(2008). The mean of fluorescence intensity of stained cells was acquired using a Small molecule library BD FACSCalibur flow cytometer (BD Biosciences, Mississauga, ON, Canada) and data analyzed with CellQuest software (BD Biosciences, Mississauga, ON, Canada). All values were expressed as mean ± SEM. Parametric data were evaluated using analysis of variance, followed by the Tukey test for multiple comparisons. Non-parametric data were assessed using the Mann–Whitney test. Differences were considered statistically significant at p < 0.05. The SPSS statistical package (Release 8.0, Standard Version, 1997) was employed. First we identified the ability of natterins and nattectin to bind extracellular matrix proteins (laminin,

types I and IV collagen). In Fig. 1A–C, we only observed high recognition of untreated ECM components by antibodies direct against type I collagen, laminin or type IV collagen; and GSK126 datasheet an insignificant binding was reached by anti-venom, anti-natterins or anti-nattectin antibodies. After treatment of ECM components with T. nattereri venom (3 h, 37 °C), high levels of binding of natterins and nattectin were demonstrated to type I collagen ( Fig. 1D), and of nattectin to type IV collagen ( Fig. 1F). Natterins or nattectin showed weak binding

affinity to laminin ( Fig. 1E). To determine whether binding of toxins to ECM components altered the adherent properties, HeLa cells were incubated in dishes coated with types I and IV collagen and laminin,

all previously treated with venom or toxins. Decitabine solubility dmso HeLa cells that exhibit anchorage-independent cell growth (Aplin et al., 1998) showed similar adhesion levels to types I or IV collagen and laminin-coated dishes, which did not differ to adhesion levels of HeLa cells to plastic (the first two columns on the left in Fig. 2A–C). As shown in the last column of Fig. 2A and C, adhesion of HeLa cells was not hampered by binding of nattectin to types I or IV collagen, while venom and mainly natterins treatments inhibited the adhesion of cells on dishes coated with types I and IV collagen (third and fourth columns in the Fig. 2A and C). In Fig. 2B, adhesion levels of HeLa cells to laminin were similar after venom or toxins treatments. Based on the results that show natterins posses protease activity (Lopes-Ferreira et al., 2004) we investigate whether treatment of natterins directly degrade ECM components. For this, SDS-polyacrylamide gel electrophoresis after 24 h at 37 °C of incubation with natterins was carried out. Under reducing conditions, soluble type IV collagen appears in two forms, full-length (>250 kDa) and a 120 kDa form, which was degraded by natterins (Fig. 3, lanes 7–8). The high molecular forms of type I collagen above 250 kDa were also cleavage by natterins (Fig. 3, lanes 3–4). No proteolytic activity of natterins was observed to laminin (Fig. 3, lane 5–6).

, 2009). This trend is set to continue, with general circulation models

predicting particularly rapid warming at polar latitudes (Convey et al., 2009 and Kattenberg et al., 1996). In addition, specific microhabitats, such as the surfaces of rocks and bryophyte clumps, can experience maximum temperatures approaching or exceeding 30 °C (Convey, CYC202 solubility dmso 1996, Everatt et al., 2013 and Smith, 1988). Climate warming may increase the prevalence and duration of these exposures (Bokhorst et al., 2011 and Nielsen and Wall, 2013). The ability of polar terrestrial invertebrates to remain active at high temperatures has only as yet been explored in three continental Antarctic Collembola, and all show a remarkable capacity to remain active above 30 °C (Sinclair et al., 2006). The vast majority of polar terrestrial invertebrates express seasonal and shorter term thermal tolerance strategies to enable survival of shifts in temperature (Cannon et al., 1988, Worland, 2001 and Denlinger and Lee, 2010). However, the ability of polar terrestrial invertebrates to acclimate or acclimatise their thermal activity thresholds is less well known. Only

two polar species, the aphid, Myzus polaris, and the collembolan, Isotoma klovstadi, have been demonstrated to have this ability, with a depression in the CTmin of individuals reared at, or taken from, lower temperatures ( Hazell et al., 2010 and Sinclair et al., 2006). In the current study, the lower and upper thermal activity thresholds are characterised Ganetespib cell line in three common polar invertebrates widely regarded as ‘model’ species in their respective ecosystems: Cryptopygus antarcticus ( Block et al., 2009 and Tilbrook, 1967) and Alaskozetes antarcticus ( Block and Convey, 1995 and Burn, 1986) from the maritime Antarctic, and Megaphorura arctica ( Fjellberg, 1994) from the High Arctic.

