Atypical “seizure-like” activity in cortical reverberating networks in vitro can be caused by LPS-induced inflammation: a multi-electrode array study from a hundred neurons

We show here that a mild sterile inflammation induced by the endotoxin lipopolysaccharide (LPS), in a neuron/astrocyte/microglial cortical network, modulates neuronal excitability and can initiate long-duration burst events resembling epileptiform seizures, a recognized feature of various central nervous neurodegenerative, neurological and acute systemic diseases associated with neuroinflammation. To study this action, we simultaneously analyzed the reverberating bursting activity of a hundred neurons by using in vitro multi-electrode array (MEA) methods. ~5 hours after LPS application, we observed a net increase in the average number of spikes elicited in engaged cells and within each burst, but no changes neither in spike waveforms nor in burst rate. This effect was characterized by a slow, two-fold exponential increase of the burst duration and the appearance of rarely occurring long-burst events that were never seen during control recordings. These changes and the time-course of microglia-released proinflammatory cytokine, tumor necrosis factor-alpha (TNF-α), were blocked by pre-treatment with 50 nM minocycline, an established anti-inflammatory agent which was inactive when applied alone. Assay experiments also revealed that application of 60 pM exogenous TNF-α after 12-15 h, produced non-washable changes of neuronal excitability, completely different from those induced by LPS, suggesting that TNF-α release alone was not responsible for our observed findings. Our results indicate that the link between neuroinflammation and hyperexcitability can be unveiled by studying the long-term activity of in vitro neuronal/astrocyte/microglial networks

The Trifluoromethyl Group as a Bioisosteric Replacement of the Aliphatic Nitro Group in CB1 Receptor Positive Allosteric Modulators (PAMs)

The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jmedchem.9b00252. Experimental procedures, characterization of all intermediates and target compounds, and copies of NMR spectra of compounds 1, 39-57. Molecular formula strings of target compounds are available. ACKNOWLEDGEMENTS. We gratefully thank Signal Pharma and the Canadian Institutes of Health Research Proof of Principle grants PPP-125784 and PP2-139101 for financial support and fellowship (C.C.T), NIH grants R01DA039942, P30DA033934 and VCU School of Pharmacy start-up funds (A.H.L.). We thank the EPSRC National Crystallography Service (University of Southampton) for the X-ray data collection. We are grateful to Dr Monica Sani (CNR-ICRM, Milan, Italy) and Mr Massimo Frigerio (Politecnico di Milano, Italy) for the synthesis of two tetrazole-substituted indoles (Het-1 and Het-2)Peer reviewedPostprin

The interaction between alpha 7 nicotinic acetylcholine receptor and nuclear peroxisome proliferator-activated receptor-alpha represents a new antinociceptive signaling pathway in mice

Recently, alpha 7 nicotinic acetylcholine receptors (nAChRs), primarily activated by binding of orthosteric agonists, represent a target for anti-inflammatory and analgesic drug development. These receptors may also be modulated by positive allosteric modulators (PAMs), ago-allosteric ligands (ago-PAMs), and alpha 7-silent agonists. Activation of 00 nAChRs has been reported to increase the brain levels of endogenous ligands for nuclear peroxisome proliferator-activated receptors type-alpha (PPAR-alpha), palmitoylethanolamide (PEA) and oleoylethanolamide (OEA), in a Ca2+-dependent manner. Here, we investigated potential crosstalk between alpha 7 nAChR and PPAR-alpha, using the formalin test, a mouse model of tonic pain. Using pharmacological and genetic approaches, we found that PNU282987, a full alpha 7 agonist, attenuated formalin-induced nociceptive behavior in alpha 7 -dependent manner. Interestingly, the selective PPAR-alpha antagonist GW6471 blocked the antinociceptive effects of PNU282987, but did not alter the antinociceptive responses evoked by the alpha 7 nAChR PAM PNU120596, ago-PAM GAT107, and silent agonist NS6740. Moreover, GW6471 administered systemically or spinally, but not via the intraplantar surface of the formalin-injected paw blocked PNU282987-induced antinociception. Conversely, exogenous administration of the naturally occurring PPAR-alpha agonist PEA potentiated the antinociceptive effects of PNU282987. In contrast, the cannabinoid 031 antagonist rimonabant and the CB2 antagonist SR144528 failed to reverse the antinociceptive effects of PNU282987. These findings suggest that PPAR-alpha plays a key role in a putative antinociceptive alpha 7 nicotinic signaling pathway.United States Department of Health & Human Services National Institutes of Health (NIH) - USA - GM57481 - R01 CA206028United States Department of Health & Human Services National Institutes of Health (NIH) - USA NIH National Cancer Institute (NCI) - R01CA206028United States Department of Health & Human Services National Institutes of Health (NIH) - USA NIH National Institute of General Medical Sciences (NIGMS) - R01GM057481United States Department of Health & Human Services National Institutes of Health (NIH) - USA NIH National Institute on Drug Abuse (NIDA) European Commission - T32DA00702

The Endogenous Cannabinoid System: A Budding Source of Targets for Treating Inflammatory and Neuropathic Pain

