Proteomics analysis of vesicles isolated from plasma and urine of prostate cancer patients using a multiplex, aptamer-based protein array

Proteomics analysis of biofluid-derived vesicles holds enormous potential for discovering non-invasive disease markers. Obtaining vesicles of sufficient quality and quantity for profiling studies has, however, been a major problem, as samples are often replete with co-isolated material that can interfere with the identification of genuine low abundance, vesicle components. Here, we used a combination of ultracentrifugation and size-exclusion chromatography to isolate and analyse vesicles of plasma or urine origin. We describe a sample-handling workflow that gives reproducible, quality vesicle isolations sufficient for subsequent protein profiling. Using a semi-quantitative aptamer-based protein array, we identified around 1,000 proteins, of which almost 400 were present at comparable quantities in plasma versus urine vesicles. Significant differences were, however, apparent with elements like HSP90, integrin αVβ5 and Contactin-1 more prevalent in urinary vesicles, while hepatocyte growth factor activator, prostate-specific antigen–antichymotrypsin complex and many others were more abundant in plasma vesicles. This was also applied to a small set of specimens collected from men with metastatic prostate cancer, highlighting several proteins with the potential to indicate treatment refractory disease. The study provides a practical platform for furthering protein profiling of vesicles in prostate cancer, and, hopefully, many other disease scenarios

Fundam Clin Pharmacol

BACKGROUND: Due to its psychoactive effects, ketamine has become a drug used for non-medical purpose. OBJECTIVES: To assess the latest trends in ketamine use among people with substance use disorder and to characterize its clinical complications using complementary health data sources of the French Addictovigilance Network. METHODS: First, we extracted all reports involving ketamine from 2012 to 2021 from the database of the OPPIDUM program (i.e., a multicentric program conducted in collaboration with hundreds of substance abuse treatment facilities that collects data on drugs used by subjects with substance use disorders). We described the reports globally and the changes from 2012 to 2021. Second, we extracted all cases involving ketamine from July 2020 to December 2022 from the French National Pharmacovigilance Database (BNPV). We identified the cases related to ketamine use among people with substance use disorder and described them. RESULTS: There was a 2.5-fold increase in the number of ketamine users with substance use disorder in the OPPIDUM program, from 35 (0.7%) subjects in 2012 to 89 (1.7%) subjects in 2021. There was an increase in the proportion of subjects who were daily users, had distress upon discontinuation, and presented addiction. There were 238 cases related to ketamine use among people with substance use disorder in the French National Pharmacovigilance Database from July 2020 to December 2022. Among them, 94 (39.5%) cases involved ketamine use disorder, 20 (8.4%) cases involved urinary tract and kidney symptoms, and 13 (5.5%) cases involved hepatobiliary symptoms. CONCLUSION: The trend observed over 10 years reflects the growth in ketamine use among people with substance use disorder, although it does not allow to estimate the rates of non-medical use of ketamine in the general population. Ketamine-induced uropathy and cholangiopathy are reported in ketamine users with substance use disorder, especially in case of repeated and/or prolonged use of high doses

Comportement mécanique des tranches fines de silicium pour applications photovoltaïques : Influence de la qualité du matériau et de sa découpe en tranche

