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cis-Aconitic acid is a constituent from the leaves of Echinodorus grandiflorus, a medicinal plant traditionally used in Brazil to treat inflammatory conditions, including arthritic diseases. The present study aimed to investigate the anti-arthritic effect of cis-aconitic acid in murine models of antigen-induced arthritis and monosodium urate-induced gout. The possible underlying mechanisms of action was evaluated in THP-1 macrophages. Oral treatment with cis-aconitic acid (10, 30, and 90 mg/kg) reduced leukocyte accumulation in the joint cavity and C-X-C motif chemokine ligand 1 and IL-1β levels in periarticular tissue. cis-Aconitic acid treatment reduced joint inflammation in tissue sections of antigen-induced arthritis mice and these effects were associated with decreased mechanical hypernociception. Administration of cis-aconitic acid (30 mg/kg p. o.) also reduced leukocyte accumulation in the joint cavity after the injection of monosodium urate crystals. cis-Aconitic acid reduced in vitro the release of TNF-α and phosphorylation of IκBα in lipopolysaccharide-stimulated THP-1 macrophages, suggesting that inhibition of nuclear factor kappa B activation was an underlying mechanism of cis-aconitic acid-induced anti-inflammatory effects. In conclusion, cis-aconitic acid has significant anti-inflammatory effects in antigen-induced arthritis and monosodium urate-induced arthritis in mice, suggesting its potential for the treatment of inflammatory diseases of the joint in humans. Additionally, our findings suggest that this compound may contribute to the anti-inflammatory effect previously reported for E. grandiflorus extracts.

Bone protective effects of TNFα inhibition in rheumatoid arthritis are thought to be mediated by inhibiting synovial osteoclast differentiation and activity. However, it has not been addressed, if TNFα inhibitors alter the pool of peripheral osteoclast precursor cells (OPCs). Here, we blocked TNFα function in C57BL/6 mice with antigen induced arthritis (AIA) using the soluble TNFα receptor etanercept. Synovial bone lesions and osteoclasts were markedly reduced upon Etanercept in the early chronic phase of AIA. Unexpectedly this was not associated with a reduced recruitment of circulating OPCs to the arthritic joint nor to reduced synovial inflammation. In contrast we found that OPC numbers in bone marrow and blood were significantly reduced. Overall our study suggests that arrest of osteoclast mediated bone lesions upon inhibition of TNFα is, at least initially, based on reduced OPC availability in the periphery, and not on OPC recruitment or local anti-inflammatory effects in the arthritic joint.

The leaves of Echinodorus grandiflorus are traditionally used in Brazil to treat several inflammatory conditions, including arthritis. This study aimed to investigate the antiarthritis activity of the 70 % ethanol extract of E. grandiflorus leaves and a standardized flavonoid-rich fraction in an antigen-induced arthritis model in mice. Previously immunized mice were treated per os with saline (control group), 70 % ethanol extract (100–1000 mg/kg), or a flavonoid-rich fraction (0.7–7.2 mg/kg) 40 minutes before and 3 and 6 hours after the challenge with antigen into the knee joint. The administration of the 70 % ethanol extract and flavonoid-rich fraction to mice significantly reduced neutrophil recruitment to the joint cavity and in periarticular tissue. The levels of chemokine (C–X-C motif) ligand 1, tumor necrosis factor-α, and interleukin-1β quantified by the enzyme-linked immunosorbent assay (ELISA) in the periarticular tissue were also diminished in mice treated with the 70 % ethanol extract and flavonoid-rich fraction, as well as mechanical hypernociception. Histological analysis confirmed that both the 70 % ethanol extract and flavonoid-rich fraction suppressed joint inflammation and inhibited cartilage and bone destruction when compared to the control group. Our results demonstrate, for the first time, that E. grandiflorus has anti-inflammatory activity in an experimental arthritis model and highlights the role of flavonoids in the observed response.

OBJECTIVE: Ischemic stroke may result from transient or permanent reductions of regional cerebral blood flow. Polymorphonuclear neutrophils have been described as the earliest inflammatory cells to arrive in ischemic tissue. CXCR1/2 receptors are involved in the recruitment of these cells. However, the contribution of these chemokine receptors during transient brain ischemia in mice remains poorly understood. In this work, we investigated the effects of reparixin, an allosteric antagonist of CXCR1/2 receptors, in a model of middle cerebral artery occlusion and reperfusion in mice. METHODS: C57BL/6J male mice treated with reparixin or vehicle were subjected to a middle cerebral artery occlusion procedure 1 h after the treatment. Ninety minutes after ischemia induction, the monofilament that prevented blood flow was removed. Twenty-four hours after the reperfusion procedure, behavioral changes, including motor signs, were analyzed with the SmithKline/Harwell/lmperial College/Royal Hospital/Phenotype Assessment (SHIRPA) battery. The animals were sacrificed, and brain tissue was removed for histological and biochemical analyses. Histological sections were stained with hematoxylin and eosin, neutrophil infiltration was estimated by myeloperoxidase activity and the inflammatory cytokine IL-iβ was measured by ELISA. RESULTS: Pre-treatment with reparixin reduced the motor deficits observed in this model of ischemia and reperfusion. Myeloperoxidase activity and IL-iβ were reduced in the reparixin-treated group. Histological analysis revealed that ischemic injury was also attenuated by reparixin pre-treatment. CONCLUSIONS: Our results suggest that the blockade of the CXCR1/2 receptors by reparixin promotes neuroprotective effects by reducing the levels of polymorphonuclear infiltration in the brain and the tissue damage associated with middle cerebral artery occlusion and reperfusion.

