NOSH-Aspirin (NBS-1120), a Dual Nitric Oxide and Hydrogen Sulfide-Releasing Hybrid, Reduces Inflammatory Pain

The development of nitric oxide (NO)- and hydrogen sulfide (H2S)-releasing nonsteroidal anti-inflammatory drugs (NSAIDs) has generated more potent anti-inflammatory drugs with increased safety profiles. A new hybrid molecule incorporating both NO and H2S donors into aspirin (NOSH-aspirin) was recently developed. In the present study, the antinociceptive activity of this novel molecule was compared with aspirin in different models of inflammatory pain. It was found that NOSH-aspirin inhibits acetic acid-induced writhing response and carrageenan (Cg)-induced inflammatory hyperalgesia in a dosedependent (5–150 lmol/kg, v.o.) manner, which was superior to the effect of the same doses of aspirin. NOSH-aspirin’s antinociceptive effect was also greater and longer compared to aspirin upon complete Freund’s adjuvant (CFA)-induced inflammatory hyperalgesia. Mechanistically, NOSH-aspirin, but not aspirin, was able to reduce the production/release of interleukin-1 beta (IL-1b) during Cg-induced paw inflammation. Furthermore, NOSH-aspirin, but not aspirin, reduced prostaglandin E2-induced hyperalgesia, which was prevented by treatment with a ATP-sensitive potassium channel (KATP) blocker (glibenclamide; glib.). Noteworthy, the antinociceptive effect of NOSH-aspirin was not associated with motor impairment. The present results indicate that NOSH-aspirin seems to present greater potency than aspirin to reduce inflammatory pain in several models. The enhanced effects of NOSH-aspirin seems to be due to its ability to reduce the production of pronociceptive cytokines such as IL-1 b and directly block hyperalgesia caused by a directly acting hyperalgesic mediator in a mechanism dependent on modulation of KATP channels. In conclusion, we would like to suggest that NOSH-aspirin represents a prototype of a new class of analgesic drugs with more potent effects than the traditional NSAID, aspirin

Stimulation of peripheral Kappa opioid receptors inhibits inflammatory hyperalgesia via activation of the PI3Kγ/AKT/nNOS/NO signaling pathway

<p>Abstract</p> <p>Background</p> <p>In addition to their central effects, opioids cause peripheral analgesia. There is evidence showing that peripheral activation of kappa opioid receptors (KORs) inhibits inflammatory pain. Moreover, peripheral μ-opioid receptor (MOR) activation are able to direct block PGE₂-induced ongoing hyperalgesia However, this effect was not tested for KOR selective activation. In the present study, the effect of the peripheral activation of KORs on PGE₂-induced ongoing hyperalgesia was investigated. The mechanisms involved were also evaluated.</p> <p>Results</p> <p>Local (paw) administration of U50488 (a selective KOR agonist) directly blocked, PGE₂-induced mechanical hyperalgesia in both rats and mice. This effect was reversed by treating animals with L-NMMA or N-propyl-L-arginine (a selective inhibitor of neuronal nitric oxide synthase, nNOS), suggesting involvement of the nNOS/NO pathway. U50488 peripheral effect was also dependent on stimulation of PI3Kγ/AKT because inhibitors of these kinases also reduced peripheral antinociception induced by U50488. Furthermore, U50488 lost its peripheral analgesic effect in PI3Kγ null mice. Observations made in vivo were confirmed after incubation of dorsal root ganglion cultured neurons with U50488 produced an increase in the activation of AKT as evaluated by western blot analyses of its phosphorylated form. Finally, immunofluorescence of DRG neurons revealed that KOR-expressing neurons also express PI3Kγ (≅ 43%).</p> <p>Conclusions</p> <p>The present study indicates that activation of peripheral KORs directly blocks inflammatory hyperalgesia through stimulation of the nNOS/NO signaling pathway which is probably stimulated by PI3Kγ/AKT signaling. This study extends a previously study of our group suggesting that PI3Kγ/AKT/nNOS/NO is an important analgesic pathway in primary nociceptive neurons.</p

