Physical activity in the post-COVID-19: AEROBICOVID project design and perspectives

Durante a pandemia de COVID-19 estão sendo observados vários efeitos tardios, afetando a saúde e a funcionalidade dos acometidos. Por outro lado, foram observadas menores taxas de infecção pelo SARS-CoV-2 e gravidade dos sintomas em cidades de elevada altitude. Neste sentido, o projeto AEROBICOVID foi desenvolvido com a hipótese de que o exercício seria uma proposta importante para a melhoria da saúde e que a hipóxia promoveria benefícios adicionais no processo de recuperação. Participaram 84 pessoas com aproximadamente 30 dias desde a recuperação dos sintomas da COVID-19, 25 no grupo de controle e 59 divididos em três grupos de treinamento físico moderado. O projeto teve bons resultados no ensino, pesquisa e extensão, mas também enfrentou dificuldades na operacionalização. Estas experiências são a base para propostas futuras através de um projeto de extensão na Universidade de São Paulo e em uma Unidade de Saúde da Família, além de um projeto de pesquisa que desenvolverá uma nova tecnologia de hipóxia de baixo custo.During the COVID-19 pandemic, several late-onset impairments have been observed, affecting the health and functionality of those involved. On the other hand, lower SARS-CoV-2 infection rates and severity of symptoms were observed in high-altitude cities. In this sense, the AEROBICOVID project was developed with the hypothesis that exercise would be an important opportunity for health improvement and that hypoxia would promote additional benefits in the recovery process. The cohort was about 84 participants with approximately 30 days since the COVID-19 symptoms recovery, 25 in the control group, and 59 divided into three moderate physical training groups. The project had good results in teaching, research, and extension, but also faced difficulties in operationalization. This experience is the basis for future proposals through an extension project at the University of São Paulo and in a Family Health Unit, besides a research project that will develop a new low-cost hypoxia technology

GM-CSF Priming Drives Bone Marrow-Derived Macrophages to a Pro-Inflammatory Pattern and Downmodulates PGE(2) in Response to TLR2 Ligands

In response to pathogen recognition by Toll-like receptors (TLRs) on their cell surface, macrophages release lipid mediators and cytokines that are widely distributed throughout the body and play essential roles in host responses. Granulocyte macrophage colony-stimulating factor (GM-CSF) is important for the immune response during infections to improve the clearance of microorganisms. In this study, we examined the release of mediators in response to TLR2 ligands by bone marrow-derived macrophages (BMDMs) primed with GM-CSF. We demonstrated that when stimulated with TLR2 ligands, non-primed BMDMs preferentially produced PGE(2) in greater amounts than LTB4. However, GM-CSF priming shifted the release of lipid mediators by BMDMs, resulting in a significant decrease of PGE(2) production in response to the same stimuli. The decrease of PGE(2) production from primed BMDMs was accompanied by a decrease in PGE-synthase mRNA expression and an increase in TNF-alpha and nitric oxide (NO) production. Moreover, some GM-CSF effects were potentiated by the addition of IFN-gamma. Using a variety of TLR2 ligands, we established that PGE(2) release by GM-CSF-primed BMDMs was dependent on TLR2 co-receptors (TLR1, TLR6), CD14, MyD88 and the nuclear translocation of NF kappa B but was not dependent on peroxisome proliferator-activated receptor-gamma (PPAR-gamma) activation. Indeed, GM-CSF priming enhanced TLR2, TLR4 and MyD88 mRNA expression and phospho-I kappa B alpha formation. These findings demonstrate that GM-CSF drives BMDMs to present a profile relevant to the host during infections.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)FAPESP - Fundacao de Amparo a Pesquisa do Estado de Sao PauloConselho Nacional de Pesquisa (CNPq)CNPq - Conselho Nacional de Pesquis

Leukotrienes Are Upregulated and Associated with Human T-Lymphotropic Virus Type 1 (HTLV-1)-Associated Neuroinflammatory Disease

