However, on UM clearfelled sites desired invader species such as

However, on UM clearfelled sites desired invader species such as Oxalis acetosella (woodsorrel), Anemone nemorosa (wood anemone), Conopodium majus (pignut) and Primula vulgaris (primrose) were not found, while bluebell was seen on only 15 quadrats and Teucrium scorodonia (wood sage) on just 2. The solitary PAWS site that was examined had a considerably richer ground flora with wood sorrel, wood sage and bluebell seen on 21%, 29% and 79% of quadrats respectively. We found that the sites which had been clearfelled 10 years PF-01367338 cell line ago had significantly

greater vascular plant coverage (111%) compared to sites that had been clearfelled 2 years ago (11.7%, p = 0.001). The % mean woody debris on spruce clearfell sites declined from 51% 2 years after felling to 12.7% and 5.1% at 5 and 10 years post-felling respectively. We have

explored the regeneration density of native broadleaved species on clearfelled conifer sites in upland Britain. We compared regeneration on clearfelled sites to control sites that had neither been planted with conifers or clearfelled. We restricted our analysis to a subset of sites with similar BGB324 order time since clearfelling and soil type. Mean regeneration density on this subset of clearfelled upland moorland sites (3392 individuals/ha) was significantly greater than on upland moorland (64 individuals/ha) or improved farmland (14 individuals/ha) sites. Availability of data meant that in this analysis we combined sites across regions (Lake District and eastern

Scotland) and were unable to account for site location as a covariate. Regeneration density on all clearfelled upland moorland sites (3515 individuals/ha) was at the lower end of that recorded by Harmer and Morgan (2009) (3000–11,000 individuals/ha) in a storm damaged lowland conifer site in south-east England that had been allowed to naturally regenerate. The regeneration density we recorded was lower than conifer regeneration Liothyronine Sodium within small windthrows (Jonásová et al., 2010) or clearfells (Modrý et al., 2004 and Holgén and Hånell, 2000) where sapling densities as great as 160,000 individuals/ha have been recorded (Modrý et al., 2004, Holgén and Hånell, 2000 and Jonásová et al., 2010). The high regeneration density in these studies was likely due to an ample seed source due to the surrounding woodland whereas in our study the seed source was limited to individual mature trees. Nevertheless, the regeneration density on clearfelled upland moorland sites and a clearfelled PAWS site (5790 stems/ha) exceeded the suggested sapling stocking densities for new native woodland in Britain of between 500 and 2000 stems/ha (Forestry Commission, 2010). The diversity of regenerating species was usually lower than that of the adjacent seed sources with regeneration dominated by birch on all but one clearfelled site, as has been found previously at storm damaged lowland sites in Britain (Harmer and Morgan, 2009 and Harmer et al.

pylori-associated gastric disease The Mongolian gerbil model is

pylori-associated gastric disease. The Mongolian gerbil model is the best animal model for this purpose because H. pylori infection induces chronic gastritis, gastric ulcers, and intestinal metaplasia in these animals. Mongolian gerbils develop gastric neoplasia and gastric cancer after chronic infection by H. pylori strain 7.13 [28] and [29], as used in the present study. After the infection of gerbils with H. pylori, we determined: the changes in LPO level, which is an index of oxidative membrane damage; the activity of MPO, a biomarker of neutrophil infiltration; the induction of inflammatory mediator keratinocyte chemoattractant

factor (KC), an IL-8 homolog in rodents [30]; IL-1β; iNOS; and the phosphorylation of selleck kinase inhibitor IκBα, which reflects the activation of NF-κB. In addition,

Microbiology inhibitor viable H. pylori colonization in the stomach, changes in food intake and body weight, stomach weight/total body weight, and histological analysis of gastric mucosa were compared between animals that received RGE and those that did not. Five-wk-old male specific-pathogen-free Mongolian gerbils (MGS/Sea) with an average weight of approximately 40 g were purchased from Charles River Laboratories (Wilmington, MA, USA). Gerbils were housed in polypropylene cages on hard wood chip bedding in groups of five/cage. Food and water were provided ad libitum. The animals were maintained in a temperature-controlled room (22 ± 2°C) with a 12-h light–dark cycle. The