In particular, how the thermal activity thresholds of these species respond to acclimation is explored. Summer acclimatised individuals of M. arctica were collected (-)-p-Bromotetramisole Oxalate from moss-covered slopes at Krykkefjellet and Stuphallet, near Ny-Ålesund, Spitsbergen, Svalbard (78°55′N, 11°56′E) in August 2011. Summer acclimatised individuals of C. antarcticus and A. antarcticus were collected from moss and algae, and the underside of rocks, on Lagoon Island (67°35′S, 68°16′W) and Léonie Island (67°36′S, 68°21′W), near to Rothera Research Station, Adelaide Island (western Antarctic Peninsula, maritime Antarctic), between January and March 2012. Samples of C. antarcticus and A. antarcticus were held at +4 °C (24:0 L:D) in plastic bags or boxes containing substratum from the sites at which they were found whilst at Rothera Research Station and were used shortly after collection in experiments 2.3, 2.4 and 2.6. These individuals were designated as the “summer acclimatised” group. Following each respective field season, samples of M. arctica, and C. antarcticus and A.

In case of Hamburg, climatically induced changes have to be combined with other changes, which may result from further modifications of the Elbe estuary (see Section 2). find more An important facet of these scenarios is the perspective of different time horizons, which will be associated with different geophysical changes. While not quantifiable, it is clear that also the uncertainty of future projections will be diminishing. A scenario for a certain time window constructed with the knowledge of 2030 will be less uncertain than a scenario

for the same time window constructed with the knowledge available in 2010. Natural science is generating knowledge about the sensitivity of coastal processes to natural and human influences and about possible pathways of future developments. However, transforming these insights into Francis Bacon’s knowledge, scientia est potentia = capacity to set something in motion ( Stehr, 2012), needs more than just “good science”. When it comes to decisions, the role of science BYL719 chemical structure diminishes, and the responsibility is with stakeholders representing political, economic or social interests. Decisions are not scientific,

but follow power structures, political and economic priorities and societal developments. Scientifically produced decision support systems can support decisions by providing specific sets of information and supply evidence-based decision support. Decisions themselves are in most cases normative and interest driven. When scientific actors try to interact with stakeholders, including media and public at large, they often follow

simplistic worldviews – in particular the “linear model” according to which scientifically constructed knowledge is superior und “true” (van der Sluijs, 2010). Therefore, in this naïve view, science is legitimized in determining what is a “right” or a “wrong” decision. The other model is that of the “empty vessel”, according to which stakeholders and public are simply uneducated and do not understand (like small children). Thus, they need to be taught by scientists. As soon as these so far uneducated people understand the considered system, they will opt for the “right” decision. Philosophy of science informs us that heptaminol science is not providing “truth” but “best explanations” for the time being, consistent with empirical evidence and with generally accepted theories (e.g., Fleck, 1980). Attempting falsification is important, because it represents a permanent testing if an explanation is still the “best” for the time being. According to social science models like the linear one or the empty vessel are not realistically describing social reality. Stakeholders hold their own knowledge, which often enough is not really science-based but rooted in cultural constructions or economic or political interests (von Storch and Stehr, 2014).

“
“The etiology of GI neuromuscular diseases, including functional GI disorders, remains largely unknown. There is recent evidence to

support underlying neuromuscular pathological changes that are heterogeneous and include the loss of interstitial cells of Cajal (ICC) and enteric nerves and the presence of inflammatory infiltrates.1, 2, 3, 4 and 5 For example, surgically obtained full-thickness gastric biopsy (FTGB) samples from patients with gastroparesis show a decrease in ICC in 50% of patients, an immune infiltrate in 45%, and a decrease in nerve fibers.6 The presence of an immune infiltrate correlated with nausea and vomiting.7 Nonsurgically obtained FTGB samples that include the muscularis propria to evaluate the enteric nervous system, ICC, immune cells, and other related cells are essential to further our understanding of the pathophysiology find more of these

disorders and intervene Dasatinib in vitro earlier in the disease process. Mucosa-based biopsies are insufficient as they do not allow evaluation of the deep muscle layers as well as the myenteric plexus present between the inner circular and outer longitudinal muscle layers. Our earlier work with experimental endoscopic techniques was limited by a combination of poor safety data and inadequate tissue sampling.8 and 9 Endoscopic acquisition of FTGB samples that is safe, effective, and minimally invasive would contribute to accurate diagnosis and identification Glutamate dehydrogenase of patients who would benefit from targeted therapy. Full-thickness gastric biopsy by using the submucosal endoscopy with mucosal flap technique with endoscopic suturing is feasible, reproducible, and safe. Ample tissue samples can be obtained

