NLRP3 inflammasome-mediated neutrophil recruitment and hypernociception depend on leukotriene B4 in a murine model of gout

Objective Deposition of monosodium urate monohydrate (MSU) crystals in the joints promotes an intense inflammatory response and joint dysfunction. This study evaluated the role of the NLRP3 inflammasome and 5-lipoxygenase (5-LOX)derived leukotriene B4 (LTB4) in driving tissue inflammation and hypernociception in a murine model of gout. Methods. Gout was induced by injecting MSU crystals into the joints of mice. Wild-type mice and mice deficient in NLRP3, ASC, caspase 1, interleukin-1 beta (IL-1 beta), IL-1 receptor type I (IL-1RI), IL-18R, myeloid differentiation factor 88 (MyD88), or 5-LOX were used. Evaluations were performed to assess neutrophil influx, LTB4 activity, cytokine (IL-1 beta, CXCL1) production (by enzyme-linked immunosorbent assay), synovial microvasculature cell adhesion (by intravital microscopy), and hypernociception. Cleaved caspase 1 and production of reactive oxygen species (ROS) were analyzed in macrophages by Western blotting and fluorometric assay, respectively. Results. Injection of MSU crystals into the knee joints of mice induced neutrophil influx and neutrophildependent hypernociception. MSU crystal-induced neutrophil influx was CXCR2-dependent and relied on the induction of CXCL1 in an NLRP3/ASC/caspase 1/IL-1 beta/MyD88-dependent manner. LTB4 was produced rapidly after injection of MSU crystals, and this was necessary for caspase 1-dependent IL-1 beta production and consequent release of CXCR2-acting chemokines in vivo. In vitro, macrophages produced LTB4 after MSU crystal injection, and LTB4 was relevant in the MSU crystalinduced maturation of IL-1 beta. Mechanistically, LTB4 drove MSU crystal-induced production of ROS and ROS-dependent activation of the NLRP3 inflammasome. Conclusion. These results reveal the role of the NLRP3 inflammasome in mediating MSU crystalinduced inflammation and dysfunction of the joints, and highlight a previously unrecognized role of LTB4 in driving NLRP3 inflammasome activation in response to MSU crystals, both in vitro and in vivo.

Taxonomic and distributional notes on Telebasis Selys, 1865, with a redescription of T. pallida Machado, 2010, and an evaluation of the T. racenisi Bick & Bick, 1995 "complex" of species (Odonata, Coenagrionidae)

A full checklist of the species of Telebasis Selys, 1865, housed in the Brazilian collections Colecao Entomologica Prof. Jose Alfredo Pinheiro Dutra, Departamento de Zoologia, Instituto de Biologia, Universidade Federal do Rio do Janeiro (DZRJ), and Museu de Zoologia, Universidade de Sao Paulo (MZSP) is presented. A total of 325 specimens representing 19 species were studied. Ten new records for Brazilian States were found for T. carmesina Calvert, 1909 (Rio de Janeiro and Rio Grande do Sul), T. corallina (Selys, 1876) (Pernambuco), T. demarara (Williamson, 1917) (Maranhao), T. filiola (Perty, 1834) (Paraiba and Santa Catarina), T. gigantea Daigle, 2002 (Sao Paulo), T. inalata (Calvert, 1961) (Mato Grosso do Sul), T. pallida Machado, 2010 (Goias) and T. obsoleta (Selys, 1876) (Mato Grosso do Sul), as well as a new record of T. carminita Calvert, 1909 for Suriname. Telebasis pallida Machado, 2010 is redescribed and diagnosed based on 14 males collected near the type locality, and its genital ligula is described and illustrated for the first time. Furthermore, the status of the three species of the Telebasis racenisi Bick & Bick, 1995 complex is evaluated. Of these, Telebasis pareci Machado, 2010 syn. n. is proposed as junior subjective synonym of Telebasis lenkoi Machado, 2010, and a possible synonymy among the three species is discussed under T. racenisi. ((c) 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Leukotriene B4-loaded microspheres: a new therapeutic strategy to modulate cell activation

Abstract Background Leukotriene B4 (LTB4) is a potent inflammatory mediator that also stimulates the immune response. In addition, it promotes polymorphonuclear leukocyte phagocytosis, chemotaxis, chemokinesis and modulates cytokines release. Regarding chemical instability of the leukotriene molecule, in the present study we assessed the immunomodulatory activities conferred by LTB4 released from microspheres (MS). A previous oil-in-water emulsion solvent extraction-evaporation method was chosen to prepare LTB4-loaded MS. Results In the mice cremasteric microcirculation, intraescrotal injection of 0.1 ml of LTB4-loaded MS provoked significant increases in leukocyte rolling flux, adhesion and emigration besides significant decreases in the leukocyte rolling velocity. LTB4-loaded MS also increase peroxisome proliferator-activated receptor-α (PPARα) expression by murine peritoneal macrophages and stimulate them to generate nitrite levels. Monocyte chemoattractant protein-1 (MCP-1) and nitric oxide (NO) productions were also increased when human umbilical vein and artery endothelial cells (HUVECs and HUAECs, respectively) were stimulated with LTB4-loaded MS. Conclusion LTB4-loaded MS preserve the biological activity of the encapsulated mediator indicating their use as a new strategy to modulate cell activation, especially in the innate immune response.