It is not known how naive B cells compute divergent chemoattractant signals of the T-cell area and B-cell follicles during in vivo migration. Here, we used two-photon microscopy of peripheral lymph nodes (PLNs) to analyze the prototype G-protein-coupled receptors (GPCRs) CXCR4, CXCR5, and CCR7 during B-cell migration, as well as the integrin LFA-1 for stromal guidance. CXCR4 and CCR7 did not influence parenchymal B-cell motility and distribution, despite their role during B-cell arrest in venules. In contrast, CXCR5 played a nonredundant role in B-cell motility in follicles and in the T-cell area. B-cell migration in the T-cell area followed a random guided walk model, arguing against directed migration in vivo. LFA-1, but not α4 integrins, contributed to B-cell motility in PLNs. However, stromal network guidance was LFA-1 independent, uncoupling integrin-dependent migration from stromal attachment. Finally, we observed that despite a 20-fold reduction of chemokine expression in virus-challenged PLNs, CXCR5 remained essential for B-cell screening of antigen-presenting cells. Our data provide an overview of the contribution of prototype GPCRs and integrins during naive B-cell migration and shed light on the local chemokine availability that these cells compute.

HIV-1 negative factor (Nef) elevates virus replication and contributes to immune evasion in vivo. As one of its established in vitro activities, Nef interferes with T-lymphocyte chemotaxis by reducing host cell actin dynamics. To explore Nef’s influence on in vivo recirculation of T lymphocytes, we assessed lymph-node homing of Nef-expressing primary murine lymphocytes and found a drastic impairment in homing to peripheral lymph nodes. Intravital imaging and 3D immunofluorescence reconstruction of lymph nodes revealed that Nef potently impaired T-lymphocyte extravasation through high endothelial venules and reduced subsequent parenchymal motility. Ex vivo analyses of transendothelial migration revealed that Nef disrupted T-lymphocyte polarization and interfered with diapedesis and migration in the narrow subendothelial space. Consistently, Nef specifically affected T-lymphocyte motility modes used in dense environments that pose high physical barriers to migration. Mechanistically, inhibition of lymph node homing, subendothelial migration and cell polarization, but not diapedesis, depended on Nef’s ability to inhibit host cell actin remodeling. Nef-mediated interference with in vivo recirculation of T lymphocytes may compromise T-cell help and thus represents an important mechanism for its function as a HIV pathogenicity factor.

BACKGROUND: Frailty syndrome in elderly people is characterized by a reduction of energy reserves and also by a decreased of resistance to stressors, resulting in an increase of vulnerability. OBJECTIVE: The aim of this study was to verify the effect of a muscle-strengthening program with load in pre-frail elder women with regards to the functional capacity, knee extensor muscle strength and their correlation. METHODS: Thrity-two pre-frail community-dwelling women participated in this study. Potential participants with cognitive impairment (MEEM), lower extremities orthopedic surgery, fractures, inability to walk unaided, neurological diseases, acute inflammatory disease, tumor growth, regular physical activity and current use of immunomodulators were excluded. All partcipants were evaluated by a blinded assessor using: Timed up and go (TUG), 10-Meter Walk Test (10MWT) and knee extensor muscle strength (Byodex System 3 Pro® isokinetic dynamometer at angular speeds of 60 and 1800/s). The intervention consisted of strengthening exercises of the lower extremities at 70% of 1RM, three times/ week for ten weeks. The statistical analysis was performed using the ANOVA and Spearman tests RESULTS: After the intervention, it was observed statistical significance on the work at 1800/s (F=12.71, p=0.02), on the power at 1800/s (F=15.40, p=0.02) and on the functional capacity (TUG, F=9.54, p=0.01; TC10, F=3.80, p=0.01). There was a good negative and statistically significant correlation between the TUG and work at 600/s, such as the TUG and work at 1800/s (r=-0.65, p=0.01; r=-0.72, p=0.01). CONCLUSION: The intervention improved the muscular power and the functional capacity. The increase of the power correlated with function, which is an important variable of the quality of life in the pre-frail elders. Article registered in the ISRCT register under number ISRCTN62824599.

