine (Cr) were quantified and compared to with controls. Metabolites of remethylation cycle and creatine biosynthesis pathway were measured in plasma and urine. RESULTS: MRSI revealed isolated Cho deficiency in all regions examined (mean concentration units +/- SD, patients vs controls): frontal white matter (0.051 +/- 0.010 vs 0.064 +/- 0.010; p = 0.001), lenticular nucleus (0.056 +/- 0.011 vs 0.069 +/- 0.009; p < 0.001), and thalamus (0.063 +/- 0.010 vs 0.071 +/- 0.007; p = 0.006). In contrast to controls, the Cho/Cr ratio decreased with age in patients in the three brain regions examined. Low creatine urinary excretion (p < 0.005), normal urine and plasma guanidinoacetate, and a paradoxical increase in plasma S-adenosylmethionine (p < 0.005) concentrations were observed. CONCLUSION: Patients with homocystinuria due to remethylation defects have an isolated brain choline deficiency, probably secondary to depletion of labile methyl groups produced by the transmethylation pathway. Although biochemical studies suggest mild peripheral creatine deficiency, brain creatine is in the reference range, indicating a possible compartmentation phenomenon. Paradoxical increase of S-adenosylmethionine suggests that secondary inhibition of methylases contributes to the transmethylation defect in these conditions.

Monitoring the effects of mobile phone use on the brain by proton magnetic resonance spectroscopy.

PURPOSE: To determine whether extensive use of mobile phones affects brain metabolites detectable by proton magnetic resonance spectroscopy (1H MRS). MATERIALS AND METHODS: Twenty-one extensive mobile phone users (average use = 5.5 +/- 2.2 years at 2.4 +/- 1.1 hours/day) and 15 control subjects were recruited and submitted to a 1H MRS brain examination at 1.5 Tesla. Data were recorded in the most exposed right temporal and pontobulbar areas as well as in the contralateral left temporal area. The ratios of N-acetylaspartate (NAA), choline (Cho) and myo-inositol (mI) to creatine/phosphocreatine (Cr) were measured. RESULTS: No statistically significant changes in the NAA/Cr, Cho/Cr and mI/Cr ratios were measured between mobile phone users and control subjects and between the exposed and contralateral temporal areas. CONCLUSION: These results indicate that extensive exposition to mobile phone radiation does not cause MRS-detectable brain metabolic changes.

Influence of post-treatment delay on the evaluation of the response to focused ultrasound surgery of breast cancer by dynamic contrast enhanced MRI.

The assessment of the effectiveness of MRI-guided focused ultrasound surgery (MRIgFUS) of breast carcinomas can be performed by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) parameters which monitor the presence of residual tumour. The aim of this study was to evaluate the effect of the post-treatment delay on this assessment. DCE-MRI data were acquired immediately and 3-14 days after MRIgFUS treatment of 26 tumours (<7 days, n = 6; = or > ge;7 days, n = 20). The percentage of residual tumour was determined histologically on the resected mass and correlated with two DCE-MRI parameters: increase in signal intensity (ISI) and positive enhancement integral (PEI). No correlation could be found between DCE-MRI data acquired immediately after treatment and the percentage of residual tumour. Good correlation coefficients were found for data acquired several days after treatment (ISI, r = 0.749; PEI, r = 0.778). However, they were higher when the post-treatment time interval was 7 days or more (ISI, r = 0.962; PEI, r = 0.934). These results suggest that a post-treatment delay of 7 days is necessary for the accurate assessment of the presence of residual tumour by DCE-MRI parameters.

Brain metabolic differences as a function of hemisphere, writing hand preference, and gender.