Neovestitol, an isoflavonoid isolated from brazilian red propolis, reduces acute and chronic inflammation: involvement of nitric oxide and IL-6

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOIsoflavonoids have been largely studied due to their distinct biological activities identified thus far. Herein, we evaluated the activity of neovestitol, an isoflavonoid isolated from Brazilian red propolis, in acute and chronic inflammation. As for acute inflammation, we found that neovestitol reduced neutrophil migration, leukocyte rolling and adhesion, as well as expression of ICAM-1 in the mesenteric microcirculation during lipopolysaccharide-induced acute peritonitis. No changes were observed in the levels of TNF-alpha, CXCL1/KC and CXCL2/MIP-2 upon pretreatment with neovestitol. The administration of an inducible nitric oxide synthase (iNOS) inhibitor abolished the inhibitory effects of neovestitol in neutrophil migration and ICAM-1 expression. Nitrite levels increased upon treatment with neovestitol. No effects of neovestitol were observed on the chemotaxis of neutrophils in vitro. As for chronic inflammation, neovestitol also reduced the clinical score and joint damage in a collagen-induced arthritis model. There was no change in the frequency of IL-17-producing TCD4+ cells. In addition, pretreatment with neovestitol reduced the levels of IL-6. These results demonstrate a potential anti-inflammatory activity of neovestitol, which may be useful for therapeutic purposes and/or as a nutraceutical.Isoflavonoids have been largely studied due to their distinct biological activities identified thus far. Herein, we evaluated the activity of neovestitol, an isoflavonoid isolated from Brazilian red propolis, in acute and chronic inflammation. As for acute inflammation, we found that neovestitol reduced neutrophil migration, leukocyte rolling and adhesion, as well as expression of ICAM-1 in the mesenteric microcirculation during lipopolysaccharide-induced acute peritonitis. No changes were observed in the levels of TNF-alpha, CXCL1/KC and CXCL2/MIP-2 upon pretreatment with neovestitol. The administration of an inducible nitric oxide synthase (iNOS) inhibitor abolished the inhibitory effects of neovestitol in neutrophil migration and ICAM-1 expression. Nitrite levels increased upon treatment with neovestitol. No effects of neovestitol were observed on the chemotaxis of neutrophils in vitro. As for chronic inflammation, neovestitol also reduced the clinical score and joint damage in a collagen-induced arthritis model. There was no change in the frequency of IL-17-producing TCD4+ cells. In addition, pretreatment with neovestitol reduced the levels of IL-6. These results demonstrate a potential anti-inflammatory activity of neovestitol, which may be useful for therapeutic purposes and/or as a nutraceutical6FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2012/01365-0; 2012/22378-

Behavior of the training load, recovery and well-being in volleyball professional athletes in weeks with and without matches

O objetivo deste estudo foi comparar a carga interna de treinamento, o estado de recuperação e o bem-estar em atletas profissionais de Voleibol de acordo com a frequência semanal de jogos. A recuperação foi quantificada pela Escala de TQR, a carga pela PSE da sessão e o bem-estar pela escala de QBE. Para comparar os valores da Carga de Treinamento Semanal Total, o delta da Qualidade Total de Recuperação (ΔTQR) e o delta da Qualidade de bem-estar (ΔQBE) foi aplicado o teste ANOVA para medidas repetidas. Não foram encontradas diferenças significativas ao comparar a carga de treinamento com a frequência de jogos semanais, porém o tamanho do efeito foi muito elevado quando se comparou a diferença entre um e dois jogos semanais, assim como de nenhum a dois jogos na semana. Houve diferença entre o ΔTQR entre as semanas sem jogos e com dois jogos assim como entre as semanas com um e dois jogos. Já o ΔQBE foi diferente para as semanas que não havia a realização de jogos e aquelas com jogos. Concluise que, mesmo que não haja uma variação das cargas com diferentes frequências semanais de jogos, a recuperação assim como o bem-estar dos atletas foi alterada.The aim of this study was to compare the internal training load, recovery state and the well-being of volleyball professional athletes, according to the number of matches during the week. The recovery was quantified with TQR Scale, the training load with sRPE and the well-being with QBE Scale. The ANOVA was applied for repeated measures in order to compare the values of Total weekly training load, the delta of Total Quality Recovery (ΔTQR) and the delta of the quality of well-being (ΔQBE),. There was no significant difference among the training load and the frequency of weekly games, but the effect size was very high when it was compared to the difference between one and two weekly games, as well as any and two games in a week. There were differences on the ΔTQR between weeks without games and with two games, as well as weeks with one and two matches. The ΔQBE was different for weeks in which there was no game and those with matches, regardless of the frequency. It was concluded that, even if there is no variation of loads in different weekly frequency of matches, the athletes' recovery and well-being change.Facultad de Humanidades y Ciencias de la Educació