CONTEXTUALIZAÇÃO: Na síndrome de fragilidade em idosos, há diminuição das reservas de energia e resistência aos estressores, com aumento da vulnerabilidade. OBJETIVO: Verificar o efeito do treinamento de força muscular com carga na capacidade funcional e força muscular dos extensores do joelho e sua associação, após treinamento, em idosas pré-frágeis da comunidade. MÉTODOS:Participaram 32 idosas, pré-frágeis, da comunidade. Excluíram-se aquelas com Miniexame do Estado Mental (MEEM) incompatível; cirurgias ortopédicas dos membros inferiores; fraturas; doenças neurológicas; doenças inflamatórias agudas; neoplasias; atividade física regular; uso de medicamento com ação no sistema imunológico e sem marcha independente. Avaliou-se a capacidade funcional (Timed Up and Go - TUG e velocidade de marcha - TC10) e a força muscular dos extensores do joelho (Byodex System 3 Pro®) nas velocidades angulares de 60 e 180(0)/s. Para o fortalecimento muscular, utilizou-se carga de 75% de resistência máxima (1RM), durante dez semanas, três vezes/semana. A análise estatística foi feita pela ANOVA e Spearman (α=5%). RESULTADOS: Após o treinamento, houve melhora estatística do trabalho normalizado em 180(0)/s (F=12,71, p=0,02), na potência, em 180(0)/s (F=15,40, p=0,02) e na capacidade funcional (TUG, F=9,54, p=0,01; TC10, F=3,80, p=0,01). Houve boa correlação negativa significativa do TUG com as medidas de trabalho normalizado em 60 e 180(0)/s (r=-0,65, p=0,01; r=-0,72, p=0,01). CONCLUSÃO: O treinamento produziu melhora da potência muscular e capacidade funcional. A melhora da potência associou-se à melhora funcional, importante variável para a qualidade de vida de idosas pré-frágeis. Artigo registrado no ISRCT register sob o número ISRCTN62824599.BACKGROUND: Frailty syndrome in elderly people is characterized by a reduction of energy reserves and also by a decreased of resistance to stressors, resulting in an increase of vulnerability. OBJECTIVE: The aim of this study was to verify the effect of a muscle-strengthening program with load in pre-frail elder women with regards to the functional capacity, knee extensor muscle strength and their correlation. METHODS: Thrity-two pre-frail community-dwelling women participated in this study. Potential participants with cognitive impairment (MEEM), lower extremities orthopedic surgery, fractures, inability to walk unaided, neurological diseases, acute inflammatory disease, tumor growth, regular physical activity and current use of immunomodulators were excluded. All partcipants were evaluated by a blinded assessor using: Timed up and go (TUG), 10-Meter Walk Test (10MWT) and knee extensor muscle strength (Byodex System 3 Pro® isokinetic dynamometer at angular speeds of 60 and 180(0)/s). The intervention consisted of strengthening exercises of the lower extremities at 70% of 1RM, three times/ week for ten weeks. The statistical analysis was performed using the ANOVA and Spearman tests RESULTS: After the intervention, it was observed statistical significance on the work at 180(0)/s (F=12.71, p=0.02), on the power at 180(0)/s (F=15.40, p=0.02) and on the functional capacity (TUG, F=9.54, p=0.01; TC10, F=3.80, p=0.01). There was a good negative and statistically significant correlation between the TUG and work at 60(0)/s, such as the TUG and work at 180(0)/s (r=-0.65, p=0.01; r=-0.72, p=0.01). CONCLUSION: The intervention improved the muscular power and the functional capacity. The increase of the power correlated with function, which is an important variable of the quality of life in the pre-frail elders. Article registered in the ISRCT register under number ISRCTN62824599.

The aim of this study is to compare the response of interleukin-6 (IL-6) and soluble tumor necrosis factor alpha receptor 1 (s-TNFr1) to two submaximal intensities of exercise in individuals with heart failure (HF). Thirty-two HF individuals aged 45.53 ± 9.41 years, classes II and III of the New York Heart Association (NYHA) classification underwent two sessions of exercise at low and moderate intensities with blood analysis at baseline, exercise and after exercise. The differences were evaluated by Friedman test and factorial ANOVA. Alpha = 5% was considered. No difference in IL-6 was detected for low intensity. At moderate intensity, there was a significant increase after exercise. The s-TNFr1 increased in moderate-intensity exercise and went back to baseline levels after it. A session of moderate-intensity exercise is better than low-intensity exercise at promoting positive immediate inflammatory responses in individuals with HF class II and III of the NYHA.

BACKGROUND AND PURPOSE Chronic joint inflammation and pain are the hallmarks of disease in patients with inflammatory arthritis, notably rheumatoid arthritis. The aim of the present study was to investigate the relative contribution of tumour necrosis factor (TNF)-α, interleukin (IL)-1β and neutrophil influx for joint inflammation and nociception in a novel murine model of antigen-induced arthritis (AIA). EXPERIMENTAL APPROACH AIA was induced by administration of antigen into knee joint of previously immunized mice. Neutrophil accumulation was determined by counting neutrophils in the joints and assessing myeloperoxidase activity in tissues surrounding the joints. TNF-α, IL-1β and CXCL-1 were measured by elisa. Mechanical hypernociception was assessed in parallel, using an electronic pressure meter. KEY RESULTS Hypernociception was dependent on antigen dose and the time after its administration; it was prevented by treatment with morphine and associated with neutrophil infiltration and local production of TNF-α, IL-1β and CXCL-1. Administration of a chimeric monoclonal antibody to TNF-α (infliximab) or IL-1receptor antagonist prevented neutrophil influx and hypernociception, and this was comparable to the effects of dexamethasone. Treatment with fucoidin (a leucocyte adhesion inhibitor) greatly suppressed neutrophil influx and local production of TNF-α and IL-1β, and hypernociception. CONCLUSIONS AND IMPLICATIONS In conclusion, the present study describes a new model that allows for the concomitant evaluation of articular hypernociception and inflammation. Using this system, we demonstrated that a positive feedback loop involving neutrophil influx and the pro-inflammatory cytokines TNF-α and IL-1β is necessary for articular hypernociception after antigen challenge of immunized mice.

Sarcopenia is a loss of muscle mass related to aging and leads to muscle performance decline. An increase in inflammatory mediator levels, especially of IL-6, has been associated to reduced muscle strength in the elderly. The aim of the present cross-sectional study was to correlate IL-6 plasma levels with manual muscle strength (MMS) in 63 community-dwelling elderly women. (71.2+/-7.4years). IL-6 was measured using enzyme-linked immunosorbent assay (ELISA) and MMS was measured using the JAMAR dynamometer. Pearson's test was used to explore the relationship between the outcomes at the significance level of alpha=0.05. IL-6 levels (2.56+/-3.44pg/ml) and MMS (22.86+/-4.62kgf) exhibited an inverse correlation (r=-0.2673 and p=0.0373). The increase in IL-6 plasma levels possibly contributed toward the reduction in manual muscle strength among the elderly women studied.