The crystalline silicon wafer is the key component of the solar cell and accounts for a significant portion of the total photovoltaic (PV) module cost. Reducing wafer thickness is therefore a privileged pathway to decrease solar energy production costs. Maintaining low breakage rates when processing such thin samples remains however challenging. In this context, it is essential to improve our understanding of the mechanisms responsible for wafer embrittlement and failure. This work investigates the mechanical properties of silicon wafers obtained using diamond wire sawing. We developed a mechanical characterization methodology suited for these thin, brittle samples, combining destructive tests with 4-line bending, biaxial bending and dynamic impacts. In parallel, finite element simulations were implemented to better understand the underlying phenomena. Tests performed on as-cut, chemically etched and annealed samples revealed that the most critical damage regarding mechanical failure is located within a thin subsurface layer (less than 3 µm), which properties are controlled by the sawing step. Through an extensive characterization campaign on wafers with different thicknesses (from 180 to 100 µm), we demonstrated that thinner samples exhibit an increased bending flexibility without alteration of their intrinsic mechanical strength, accompanied however by a higher risk of failure following an impact. Finally, we highlighted that the presence of structural defects in multicrystalline and mono-like silicon is indirectly responsible for the lower fracture strength of the wafers: the increased suffering of the diamond wire when cutting through these defects generates indeed deeper microcracks.Le wafer de silicium cristallin est le composant clé de la cellule solaire et représente une part significative du prix du module photovoltaïque. La réduction de l’épaisseur des wafers offre donc une voie privilégiée pour diminuer les coûts de production de l’énergie solaire. Le maintien de faibles taux de casse lors de la manipulation de ces fines plaquettes reste cependant un obstacle majeur. Dans ce contexte, il est primordial d’améliorer notre compréhension des mécanismes de fragilisation et de rupture des wafers. Ce travail étudie les propriétés mécaniques des wafers de silicium obtenus par découpe au fil diamanté. Nous avons développé une méthodologie de caractérisation mécanique adaptée à l’extrême fragilité de ces échantillons, en combinant des essais de rupture en flexion 4-lignes, biaxiale ainsi que des sollicitations dynamiques par chocs. En parallèle, des simulations par la méthode des éléments finis ont été implémentées afin de mieux comprendre les phénomènes en jeu. Des essais réalisés sur des échantillons bruts de découpe, attaqués chimiquement et recuits thermiquement ont révélé que l’endommagement le plus critique pour la défaillance mécanique se situe dans une couche de faible épaisseur inférieur à 3 µm) sous la surface, dont les propriétés sont contrôlées par l’étape de découpe. Au travers d’une vaste campagne de caractérisation sur des wafers de différentes épaisseurs (de 180 à 100 µm), nous avons montré que l’amincissement des plaquettes permet un gain de flexibilité sans diminution de la résistance mécanique intrinsèque, mais qui s’accompagne d’un risque plus élevé de rupture suite à un impact sur la tranche. Enfin, nous avons mis en évidence que les défauts structurels dans le silicium multicristallin et mono-like sont indirectement responsables de la diminution de la résistance à rupture des wafers : la difficulté accrue du fil à traverser ces défauts se traduit par des microfissures plus profondes

Impact on memory of chronic and acute benzodiazepine and/or ethanol intoxication : from rodents models

