WIN-induced vesiculation cooperates to the inhibition of osteosarcoma cell migration

Introduction. Animal cells release vesicles that mediate the secretion of a variety of factors in the surrounding environment affecting neighboring cells. There is increasing evidence that secreted vesicles play an important role as vehicle of intercellular communication in different biological systems and are able to influence both physiological and pathological processes. Recently, we have reported that the synthetic cannabinoid WIN55,512 is able to induce osteosarcoma MG63 cell death and negatively affect cell migration. Here, we study the effects of WIN on the induction of vesicle secretion and their possible role in WIN-dependent reduction of osteosarcoma cell migratory ability. Methods. Vesicles from MG63 cells were obtained by ultracentrifuging at 140,000g media derived from cell cultures untreated and treated for 24 h with 5 uM WIN. Purified vesicles were quantified by cytofluorimetry and by detecting acetilcholinesterase activity according to established criteria. Scratch wound healing assay was employed to monitor cell migration toward the center of a gap created in a cell monolayer. Zymographic analysis was used to evaluate metalloproteinase activities in the vesicles. Results. WIN treatment induced a significant increase (about 4-fold) in the number of vesicles released by osteosarcoma cells. Wound healing assay showed that in the presence of vesicles from WIN-treated cultures, cells only partially filled the gap with respect to those conditioned with vesicles isolated from control cells which closed the gap within about 24 h. Furthermore, zymography assay showed a reduced activity of MMP-2 and MMP-9 in the vesicles obtained from WIN-treated cells. Conclusion. Data indicate that the increase in the number of vesicles released after WIN treatment and/or their probable different composition can be responsible for the relevant inhibition of MG63 cell migration induced by the cannabinoid

Predictions as a window into learning: Anticipatory fixation offsets carry more information about environmental statistics than reactive stimulus-responses

published February 19, 2019A core question underlying neurobiological and computational models of behavior is how individuals learn environmental statistics and use them to make predictions. Most investigations of this issue have relied on reactive paradigms, in which inferences about predictive processes are derived by modeling responses to stimuli that vary in likelihood. Here we deployed a novel anticipatory oculomotor metric to determine how input statistics impact anticipatory behavior that is decoupled from target-driven-response. We implemented transition constraints between target locations, so that the probability of a target being presented on the same side as the previous trial was 70% in one condition (pret70) and 30% in the other (pret30). Rather than focus on responses to targets, we studied subtle endogenous anticipatory fixation offsets (AFOs) measured while participants fixated the screen center, awaiting a target. These AFOs were small (<0.4° from center on average), but strongly tracked global-level statistics. Speaking to learning dynamics, trial-by-trial fluctuations in AFO were well-described by a learning model, which identified a lower learning rate in pret70 than pret30, corroborating prior suggestions that pret70 is subjectively treated as more regular. Most importantly, direct comparisons with saccade latencies revealed that AFOs: (a) reflected similar temporal integration windows, (b) carried more information about the statistical context than did saccade latencies, and (c) accounted for most of the information that saccade latencies also contained about inputs statistics. Our work demonstrates how strictly predictive processes reflect learning dynamics, and presents a new direction for studying learning and prediction.We thank Leonardo Chelazzi for his comments. UH's work was conducted in part while serving at and with support of the National Science Foundation. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the NSF. The study was partially funded by a European Research Council grant to UH (ERC-STG 263318)

Involvement of PAR-4 in Cannabinoid-Dependent Sensitization of Osteosarcoma Cells to TRAIL-Induced Apoptosis