Behavior of the training load, recovery and well-being in volleyball professional athletes in weeks with and without matches

O objetivo deste estudo foi comparar a carga interna de treinamento, o estado de recuperação e o bem-estar em atletas profissionais de Voleibol de acordo com a frequência semanal de jogos. A recuperação foi quantificada pela Escala de TQR, a carga pela PSE da sessão e o bem-estar pela escala de QBE. Para comparar os valores da Carga de Treinamento Semanal Total, o delta da Qualidade Total de Recuperação (ΔTQR) e o delta da Qualidade de bem-estar (ΔQBE) foi aplicado o teste ANOVA para medidas repetidas. Não foram encontradas diferenças significativas ao comparar a carga de treinamento com a frequência de jogos semanais, porém o tamanho do efeito foi muito elevado quando se comparou a diferença entre um e dois jogos semanais, assim como de nenhum a dois jogos na semana. Houve diferença entre o ΔTQR entre as semanas sem jogos e com dois jogos assim como entre as semanas com um e dois jogos. Já o ΔQBE foi diferente para as semanas que não havia a realização de jogos e aquelas com jogos. Concluise que, mesmo que não haja uma variação das cargas com diferentes frequências semanais de jogos, a recuperação assim como o bem-estar dos atletas foi alterada.The aim of this study was to compare the internal training load, recovery state and the well-being of volleyball professional athletes, according to the number of matches during the week. The recovery was quantified with TQR Scale, the training load with sRPE and the well-being with QBE Scale. The ANOVA was applied for repeated measures in order to compare the values of Total weekly training load, the delta of Total Quality Recovery (ΔTQR) and the delta of the quality of well-being (ΔQBE),. There was no significant difference among the training load and the frequency of weekly games, but the effect size was very high when it was compared to the difference between one and two weekly games, as well as any and two games in a week. There were differences on the ΔTQR between weeks without games and with two games, as well as weeks with one and two matches. The ΔQBE was different for weeks in which there was no game and those with matches, regardless of the frequency. It was concluded that, even if there is no variation of loads in different weekly frequency of matches, the athletes' recovery and well-being change.Facultad de Humanidades y Ciencias de la Educació

Behavior of the training load, recovery and well-being in volleyball professional athletes in weeks with and without matches

O objetivo deste estudo foi comparar a carga interna de treinamento, o estado de recuperação e o bem-estar em atletas profissionais de Voleibol de acordo com a frequência semanal de jogos. A recuperação foi quantificada pela Escala de TQR, a carga pela PSE da sessão e o bem-estar pela escala de QBE. Para comparar os valores da Carga de Treinamento Semanal Total, o delta da Qualidade Total de Recuperação (ΔTQR) e o delta da Qualidade de bem-estar (ΔQBE) foi aplicado o teste ANOVA para medidas repetidas. Não foram encontradas diferenças significativas ao comparar a carga de treinamento com a frequência de jogos semanais, porém o tamanho do efeito foi muito elevado quando se comparou a diferença entre um e dois jogos semanais, assim como de nenhum a dois jogos na semana. Houve diferença entre o ΔTQR entre as semanas sem jogos e com dois jogos assim como entre as semanas com um e dois jogos. Já o ΔQBE foi diferente para as semanas que não havia a realização de jogos e aquelas com jogos. Concluise que, mesmo que não haja uma variação das cargas com diferentes frequências semanais de jogos, a recuperação assim como o bem-estar dos atletas foi alterada.The aim of this study was to compare the internal training load, recovery state and the well-being of volleyball professional athletes, according to the number of matches during the week. The recovery was quantified with TQR Scale, the training load with sRPE and the well-being with QBE Scale. The ANOVA was applied for repeated measures in order to compare the values of Total weekly training load, the delta of Total Quality Recovery (ΔTQR) and the delta of the quality of well-being (ΔQBE),. There was no significant difference among the training load and the frequency of weekly games, but the effect size was very high when it was compared to the difference between one and two weekly games, as well as any and two games in a week. There were differences on the ΔTQR between weeks without games and with two games, as well as weeks with one and two matches. The ΔQBE was different for weeks in which there was no game and those with matches, regardless of the frequency. It was concluded that, even if there is no variation of loads in different weekly frequency of matches, the athletes' recovery and well-being change.Facultad de Humanidades y Ciencias de la Educació