La prise concomitante de benzodiazépines et d’alcool est fréquemment décrite, que ce soit lors de situationsaiguës ou au long cours. Une des préoccupations majeures liée à l’usage de ces substances est l’impact sur lamémoire, ces deux substances ayant en commun une facilitation de la transmission GABAergique, impliquéedans les processus mnésiques. Plusieurs études épidémiologiques ont suggéré un lien entre l’utilisationchronique de benzodiazépines et le développement de troubles cognitifs, mais ces données sont encorecontroversées. De même, l’effet sur la mémoire d’une prise conjointe de benzodiazépines/alcool, chronique ouaiguë, est peu connu. Dans ce contexte, l’étude de modèles animaux prend tout son sens pour surmonter leslimites des études cliniques (biais de confusion, déclaration, attrition, limites éthiques des études prospectives).L’objectif de cette thèse était d’évaluer l’impact sur la mémoire d’une intoxication aiguë et chronique auxbenzodiazépines et/ou à l'éthanol à partir de modèles murins.L’impact de la prise au long cours de diazépam (benzodiazépine à demi-vie longue) pendant 16 semaines surla mémoire a d’abord été évalué chez la souris mâle en prenant en compte des facteurs liés à l’âge (souris de 6et 12 mois) et à la posologie de diazépam administré (15mg/kg/j et 30mg/kg/j). L’évaluation a été réalisée àl’aide de tests comportementaux à huit semaines de traitement puis une semaine après l’arrêt. Malgré un impactdu diazépam sur la mémoire de travail pendant le traitement, l'absence d’altération rémanente dans lesdifférents types de mémoires évaluées suggère que le diazépam seul n'entraîne pas d'effets délétèresirréversibles. L’impact du diazépam associé à l’éthanol a ensuite été étudié chez des souris âgées de 12 mois enutilisant les mêmes tests comportementaux que dans l’étude précédente, associés à des données despectroscopie par résonance magnétique (SRM) permettant d’évaluer les niveaux de certains métabolitescérébraux dans l’hippocampe. Cette étude n’a pas mis en évidence d’effet additif des benzodiazépines et del’alcool sur la mémoire. Seule une altération de l’apprentissage au cours de la phase d’acquisition d’un testévaluant la mémoire de référence spatiale a été mise en évidence semblant affecter les groupes traités paréthanol par rapport au groupe contrôle, et plus particulièrement le groupe diazépam + éthanol. Une modificationdes métabolites a été constatée en cours de traitement, notamment chez les souris traitées par éthanol, sanseffet additif significatif et avec une réversibilité trois mois après arrêt du traitement.L’évaluation d’une intoxication aiguë a été réalisée chez le rat mâle adulte. Tout d’abord, l’évolution desmétabolites cérébraux après intoxication à l’éthanol a été évaluée dans le cortex préfrontal en utilisant la SRM.Une évolution dose dépendante des métabolites préfrontaux a été mise en évidence, avec une diminution desniveaux de GABA après une intoxication à 1g/kg d’éthanol et une diminution des niveaux de choline après uneintoxication à 2g/kg d’éthanol. L’effet additif d’une intoxication aiguë à l’éthanol et ou aux benzodiazépines, à lafois au décours mais également à distance de la prise, a ensuite été évalué en utilisant des données de SRMhippocampiques et en les couplant à des données comportementales. Si cette étude a bien mis en évidence uneimprégnation éthylique hippocampique au décours immédiat de l’intoxication et un niveau de GABA et deglutamate abaissés 4 semaines après dans le groupe éthanol seul, elle n’a pas montré d’effet additif de ces deuxsubstances au niveau comportemental ou spectroscopique lors de l’évaluation à distance.Au total, ce travail n’a pas permis de démontrer un effet additif de l’éthanol et du diazépam sur lesdifférentes mémoires évaluées. Cependant, l’hypothèse d’un tel effet additif ne peut être écartée au vu desmodifications comportementales et spectroscopiques induites par ces substances dans notre étude.The concomitant use of benzodiazepines and alcohol is frequently described, whether in acute or long-termsituations. One of the major concerns related to the use of these substances is the impact on memory, as bothsubstances have in common a facilitation of GABAergic transmission, involved in memory processes. Severalepidemiological studies have suggested a link between chronic benzodiazepine use and the development ofcognitive disorders, but these data are still controversial. Similarly, little is known about the effect on memory ofchronic or acute benzodiazepine/alcohol co-administration. In this context, the study of animal models makessense to overcome the limitations of clinical studies (confounding bias, reporting, attrition, ethical limitations ofprospective studies). The objective of this thesis was to evaluate the impact on memory of acute and chronicintoxication with benzodiazepines and/or ethanol using mouse models.The impact of long-term administration of diazepam (long half-life benzodiazepine) for 16 weeks on memorywas first evaluated in male mice considering factors related to age (6- and 12-months old mice) and the dosageof diazepam administered (15 mg/kg/d and 30 mg/kg/d). The evaluation was performed using behavioral testsat eight weeks of treatment and one week after discontinuation. Despite an impact of diazepam on workingmemory during treatment, the absence of persistent impairment in the different types of memory assessedsuggests that diazepam alone does not cause irreversible deleterious effects. The impact of diazepam combinedwith ethanol was then studied in 12-month-old mice using the same behavioral tests as in the previous study,combined with magnetic resonance spectroscopy (MRS) data to assess levels of certain brain metabolites in thehippocampus. This study did not show an additive effect of benzodiazepines and alcohol on memory. Only animpairment of learning during the acquisition phase of a test evaluating spatial reference memory wasdemonstrated, which seemed to affect the ethanol-treated groups compared to the control group, and moreparticularly the diazepam + ethanol group. A change in metabolites was observed during treatment, particularlyin the ethanol-treated mice, without significant additive effect and with reversibility three months after cessationof treatment.The evaluation of acute intoxication was performed in adult male rats. First, the evolution of brainmetabolites after ethanol intoxication was assessed in the prefrontal cortex using MRS. A dose-dependentevolution of prefrontal metabolites was demonstrated, with a decrease in GABA levels after 1g/kg ethanolintoxication and a decrease in choline levels after 2g/kg ethanol intoxication. The additive effect of acute ethanoland/or benzodiazepine intoxication, both during and at a distance from the intake, was then evaluated usinghippocampal MRS data and coupling them to behavioral data. Although this study showed a hippocampal ethylimpregnation immediately after intoxication and a lowered level of GABA and glutamate 4 weeks later in theethanol alone group, it did not show an additive effect of these two substances at the behavioral or spectroscopiclevel during the remote evaluation.In total, this work did not demonstrate an additive effect of ethanol and diazepam on the different memoriesevaluated. However, the hypothesis of such an additive effect cannot be ruled out in view of the behavioral andspectroscopic changes induced by these substances in our study

