RESUMO
Cytotoxic agents synergize with immune checkpoint inhibitors and improve outcomes for patients with several cancer types. Nonetheless, a parallel increase in the incidence of dose-limiting side effects, such as peripheral neuropathy, is often observed. Here, we investigated the role of the programmed cell death-1 (PD-1)/programmed death-ligand 1 (PD-L1) axis in the modulation of paclitaxel-induced neuropathic pain. We found that human and mouse neural tissues, including the dorsal root ganglion (DRG), expressed basal levels of PD-1 and PD-L1. During the development of paclitaxel-induced neuropathy, an increase in PD-L1 expression was observed in macrophages from the DRG. This effect depended on Toll-like receptor 4 activation by paclitaxel. Furthermore, PD-L1 inhibited pain behavior triggered by paclitaxel or formalin in mice, suggesting that PD-1/PD-L1 signaling attenuates peripheral neuropathy development. Consistent with this, we observed that the combined use of anti-PD-L1 plus paclitaxel increased mechanical allodynia and chronic neuropathy development induced by single agents. This effect was associated with higher expression of inflammatory markers (Tnf, Il6, and Cx3cr1) in peripheral nervous tissue. Together, these results suggest that PD-1/PD-L1 inhibitors enhance paclitaxel-induced neuropathic pain by suppressing PD-1/PD-L1 antinociceptive signaling.
Assuntos
Antineoplásicos Fitogênicos , Neuralgia , Ratos , Humanos , Camundongos , Animais , Receptor de Morte Celular Programada 1 , Antineoplásicos Fitogênicos/efeitos adversos , Ratos Sprague-Dawley , Neuralgia/induzido quimicamente , Neuralgia/metabolismo , Paclitaxel , Analgésicos/efeitos adversosRESUMO
Cannabidiol (CBD) is a natural compound with psychoactive therapeutic properties well described. Conversely, the immunological effects of CBD are still poorly explored. In this study, the potential anti-inflammatory effects and underlying mechanisms of CBD and its analog Dimethyl-Heptyl-Cannabidiol (DMH-CBD) were investigated using RAW 264.7 macrophages. CBD and DMH-CBD suppressed LPS-induced TNF production and NF-kB activity in a concentration-dependent manner. Both compounds reduced the NF-kB activity in a µM concentration range: CBD (IC50â¯=â¯15⯵M) and DMH-CBD (IC50â¯=â¯38⯵M). However, the concentrations of CBD that mediated NF-kB inhibition were similar to those that cause cytotoxicity (LC50â¯=â¯58⯵M). Differently, DMH-CBD inhibited the NF-kB activation without cytotoxic effects at the same concentrations, although it provokes cytotoxicity at long-term exposure. The inhibitory action of the DMH-CBD on NF-kB activity was not related to the reduction in IkBα degradation or either p65 (NF-kB) translocation to the nucleus, although it decreased p38 MAP kinase phosphorylation. Additionally, 8-(3-Chlorostyryl) caffeine (CSC), an A2A antagonist, reversed the effect of DMH-CBD on NF-kB activity in a concentration-dependent manner. Collectively, our results demonstrated that CBD reduces NF-kB activity at concentrations intimately associated with those that cause cell death, whereas DMH-CBD decreases NF-kB activity at non-toxic concentrations in an A2A receptor dependent-manner.
Assuntos
Agonistas do Receptor A2 de Adenosina/farmacologia , Canabidiol/análogos & derivados , Canabidiol/farmacologia , Macrófagos/efeitos dos fármacos , NF-kappa B/metabolismo , Receptor A2A de Adenosina/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , Agonistas do Receptor A2 de Adenosina/toxicidade , Animais , Canabidiol/química , Canabidiol/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Concentração Inibidora 50 , Macrófagos/metabolismo , Macrófagos/patologia , Camundongos , Fosforilação , Células RAW 264.7 , Receptor A2A de Adenosina/metabolismo , Via Secretória , Transdução de Sinais , Células THP-1 , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
BJ-32 (also known as BjcuL) is a C-type lectin from the venom of Bothrops jararacussu with specificity for beta-galactosides and a remarkable ability to agglutinate several species of trypanosomatids. Our objective was to study the oligomerization state of native BJ-32 by using different biophysical and computational methods. Small-angle X-ray light scattering (SAXS) experiments disclosed a compact, globular protein with a radius of gyration of 36.72+/-0.04A and molecular weight calculated as 147.5+/-2.0kDa. From analytical ultracentrifugation analysis, it was determined that the BJ-32 sedimentation profile fits nicely to a decamer model. The analysis of the intrinsic emitted fluorescence spectra for BJ-32 solutions indicated that association of subunits in the decamer is accompanied by changes in the environment of Tryptophan residues. Both ab initio and comparative models of BJ-32 supported the resemblance of the decamer in the crystallographic structure from a close homologue, the rattlesnake venom lectin (RSL) from Crotalus atrox.