A total of 35 university-educated normal men (24 right handwriters and 11 left handwriters) and 36 age- and education-matched women (25 right handwriters and 11 left handwriters) underwent a proton magnetic resonance spectroscopy examination in seven 8 cm(3) voxels including the right and left frontal lobe tips, the right and left mid-temporal lobes, the right and left thalami, and the hypothalamus. Dependent measures were N-acetylaspartate (NAA), choline-containing compounds (Cho) and creatine/phosphocreatine (Cr) metabolite peak area ratios relative to total H(2)O. As expected, thalamic grey matter contained higher NAA ratios than telencephalic voxels (containing white and grey matter) (p < .001). The thalamic Cr/ H(2)O ratio was higher on the right, but the opposite asymmetry was observed for the temporal lobe (p < .05). Women had a higher left frontal NAA/ H(2)O ratio than men, but men had a higher hypothalamic NAA/ H(2)O ratio than women. Right-handers had a higher temporal lobe NAA/H(2)O ratio than left-handers, particularly in the left hemisphere. In addition, several significant 2- and 3-way interactions between writing hand preference, gender, and hemisphere were observed, but only in the frontal lobe.

Long-term brain metabolic alterations in exogenous Cushing's syndrome as monitored by proton magnetic resonance spectroscopy.

The effects of exogenous Cushing's syndrome on the brain metabolism were investigated by proton magnetic resonance spectroscopy (MRS). Thirteen patients having been treated for 2 to 22 years with prednisone were recruited. On the average, none of the metabolites (NAA, Cr, Cho and mI) were significantly different from those of 40 normal subjects in any of the three regions studied: frontal area, thalamus and temporal area. However, the Cho/H(2)O ratios were found to decrease significantly in the thalamic area as a function of treatment period (-1.3%/year). In the frontal and temporal areas, decreases of the Cho/H(2)O ratios were measured with treatment period but they did not reach statistical significance. Effects on Cho levels can be related to those observed for patients with endogenous Cushing's syndrome and suggest an impairment at the membrane level. The Cho/H(2)O reductions were not found to be dose- or age-dependent. Other metabolite ratios did not vary with treatment period, dose or age.

Comparison of metabolite levels and water diffusion between cortical and subcortical strokes as monitored by MRI and MRS.

UNLABELLED: Labelle M, Khiat A, Durocher A, et al. Comparison of metabolite levels and water diffusion between cortical and subcortical strokes as monitored by MRI and MRS. Invest Radiol 2001;36:155-163. RATIONALE AND OBJECTIVES: Proton magnetic resonance spectroscopy (MRS) and functional imaging techniques are increasingly recognized as useful tools for the characterization of strokes. The aim of this study was to compare cortical and subcortical (lacunar) strokes by MRS and diffusion-weighted imaging (DWI) experiments as a function of time. METHODS: Single-voxel MRS, DWI, and perfusion-weighted imaging data were recorded on patients with cortical (n = 7) or subcortical (n = 7) strokes in the acute, subacute, and chronic periods. Magnetic resonance spectra were acquired in three regions: hyperintense DWI area, adjacent area with normal DWI intensity, and contralateral area. Neurological deficits were estimated by the National Institutes of Health Stroke Scale. RESULTS: Decreases in N-acetylaspartate, choline-containing compounds, and creatine/phosphocreatine signal intensity as well as the presence of lactate were observed at all times in the hyperintense DWI area of all lesions. Small decreases were measured in the subacute and chronic phases for the adjacent area of cortical strokes but not for the adjacent area of subcortical strokes. The existence of a surrounding affected area in subcortical strokes is deduced from a combination of MRS and DWI results, possibly corresponding to the ischemic penumbra. Differences were found between the two types of lesion, especially an increased time variability of apparent diffusion coefficients in subcortical strokes. CONCLUSIONS: Magnetic resonance spectroscopy provides evidence for the existence of affected tissue outside the hyperintense DWI regions in subcortical strokes. Cortical and subcortical strokes display different DWI and MRS characteristics.

Conformational analysis of endothelin-1 analogs with indolizidinone amino acids incorporated at the C-terminus.