The pentraxin superfamily is a group of evolutionarily conserved proteins that play important roles in the immune system. The long pentraxin PTX3 protein was originally described as able to be induced by pro-inflammatory stimuli in a variety of cell types. In this study, we evaluated the phenotype of Ptx3(-/-) mice subjected to ischemia followed by reperfusion of the superior mesenteric artery. In reperfused wild-type mice, there was significant local and remote injury as demonstrated by increases in vascular permeability, neutrophil influx, nuclear factor-kappaB activation, and production of CXCL1 and tumor necrosis factor-alpha. PTX3 levels were elevated in both serum and intestine after reperfusion. In Ptx3(-/-) mice, local and remote tissue injury was inhibited, and there were decreased nuclear factor-kappaB translocation and cytokine production. Intestinal architecture was preserved, and there were decreased neutrophil influx and significant prevention of lethality in Ptx3(-/-) mice as well. PTX3 given intravenously before reperfusion reversed the protection observed in Ptx3(-/-) mice in a dose-dependent manner, and PTX3 administration significantly worsened tissue injury and lethality in wild-type mice. In conclusion, our studies demonstrate a major role for PTX3 in determining acute reperfusion-associated inflammation, tissue injury, and lethality and suggest the soluble form of this molecule is active in this system. Therapeutic blockade of PTX3 action may be useful in the control of the injuries associated with severe ischemia and reperfusion syndromes.

Strongyloides venezuelensis migrates through the lungs and induces airway hyperresponsiveness (AHR). The present study evaluated the role of C-fibers in mediating airway inflammation and AHR after infection of rats with S. venezuelensis. Neonatal treatment with capsaicin effectively depleted sensory nerves. This was accompanied by inhibition of the AHR induced by S. venezuelensis infection. In contrast, capsaicin treatment greatly enhanced pulmonary inflammation, eosinophil influx and the local production of TNF-alpha. In conclusion, this is the first demonstration that, akin to viral and allergic AHR, permanent loss of sensory nerve C-fibers also reduces AHR induced by infection with a helminth.

Background and purpose: Neutrophil migration into tissues is involved in the genesis of inflammatory pain. Here, we addressed the hypothesis that the effect of CXC chemokines on CXCR1/2 is important to induce neutrophil migration and inflammatory hypernociception. Experimental approach: Mice were treated with a non-competitive allosteric inhibitor of CXCR1/2, DF 2162, and neutrophil influx and inflammatory hypernociception were assessed by myeloperoxidase assay and electronic pressure meter test, respectively, in various models of inflammation. Key results: DF 2162 inhibited neutrophil chemotaxis induced by CXCR1/2 ligands but had no effect on CXCL8 binding to neutrophils. A single mutation of the allosteric site at CXCR1 abrogated the inhibitory effect of DF 2162 on CXCL-8-induced chemotaxis. Treatment with DF 2162 prevented influx of neutrophils and inflammatory hypernociception induced by CXCL1 in a dose-dependent manner. The compound inhibited neutrophil influx and inflammatory hypernociception induced by carrageenan, lipopolysaccharide and zymosan, but not hypernociception induced by dopamine and PGE2. DF 2162 had a synergistic effect with indomethacin or the absence of TNFR1 to abrogate carrageenan-induced hypernociception. Treatment with DF 2162 diminished neutrophil influx, oedema formation, disease score and hypernociception in collagen-induced arthritis. Conclusions and implications: CXCR1/2 mediates neutrophil migration and is involved in the cascade of events leading to inflammatory hypernociception. In addition to modifying fundamental pathological processes, non-competitive allosteric inhibitors of CXCR1/2 may have the additional benefit of providing partial relief for pain and, hence, may be a valid therapeutic target for further studies aimed at the development of new drugs for the treatment of rheumatoid arthritis. British Journal of Pharmacology (2008) 154, 460–470; doi:10.1038/bjp.2008.94; published online 24 March 2008

Bloodsucking parasites such as ticks have evolved a wide variety of immunomodulatory proteins that are secreted in their saliva, allowing them to feed for long periods of time without being detected by the host immune system. One possible strategy used by ticks to evade the host immune response is to produce proteins that selectively bind and neutralize the chemokines that normally recruit cells of the innate immune system that protect the host from parasites. We have identified distinct cDNAs encoding novel chemokine binding proteins (CHPBs), which we have termed Evasins, using an expression cloning approach. These CHBPs have unusually stringent chemokine selectivity, differentiating them from broader spectrum viral CHBPs. Evasin-1 binds to CCL3, CCL4, and CCL18; Evasin-3 binds to CXCL8 and CXCL1; and Evasin-4 binds to CCL5 and CCL11. We report the characterization of Evasin-1 and -3, which are unrelated in primary sequence and tertiary structure, and reveal novel folds. Administration of recombinant Evasin-1 and -3 in animal models of disease demonstrates that they have potent antiinflammatory properties. These novel CHBPs designed by nature are even smaller than the recently described single-domain antibodies (Hollinger, P., and P.J. Hudson. 2005. Nat. Biotechnol. 23:1126–1136), and may be therapeutically useful as novel antiinflammatory agents in the future.

Rheumatoid Arthritis (RA) is a chronic disease characterized by persistent inflammation and pain. Alternative therapies to reduce these symptoms are needed. Marine algae are valuable sources of diverse bioactive compounds. Lithothamnion muelleri (Hapalidiaceae) is a marine algae with anti-inflammatory, antitumor, and immunomodulatory properties. Here, we investigated the potential anti-inflammatory and analgesic activities of L. muelleri in a murine model of antigen-induced arthritis (AIA) in mice. Our results demonstrate that treatment with L. muelleri prevented inflammation and hypernociception in arthritic mice. Mechanistically, the crude extract and the polysaccharide-rich fractions of L. muelleri may act impairing the production of the chemokines CXCL1 and CXCL2, and consequently inhibit neutrophil influx to the knee joint by dampening the adhesion step of leukocyte recruitment in the knee microvessels. Altogether our results suggest that treatment with L.muelleri has a potential therapeutic application in arthritis treatment.