The synthetic cannabinoid WIN 55,212-2 is a potent cannabinoid receptor agonist with anticancer potential. Experiments were performed to determine the effects of WIN on proliferation, cell cycle distribution, and programmed cell death in human osteosarcoma MG63 and Saos-2 cells. Results show that WIN induced G2/M cell cycle arrest, which was associated with the induction of the main markers of ER stress (GRP78, CHOP and TRB3). In treated cells we also observed the conversion of the cytosolic form of the autophagosome marker LC3-I into LC3-II (the lipidated form located on the autophagosome membrane) and the enhanced incorporation of monodansylcadaverine and acridine orange, two markers of the autophagic compartments such as autolysosomes. WIN also induced morphological effects in MG63 cells consisting in an increase in cell size and a marked cytoplasmic vacuolization. However, WIN effects were not associated with a canonical apoptotic pathway, as demonstrated by the absence of specific features, and only the addition of TRAIL to WIN-treated cells led to apoptotic death probably mediated by up-regulation of the tumor suppressor factor PAR-4, whose levels increased after WIN treatment, and by the translocation of GRP78 on cell surface

Predictions as a window into learning:Anticipatory fixation offsets carry more information about environmental statistics than reactive stimulus-responses

published February 19, 2019A core question underlying neurobiological and computational models of behavior is how individuals learn environmental statistics and use them to make predictions. Most investigations of this issue have relied on reactive paradigms, in which inferences about predictive processes are derived by modeling responses to stimuli that vary in likelihood. Here we deployed a novel anticipatory oculomotor metric to determine how input statistics impact anticipatory behavior that is decoupled from target-driven-response. We implemented transition constraints between target locations, so that the probability of a target being presented on the same side as the previous trial was 70% in one condition (pret70) and 30% in the other (pret30). Rather than focus on responses to targets, we studied subtle endogenous anticipatory fixation offsets (AFOs) measured while participants fixated the screen center, awaiting a target. These AFOs were small (<0.4° from center on average), but strongly tracked global-level statistics. Speaking to learning dynamics, trial-by-trial fluctuations in AFO were well-described by a learning model, which identified a lower learning rate in pret70 than pret30, corroborating prior suggestions that pret70 is subjectively treated as more regular. Most importantly, direct comparisons with saccade latencies revealed that AFOs: (a) reflected similar temporal integration windows, (b) carried more information about the statistical context than did saccade latencies, and (c) accounted for most of the information that saccade latencies also contained about inputs statistics. Our work demonstrates how strictly predictive processes reflect learning dynamics, and presents a new direction for studying learning and prediction.We thank Leonardo Chelazzi for his comments. UH's work was conducted in part while serving at and with support of the National Science Foundation. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the NSF. The study was partially funded by a European Research Council grant to UH (ERC-STG 263318)

Direct Scaling of Measure on Vortex Shedding through a Flapping Flag Device in the Open Channel around a Cylinder at Re ∼ 10^3: Taylor’s Law Approach.

none8noThe problem of vortex shedding, which occurs when an obstacle is placed in a regular flow, is governed by Reynolds and Strouhal numbers, known by dimensional analysis. The present work aims to propose a thin films-based device, consisting of an elastic piezoelectric flapping flag clamped at one end, in order to determine the frequency of vortex shedding downstream an obstacle for a flow field at Reynolds number Re∼103 in the open channel. For these values, Strouhal number obtained in such way is in accordance with the results known in literature. Moreover, the development of the voltage over time, generated by the flapping flag under the load due to flow field, shows a highly fluctuating behavior and satisfies Taylor’s law, observed in several complex systems. This provided useful information about the flow field through the constitutive law of the device.openSamuele De Bartolo, Massimo De Vittorio, Antonio Francone, Francesco Guido, Elisa Leone, Vincenzo Mariano Mastronardi, Andrea Notaro, Giuseppe Roberto TomasicchioDE BARTOLO, Samuele; DE VITTORIO, Massimo; Francone, Antonio; Guido, Francesco; Leone, Elisa; Mariano Mastronardi, Vincenzo; Notaro, Andrea; Tomasicchio, Giusepp

Parthenolide and Its Soluble Analogues: Multitasking Compounds with Antitumor Properties