by using this technique to allow analysis of multiple cell types including myenteric ganglia and interstitial cells of Cajal. The aims of this study were to determine the technical feasibility, reproducibility, and safety of performing an FTGB by using a submucosal endoscopy with mucosal flap (SEMF) technique; reliable tissue closure by using endoscopic suturing; the ability to identify myenteric ganglia in resected specimens; and long-term safety. This preclinical survival study in a pig model was approved by the Institutional Animal Care and Use Committee. Twelve pigs were studied. Each animal underwent an SEMF procedure with an FTGB followed by closure of the offset mucosal entry point by using an endoscopic suturing device. Animals were kept alive for 2 weeks at which time a repeat endoscopy was performed, followed by necropsy. The main study outcome measurements were the clinical course of animals, technical feasibility, reproducibility, and short- and long-term (2 weeks) safety of the procedure. Data on the procedure, clinical course, and follow-up endoscopy with necropsy were recorded. Data analysis was descriptive for this feasibility study.

The norms cover a maximum age of 12 years, 11 months; therefore, we used data from a larger control sample to convert raw scores to standard scores (see Barry et al., 2007). Handedness was assessed using a brief demonstration hand preference (based

upon the Edinburgh Handedness Inventory; Oldfield, 1971). Children were asked to demonstrate how they would perform each of 10 actions; write, this website draw, throw, use scissors, brush their teeth, cut with a knife, use a spoon, sweep with a broom (upper hand), take the lid of a box, and deal cards. Left, Right, or either (if child indicated both) hand was recorded in each case. The number of right hand preferences was taken as a measure of hand dominance. MRI data were obtained using a 1.5-T Siemens Sonata scanner with a single-channel head coil. Participants wore noise-attenuating headphones and padding was inserted around the head to restrict movement. They watched a DVD of their choice via a mirror on the head coil during acquisition of the structural data. A T1-weighted image was acquired in each participant Linsitinib nmr for structural analysis and image registration (3D

FLASH; TR = 12 ms; TE = 5.6 ms; 1 mm isotropic voxels; matrix = 256 × 256 × 208; elliptical sampling; orientation = coronal). One acquisition of this T1-weighted sequence took five minutes. At the end of these five minutes, the image was inspected for motion artefacts and, if necessary, children were reminded to keep still for a further five minutes. Three artefact-free images were successfully acquired in each participant. The first and third images were registered (rigid-body transformation; 6 degrees of freedom; trilinear interpolation) to the second image to correct for movement CYTH4 between acquisitions and summed to create a single T1-weighted

image in each participant. Before the functional task, participants were removed from the scanner for a break if necessary. For the functional scan, whole-head T2∗-weighted echo-planar images (TR = 3s; TE = 50 ms; 120 volumes, 6 min), were acquired. Each volume comprised 35 4-mm axial slices (in-plane resolution 3 mm × 3 mm). Stimuli were presented over MRI compatible headphones (MR Confon: http://www.mr-confon.de) at a comfortable listening level (estimated ∼70 dB). Participants were asked to keep their eyes closed. The task used for functional imaging was based on the Auditory Responsive Naming task previously used with PET (Bookheimer et al., 1998). It was chosen because it was thought to be engaging for children, easy enough for them to comply with and known to produce activation in both posterior and anterior language areas (Wernicke’s and Broca’s area, respectively). In the Speech condition, participants heard simple three-word auditory definitions of a high frequency word and were required to silently generate an appropriate word; for example, ‘wear on head’ > silently generate ‘hat’.

We also used the studentized version of the test, which is more robust to non-Gaussian variation (Koenker, 1981), and the results remained identical to at least two decimal places. Once the power transformation has been selected, the regression is not used further. For the 20 pools, the selected powers ranged from 0.23 to 0.31, mean 0.27. In other words, the optimal transformations were close to fourth root (power = 1/4). Fig. 1 this website shows the Bland and Altman plots for the first haemagglutinin pool, and the second neuraminidase pool. These plots

also show i) the test wells positive on the T-SPOT criteria (see Introduction), and ii) the control wells which would have been positive on the same criteria, had the test and control status been reversed, hereafter referred to as pseudo-positive. For haemagglutinin, the T-SPOT-positive test wells greatly outnumber the pseudo-positive control wells (247:46), but this is not the case for neuraminidase (58:59). By quartile on the horizontal axis, the proportions positive on the T-SPOT criteria are: 0, 23, 26 and 32% for haemagglutinin and 0, 0, 6 and 16% for neuraminidase. To select a threshold