Angiotensin (Ang) II and its AT1 receptors have been implicated in the pathogenesis of rheumatoid arthritis. Activation of the counter-regulatory Ang-(1-7)-Mas receptor axis may contribute to some of the effects of AT₁ receptor blockers (ARBs). In this study, we have used losartan, an ARB, to investigate the role of and the mechanisms by which AT₁ receptors participated in two experimental models of arthritis: antigen-induced arthritis (AIA) in mice and adjuvant-induced arthritis (AdIA) in rats. Treatment with losartan decreased neutrophil recruitment, hypernociception and the production of TNF-α, IL-1β and chemokine (C-X-C motif) ligand 1 in mice subjected to AIA. Histopathological analysis showed significant reduction of tissue injury and inflammation and decreased proteoglycan loss. In addition to decreasing cytokine production, losartan directly reduced leukocyte rolling and adhesion. Anti-inflammatory effects of losartan were not associated to Mas receptor activation and/or Ang-(1-7) production. Anti-inflammatory effects were reproduced in rats subjected to AdIA. This study shows that ARBs have potent anti-inflammatory effects in animal models of arthritis. Mechanistically, reduction of leukocyte accumulation and of joint damage was associated with local inhibition of cytokine production and direct inhibition of leukocyte-endothelium interactions. The anti-inflammatory actions of losartan were accompanied by functional improvement of the joint, as seen by reduced joint hypernociception. These findings support the use of ARBs for the treatment of human arthritis and provide potential mechanisms for the anti-inflammatory actions of these compounds.

Objectives To investigate the effect of 2 standardized exercise programs, muscle strength exercises (SE) and aerobic exercises (AE), on the plasma levels of brain-derived neurotrophic factor (BDNF) and depressive symptoms in 451 elderly women. Design A randomized controlled trial. Setting Belo Horizonte/MG–Brazil. Participants Community-dwelling older women (N=451; age, 65–89y). Intervention The participants were divided into 2 groups: SE and AE. Both protocols lasted 10 weeks, and 30 sessions (1-h sessions) in total were performed 3 times a week under the direct supervision of physical therapists. Main Outcome Measures Plasma levels of BDNF (enzyme-linked immunosorbent assay) and depressive symptoms (Geriatric Depression Scale). Results There was a significant difference for BDNF plasma levels between the SE and AE groups ( P =.009). Post hoc analysis revealed a pre-post intervention difference in BDNF levels only for the SE group ( P =.008). A statistically significant difference was found for the pre- and postintervention Geriatric Depression Scale scores in both groups ( P =.001), showing that the effects of both exercise protocols were comparable regarding depressive symptoms ( P =.185). Conclusions The present findings have demonstrated the positive effect of muscle strengthening and aerobic intervention on depressive symptoms in community-dwelling elderly women. Interestingly, only SE significantly increased the plasma levels of BDNF in our sample. The positive effects of physical exercise on depressive symptoms in the elderly were not mediated by BDNF.

Objective To examine the impact of a muscle resistance program (MRP) on muscular and functional performance and on interleukin 6 (IL-6) and soluble tumor necrosis factor receptor-1 (sTNFr1) plasma levels in prefrail community-dwelling women. Design Randomized controlled trial crossover design with a postintervention and short-term follow-up. Setting University hospital. Participants Prefrail community-dwelling women (N=32; ≥65y). Intervention The MRP was designed based on the exercise at 75% of each participant's maximum load (10wk, 3 times/wk). Main Outcome Measures Plasma concentrations of IL-6 and sTNFr1 (high-sensitivity enzyme-linked immunosorbent assay kits), muscle strength of the knee extensors (isokinetic), and functional performance (Timed Up & Go [TUG] test and 10-meter walk test [10MWT]). Results There were significant differences in functional and muscular performance between the pre-MRP, post-MRP, and 10-week follow-up period. After the MRP, both functional (TUG, pre-MRP=11.1s vs post-MRP=10.4s, P =.00; 10MWT, pre-MRP=4.9s vs post-MRP, 4.4s, P =.00) and muscular performances (pre-MRP=77.8% and post-MRP=83.1%, P =.02) improved. After cessation of the MRP (follow-up period), sTNFr1 plasma levels increased by 21.4% at 10-week follow-up (post-MRP, 406.4pg/mL; 10-week follow-up, 517.0pg/mL; P =.03). There were significant differences in sTNFr1 ( P =.01). Conclusions The MRP was effective in improving functional and muscular performances, although alterations in plasma levels of IL-6 and sTNFr1 could not be identified after the MRP. Cessation of the MRP after 10 weeks resulted in increased plasma levels of sTNFr1.

Aim This study aimed to investigate whether chronic antigen-induced arthritis (AIA) influences infection-induced periodontitis (PD) in mice and whether PD modifies the clinical course of AIA. The contribution of anti-TNF-α therapy was also evaluated. Materials and methods The PD was induced in C57BL/6 mice by oral infection with Aggregatibacter actinomycetemcomitans. AIA was induced after infection. Anti-TNF-α and chlorhexidine therapies were used to investigate the role of TNF-α and oral infection on PD and AIA interaction. Maxillae, knee joints, lymph nodes and serum samples were used for histomorphometric, immunoenzymatic and/or real time-PCR analyses. Results Antigen-induced arthritis exacerbated alveolar bone loss triggered by PD infection. In contrast, PD did not influence AIA in the evaluated time-points. PD exacerbation was associated with enhanced production of IFN-γ in maxillae and expression of the Th1 transcription factor tBET in submandibular lymph nodes. Increased serum levels of IL-6 and C-reactive protein were also detected. Anti-TNF-α and antiseptic therapies prevented the development and exacerbation of infectious-PD. Anti-TNF-α therapy also resulted in reduced expression of IFN-γ, TNF-α and IL-17 in maxillae. Conclusions Altogether, the current results indicate that the exacerbation of infection-induced PD by arthritis is associated with an alteration in lymphocyte polarization pattern and increased systemic immunoreactivity. This process was ameliorated by anti-TNF-α and antiseptic therapies.