Toll-like receptor 2/MyD88 signaling mediates zymosan-induced joint hypernociception in mice: Participation of TNF-alpha, IL-1 beta and CXCL1/KC

Arthritic pain is a serious health problem that affects a large number of patients. Toll-like receptors (TLRs) activation within the joints has been implicated in pathophysiology of arthritis. However, their role in the genesis of arthritic pain needs to be demonstrated. In the present study, it was addressed the participation of TLR2 and TLR4 and their adaptor molecule MyD88 in the genesis of joint hypernociception (a decrease in the nociceptive threshold) during zymosan-induced arthritis. Zymosan injected in the tibio-tarsal joint induced mechanical hypernociception in C57BL/6 wild type mice that was reduced in TLR2 and MyD88 null mice. On the other hand, zymosan-induced hypernociception was similar in C3H/HePas and C3H/Hej mice (TLR4 mutant mice). Zymosan-induced joint hypernociception was also reduced in TNFR1 null mice and in mice treated with IL-1 receptor antagonist or with an antagonist of CXCR1/2. Moreover, the joint production of TNF-alpha, IL-1 beta and CXCL1/KC by zymosan was dependent on TLR2/MyD88 signaling. Investigating the mechanisms by which TNF-alpha, IL-1 beta and CXCL1/KC mediate joint hypernociception, joint administration of these cytokines produced mechanical hypernociception, and they act in an interdependent manner. In last instance, their hypernociceptive effects were dependent on the production of hypernociceptive mediators, prostaglandins and sympathetic amines. These results indicate that in zymosan-induced experimental arthritis, TLR2/MyD88 is involved in the cascade of events of joint hypernociception through a mechanism dependent on cytokines and chemokines production. Thus, TLR2/MyD88 signaling might be a target for the development of novel drugs to control pain in arthritis. (C) 2011 Elsevier B.V. All rights reserved.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Pesquisa (CNPq)Coordenadoria de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Programa de Nucleos de Excelencia (PRONEX), Brazi

Coagulant and antibacterial activities of the water-soluble seed lectin from Moringa oleifera

Aims: The aim of this work was to analyse the coagulant and antibacterial activities of lectin isolated from Moringa oleifera seeds that are used for water treatment. Methods and Results:  The water-soluble M. oleifera lectin (WSMoL) was separated from nonhemagglutinating components (NHC) by chitin chromatography. WSMoL fluorescence spectrum was not altered in the presence of ions that are often present in high concentrations in polluted waters. Seed extract, NHC and WSMoL showed coagulant activity on a turbid water model. Both NHC and WSMoL reduced the growth of Staphylococcus aureus, but only WSMoL caused a reduction in Escherichia coli. WSMoL was also more effective in reducing the growth of ambient lake water bacteria. Conclusions:  Data obtained from this study indicate that WSMoL is a potential natural biocoagulant for water, reducing turbidity, suspended solids and bacteria. Significance and Impact of the Study: Moringa oleifera seeds are a material effective in the treatment of water.The authors express their gratitude to the Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) for research grants and fellowship (LCBBC, MLVO and PMGP), the Fundacao de Amparo a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE) and the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) for financial support. Authors are grateful to Maria Barbosa Reis da Silva for the technical assistance and to David Pillard and Felix Nonnenmacher for English editing