Towards personalized pharmacology: Antipsychotics and schizophrenia

International audienc

Comparative analysis of mechanical strength of diamond-sawn silicon wafers depending on saw mark orientation, crystalline nature and thickness

International audienc

What Is the Potential for Abuse of Lisdexamfetamine in Adults? A Preclinical and Clinical Literature Review and Expert Opinion

INTRODUCTION: Lisdexamfetamine dimesylate (LDX) is a prodrug approved for attention deficit/hyperactivity disorder and for moderate-to-severe binge eating disorder in adults in some countries. AREA COVERED: We aimed to specify the abuse potential of LDX in adults, using a review of pharmacokinetic/pharmacodynamic (PK/PD), animal, clinical, and pharmaco-epidemiological studies, through a PubMed search since inception until May 2021 using the following keywords: "lisdexamfetamine AND ('misuse' OR 'abuse' OR 'diversion' OR 'addiction')". EXPERT OPINION: Most of the studies highlighted a longer Tmax than dexamphetamine leading to a delayed onset of effects and a decreased Cmax. These PK parameters were often associated with a diminished feeling of euphoria, in comparison to immediate-release dexamphetamine. The potential for abuse was also limited by the prodrug property of LDX, thus reducing the risk of misuse. Nevertheless, all the data were not convergent, as some authors reported similar Cmax for LDX and dexamphetamine and reinforcing properties with a dose-dependent effect. Epidemiological studies found that abuse rates of LDX were substantially lower than those of immediate-release dexamphetamine. Overall, although LDX abuse seems possible, we did not find evidence concerning current safety signal. However, more long-term pharmaco-epidemiological studies are still needed to confirm this finding

Acute psychosis related to baclofen in a patient treated for binge eating disorder highlights the urgent need to regulate off-label prescriptions

International audienc

Pharmaceutical cognitive doping in students: a chimeric way to get-a-head?

International audienceFor students, the pressing demands for memorization, top-level performance, and peer competition create an environment favorable for pharmaceutical cognitive doping behavior. We aimed to describe recent practices and the benefit/risk ratio of such behavior and to discuss the issues at stake. The prevalence of pharmaceutical cognitive doping among students has been reported from 1.3% to 33% across studies, with variations depending on country and definition of pharmaceutical cognitive doping. The therapeutic classes most frequently cited as being diverted for doping purposes are psychostimulants and nootropics (methylphenidate, modafinil, piracetam), corticosteroids, sedative drugs and beta-blockers. Some illegal substances such as cannabis, amphetamines and cocaine are also consumed in order to boost mental function. Finally, over-the-counter products, such as caffeine-based tablets or energy drinks, or alcohol, are also widely used by students whose motivations involve enhanced performance, concentration, memory, and staying awake during the revision and exam period. However, the expected (often fantasized) effectiveness of these products does not correspond to the reality of a modest controversial impact on cognitive performance. There appears to be an emerging profile of the student more inclined to doping behavior. Cognitive doping thus raises the question of its regulation, opening a debate opposing, on one hand, individual freedom and supposed collective benefits and, on the other hand, health consequences, educational (in)equality, and the risk of tarnished academic success. Strengthening school and university medicine, through prevention campaigns and the identification of subjects at risk, is essential to limit the extent, risk, and damages associated with such practices