Leukotrienes (LTs) are lipid mediators involved in several inflammatory disorders. We investigated the LT pathway in human T-lymphotropic virus type 1 (HTLV-1) infection by evaluating LT levels in HTLV-1-infected patients classified according to the clinical status as asymptomatic carriers (HACs) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients. Bioactive LTB4 and CysLTs were both increased in the plasma and in the supernatant of peripheral blood mononuclear cell cultures of HTLV-1-infected when compared to non-infected. Interestingly, CysLT concentrations were increased in HAM/TSP patients. Also, the concentration of plasma LTB4 and LTC4 positively correlated with the HTLV-1 proviral load in HTLV-1-infected individuals. The gene expression levels of LT receptors were differentially modulated in CD4(+) and CD8(+) T cells of HTLV-1-infected patients. Analysis of the overall plasma signature of immune mediators demonstrated that LT and chemokine amounts were elevated during HTLV-1 infection. Importantly, in addition to CysLTs, IP-10 was also identified as a biomarker for HAM/TSP activity. These data suggest that LTs are likely to be associated with HTLV-1 infection and HAM/TSP development, suggesting their putative use for clinical monitoring.Fundacao de Amparo a Pesquisa do Estado de Sao PauloFundacao de Amparo a Pesquisa do Estado de Sao PauloFundacao de Amparo a Pesquisa do Estado de Minas GeraisFundacao de Amparo a Pesquisa do Estado de Minas GeraisConselho Nacional de Desenvolvimento Cientifico e TecnologicoConselho Nacional de Desenvolvimento Cientifico e TecnologicoFundacao Hemocentro de Ribeirao PretoFundacao Hemocentro de Ribeirao PretoCNPqCNP

Amblyomma sculptum Salivary PGE2 Modulates the Dendritic Cell-Rickettsia rickettsii Interactions in vitro and in vivo

Amblyomma sculptum is an important vector of Rickettsia rickettsii, causative agent of Rocky Mountain spotted fever and the most lethal tick-borne pathogen affecting humans. To feed on the vertebrate host's blood, A. sculptum secretes a salivary mixture, which may interact with skin resident dendritic cells (DCs) and modulate their function. The present work was aimed at depicting the A. sculptum saliva-host DC network and the biochemical nature of the immunomodulatory component(s) involved in this interface. A. sculptum saliva inhibits the production of inflammatory cytokines by murine DCs stimulated with LPS. The fractionation of the low molecular weight salivary content by reversed-phase chromatography revealed active fractions eluting from 49 to 55% of the acetonitrile gradient. Previous studies suggested that this pattern of elution matches with that observed for prostaglandin E2 (PGE2) and the molecular identity of this lipid mediator was unambiguously confirmed by a new high-resolution mass spectrometry methodology. A productive infection of murine DCs by R. rickettsii was demonstrated for the first time leading to proinflammatory cytokine production that was inhibited by both A. sculptum saliva and PGE2, a result also achieved with human DCs. The adoptive transfer of murine DCs incubated with R. rickettsii followed by treatment with A. sculptum saliva or PGE2 did not change the cytokine profile associated to cellular recall responses while IgG2a-specific antibodies were decreased in the serum of these mice. Together, these findings emphasize the role of PGE2 as a universal immunomodulator of tick saliva. In addition, it contributes to new approaches to explore R. rickettsii-DC interactions both in vitro and in vivo

Silencing of mitochondrial alternative oxidase gene of Aspergillus fumigatus enhances reactive oxygen species production and killing of the fungus by macrophages

We previously demonstrated that conidia from Aspergillus fumigatus incubated with menadione and paraquat increases activity and expression of cyanide-insensitive alternative oxidase (AOX). Here, we employed the RNA silencing technique in A. fumigatus using the vector pALB1/aoxAf in order to down-regulate the aox gene. Positive transformants for aox gene silencing of A. fumigatus were more susceptible both to an imposed in vitro oxidative stress condition and to macrophages killing, suggesting that AOX is required for the A. fumigatus pathogenicity, mainly for the survival of the fungus conidia during host infection and resistance to reactive oxygen species generated by macrophages.FAPESP[06/04909-0]FAPESP[03/09781-3]CNP

Tityus serrulatus venom and toxins Ts1, Ts2 and Ts6 induce macrophage activation and production of immune mediators

Scorpion envenomation induces a systemic immune response, and neurotoxins of venom act on specific ion channels, modulating neurotransmitter release or activity. However, little is known about the immunomodulatory effects of crude venom from scorpion Tityus serrulatus (TsV) or its toxins (Ts1, Ts2 and Ts6) in combination with lipopolysaccharide (LPS). To investigate the immunomodulatory effects of TsV and its toxins (Ts1, Ts2 and Ts6), J774.1 cells were stimulated with different concentrations (25, 50 and 100 mu g/mL) of venom or toxins pre-stimulated or not with LPS (0.5 mu g/mL). Macrophage cytotoxicity was assessed, and nitric oxide (NO) and cytokine production were analyzed utilizing the culture supernatants. TsV and its toxins did not produce cytotoxic effects. Depending on the concentrations used, TsV, Ts1 and Ts6 stimulated the production of NO, interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha in J774.1 cells, which were enhanced under LPS co-stimulation. However, LPS + Ts2 inhibited NO, IL-6 and TNF-alpha production, and Ts2 alone stimulated the production of IL-10, suggesting an anti-inflammatory activity for this toxin. Our findings are important for the basic understanding of the mechanisms involved in macrophage activation following envenomation: additionally, these findings may contribute to the discovery of new therapeutic compounds to treat immune-mediated diseases. (C) 2011 Elsevier Ltd. All rights reserved.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2005/54855-0]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2009/09829-2]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[2009/07169-5]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq

