Identificação de Neutrophil Extracellular Traps em lesões tegumentares de pacientes com Paracoccidiodomicose e análise da formação de NETs in vitro por neutrófilos humanos desafiados com Paracoccidioides brasiliensis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Monocyte migration driven by galectin-3 occurs through distinct mechanisms involving selective interactions with the extracellular matrix

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nitric oxide and brazilian propolis combined accelerates tissue repair by modulating cell migration, cytokine production and collagen deposition in experimental leishmaniasis

The fact that drugs currently used in the treatment of Leishmania are highly toxic and associated with acquired resistance has promoted the search for new therapies for treating American tegumentary leishmaniasis (ATL). In this study, BALB/c mice were injected in the hind paw with Leishmania (Leishmania) amazonensis and subsequently treated with a combination of nitric oxide (NO) donor (cis-[Ru(bpy)(2)imN(NO)](PF6)(3)) (Ru-NO), given by intraperitoneal injection, and oral Brazilian propolis for 30 days. Ru-NO reached the center of the lesion and increased the NO level in the injured hind paw without lesion exacerbation. Histological and immunological parameters of chronic inflammation showed that this combined treatment increased the efficacy of macrophages, determined by the decrease in the number of parasitized cells, leading to reduced expression of proinflammatory and tissue damage markers. In addition, these drugs in combination fostered wound healing, enhanced the number of fibroblasts, pro-healing cytokines and induced collagen synthesis at the lesion site. Overall, our findings suggest that the combination of the NO donor Ru-NO and Brazilian propolis alleviates experimental ATL lesions, highlighting a new therapeutic option that can be considered for further in vivo investigations as a candidate for the treatment of cutaneous leishmaniasis.

The evolution of a Pluto-like system during the migration of the ice giants

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Redox-active biflavonoids from Garcinia brasiliensis as inhibitors of neutrophil oxidative burst and human erythrocyte membrane damage

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Neutrophil extracellular traps identification in tegumentary lesions of patients with paracoccidioidomycosis and different patterns of NETs generation in vitro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of gamma irradiation on caprolactam migration from multilayer polyamide 6 films into food simulants: development and validation of a gas chromatographic method

A GC method to determine caprolactam in water, 15 ethanol, and olive oil food simulants was developed and validated. Linear ranges varied from 0.96 to 642.82 g/mL for water, 0.64 to 800.32 g/mL for 15 ethanol, and 1.06 to 1062.34 g/g for olive oil, with correlation coefficients higher than 0.999. Method precision studies showed RSD values lower than 5.45, while method accuracy studies showed recovery from 72 to 111 for all simulants. The effect of gamma irradiation on caprolactam migration from multilayer polyamide 6 (PA-6) films intended for cheese into water, 15 ethanol, olive oil, and 3 acetic acid simulants was also studied. For migration assay, non-irradiated and irradiated (12 kGy) films were placed in contact with the simulant and exposed at 40C for 10 days. The validated method was used to quantify caprolactam migration from multilayer PA-6 films into the simulants, which ranged from 1.03 to 7.59 mg/kg for non-irradiated films, and from 4.82 to 11.32 mg/kg for irradiated films. Irradiation caused almost no changes in caprolactam levels, with the exception of olive oil, which showed an increase in the caprolactam level. All multilayer PA-6 films were in accordance with the requirements of the legislation for caprolactam migration.