Trypanosoma cruzi Adjuvants Potentiate T Cell-Mediated Immunity Induced by a NY-ESO-1 Based Antitumor Vaccine

Immunological adjuvants that induce T cell-mediate immunity (TCMI) with the least side effects are needed for the development of human vaccines. Glycoinositolphospholipids (GIPL) and CpGs oligodeoxynucleotides (CpG ODNs) derived from the protozoa parasite Trypanosoma cruzi induce potent pro-inflammatory reaction through activation of Toll-Like Receptor (TLR)4 and TLR9, respectively. Here, using mouse models, we tested the T. cruzi derived TLR agonists as immunological adjuvants in an antitumor vaccine. For comparison, we used well-established TLR agonists, such as the bacterial derived monophosphoryl lipid A (MPL), lipopeptide (Pam3Cys), and CpG ODN. All tested TLR agonists were comparable to induce antibody responses, whereas significant differences were noticed in their ability to elicit CD4+ T and CD8+ T cell responses. In particular, both GIPLs (GTH, and GY) and CpG ODNs (B344, B297 and B128) derived from T. cruzi elicited interferon-gamma (IFN-γ) production by CD4+ T cells. On the other hand, the parasite derived CpG ODNs, but not GIPLs, elicited a potent IFN-γ response by CD8+ T lymphocytes. The side effects were also evaluated by local pain (hypernociception). The intensity of hypernociception induced by vaccination was alleviated by administration of an analgesic drug without affecting protective immunity. Finally, the level of protective immunity against the NY-ESO-1 expressing melanoma was associated with the magnitude of both CD4+ T and CD8+ T cell responses elicited by a specific immunological adjuvant

Smoking-induced aggravation of experimental arthritis is dependent of aryl hydrocarbon receptor activation in Th17 cells.

Background: Epidemiologic studies have highlighted the association of environmental factors with the development and progression of autoimmune and chronic inflammatory diseases. Among the environmental factors, smoking has been associated with increased susceptibility and poor prognosis in rheumatoid arthritis (RA). However, the immune and molecular mechanism of smoking-induced arthritis aggravation remains unclear. The transcription factor aryl hydrocarbon receptor (AHR) regulates the generation of Th17 cells, CD4 T cells linked the development of autoimmune diseases. AHR is activated by organic compounds including polycyclic aromatic hydrocarbons (PAHs), which are environmental pollutants that are also present in cigarette smoke. In this study, we investigated the role of AHR activation in the aggravation of experiment arthritis induced by exposure to cigarette smoke. Methods: Mice were exposed to cigarette smoke during the developmental phase of antigen-induced arthritis and collagen-induced arthritis to evaluate the effects of smoking on disease development. Aggravation of articular inflammation was assessed by measuring neutrophil migration to the joints, increase in articular hyperalgesia and changes in the frequencies of Th17 cells. In vitro studies were performed to evaluate the direct effects of cigarette smoke and PAH on Th17 differentiation. We also used mice genetically deficient for AHR (Ahr KO) and IL-17Ra (Il17ra KO) to determine the in vivo mechanism of smoking-induced arthritis aggravation. Results: We found that smoking induces arthritis aggravation and increase in the frequencies of Th17 cells. The absence of IL-17 signaling (Il17ra KO) conferred protection to smoking-induced arthritis aggravation. Moreover, in vitro experiments showed that cigarette smoke can directly increase Th17 differentiation of T cells by inducing AHR activation. Indeed, Ahr KO mice were protected from cigarette smoke-induced arthritis aggravation and did not display increase in TH17 frequencies, suggesting that AHR activation is an important mechanism for cigarette smoke effects on arthritis. Finally, we demonstrate that PAHs are also able to induce arthritis aggravation. Conclusions: Our data demonstrate that the disease-exacerbating effects of cigarette smoking are AHR dependent and environmental pollutants with AHR agonist activity can induce arthritis aggravation by directly enhancing Th17 cell development