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-1β (IL-1β), 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-1β, 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 neutrophil-dependent hypernociception. MSU crystal–induced neutrophil influx was CXCR2-dependent and relied on the induction of CXCL1 in an NLRP3/ASC/caspase 1/IL-1β/MyD88–dependent manner. LTB4 was produced rapidly after injection of MSU crystals, and this was necessary for caspase 1–dependent IL-1β 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 crystal–induced maturation of IL-1β. 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 crystal–induced 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.

Rheumatoid arthritis (RA) and periodontal disease (PD) are prevalent chronic inflammatory disorders that affect bone structures. Individuals with RA are more likely to experience PD, but how disease in joints could induce PD remains unknown. This study aimed to experimentally mimic clinical parameters of RA-induced PD and to provide mechanistic findings to explain this association. Chronic Ag-induced arthritis (AIA) was triggered by injection of methylated BSA in the knee joint of immunized mice. Anti–TNF-α was used to assess the role of this cytokine. Intra-articular challenge induced infiltration of cells, synovial hyperplasia, bone resorption, proteoglycan loss, and increased expression of cytokines exclusively in challenged joints. Simultaneously, AIA resulted in severe alveolar bone loss, migration of osteoclasts, and release of proinflammatory cytokines in maxillae. Anti–TNF-α therapy prevented the development of both AIA and PD. AIA did not modify bacterial counts in the oral cavity. PD, but not AIA, induced by injection of Ag in immunized mice was decreased by local treatment with antiseptic, which decreased the oral microbiota. AIA was associated with an increase in serum C-reactive protein levels and the expression of the transcription factors RORγ and Foxp3 in cervical lymph nodes. There were higher titers of anti-collagen I IgG, and splenocytes were more responsive to collagen I in AIA mice. In conclusion, AIA-induced PD was dependent on TNF-α and the oral microbiota. Moreover, PD was associated with changes in expression of lymphocyte transcription factors, presence of anti-collagen Abs, and increased reactivity to autoantigens.

Objectives Among several other factors, the neuro-toxic β-amyloid peptide (βAP)-induced inflammatory mechanisms have also been implicated in the pathogenesis of Alzheimer's dementia (AD). Cytokines have recently emerged as prime candidates underlying this immune reaction. The purpose of this study was to evaluate the inflammatory response of peripheral blood mono-nuclear cells (PBMC) in AD. Design Cross-sectional (observational) study. Setting Behavioral and cognitive neurology clinic of the Universidade Federal de Minas Gerais in Belo Horizonte, Brazil. Participants AD patients (n = 19), healthy elderly (n = 19) and young (n = 14) individuals. Measurements Cytokine levels were assessed by enzyme-linked immuno-sorbent assay (ELISA) after exposing cells to a broad range of βAP concentrations (10 − 4 –10 − 10 M) as a stimulus. AD samples were weighed against leukocytes harvested from non-demented young and elderly subjects. Results Cytokine production of PBMCs in the youth was characterized by low baseline levels when compared to cells from the older generation. In the aging population, AD cells were distinguished from the healthy elderly sub-group by an even higher basal cytokine secretion. The low resting concentration in young individuals was markedly increased after treatment with βAP, however cells from the elderly, irrespective of their disease status, showed unchanged cytokine release following βAP administration. Non-specific activation of PBMCs with anti-CD3/CD28 antibodies resulted in elevated interleukin (IL)-1β concentrations in AD. Conclusions These results demonstrate a general over-production of cytokines and resistance to βAP in the old comparison group, with a more pronounced disruption/boosted pattern in AD. Our findings are in line with the hypothesis of “inflammaging”, i.e. an enhanced inflammatory profile with normal aging and a further perturbed environment in AD. The observed cytokine profiles may serve as diagnostic biomarkers in dementia.

The intravital microscopy is a valuable tool to capture images of cells in living organisms and to make studies of molecular determinants of leukocyte trafficking easier. Using this technique, we can directly visualize and measure each step of the leukocyte recruitment paradigm, including leukocyte rolling flux, rolling velocity, adhesion, and emigration. Thus, it is possible to understand the process involved in leukocyte homing as well as the cell recruitment to inflammatory tissues. Nowadays, two types of intravital microscopy are used routinely. The light microscopy is used to assess migration of intravascular cells in thin, tissues which must be sufficiently translucent. Epifluorescence microscopy allows the visualization of the microcirculation while permitting the distinction of leukocyte subpopulations in solid organs.