Estudo do efeito modulador do macrolídeo tilosina na reação inflamatória aguda de tilápias do Nilo (Oreochromis niloticus), vacinadas e desafiadas com Streptococcus agalactiae

Pós-graduação em Medicina Veterinária - FCAV

Segregation of migration by feeding ground origin in North Atlantic humpback whales (Megaptera novaeangliae)

Results from a large-scale, capture–recapture study of humpback whales Megaptera novaeangliae in the North Atlantic show that migration timing is influenced by feeding ground origin. No significant differences were observed in the number of individuals from any feeding area that were re-sighted in the common breeding area in the West Indies. However, there was a relationship between the proportion (logit transformed) of West Indies sightings and longitude (r2 = 0.97, F1,3 = 98.27, P = 0.0022) suggesting that individuals feeding farther to the east are less likely to winter in the West Indies. A relationship was also detected between sighting date in the West Indies and feeding area. Mean sighting dates in the West Indies for individuals identified in the Gulf of Maine and eastern Canada were significantly earlier than those for animals identified in Greenland, Iceland and Norway (9.97 days, t179 = 3.53, P = 0.00054). There was also evidence for sexual segregation in migration; males were seen earlier on the breeding ground than were females (6.63 days, t105 = 1.98, P = 0.050). This pattern was consistently observed for animals from all feeding areas; a combined model showed a significant effect for both sex (F1 = 5.942, P = 0.017) and feeding area (F3 =4.756, P=0.0038). The temporal difference in occupancy of the West Indies between individuals from different feeding areas, coupled with sexual differences in migratory patterns, presents the possibility that there are reduced mating opportunities between individuals from different high latitude areas.

Avian Use of Harvested Crop Fields in North Dakota During Spring Migration

Broad-spectrum herbicide applications and improved harvesting efficiency of crops have reduced the availability of weed seeds and waste grains for game and nongame wildlife. Over the last decade, corn and soybean plantings have steadily increased in the Prairie Pothole Region (PPR) of North Dakota, while sunflower plantings have declined. The PPR is an important corridor for migratory birds, and changes in food availabilities at stopover habitats may affect how food resources are used. In early spring 2003 and 2004, we compared bird use of harvested fields of sunflower, soybeans, small grains, and corn in the PPR of North Dakota. Across both years and all crop types, we observed 20,400 birds comprising 29 species. Flocks of Lapland Longspurs (Calcarius lapponicus) and Horned Larks (Eremophila alpestris) and flocks of Red-winged Blackbirds (Agelaius phoeniceus) made up 60% and 15%, respectively, of the bird counts. We found that species richness and bird densities were higher in harvested sunflower fields and cornfields than in harvested small-grain and soybean fields, with soybean fields harboring the fewest species and lowest bird density. Blackbird densities tended to be lower in fields tilled after fall harvest than in fields not tilled. These results suggest that some granivorous bird populations in the Northern Great Plains could be positively affected by planting of row crops with postharvest vertical structure (e.g., sunflower, corn) and use of no-till land management practices.

Annexin A1 regulates neutrophil clearance by macrophages in the mouse bone marrow

Under homeostatic conditions, a proportion of senescent CXCR4(hi) neutrophils home from the circulation back to the bone marrow, where they are phagocytosed by bone marrow macrophages. In this study, we have identified an unexpected role for the anti-inflammatory molecule annexin A1 (AnxA1) as a critical regulator of this process. We first observed that AnxA1(-/-) mice have significantly increased neutrophil numbers in their bone marrow while having normal levels of GM and G colony-forming units, monocytes, and macrophages. Although AnxA1(-/-) mice have more neutrophils in the bone marrow, a greater proportion of these cells are senescent, as determined by their higher levels of CXCR4 expression and annexin V binding. Consequently, bone marrow neutrophils from AnxA1(-/-) mice exhibit a reduced migratory capacity in vitro. Studies conducted in vitro also show that expression of AnxA1 is required for bone marrow macrophages, but not peritoneal macrophages, to phagocytose apoptotic neutrophils. Moreover, in vivo experiments indicate a defect in clearance of wild-type neutrophils in the bone marrow of AnxA1(-/-) mice. Thus, we conclude that expression of AnxA1 by resident macrophages is a critical determinant for neutrophil clearance in the bone marrow.-Dalli, J., Jones, C. P., Cavalcanti, D. M., Farsky, S. H., Perretti, M., Rankin, S. M. Annexin A1 regulates neutrophil clearance by macrophages in the mouse bone marrow. FASEB J. 26, 387-396 (2012). www.fasebj.org