animal experiments were performed in accordance with institutional guidelines. Protocols were reviewed and approved by the Institutional Animal Care and Use Committee of the Yonsei University Medical Center (Seoul, Korea; Permit No.: 10-107). Ten gerbils were included in each group. Histological observations are reported for 10 gerbils/group. All animals were maintained in the specific pathogen-free facility at Yonsei University Medical Center. H. pylori strain 7.13 was maintained as frozen stock at –80°C in brain–heart infusion medium supplemented with 20% glycerol and 10% fetal bovine serum. Bacteria were grown on horse blood agar plates containing 4% Columbia agar base (Oxoid, Basingstoke, Hampshire, UK), 5% defibrinated horse blood (HemoStat Labs, Dixon, CA, USA), Morin Hydrate 0.2% β-cyclodextrin, 10 μg/mL vancomycin, 5 μg/mL cefsulodin, 2.5 U/mL polymyxin B, 5 μg/mL trimethoprim, and 8 μg/mL amphotericin B at 37°C under microaerophilic conditions. A microaerobic atmosphere was generated using a CampyGen sachet (Oxoid) in a gas pack jar. For liquid culture, H. pylori was grown in brucella broth (Difco & BBL Diagnostics, Franklin Lakes, NJ, USA) containing 10% FBS (Gibco-BRL, Grand Island, NY, USA). Cultures were shaken in a microaerobic environment. According to the growth curve, 108 bacteria were collected and resuspended in 500 μL of brucella broth for the infection of each animal.


on virus neutralization in monocytes were support


on virus neutralization in monocytes were supported by the Singapore National Research Foundation under its Clinician-Scientist Award administered by the National Medical Research Council (NMRC/CSA/025/2010). “
“Defective interfering (DI) viruses are natural mutants that arise spontaneously, occur widely, and have a genome that has undergone at least one major deletion. As a result their replication is dependent on complementation by a genetically compatible infectious helper virus to provide any missing BGB324 order function. All DI genomes retain sequences that allow them to be packaged and replicated. The resulting DI virus particle is usually indistinguishable from that of the infectious virus. In cell culture DI viruses are not only defective but also interfering, the DI genome being the structure responsible for this property. Thus, under appropriate conditions, the presence of the DI genome reduces the amount of infectious progeny virus produced (Holland, 1990a, Holland, 1990b, Huang and Baltimore, 1970, Nayak et al., 1989 and Perrault, 1981). Some, but not all DI viruses can protect animals from clinical

disease caused by the homologous virus (Barrett and Dimmock, 1986, Dimmock, 1991, Dimmock, 1996, Dimmock et al., 2008 and Roux et al., 1991). Influenza DI 244 virus also protects against genetically unrelated (heterologous) viruses Cilengitide clinical trial (pneumonia virus of mice (PVM: Paramyxoviridae and influenza B virus) in vivo, primarily by induction of type 1 Oxalosuccinic acid interferon ( Easton et al., 2011 and Scott et al., 2011b). DI virus-induced interferon is not required for protection against a lethal challenge with influenza A viruses ( Easton et al., 2011). Influenza A DI viruses were the first to be described (von Magnus, 1954) and have been studied extensively (Nayak, 1980, Nayak et al., 1985 and Nayak et al., 1989). However, most DI influenza virus preparations contain many different defective RNA sequences, so that it was not possible to determine the relationship between a particular defective RNA sequence and its biological

properties. Recently we solved this problem using reverse genetics to make cloned DI viruses that contain one major species of DI RNA (Dimmock et al., 2008; Duhaut and Dimmock, 1998, Duhaut and Dimmock, 2000, Duhaut and Dimmock, 2002 and Duhaut and Dimmock, 2003). One such influenza virus, containing the 244 DI RNA, derived from segment 1, strongly protected mice against disease caused by several different influenza A virus subtypes when inoculated intranasally (Dimmock et al., 2008). This protection is dependent on the integrity of the 244 DI RNA and protection is lost when the DI RNA is destroyed by extensive UV irradiation. The DI influenza A virus particle retains receptor specificity and, when topically applied, targets the DI RNA to influenza virus-susceptible cells in the respiratory tract.