Frailty syndrome in elderly people is characterized by a reduction of energy reserves and also by a decreased of resistance to stressors, resulting in an increase of vulnerability.The aim of this study was to verify the effect of a muscle-strengthening program with load in pre-frail elder women with regards to the functional capacity, knee extensor muscle strength and their correlation.Thrity-two pre-frail community-dwelling women participated in this study. Potential participants with cognitive impairment (MEEM), lower extremities orthopedic surgery, fractures, inability to walk unaided, neurological diseases, acute inflammatory disease, tumor growth, regular physical activity and current use of immunomodulators were excluded. All partcipants were evaluated by a blinded assessor using: Timed up and go (TUG), 10-Meter Walk Test (10MWT) and knee extensor muscle strength (Byodex System 3 Pro® isokinetic dynamometer at angular speeds of 60 and 180(0)/s). The intervention consisted of strengthening exercises of the lower extremities at 70% of 1RM, three times/ week for ten weeks. The statistical analysis was performed using the ANOVA and Spearman testsAfter the intervention, it was observed statistical significance on the work at 180(0)/s (F=12.71, p=0.02), on the power at 180(0)/s (F=15.40, p=0.02) and on the functional capacity (TUG, F=9.54, p=0.01; TC10, F=3.80, p=0.01). There was a good negative and statistically significant correlation between the TUG and work at 60(0)/s, such as the TUG and work at 180(0)/s (r=-0.65, p=0.01; r=-0.72, p=0.01).The intervention improved the muscular power and the functional capacity. The increase of the power correlated with function, which is an important variable of the quality of life in the pre-frail elders. Article registered in the ISRCT register under number ISRCTN62824599.

IL-13 is a cytokine known to play a role in several pulmonary diseases, including asthma and fibrosis. The role of IL-13 in the context of pulmonary changes induced by helminth infection is unclear. Rats experimentally infected with Strongyloides venezuelensis and treated with anti-IL-13 neutralizing antibody were used to evaluate the role of IL-13 on functional and inflammatory changes of host lungs, and on parasite control. S. venezuelensis-induced airway hyperreactivity was IL-13-independent, but IL-13 played an essential role in driving airway mucus production and eosinophil infiltration. IL-13 was important for the control of egg production but not establishment in the intestine.

To evaluate plasma sTNFR-1 and IL-6 levels and correlate with hand grip in the institutionalized and community living Brazilian elderly.A convenience sample of 110 elderly women (71.17 + or - 7.44 years) was selected. Plasma sTNFR-1 and IL-6 levels were measured by ELISA. For the measurement of hand grip, a JAMAR dynamometer was used.Plasma concentrations of inflammatory markers were significantly higher in institutionalized elderly (sTNFR-1: 479 + or - 22 pg/mL; IL-6: 6.3 + or - 0.8 pg/mL) than in community-dwelling elderly (sTNFR-1: 329 + or - 24 pg/mL; IL-6: 2.5 + or - 0.4 pg/mL; P0.0001). Institutionalized elderly had reduced hand grip (15 + or - 0.8 Kgf) in comparison to community dwelling elderly (23 + or - 0.6 Kgf; P0.05). When individuals were subdivided in age groups, sTNFR-1 was higher in community dwelling versus institutionalized elderly in the 60-70 age range.Our results demonstrate that being institutionalized has an impact on levels of inflammatory markers.

Objective The chemokine receptors CXCR1 and CXCR2 play a role in mediating neutrophil recruitment and neutrophil-dependent injury in several models of inflammation. We undertook this study to investigate the role of these receptors in mediating neutrophil adhesion, subsequent migration, and neutrophil-dependent hypernociception in a murine model of monarticular antigen-induced arthritis (AIA). Methods AIA was induced by administration of antigen into the knee joint of previously immunized mice. Intravital microscopy studies were performed to assess leukocyte rolling and adhesion. Mechanical hypernociception was investigated using an electronic pressure meter. Neutrophil accumulation in the tissue was measured by counting neutrophils in the synovial cavity and assaying myeloperoxidase activity. Levels of tumor necrosis factor α (TNFα) and the chemokines CXCL1 and CXCL2 were quantified by enzyme-linked immunosorbent assay. Histologic analysis was performed to evaluate the severity of arthritis and leukocyte infiltration. Results Antigen challenge in immunized mice induced production of TNFα, CXCL1, and CXCL2 and also resulted in neutrophil recruitment, leukocyte rolling and adhesion, and hypernociception. Treatment with reparixin or DF2162 (allosteric inhibitors of CXCR1/CXCR2) decreased neutrophil recruitment, an effect that was associated with marked inhibition of neutrophil adhesion. Drug treatment also inhibited TNFα production, hypernociception, and the overall severity of the disease in the tissue. Conclusion Blockade of CXCR1/CXCR2 receptors inhibits neutrophil recruitment by inhibiting the adhesion of neutrophils to synovial microvessels. As a consequence, there is decreased local cytokine production and reduced hypernociception, as well as ameloriation of overall disease in the tissue. These studies suggest a potential therapeutic role for the modulation of CXCR1/CXCR2 receptor signaling in the treatment of arthritis.

Background Neuro-inflammation, triggered by β-amyloid peptide, is implicated as one of the primary contributors to Alzheimer's disease (AD) pathogenesis, and several cytokines were identified as key instigating factors. Methods To reveal the inflammatory response of lymphocytes to the neuro-toxic β-amyloid peptide, we evaluated the release of several cytokines from peripheral blood mononuclear cells with immuno-assays (ELISA). From hyper-acute to chronic effects of β-amyloid peptide were assessed at a wide range of concentrations. Results The pro-inflammatory interleukin (IL)-1β, tumor necrosis factor-α, monocyte chemotactic protein-1, and Rantes (acronym for r egulated on a ctivation, n ormal T -cell e xpressed and s ecreted) as well as the pleiotropic IL-6 showed a biphasic release pattern over time in both low and high doses of amyloid treatment: after an initial increase, their concentration gradually fell to the baseline. The suppressors IL-4 and IL-10 had a sinus-like secretion panel: an acute increase in their levels turned to a depression and later followed by their over-secretion. Interestingly, β-amyloid below 10 −8 mol/L produced no effect at all, but any molarity above this threshold caused the very same cytokine secretion pattern, the mark of an all-or-nothing response of β-amyloid peptide. Conclusions These results delineate a highly organized pro- and anti-inflammatory response of cells to the neuro-toxic peptide. This is the first study to describe how the β-amyloid–induced inflammatory processes in Alzheimer's dementia are regulated.