In light of the fact that a beta-turn conformation at the C-terminus of endothelin-1 (ET-1) could be responsible for activity at the ET(B) receptor, the incorporation of (3S, 6S, 9S)-2-oxo-3-amino-1-azabicyclo[4.3.0]nonane-9-carboxylic acid (IAA) should provide an elegant method to establish whether a formyl group on Trp21 plays a role in stabilizing a beta-turn. Eight linear ET-1 analogs, four formylated and four nonformylated, ET-1-(Leu17-Asp18-IAA-Trp21); ET-1-(Leu17-IAA-Ile20-Trp21); ET-1-(Leu17-Asp18-Pro19-Ile20-Trp21) and ET-1-(Leu17-Asp 8-Ile19-Pro20-Trp21) have been analyzed by high-resolution nuclear magnetic resonance (NMR) spectroscopy and molecular modeling. Two-dimensional double quantum filtered correlation spectroscopy (DQFCOSY), total correlation spectroscopy (TOCSY) and nuclear Overhausen enhancement spectroscopy (NOESY) were resolved and analyzed for each molecule. Interspatial distance constraints were derived from the intensity of the NOESY connectivities, The formation of hydrogen bonding was monitored from the temperature-dependence of the NH chemical shifts. Molecular models calculated by means of distance geometry, simulated annealing and energy minimization, suggested a global elongated structure for the formylated analogs and a folded arrangement for the nonformylated derivatives, but no hydrogen bonding was detected at the C-terminus of ET-1 analogs.

Role of phospholipase A(2) on the variations of the choline signal intensity observed by 1H magnetic resonance spectroscopy in brain diseases.

Phospholipase A(2) catalyzes the hydrolysis of membrane glycerophospholipids leading to the production of metabolites observable by both 1H and 31P magnetic resonance spectroscopy. The signal of choline-containing compounds (Cho) observed by 1H magnetic resonance spectroscopy is constituted of metabolites of phosphatidylcholine, especially phosphocholine (PCho) and glycerophosphocholine (GPCho). The phosphomonoester (PME) and phosphodiester (PDE) signals observed by 31P magnetic resonance spectroscopy are, respectively, precursors and catabolites of phospholipids. A large number of brain diseases have been reported to cause variations in the intensity of the Cho, PME and PDE signals. Changes in the activity of phospholipase A(2) have been measured in many brain diseases. In this review, the relationships between the results of 1H and 31P magnetic resonance spectroscopy and the phospholipase A(2) assays are analyzed. In many brain diseases, the variation in the Cho signal intensity can be correlated with a stimulation or inhibition of the phospholipase A(2) activity.

Recovery of the brain choline level in treated Cushing's patients as monitored by proton magnetic resonance spectroscopy.

In a previous study from our group [A. Khiat, C. Bard, A. Lacroix, J. Rousseau, Y. Boulanger, Brain metabolic alterations in Cushing's syndrome as monitored by proton magnetic resonance spectroscopy, NMR Biomed. 12 (1999) 357-363], proton magnetic resonance spectroscopy (1H MRS) was used to evaluate changes in cerebral metabolites in patients with Cushing's syndrome as compared to normal subjects. Data recorded in the frontal, thalamic and temporal areas demonstrated statistically significant decreases of the Cho/Cr ratios in the frontal and thalamic areas but not in the temporal area for Cushing's syndrome patients. No statistically significant changes in the NAA/Cr ratios were measured in any of the areas studied. In this follow-up study, MRS data are reported for ten patients after correction of hypercortisolism which demonstrate a statistically significant recovery of the choline levels in the frontal and thalamic areas. No variation in the NAA, Cr and mI metabolite ratios relative to H(2)O could be measured. Results are interpreted as an inhibition of the phosphatidylcholine degrading phospholipases by glucocorticoids which disappears after correction of hypercortisolism.

Brain metabolic alterations in Cushing's syndrome as monitored by proton magnetic resonance spectroscopy.

Proton magnetic resonance spectroscopy ((1)H MRS) was used to evaluate changes in cerebral metabolites in 13 patients with Cushing's syndrome (including seven with pituitary corticotroph adenomas and six with primary adrenal disease) as compared to 40 normal subjects. Data were recorded in the frontal, thalamic and temporal areas; quantification of the MRS signals demonstrated a statistically significant decrease of the Cho/Cr ratio in the frontal and thalamic areas but not in the temporal area for patients with Cushing's syndrome. The largest decrease in Cho/Cr was measured in the thalamic area of patients with a Cushing's syndrome secondary to an adrenal disease. No statistically significant changes in the NAA/Cr ratio were measured in any of the areas studied. These results suggest that the quantification of choline levels could be helpful for monitoring the cerebral metabolite alterations in patients with hypercortisolism.