1a) Of all submitted bids players bid zero points on M = 14 4, 9

1a). Of all submitted bids players bid zero points on M = 14.4, 95% CI [8%; 21%] of all trials. Surprisingly, players reduced their bids over the course of auctions in the PV± and PV+ conditions measured as the difference between the mean first five bids and the mean last five bids ( Fig. 1b and Table 1). Wide confidence intervals of effect estimates ( Table 1) indicate that the strength of reduction was not consistent across players. Indeed, these differences were, at least partly, driven by the initial difference between the bids of

the two players in the PV± and PV+ condition ( Fig. 2). Players adjusted their bids in the direction of the bids of the other player, with stronger adjustments for the player initially bidding more

(slope estimate for interactions <0.5 in Table 2). This resulted in 85% of the participants bidding initially more in the PV+ selleck inhibitor condition also winning the majority of the auctions. In the PV± condition only 52% of the players that initially bid more also won more than half of the auctions. To examine the effects of underlying dynamics on a trial-to-trial basis, we Galunisertib price focused our analysis on the effect of the two previous auction outcomes on player’s propensity to increase or decrease their bids. Player bids show a consistent pattern across all preference levels where players increased their bids when losing and decreased their bids when winning (Table 3). The positive effect on bids was slightly larger when players

first won and then lost with regard to auctions with one particular item. As final player bids did not reflect the preference for an item, we analyzed pre- and post-auction preference statements for the five auction items. A considerable number of players (66.6%) changed their preference ranking. Our main goal was to identify factors from the auction that influence player preference changes, an index for private value change. We Rucaparib research buy found that the initial difference between player bids and the evolution of bids for a particular item affected bid dynamics (see Results on dynamics during the auction). Two additional factors entered the analysis as measures for the degree of competition: sunk costs, i.e. amount points lost in auctions, and the number of wins minus the number of losses. Based on these factors, we constructed a multinomial model where we contrasted auctions with increasing and decreasing preference with auctions without a change. Two patterns emerge from this analysis. First, some model coefficient estimates for increasing and decreasing preference point in the same direction (same sign) with approximately same effect size (Fig. 3 and Table S1). This indicates that these factors influence the probability to change preference in general, i.e. not restricted to increasing or decreasing changes. The most noteworthy of these factors was the difference between the two initial bids between the two players of a pair (ID).

In view of the evidence presented, there is no need to belabor th

In view of the evidence presented, there is no need to belabor the point that in the timespan under consideration human impacts overrode natural phenomena, a distinguishing characteristic of the Anthropocene. Most UMI-77 clinical trial proponents of such a chron correlate its onset with the Industrial Revolution, but this would seem justifiable in Tlaxcala only if the effects of rows F, H, and I significantly outweighed all previous historical conjunctures. This is not the case, and land use overrode climate in determining sediment transfers since the local Neolithic Revolution in the 1st millennium BC. But, the post-Conquest era left novel and durable stratigraphic markers

exclusive of the Anthropocene, in rural areas and close to drainage divides, the places least expected by Zalasiewicz et al. (2011). The ubiquituous tepetate surfaces are erosional unconformities that persist in the stratigraphic record. Even after burial, lag deposits of sherds and architectural NLG919 order rubble distinguish them from similar

boundaries formed in pyroclastics before the advent of village life. The cover layer has all the defining attributes of a ‘legacy sediment’ (James, 2013) but is significantly older than examples named as such in the United States. This type of legacy sediment is widespread in other terraced landscapes, characterized by composite, polycyclic and spatially variable soils (Krahtopoulou and Frederick, 2009), but Tlaxcala is the only example I know where it is mapped at regional scales of 1:100,000. It allows the recognition agricultural management even after risers have been erased and the original O-methylated flavonoid slope gradient reestablished. The most predictable way of framing the discussion is in terms of the so-called Columbian debate about the positive or negative impact of indigenous vs. introduced European land use (Butzer, 1993, Crosby, 1972, Crosby, 1986 and Denevan, 1992). In Mexico, it came to be known as the Melville-Butzer controversy (Hunter, 2009), and around the time of the

quincentennial it revolved around arguments for (Melville, 1994) and against (Butzer and Butzer, 1993 and Butzer and Butzer, 1995) a ‘plague of sheep’. In Tlaxcala, the problem has been pondered since its inception. From the perspective of a 16th C. member of the local nobility like Muñoz Camargo, for whom the most valuable asset of a landed estate were its tenants, the epidemics were indeed a disaster to be decried, though he apparently had no qualms about his family’s profits from stocking the vacant land with sheep and cattle (Gibson, 1952, 152; Muñoz Camargo, 2000[1585], 88). Tlaxcala would seem a prime candidate for the ‘plague of sheep’ hypothesis, though historians disagree as to the permanent or transient nature of sheep ranching, and the reliability of Colonial head counts.