A great need exists for the development of new medications to treat pain resulting from various disease states and types of injury. Given that the endogenous cannabinoid (that is, endocannabinoid) system modulates neuronal and immune cell function, both of which play key roles in pain, therapeutics targeting this system hold promise as novel analgesics. Potential therapeutic targets include the cannabinoid receptors, type 1 and 2, as well as biosynthetic and catabolic enzymes of the endocannabinoids N-arachidonoylethanolamine and 2-arachidonoylglycerol. Notably, cannabinoid receptor agonists as well as inhibitors of endocannabinoid-regulating enzymes fatty acid amide hydrolase and monoacylglycerol lipase produce reliable antinociceptive effects, and offer opioid-sparing antinociceptive effects in myriad preclinical inflammatory and neuropathic pain models. Emerging clinical studies show that ‘medicinal’ cannabis or cannabinoid-based medications relieve pain in human diseases such as cancer, multiple sclerosis, and fibromyalgia. However, clinical data have yet to demonstrate the analgesic efficacy of inhibitors of endocannabinoid-regulating enzymes. Likewise, the question of whether pharmacotherapies aimed at the endocannabinoid system promote opioid-sparing effects in the treatment of pain reflects an important area of research. Here we examine the preclinical and clinical evidence of various endocannabinoid system targets as potential therapeutic strategies for inflammatory and neuropathic pain conditions

Liposomal Delivery of Diacylglycerol Lipase-Beta Inhibitors to Macrophages Dramatically Enhances Selectivity and Efficacy <i>in Vivo</i>

Diacylglycerol lipase-beta (DAGLβ) hydrolyzes arachidonic acid (AA)-containing diacylglycerols to produce bioactive lipids including endocannabinoids and AA-derived eicosanoids involved in regulation of inflammatory signaling. Previously, we demonstrated that DAGLβ inactivation using the triazole urea inhibitor KT109 blocked macrophage inflammatory signaling and reversed allodynic responses of mice in inflammatory and neuropathic pain models. Here, we tested whether we could exploit the phagocytic capacity of macrophages to localize delivery of DAGLβ inhibitors to these cells <i>in vivo</i> using liposome encapsulated KT109. We used DAGLβ-tailored activity-based probes and chemical proteomic methods to measure potency and selectivity of liposomal KT109 in macrophages and tissues from treated mice. Surprisingly, delivery of ∼5 μg of liposomal KT109 was sufficient to achieve ∼80% inactivation of DAGLβ in macrophages with no apparent activity in other tissues <i>in vivo</i>. Our macrophage-targeted delivery resulted in a >100-fold enhancement in antinociceptive potency compared with free compound in a mouse inflammatory pain model. Our studies describe a novel anti-inflammatory strategy that is achieved by targeted <i>in vivo</i> delivery of DAGLβ inhibitors to macrophages

RACCOMANDAZIONI PER L\u2019ASSISTENZA ALLA DONNA VITTIMA DI VIOLENZA SESSUALE

Di fronte ad una vittima di violenza sessuale la priorit\ue0 assistenziale dovr\ue0 essere la tutela della sua salute e del suo benessere. \uc8 importante restituire alla donna il suo valore di persona in ogni fase del percorso clinico. Trattare la donna con rispetto ed empatia pu\uf2 essere di aiuto nella successiva elaborazione del trauma. \u2022 L\u2019esame fisico e la raccolta delle prove dovrebbero avvenire nello stesso tempo per evitare visite ripetute e lo stress a queste correlato. La completezza dell\u2019esame comporta un inevitabile impegno di tempo e di risorse da parte del personale sanitario. \u2022 Sar\ue0 importante riservare una stanza predisposta per accogliere la vittima per tutto il tempo che rimane nella struttura (in caso di Ospedale la stanza dovr\ue0 essere preferibilmente nel punto di ac\u2011 cesso e cio\ue8 il Pronto Soccorso). Il personale dovr\ue0 rivolgersi alla vittima con voce calma, senza esprimere sorpresa o incredulit\ue0, con parole e atteggiamento assolutamente non giudicante. \u2022 Per aiutare il sanitario ad applicare correttamente le procedure diagnostiche e terapeutiche validate, \ue8 suggerito l\u2019impiego di una SCHEDA CLINICA GUIDATA che si applica alle ragazze di et\ue0 superiore ai 13 anni e alle donne adulte. \u2022 La scheda costituisce documentazione clinica da archiviare e da consegnare eventualmente alla donna in copia per gli usi che ritiene opportuni (per es. la denuncia) oltre al verbale di Pronto Soc\u2011 corso che viene abitualmente compilato. \uc8 inoltre uno strumento che favorisce la raccolta di dati epidemiologici per lo studio del fenomeno. \u2022 Si dovr\ue0 ottenere il consenso per tutta la procedura e per la comunicazione delle informazioni a terzi. Le domande e le scelte della donna saranno assecondate in ogni fase. Nel caso in cui il personale sanitario debba procedere con la denuncia d\u2019ufficio all\u2019Autorit\ue0 Giudiziaria, la donna deve esserne informata ma non \ue8 richiesto consenso. \u2022 La scheda clinica guidata dovr\ue0 inserirsi in una PROCEDURA di accoglienza pi\uf9 ampia sviluppata in modo MULTIDISCIPLINARE che preveda un iter specifico fin dal momento in cui la donna incontra la struttura (generalmente il pronto soccorso) per la risposta standardizzata a ogni problematica presente in caso di violenza sessuale. In particolare si dovr\ue0 definire un protocollo per il metodo di raccolta e conservazione delle prove forensi, un protocollo per la profilassi dell\u2019HIV, un protocollo per i test tossi\u2011 cologici dato che spesso la violenza \ue8 facilitata dall\u2019uso di sostanze, infine un protocollo per la presa in carico successiva della vittima che non pu\uf2 essere abbandonata dopo la prima valutazione. \u2022 La costituzione di una RETE multidisciplinare composta dalle varie competenze e risorse presenti nello specifico ambito territoriale garantir\ue0 la coerenza della presa in carico in fase iniziale e di quella nei tempi successivi, per una tutela della vittima non solo sanitaria ma anche psicologica e sociale nonch\ue9 legale. La legge prevede la denuncia a querela di parte entro dodici mesi, quando non sono presenti le condizioni per denuncia d\u2019ufficio