value for defining positive wells, we use the principle that test minus control values should, on average, be larger than control minus test. Otherwise, there Ruxolitinib ic50 is no evidence of a ‘signal’ over the ‘noise’ of control variation, and any positivity threshold is dubious. To select the threshold we compare the empirical cumulative distribution functions (ECDFs) of i) test–control for those plates with test > control and ii) control–test for those with control > test. The ECDF of a sample is simply the proportion of the data points which lie at or below a given value. The difference between ECDFs can be used to discriminate between a mixture of two distributions. In particular, the value which maximizes the difference in ECDFs also maximizes the probability of correct classification (Stoller, 1954). Hence, for the current purpose, we choose the threshold to be the value which maximizes the difference between the above two ECDFs. Pools whose difference over

control exceeds this value are declared positive. In principle it is possible for this maximum difference in ECDFs to occur at more than Docetaxel one value on the horizontal axis. Hence we define the threshold, more precisely, to be the lowest such value on the horizontal axis. This is shown in Fig. 2 for the two selected pools. Greater data values shift the ECDF to the right, making it lower at any given point on the horizontal axis. For haemagglutinin, the ECDFs of test-minus-control and control-minus-test are much more widely separated than for neuraminidase. For haemagglutinin, the maximum difference in ECDFs is 0.22 and occurs at a transformed test-minus-control value of 1 (i.e. a value greater than 1 is considered positive).

It has been assumed

that EDN2 would mimic the actions of its more this website abundant counterpart EDN1, but recent findings in vitro and in knockout mice underscore that EDN2 does not simply amplify or duplicate EDN1 action and imply a distinct function of EDN2 in physiological and pathophysiological processes [36]. Furthermore, EDN2, and not the more abundant EDN1, was first isolated from RCC cell lines [37]. A recent paper reported EDN2 expression to be a common and early event in patients with localized ccRCC undergoing nephrectomy and proposed a potential role in ccRCC progression [38]. An association of higher tumor expression of EDN2 with longer progression-free survival could not be confirmed after adjustment for known clinicopathologic factors and it would be interesting to compare expression levels with tumors of patients with advanced metastatic disease. Grimshaw et al. reported an important influence of EDN2 on the invasive potential of breast cancer cells and proposed a mechanism where EDN2-secreting tumor cells provide chemotactic cues to tumor-infiltrating macrophages, which in turn secrete matrix metallopeptidase (MMP)-2 and MMP-9 to facilitate tumor

cell invasion PD0332991 order and metastasis [39]. The observed effect was dependent on both endothelin receptor B and MAPK signaling, and expression of EDN2 and its receptor was stronger at the invasive margin of the tumor tissue. Of note, we observed inhibition of the MAPK signaling pathway on YAP knockdown in MZ1774 cells. Overexpression of EDN2 increases the invasive potential of breast cancer cell lines in vitro but is not sufficient to induce an invasive phenotype in benign cells, indicating the cooperation with other signaling networks [40]. Concurrently, Said et al. reported

an instrumental role of EDN1 signaling through endothelin receptor A in the development of metastatic bladder cancer and delineated a proinvasive network governed by members of the endothelin family involving direct actions like the activation of proinflammatory transcription factors such as activator protein 1 and nuclear factor kappa-light-chain-enhancer of activated B cells in human monocytes and cancer cells and the stimulation of the production of a range of proinvasive cytokines like interleukin-6, cyclooxygenase GBA3 2, chemokine (C-C motif) ligand 2 (CCL2), MMP-2, and MMP-9 as well as indirect modulation of the tumor microenvironment by influencing tumor-stroma interactions as well as tumor-associated immune cells [41]. These endothelin functions were instrumental in the process of metastatic colonization, the first step of the establishment of a filial tumor at a distant site, and pharmacologic blockade of endothelin receptor signaling inhibited metastasis significantly in an experimental animal model, despite having only modest effects on primary tumor growth.