Conformational analysis of biologically active truncated linear analogs of endothelin-1 using NMR and molecular modeling.

Some linear truncated analogs of endothelin-1 display potent agonistic activity at the ET(B) receptor, especially when the side chain of Trp21 is N-formylated. Then, the three-dimensional arrangements of six structurally reduced linear analogs, three formylated and three nonformylated, have been investigated by high resolution NMR spectroscopy and molecular modeling, in order to pinpoint the conformational features related to the biological activity. Two-dimensional double-quantum-filtered correlation spectroscopy (DQFCOSY), total correlation spectroscopy (TOCSY) and nuclear Overhauser enhancement spectroscopy (NOESY) were recorded and analyzed for each molecule. Interspatial distance constraints were derived from the intensity of the NOESY connectivities. The formation of hydrogen bonding was monitored from the temperature dependence of the NH chemical shifts. Molecular models calculated by means of distance geometry, simulated annealing and energy minimization, using the NMR constraints, strongly suggested a global elongated structure for the formylated analogs exhibiting biological activity, and a folded arrangement for the unformylated derivatives. Homology comparisons allowed the identification of a beta-turn-like folding of the C-terminal segment Asp18-Trp21 as a probable key-factor for activity.

Identification of the motilide pharmacophores using quantitative structure activity relationships.

Erythromycin A and some derivatives have been shown to act as agonists at the motilin receptor site (motilides) and a structural similarity between these molecules and the N-terminal fragment of motilin has been proposed. Conformational analysis and three-dimensional quantitative structure-activity relationship (3D-QSAR) methods have been used to determine the homology between a series of erythromycin A derivatives and motilin 1-10. A total of 18 compounds has been studied to correlate the gastrointestinal motor stimulating (GMS) activity with the structure-related parameters determined by 3D-QSAR. Two models with good predictive power of the GMS activity are presented, leading to the prediction of motilin 1-10 activity. The models are consistent with the majority of the data available. The most significant parameters for GMS activity are a favorable dispersion interaction from the quaternary ammonium group of the desosamine ring. In motilin 1-10, the aromatic side chains of Phe1 and Tyr7 seem to play the same role as the quaternary ammonium group in models 1 and 2, respectively. Some hydroxyl groups of erythromycin A derivatives and hydrophobic groups of the Val2 and lle4 side chains of motilin also contribute to the GMS activity. The experimental GMS activities measured are in good agreement with the predicted values, with correlation coefficient values of 0.98 and 0.94 in models 1 and 2, respectively.

Three-dimensional structure of the Y1 receptor agonist [Leu31, Pro34]NPY as determined by NMR and molecular modeling.

The solution structure of the Y1 receptor agonist, porcine [Leu31, Pro34]NPY, has been investigated by two-dimensional NMR and molecular modeling. A complete assignment of the NMR resonances was achieved and 201 inter-residue nuclear Overhauser enhancement spectroscopy (NOESY) connectivities could be identified, comprising several connectivities between the N- and C-terminal segments. A molecular model was calculated by distance geometry, simulated annealing and conjugate gradients energy minimization using the NOE constraints. The results indicate that, like NPY and other peptides of the family, [Leu31, Pro34]NPY adopts a folded hairpin structure with the terminal segments in close proximity. Analysis of the secondary chemical shifts for the CH(alpha)'s and of the temperature dependence of the NH chemical shifts combined with the NOE constraints indicates a tendency toward helix structure for the segment 18-30 and the presence of turn structure for the C-terminal segment (residues 31-36). Native NPY and [Leu31, Pro34]NPY have most of their structures in common but differ slightly in their C-terminal portion. Based on the structures of NPY and of its specific agonists, [Leu31, Pro34]NPY and NPY 13-36, conclusions can be drawn about the structural requirements for binding to the Y1 and Y2 receptor subtypes.