Histoplasma capsulatum Cell Wall beta-Glucan Induces Lipid Body Formation through CD18, TLR2, and Dectin-1 Receptors: Correlation with Leukotriene B(4) Generation and Role in HIV-1 Infection

Histoplasma capsulatum (Hc) is a facultative, intracellular parasite of worldwide significance. Infection with Hc produces a broad spectrum of diseases and may progress to a life-threatening systemic disease, particularly in individuals with HIV infection. Resolution of histoplasmosis is associated with the activation of cell-mediated immunity, and leukotriene B(4) plays an important role in this event. Lipid bodies (LBs) are increasingly being recognized as multifunctional organelles with roles in inflammation and infection. In this study, we investigated LB formation in histoplasmosis and its putative function in innate immunity. LB formation in leukocytes harvested from Hc-infected C57BL/6 mice peaks on day 2 postinfection and correlates with enhanced generation of lipid mediators, including leukotriene B(4) and PGE(2). Pretreatment of leukocytes with platelet-activating factor and BLT1 receptor antagonists showed that both lipid mediators are involved in cell signaling for LB formation. Alveolar leukocytes cultured with live or dead Hc also presented an increase in LB numbers. The yeast alkali-insoluble fraction 1, which contains mainly beta-glucan isolated from the Hc cell wall, induced a dose- and time-dependent increase in LB numbers, indicating that beta-glucan plays a signaling role in LB formation. In agreement with this hypothesis, beta-glucan-elicited LB formation was inhibited in leukocytes from 5-LO(-/-), CD18(low) and TLR2(-/-) mice, as well as in leukocytes pretreated with anti-Dectin-1 Ab. Interestingly, human monocytes from HIV-1-infected patients failed to produce LBs after beta-glucan stimulation. These results demonstrate that Hc induces LB formation, an event correlated with eicosanoid production, and suggest a role for these lipid-enriched organelles in host defense during fungal infection. The Journal of Immunology, 2009, 182: 4025-4035.FAPESP Fundacao de Amparo a Pesquisa do Estado de Sao Paulo[02/12856-2]CNPq Conselho Nacional de Desenvolvimento Cientifico e TccnologicoFundacao de Apoio Ensino (FAEPA)Pesquisa e Assistencia (FAEPA)Faculdade de Medicina de Ribeirao PretoUniversidade de Sao Paulo (USP

GM-CSF-modified lipid mediators released by BMDM in response to bacterial TLRs-ligands.

<p>BMDM from WT mice were primed (or not) with GM-CSF for 24 h and then incubated with AraLAM (5000 ng/mL), LM (5000 ng/mL), Malp2 (300 ng/mL), Pam₃-CSK₄ (5000 ng/mL) or LPS (500 ng/mL) for 24 h. PGE₂ (<b>A</b>) and LTB₄ (<b>B</b>) release were measured in the supernatant by ELISA. Medium alone was used as a negative control. Results are the average ± S.E.M from mice cells (<i>n</i> = 6 per group), from two independent experiments; *, <i>p</i><<i>0.05</i> compared with unstimulated cells; <b>^#</b>, <i>p<0.05</i> compared with non-priming GM-CSF cells.</p

Effect of GM-CSF priming on BMDM mRNA expression.

<p>mRNA was prepared from 5×10⁶ BMDMs. Macrophages were treated with or without GM-CSF for 4 (<b>A</b>), 8 (<b>B</b>) or 18 h (<b>C</b>). qRT-PCR was used to determine the relative expression of transcripts encoding for lipid metabolism enzymes, TLRs and adaptor proteins. The results were normalized to the expression of the endogenous internal controls <i>Actb</i> and <i>Gapdh</i>. The 2–ΔΔCt method was used in the analysis of the qRT-PCR data. Dotted lines show mRNA expression in untreated cells. The results are presented as the average ± S.E.M from three independent experiments in duplicate (n = 3); *, <i>p</i><0.05 compared with non-GM-CSF-treated cells.</p