Activation of the cytokine systems may be involved in the neuropathological changes occurring in the central nervous systems of schizophrenic patients. However, associations between the levels of cytokines and the severity of symptoms have not been completely established. Objective: It was the aim of this study to evaluate serum levels of tumor necrosis factor (TNF)-α and their soluble receptors (sTNFR) in schizophrenic patients and healthy controls. Methods: Forty male institutionalized schizophrenic patients (mean age ± SD, 52.3 ± 9.9 years) and 20 asymptomatic matched controls were recruited. The severity of symptoms was assessed using the Brief Psychiatric Rating Scale, the Positive and Negative Syndrome Scale and the Abnormal Involuntary Movement Scale. Serum levels of cytokines were measured by ELISAs. Results: Serum levels of sTNFR1 and sTNFR2 were increased in schizophrenic patients when compared with controls (all p < 0.05), but there was no difference in TNF-α levels. There was no correlation between the length of disease/hospitalization or the severity of symptoms and the serum levels of these molecules. Conclusion: Inflammatory markers are increased in schizophrenia but they do not correlate with symptom severity.

Soon after birth the mammalian gut microbiota forms a permanent and collectively highly resilient consortium. There is currently no robust method for re-deriving an already microbially colonized individual again-germ-free. We previously developed the in vivo growth-incompetent E. coli K-12 strain HA107 that is auxotrophic for the peptidoglycan components D-alanine (D-Ala) and meso-diaminopimelic acid (Dap) and can be used to transiently associate germ-free animals with live bacteria, without permanent loss of germ-free status. Here we describe the translation of this experimental model from the laboratory-adapted E. coli K-12 prototype to the better gut-adapted commensal strain E. coli HS. In this genetic background it was necessary to complete the D-Ala auxotrophy phenotype by additional knockout of the hypothetical third alanine racemase metC. Cells of the resulting fully auxotrophic strain assembled a peptidoglycan cell wall of normal composition, as long as provided with D-Ala and Dap in the medium, but could not proliferate a single time after D-Ala/Dap removal. Yet, unsupplemented bacteria remained active and were able to complete their cell cycle with fully sustained motility until immediately before autolytic death. Also in vivo, the transiently colonizing bacteria retained their ability to stimulate a live-bacteria-specific intestinal Immunoglobulin (Ig)A response. Full D-Ala auxotrophy enabled rapid recovery to again-germ-free status. E. coli HS has emerged from human studies and genomic analyses as a paradigm of benign intestinal commensal E. coli strains. Its reversibly colonizing derivative may provide a versatile research tool for mucosal bacterial conditioning or compound delivery without permanent colonization. ISSN:1932-6203

In recent years, intravital twophoton microscopy (2PM) has emerged as the appropriate technique for direct in situ imaging of immune cell dynamics inside peripheral lymph nodes (PLNs) of live, anesthetized mice, yielding important insights into the regulation of immune responses. However, most current 2PM approaches are limited by the scarce availability of near-infrared (NIR) probes for multispectral time-lapse imaging, and by the use of a single excitation wavelength for multiple fluorophores. The recent availability of quantum dots (QDs) nanoparticles displaying unique optical properties have the potential to overcome this limitation but their suitability has not been yet comprehensively tested for 2PM imaging in vivo. In this study, we explored the use and delivery of NIR-emitting QDs into dendritic cells. Furthermore, we functionalized the surface of these nanoparticles with antibodies that recognize specific antigens expressed on the endothelium of the PLN microvasculature or their use as NIR plasma...

Background With the increase in the elderly population, a growing number of chronic degenerative diseases and a greater dependency on caregivers have been observed. Elderly persons in states of frailty remain more susceptible to significant health complications. There is evidence of an inverse relationship between plasma levels of inflammatory mediators and levels of functionality and muscle strength, suggesting that muscle-strengthening measures can aid in inflammatory conditions. The purpose of this study will be verified the effect of a muscle-strengthening program with load during a ten-week period in pre-frail elderly women with attention to the following outcomes: (1) plasma levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α), (2) functional capacity and (3) knee extensor muscle strength. Methods/Design The study design is a randomized crossover clinical trial evaluating 26 elderly women (regardless of their race and/or social condition) who are community residents, older than 65, and classified as pre-frail according to the criteria previously described by Fried et al. (2004). All subjects will be assessed using the Timed up and go and 10-Meter Walk Test functional tests. The plasma levels of IL-6 and TNF-α will be assessed by ELISA (enzyme-linked immunosorbent assay) with high sensitivity kits (Quantikine®HS, R&D Systems Minneapolis, MN, U.S.). Knee extensor muscle strength will be assessed using the Byodex System 3 Pro ® isokinetic dynamometer at angular speeds of 60 and 180°/s. The intervention will consist of strengthening exercises of the lower extremities at 50 to 70% of 1RM (maximal resistance) three times per week for ten weeks. The volunteers will be randomized into two groups: group E, the intervention group, and group C, the control group that did not initiate any new activities during the initial study period (ten weeks). After the initial period, group C will begin the intervention and group E will maintain everyday activities without exercising. At the end of the total study period, all volunteers will be reassessed. Discussion To demonstrate and discuss possible influences of load-bearing exercises on the modification of plasma levels of IL-6 and TNF-α and in the functional performance of pre-frail elderly women. Trial Registration ISRCTN62824599