The definition of the main sedimentary facies in the cores (indic

The definition of the main sedimentary facies in the cores (indicated with different colors in Fig. 2) is useful for interpreting the acoustic profile, identifying the sedimentary features, as well as allowing a comparison with similar environments. Most of the alluvial facies

A are located below the caranto paleosol and belong to the Pleicestocene continental succession. The sediments of the facies Ac in cores SG28 e SG27 are more recent and correspond to the unit H2a (delta plain and adjacent alluvial and lagoonal deposits) of the Holocene succession defined by Zecchin et al. (2009). In the southern Venice Lagoon they define also the unit H1 (transgressive back-barrier and shallow marine deposits) and the unit H2b (prograding delta front/prodelta, shoreface and beach selleck inhibitor ridge deposits). In the study area, however, the units H1 and H2b are not present: the lagoonal facies L (i.e. the unit H3 of tidal channels and modern lagoon deposit in Zecchin et al.

(2009)) overlies the H2a. A similar succession of seismic units is also found in the Languedocian lagoonal environment in the Gulf of Lions (unit U1 – Ante-Holocene Enzalutamide ic50 deposits and units U3F and U3L, filling channel deposits and lagoonal deposits, respectively) in Raynal et al. (2010), showing similar lagoon environmental behavior related to the sea-level rise during the Flandrian marine transgression ( Storms et al., 2008 and Antonioli et al., 2009). The micropalaeontological analyses

( Albani et al., 2007) further characterize the facies L in different environments: salt-marsh facies Lsm, mudflat facies Lm, PRKD3 tidal channel laminated facies Lcl and tidal channel sandy facies Lcs. As described by Madricardo et al. (2012), the correlation of the sedimentary and acoustic facies identifies the main sedimentary features of the area (shown in vertical section in Fig. 2 and in 2D map in Fig. 3). With this correlation and the 14C ages we could: (a) indicate when the lagoon formed in the area and map the marine-lagoon transition (caranto); (b) reconstruct the evolution of an ancient salt marsh and (c) reconstruct the evolution of three palaeochannels (CL1, CL2 and CL3). The core SG26 (black vertical line in Fig. 2a) intersects two almost horizontal high amplitude reflectors (1) and (2), interpreted as palaeosurfaces (Fig. 2a). A clear transition from the weathered alluvial facies Aa to the lagoonal salt marsh facies Lsm (in blue and violet respectively) in SG26 suggests that the palaeosurface (1) represents the upper limit of the Pleistocene alluvial plain (caranto). The 14C dating of plant remains at 2.44 m below mean sea level (m.s.l.

The effect of the association between variables and outcome (cons

The effect of the association between variables and outcome (consumption or non-consumption of at least a daily portion of each food category) was assessed

by bivariate analysis using Poisson regression with robust estimation for variance adjustment. Variables that showed an association with lower level of statistical significance or equal to 20% (p ≤ 0.20) were included in the multivariate analyses. The magnitude of association between the investigated factors and feeding practices was estimated using prevalence ratios and confidence intervals (95% CI) in robust Poisson regression bivariate and multivariate analyses. The study was approved by the Ethics Committee in Research of the Universidade Federal de Ciências da Saúde de Porto Alegre, and approved by the Porto Alegre City Hall Ethics Committee. Children with unfavorable clinical situations were referred to health services. Fig. 1 presents the flowchart of data collection, detailing

the losses, refusals, and exclusions that occurred in each phase. It is noteworthy that 149 families (20.8%) were not located for follow-up, considering all phases of data collection. In addition, 87 children (12%) needed to be excluded from analysis because they did not have 24-hour recalls at 12-16 months and/or 2-3 years. Among the children studied, the distribution between genders occurred homogeneously: 51.8% (n = 201) were male.

The characterization of the sample regarding the maternal and family data is described in Table 1. The prevalence Selleck AZD2281 of children aged 2-3 years who did not consume one serving of fruit or one serving of vegetables, considering the mean consumption obtained by two 24-hour recalls, was 58.0% (n = 225) and 87.4% (n = 340), respectively. The median MycoClean Mycoplasma Removal Kit intake of fruits and vegetables a day among children was 55 g (0-130 g) and 5 g (0-30 g), respectively. When assessing how many children consumed the recommended three servings of fruits and vegetables in at least one of the dietary surveys, it was observed that only 35 children (9%) achieved this recommendation for fruit and only one (0.2%) for vegetables. Mothers who reported offering fruits and vegetables daily to their children when responding the 12-16 month questionnaire accounted for 50.4% and 42.4% of the sample, respectively. The energy consumption of non-recommended foods at 12-16 months was on average 13.61% of total energy intake; for 9.2% of the children, these foods accounted for over 30% of energy consumption. Mothers who reported having offered soft drinks or artificial juices and at least three different types of non-recommended foods during the month before the 12-16 month data collection accounted for 83.3% and 68.3% of the sample, respectively. Table 2 shows the variables associated with the consumption of fruit at 2-3 years.