Assuntos
Bothrops , Venenos de Crotalídeos/química , Lectinas Tipo C/química , Sequência de Aminoácidos , Animais , Dicroísmo Circular , Simulação por Computador , Venenos de Crotalídeos/análise , Venenos de Crotalídeos/isolamento & purificação , Lectinas Tipo C/análise , Lectinas Tipo C/isolamento & purificação , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Quaternária de Proteína , Espalhamento a Baixo Ângulo , Alinhamento de Sequência , Espectrometria de Massas por Ionização por Electrospray , UltracentrifugaçãoRESUMO
In the catalytic cycle of skeletal muscle, myosin alternates between strongly and weakly bound cross-bridges, with the latter contributing little to sustained tension. Here we describe the action of DMSO, an organic solvent that appears to increase the population of weakly bound cross-bridges that accumulate after the binding of ATP, but before P(i) release. DMSO (5-30%, v/v) reversibly inhibits tension and ATP hydrolysis in vertebrate skeletal muscle myofibrils, and decreases the speed of unregulated F-actin in an in vitro motility assay with heavy meromyosin. In solution, controls for enzyme activity and intrinsic tryptophan fluorescence of myosin subfragment 1 (S1) in the presence of different cations indicate that structural changes attributable to DMSO are small and reversible, and do not involve unfolding. Since DMSO depresses S1 and acto-S1 MgATPase activities in the same proportions, without altering acto-S1 affinity, the principal DMSO target apparently lies within the catalytic cycle rather than with actin-myosin binding. Inhibition by DMSO in myofibrils is the same in the presence or the absence of Ca(2+) and regulatory proteins, in contrast with the effects of ethylene glycol, and the Ca(2+) sensitivity of isometric tension is slightly decreased by DMSO. The apparent affinity for P(i) is enhanced markedly by DMSO (and to a lesser extent by ethylene glycol) in skinned fibres, suggesting that DMSO stabilizes cross-bridges that have ADP.P(i) or ATP bound to them.
Assuntos
Proteínas Contráteis/metabolismo , Dimetil Sulfóxido/farmacologia , Contração Isométrica/fisiologia , Músculo Esquelético/fisiologia , Miofibrilas/fisiologia , Fosfatos/farmacologia , Trifosfato de Adenosina/metabolismo , Animais , ATPase de Ca(2+) e Mg(2+)/metabolismo , Cálcio/metabolismo , ATPases Transportadoras de Cálcio/metabolismo , Galinhas , Proteínas Contráteis/efeitos dos fármacos , Ácido Edético/farmacologia , Etilenoglicol/farmacologia , Técnicas In Vitro , Contração Isométrica/efeitos dos fármacos , Cinética , Magnésio/metabolismo , Músculo Esquelético/efeitos dos fármacos , Miofibrilas/efeitos dos fármacos , Subfragmentos de Miosina/metabolismo , Miosinas/metabolismo , CoelhosRESUMO
Serology has been the most popular method to diagnose toxoplasmosis. Accordingly, this study standardizes an enzyme-linked immunosorbent assay (ELISA) and compares its results with the IFI technique. In the IgG detection test, the standardized technique presented a sensibility (S) of 96.77%, a specificity (SP) of 75%, with a positive predictive value (PPV) of 83.33%, a negative predictive value (NPV) of 94.74%, and an adjusted concordance (K) of 73.50%. The IFI exhibited 83.87% for S, 79.16% for SP, 83.81% for PPV, 79. 16% for NPV, and 63% for K. The rough concordance between these two tests (ELISA/IFI) was 88.35% for the IgG detection test and 81.55% for the IgM detection test. K was 70.82% and 1.31% for IgG and IgM, respectively, the correlation index (r) being 0.556 for IgG and -0. 023 for IgM. We can conclude that standardized ELISA-IgG is indicated in serologic selection processes, whereas the ELISA-IgM is not recommended for presenting low values for the adjusted concordance with the reference technique, which suggests not very reliable results.