Due to its chemical properties and multiple molecular effects on different tumor cell types, the sesquiterpene lactone parthenolide (PN) can be considered an effective drug with significant potential in cancer therapy. PN has been shown to induce either classic apoptosis or alternative caspase-independent forms of cell death in many tumor models. The therapeutical potential of PN has been increased by chemical design and synthesis of more soluble analogues including dimethylaminoparthenolide (DMAPT). This review focuses on the molecular mechanisms of both PN and analogues action in tumor models, highlighting their effects on gene expression, signal transduction and execution of different types of cell death. Recent findings indicate that these compounds not only inhibit prosurvival transcriptional factors such as NF-κB and STATs but can also determine the activation of specific death pathways, increasing intracellular reactive oxygen species (ROS) production and modifications of Bcl-2 family members. An intriguing property of these compounds is its specific targeting of cancer stem cells. The unusual actions of PN and its analogues make these agents good candidates for molecular targeted cancer therapy

Autophagic cell death induced by Litchi fruit extracts in human colon cancer cells.

Litchi chinensis is a tropical fruit which cultivation has been recently introduced in Sicily. Some findings have shown that Litchi extracts display antitumor effects but the underlying mechanisms have not been elucidated. This study focuses on the effects of Litchi hydro-alcoholic extracts in colorectal cancer cells. The results indicated that Litchi exocarp (peel), mesocarp (pulp) and endocarp (seeds) extracts reduce the viability of HT-29 colon cancer cells in a dose dependent manner. This effect was accompanied with G2/M arrest of the cell cycle followed by cell death. Interestingly, exocarp and endocarp extracts triggered an autophagic response in the first phase of treatment (16-24h) with increase in the protein levels of ATG1/ Ulk kinase, p62 and LC3-II accumulation. In particular, time point evaluation indicated that with exocarp extracts the autophagic form LC3II appeared at 16h treatment whereas with endocarp extracts accumulated at 48h. ATG1/Ulk and p62 increased early at 16 h treatment in both cases and p62 remained elevated until 48h. On the other hand, autophagic effects were not observed with Litchi mesocarp extracts which directly induced caspase 3-dependent apoptosis. Polyphenol colorimetric assay showed that the Litchi extracts contain significant amounts of these compounds, particularly in exocarp and endocarp. Accordingly, treatment of cells with gallic acid, one bioactive component of polyphenols, mimicked the effects of the Litchi extracts. Taken together, these preliminary results provides in vitro evidence that litchi extracts activate the autophagic process preceding cell death and can thus be considered as potential chemopreventive agents for colorectal cancer

Hypertrophy and ER Stress Induced by Palmitate Are Counteracted by Mango Peel and Seed Extracts in 3T3-L1 Adipocytes

A diet rich in saturated fatty acids (FAs) has been correlated with metabolic dysfunction and ROS increase in the adipose tissue of obese subjects. Thus, reducing hypertrophy and oxidative stress in adipose tissue can represent a strategy to counteract obesity and obesity-related diseases. In this context, the present study showed how the peel and seed extracts of mango (Mangifera indica L.) reduced lipotoxicity induced by high doses of sodium palmitate (PA) in differentiated 3T3-L1 adipocytes. Mango peel (MPE) and mango seed (MSE) extracts significantly lowered PA-induced fat accumulation by reducing lipid droplet (LDs) and triacylglycerol (TAGs) content in adipocytes. We showed that MPE and MSE activated hormone-sensitive lipase, the key enzyme of TAG degradation. In addition, mango extracts down-regulated the adipogenic transcription factor PPARγ as well as activated AMPK with the consequent inhibition of acetyl-CoA-carboxylase (ACC). Notably, PA increased endoplasmic reticulum (ER) stress markers GRP78, PERK and CHOP, as well as enhanced the reactive oxygen species (ROS) content in adipocytes. These effects were accompanied by a reduction in cell viability and the induction of apoptosis. Interestingly, MPE and MSE counteracted PA-induced lipotoxicity by reducing ER stress markers and ROS production. In addition, MPE and MSE increased the level of the anti-oxidant transcription factor Nrf2 and its targets MnSOD and HO-1. Collectively, these results suggest that the intake of mango extract-enriched foods in association with a correct lifestyle could exert beneficial effects to counteract obesity