Reproductive migration and population dynamics of the blue crab Callinectes danae in an estuary in southeastern Brazil

Portunid crabs are an important resource in estuaries, and require appropriate management to guarantee their long-term availability. We investigated the population dynamics and reproduction of Callinectes danae in the Estuarine-Bay Complex of Sao Vicente, Sao Paulo, Brazil, to provide basic biological information for public policies for the management of this fishery. Monthly samples were obtained from March 2007 to February 2008 on eight transects, four in the estuary and four in the bay. A total of 2261 specimens (403 males, 1288 females, of which 570 were ovigerous) were collected. Males were significantly larger than females, and the size-frequency distribution was unimodal for males, females and ovigerous females. The sex ratio was nearly always skewed toward females (M:F - 1:4.6). C. danae showed seasonal-continuous reproduction, with high reproductive activity in the warmer season. C. danae breeds in the estuarine-bay complex, with males and juvenile females concentrated in the estuary. After copulation, fertilized females migrate to the estuary entrance and the bay, where ovigerous females are commonly found spawning in high-salinity areas. Therefore, to manage this important economic resource, both the estuary and the bay should be considered for protection, but special attention should be given to the estuary entrance during the summer months, when ovigerous females concentrate.

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.

INFLAMMATORY RESPONSE AND NEUTROPHIL FUNCTIONS IN PLAYERS AFTER A FUTSAL MATCH

de Moura, NR, Cury-Boaventura, MF, Santos, VC, Levada-Pires, AC, Bortolon, JR, Fiamoncini, J, Pithon-Curi, TC, Curi, R, and Hatanaka, E. Inflammatory response and neutrophil functions in players after a futsal match. J Strength Cond Res 26(9): 2507-2514, 2012-Futsal players suffer injuries resulting from muscle fatigue and contact or collision among players. Muscle lesions can be detected by measuring muscle lesion markers such as creatine kinase (CK) and lactate dehydrogenase (LDH) in plasma. After an initial lesion, there is an increase in the plasma levels of C-reactive protein (CRP) and proinflammatory cytokines. These mediators may activate neutrophils and contribute to tissue damage and increase susceptibility to invasive microorganisms. In this study, we investigated the effect of a futsal match on muscle lesion markers, cytokines, and CRP in elite players. The basal and stimulated neutrophil responsiveness after a match was also evaluated based on measurements of neutrophil necrosis, apoptosis, phagocytic capacity, reactive oxygen species (ROS) production, and cytokines (tumor necrosis factor-alpha [TNF-alpha], interleukin [IL]-8, IL-1 beta, IL-10, and IL-1ra) production. Blood samples were taken from 16 players (26.4 +/- 3.2 years, 70.2 +/- 6.9 kg, 59.7 +/- 5.1 ml.kg(-1).min(-1), sports experience of 4.4 +/- 0.9 years) before and immediately after a match. Exercise increased the serum activities of CK (2.5-fold) and LDH (1.3-fold). Playing futsal also increased the serum concentrations of IL-6 (1.6-fold) and CRP (1.6-fold). The TNF-alpha, IL-1 beta, IL-8, IL-1ra, and IL-10 serum levels were not modified in the conditions studied. The futsal match induced neutrophil apoptosis, as indicated by phosphatidylserine externalization (6.0-fold). The exercise induced priming of neutrophils by increasing ROS (1.3-fold), TNF-alpha (5.8-fold), and IL-1 beta (4.8-fold) released in nonstimulated cells. However, in the stimulated condition, the exercise decreased neutrophil function, diminishing the release of ROS by phorbol myristate acetate-stimulated neutrophils (1.5-fold), and the phagocytic capacity (1.6-fold). We concluded that playing futsal induces inflammation, primes and activates neutrophils, and reduces the efficiency of neutrophil phagocytosis immediately after a match.