5 M ethanolamine, 0.5 M NaCl pH 8.3 and PF-562271 purchase again 0.1 M AcONa, 0.5 M NaCl pH 4 were passed through the column (6 column volumes for each buffer). A syringe was used for all wash steps. Flow through and wash solutions were analysed by phenol sulphuric assay to calculate the amount of sugar linked to the resin. Blood containing anti-Salmonella antibodies was venesected from a healthy adult and left to clot at 22 °C for 4 h before separating by centrifugation at 4 °C and freezing in aliquots at − 80 °C. Ethical approval for the use

of human serum in this study was granted by the Life and Health Sciences Ethical Review Committee of the University of Birmingham. Informed written consent was obtained prior to venesection. Ammonium sulphate was added as a solid to 1 ml of human serum to give a final concentration of 0.5 g/ml and the mixture was placed on ice for 5 min. The serum was centrifuged at 4 °C, 3300 × g for

5 min and the supernatant discarded. The precipitate was washed twice with 1 ml 0.5 g/ml ammonium sulphate. The pellet was solubilized in 0.3 ml PBS and dialysed overnight against PBS at 4 °C. NHS HiTrap columns with activated OAg were equilibrated with PBS (6 column volumes) before applying the serum protein solution to the column and incubating overnight at 4 °C. Columns were then washed with PBS (6 column volumes), followed by 50 mM NaH2PO4, 500 mM NaCl pH 7.2 (6 column volumes). Bound antibodies were eluted in 5 column volumes of elution buffer, collecting fractions of 0.5 ml each. CB-839 supplier 0.1 M glycine, 0.1 M NaCl at pH 2.4, 2.6, 2.8 and 3.0; 20% ethanol; 4 M MgCl2 in 10 mM Tris base pH 7; 8 M urea; and 100 mM Tris base pH 9 were tested as elution buffers. Following elution with glycine buffers using a pH 2.4–3.0,

the pH was adjusted to 7.0 with 2 M Tris pH 9. Individual eluate fractions were analysed for protein content by measuring absorption at 280 nm. After elution, all eluates were dialysed overnight against PBS at 4 °C. Purified antibodies were stored at 4 °C. Retention of antibodies on columns was investigated by applying 1% SDS to columns and analysing SDS-eluates by SDS-PAGE. Columns were washed with PBS and stored in 0.05 M Na2HPO4, 0.1% NaN3 Phosphoglycerate kinase pH 7.0 at 4 °C. 96-well flat bottom plates (NUNC Maxisorp) were incubated with 100 μl per well of 5 μg/ml TLR-grade smooth LPS from S. Typhimurium (Alexis Biochemicals) or 15 μg/ml S. Typhimurium OAg, overnight at 4 °C. After coating, plates were washed with PBS 0.05% Tween and incubated with 200 μl blocking buffer (PBS 1% BSA) per well for 1 h at 37 °C. Plates were washed again with PBS 0.05% Tween and incubated for 1 h at 37 °C with 100 μl serial dilutions of antibody solution diluted with PBS 1% BSA 0.05% Tween. Following further washes, 100 μl of goat-antihuman alkaline-phosphatase-labelled IgG (Southern Biotech) was added to each well and plates were incubated for 1 h at 37 °C.

The covered SEMS were successfully removed from all patients within 30 days. The classification of Stapfer was used to determine the type of perforation. Table 1 Our results suggest that endoscopic management of transmural PSP can be successfully achieved in a significant

subset of patients yet at a lower cost and a shorter hospital stay. Larger studies to identify independent predictors of a successful outcome are needed. Table 1. Group I Group II P N 12 11 — Media age (years) 69.7 63.9 PD0325901 ic50 — Perforation type  I 5/12 (41.6%) 4/11 (36.3%) NS  II 7/12 (58.3%) 7/11 (63.6%) NS Success of procedure (%) 11/12(91.6%) 1 patient develop severe retroperitoneal infection and die 11/11(100%) NS Peritoneal perforation (%) 5/12 (41.6%) 3/11 (27.2%) 0.086 Retroperitoneal perforation (%) 7/12 (58.3%) 8/11 (72.7%) 0.191 Mean time of hospitalization (days; range) 4.1 [3-5] 15.2 [13-18] 0.0123 Size of perforation (mm; %)  5-10 8/12 (66.7) 6/11 (54.6%) NS  10-20 3/12 (25%) 4/11 (36.3%) NS  >20 1/12 (8.3%) 1/11 (9.1%) NS Technical procedure SEMS + clips = 12 Hepaticojejunostomy = 4/11 (36.3%) Suture of perfuration = 4/11 (36.3%) Duodenal suture = 3/11 (27.2%) — Complications