Structural comparison of alanine-substituted analogues of the calcitonin gene-related peptide 8-37. Importance of the C-terminal segment for antagonistic activity.

Replacement of specific residues of the antagonistic fragment human calcitonin gene-related peptide 8-37 (hCGRP 8-37) by alanine residues produces good antagonists to CGRP1 receptors when the replacement is made at positions 17 and 20 but a poor antagonist when the replacement is made at position 21. The solution structures of hCGRP 8-37 and of the three alanine analogues have been determined by two-dimensional 1H NMR spectroscopy and molecular modeling. Following the complete assignment of the NMR spectra, a comparison of the chemical shifts and of the temperature dependence of the amide chemical shifts showed that these parameters differed for [Ala17]-hCGRP 8-37 and [Ala20]-hCGRP 8-37 relative to hCGRP 8-37 in the N-terminal and central segments but not in the C-terminal segment (residues 31-37). In the case of [Ala21]-hCGRP 8-37, differences were observed all along the chain. Molecular modeling calculations were performed by distance geometry, simulated annealing and energy minimization using NOE distance constraints. Molecular models showed a structural homology between [Ala17]-hCGRP 8-37, [Ala20]-hCGRP 8-37 and hCGRP 8-37 in the C-terminal segment Asn31-Phe37 as well as hydrogen bonding between Val28 and Asn31. These structural similarities are not observed with [Ala21]-hCGRP 8-37. Therefore, the structure of the C-terminal segment of hCGRP 8-37 appears to be critical for antagonistic activity at CGRP1 receptors.

Structural differences between the free and bound states of the DNA-bisintercalating peptide YSPTSPSY.

The YSPTSPSY peptide is a DNA-bisintercalator that can adopt nonrandom conformations in solution. Strategies based on random conformational search and energy minimizations have been applied to generate populations of conformers characterizing YSPTSPSY. Subsequent analysis based on statistical methods and clustering allowed to determine the existence of four classes of conformers containing beta- and/or gamma-turns. NMR spectra of YSPTSPSY in solution provide evidence for such structures. Employing a Monte Carlo-based docking procedure, the YSPTSPSY peptide was docked in a DNA double-helical fragment with the sequence [d(GACGTC)]2. The peptide binds on the minor groove of DNA stacking the central CG base pairs, in a manner similar to that observed in complexes of triostin A with DNA. Upon binding, the structure of the C-terminal segment is modified into a type I beta-turn. Five intermolecular hydrogen bonds are observed, but the van der Waals interactions constitute the major stabilization factor for the complex. NMR chemical shifts, coupling constants, and NOESY connectivities are in agreement with the molecular model.

Structural effects of the selective reduction of amide carbonyl groups in motilin 1-12 as determined by nuclear magnetic resonance.

Motilin is a 22-residue peptide stimulating stomach and intestinal motility. The motilin 1-12 fragment displays biological effects similar to the native peptide. Selective reduction of the amide carbonyl groups to form CH2NH analogs leads to a significant reduction in activity for the first two N-terminal positions and to a complete loss of activity for all other positions. The structures of motilin 1-12 and ten reduced analogs were investigated using the temperature dependence of the amide NH chemical shifts. In all the analogs, the structure of the N-terminal region (residues 1-5) was different from the structure of motilin 1-12, which is characterized by hydrogen bonding between Phe1 and Ile4. The structure of the C-terminal region of analogs was similar to the structure of motilin 1-12 for the first two reduction positions only (1-2 and 2-3), indicating that the C-terminal portion of motilin 1-12 is more critical for biological activity. Complete structural characterizations of motilin 1-12, [CH2NH]1-2, and [CH2NH]4-5-motilin 1-12 were performed by two-dimensional NMR spectroscopy and molecular modeling. The structural features observed confirm the differences based on the temperature dependence of the amide NH chemical shifts. These results demonstrate that conservation of the amide bond rigidity is essential for the activity of non-hydrolyzable analogs.