Spike timing in rat somatosensory cortex contributes to behavior

The precise spike timing of sensory neurons carries more information about stimuli than the number of spikes counted over longer time windows [1-3]. However, the behavioral relevance of this extra information remains to be assessed. To investigate whether precise timing contributes to behavior, we considered spike trains recorded extracellularly in the rat somatosensory cortex during a texture discrimination task [4,5]. We analyzed the spike trains following the contact of the whiskers with the texture and we measured the information carried about texture by post-touch spike times and spike counts. This analysis is technical challenging because it is difficult to quantify the information content of precise patterns of spike that extend over relatively long time windows with the relatively little number of trials available in behavioral experiments. To address this challenge, we reduced the dimensionality of the response space by decomposing single trial spike train densities into Principal Components (PCs), selecting the most informative PC as the "temporal template" to interpret neural responses, and finally measuring the information about the presented texture carried by the projection of each trial relative to this template. We compared the so -measured spike timing information with the information carried by spike counts and we found that spike times carried more than 60% more texture information than spike counts. Next, we reasoned that if a neuronal coding mechanism contributes to behavior, then the code would be found to transmit less stimulus information in error trials, i.e. trials in which the rat made the wrong choice. We found that the texture information carried both by spike counts and spike patterns decreased significantly when we included error trials in the analysis, but the effect was much more pronounced for patterns (30% information decrease) than for counts (~15% information decrease). These differences between spike pattern and spike count codes hold for both primary and secondary somatosensory cortex, suggesting that spike timing could contribute to behavior at different stages of the sensory processing. Taken together, these results indicate that in the somatosensory cortex the time at which spikes occur, not just the number of spikes, makes a crucial contribution to tactile perception

Psychological treatments and psychotherapies in the neurorehabilitation of pain. Evidences and recommendations from the italian consensus conference on pain in neurorehabilitation

BACKGROUND: It is increasingly recognized that treating pain is crucial for effective care within neurological rehabilitation in the setting of the neurological rehabilitation. The Italian Consensus Conference on Pain in Neurorehabilitation was constituted with the purpose identifying best practices for us in this context. Along with drug therapies and physical interventions, psychological treatments have been proven to be some of the most valuable tools that can be used within a multidisciplinary approach for fostering a reduction in pain intensity. However, there is a need to elucidate what forms of psychotherapy could be effectively matched with the specific pathologies that are typically addressed by neurorehabilitation teams. OBJECTIVES: To extensively assess the available evidence which supports the use of psychological therapies for pain reduction in neurological diseases. METHODS: A systematic review of the studies evaluating the effect of psychotherapies on pain intensity in neurological disorders was performed through an electronic search using PUBMED, EMBASE, and the Cochrane Database of Systematic Reviews. Based on the level of evidence of the included studies, recommendations were outlined separately for the different conditions. RESULTS: The literature search yielded 2352 results and the final database included 400 articles. The overall strength of the recommendations was medium/low. The different forms of psychological interventions, including Cognitive-Behavioral Therapy, cognitive or behavioral techniques, Mindfulness, hypnosis, Acceptance and Commitment Therapy (ACT), Brief Interpersonal Therapy, virtual reality interventions, various forms of biofeedback and mirror therapy were found to be effective for pain reduction in pathologies such as musculoskeletal pain, fibromyalgia, Complex Regional Pain Syndrome, Central Post-Stroke pain, Phantom Limb Pain, pain secondary to Spinal Cord Injury, multiple sclerosis and other debilitating syndromes, diabetic neuropathy, Medically Unexplained Symptoms, migraine and headache. CONCLUSIONS: Psychological interventions and psychotherapies are safe and effective treatments that can be used within an integrated approach for patients undergoing neurological rehabilitation for pain. The different interventions can be specifically selected depending on the disease being treated. A table of evidence and recommendations from the Italian Consensus Conference on Pain in Neurorehabilitation is also provided in the final part of the pape