(%) Retroperitoneal abscess: 1/12 (8.3%) Fever: 3/12 (25%) Death: 1/12 (8.3%) Abscess: 1/11 (9.1%) Deiscense Epigenetics inhibitor anastomosis: 1/11 (9.1%) Wound infeccion: 1/11 (9.1%) NS Mean total cost (U\$) 14,700 ± 2835 19,872 ± 2,587 0.0103 Full-size table Table options View in workspace Download as CSV “
“Recently we reported Tau-protein kinase on the feasibility and safety of transenteric drainage of pancreatic pseudocysts and gallbladders using a newly developed lumenal apposition device (GIE 2012). We now report on the first clinical experience of creation of a transenteric

choledochoduodenostomy using the Lumenal Apposition Device (LAD). To evaluate the feasibility and, safety of transenteric biliary drainage using the LAD for palliation of obstructive jaundice. The LAD consists of braided nitinol heat-set into a dual flange configuration. Fully expanded, the stent diameter and length measure 6 mm and 8 mm, respectively. The flange diameter is 14 mm. The LAD is constrained within a 10 Fr delivery catheter. In 8 patients (3 male, mean age 61.1, range 62-99) with distal biliary obstruction and jaundice due to 4 pancreatic cancers, 3 ampullary cancers, and 1 distal bile duct cancer, the LAD was placed using a 3.7 mm channel curved array echoendoscope (Olympus). The bile duct was punctured with a 19G FNA needle under endoscopic ultrasound (EUS) guidance and a guidewire inserted. The fistula tract was primed for LAD placement with a bougie catheter and/or cautery needle and/or 4 mm non-compliant balloon. The LAD was deployed under combined EUS and endoscopic guidance. After deployment, the LAD lumen was dilated with a non-compliant balloon catheter to 6 mm to optimize drainage when indicated. Naso-biliary catheters were placed across the LAD for irrigation at the discretion of the endoscopist.

Guinea pigs have been used in experimental models to evaluate allergic airway diseases such as asthma because they are rapidly sensitized Selleckchem Sunitinib to aerolized ovalbumin without the need for intraperitoneal injections. These results in an airway response to challenge similar to that of asthmatic phenotypes, including a robust bronchoconstriction that is lacking in other rodents (Bice et al., 2000, Wenzel and Holgate, 2006 and Zosky and Sly, 2007). In addition, the pharmacological responses of guinea pig airways are very

similar to those of humans in comparison to any other animal model (Ressmeyer et al., 2006). Therefore, the aim of this study was to evaluate the effects of aerobic exercise on airway inflammation and remodeling in a model of chronic allergic airway inflammation in guinea pigs. This

study was approved by the review board for human and animal studies of the School of Medicine of the University of São Paulo (São Paulo, Brazil). All of the animals in the study received human care in compliance with the find more Guide for the Care and Use of Laboratory Animals (NHI publication 85-23, revised 1985). Thirty male Hartley guinea pigs (250–280 g) were divided into four groups: Control (non-exercised and non-sensitized; C group; n = 7); Aerobic Exercise (non-sensitized and aerobically exercised; AE group; n = 7); Ovalbumin (OVA-sensitized and non-exercised; OVA group; n = 8) and OVA + AE (sensitized and aerobically exercised; OVA + AE group; n = 8). Animals were placed filipin in an acrylic box (30 cm × 15 cm × 20 cm) coupled to an ultrasonic nebulizer (Soniclear, SP, Brazil) and received seven sessions of OVA inhalation solution diluted in sterile saline (NaCl 0.9%). The Control and AE groups (non-sensitized) received the same number of inhalation sessions with sterile saline. All

inhalation sessions lasted 15 min or until the animal displayed respiratory distress (sneezing, coryza, cough or retraction of the thoracic wall) as previously described. OVA inhalation was performed for 8 weeks (3×/week) with increasing concentrations (from 1 to 20 mg/ml) to avoid OVA tolerance (Tiberio et al., 1997). Animals were initially adapted to the treadmill for 5 days (5 min, 8% inclination, 0.3 km/h). Next, a maximal exercise treadmill test was performed to establish the intensity of AE training (low intensity corresponded to 50% of the maximal speed). The maximal exercise treadmill test consisted of a 5-min warm-up (8% inclination, 0.3 km/h) followed by a gradual increase in treadmill speed (0.3 km/h every 3 min). The maximal exercise capacity was considered to be the maximal speed that animals were able to run after receiving 10 mechanical stimuli as previously described (Vieira et al., 2007). The speed of the AE was calculated as the average of the maximal speed achieved for each animal group in the maximal exercise treadmill test.