10 Today, there is no literature available about pulmonary effect

10 Today, there is no literature available about pulmonary effects in humans after aspiration or inhalation of calcium

sulfate, while respiratory manifestations of acute inhalation and aspiration accidents of other mineral dusts like talcum and silica are extensively reported. We will present the clinical observations of other mineral dust aspiration accidents briefly below. Talcum powder consists of finely ground magnesium silicate. Talcum is therapeutically used for pleurodesis in malignant effusions.11 Commonly used for baby care before, it has lead to acute mineral dust inhalation and aspiration accidents especially in children. The described cases often occur during diaper changes, when talcum powder is accidentally spilled over the baby’s face. The children can present delayed hours after accidental inhalation VX-809 solubility dmso or aspiration with severe respiratory difficulties.12 Treatment is supportive and includes steroid administration and antibiotic treatment.12 and 13 Bronchoalveolar lavage has been advocated but remains controversial.12 Outcome can be fatal. Crystalline silica is another mineral dust leading to pulmonary diseases.14 Although silica is the most copious mineral on earth, acute silicosis is rare. In acute silicosis symptoms develop within weeks to years after exposure to crystalline

silica.14 Patients can develop respiratory failure in a short time as described in previous reviews.15 Symptoms are often present before radiographic changes occur. The later include diffuse nodular opacities, which can confluence.14 SB-3CT Treatment of acute silicosis is mainly supportive, but steroid medication can be beneficial.16 Ultimately, pulmonary transplantation has been described in one single case for a patient with aspiration-induced silicosis.17 In general, the prognosis of patients with acute silicosis remains poor.15 In analogy to these treatment strategies, and the animal studies described before, we decided to treat our patient with steroids and antibiotic

treatment after emergency bronchoscopy. Because of the risk of exothermic burns from the chemical reaction upon water of the remaining almost dry gypsum powder chunks in the tracheobronchial tree we did not perform bronchoalveolar lavage. The outcome of our patient was favorable after 3 months with no respiratory symptoms indicating bronchial hyperresponsiveness, no lung functional limitations and no radiological anomalies. This report shows the first case of accidental aspiration of a large amount of gypsum powder. The patient was treated by bronchoscopy with aspiration of remaining endobronchial gypsum deposits. To avoid a further exothermic reaction and to prevent further distribution of gypsum into the lung parenchyma, we minimized local anesthesia and did not apply any additional saline during this procedure. The outcome was favorable with no residual clinical symptoms, no lung function impairment or radiological manifestations after 3 months.

5 °C He denied purulent sputum, hemoptysis and arthronalgia Unf

5 °C. He denied purulent sputum, hemoptysis and arthronalgia. Unfortunately, the cough and shortness of breath of the patient had progressively worsened over time. Chest examination revealed absent breath sounds on the lower two thirds of the left hemithorax and a dull percussion note. No detectable peripheral lymphadenopathy was found. Laboratory results included normal creatinine, blood urea nitrogen, and serum electrolyte; lactate dehydrogenase

(LDH), 179 U/L; alanine aminotransferase (ALT), 30U/L; aspartate aminotransferase (AST), this website 25 U/L; total protein (TP), 66.3 g/L; leukocyte count, 10.3 × 109/L; hemoglobin, 16.7 g/dl; platelet count, 233 × 109/L. A peripheral blood smear examination revealed no abnormal lymphoid cells. Serum test results I-BET-762 in vitro were negative for carcinoembryonic

antigen (CEA), squamous cell carcinoma associated antigen (SCC), hepatitis B virus (HBV), human immunodeficiency virus (HIV), hepatitis C virus (HCV), Schaudinn’s bacillus. We did not carry out human herpesvirus 8 (HHV8) test in our center. Also serum test showed erythrocyte sedimentation rate (ESR), 8 mm/h; and C-reactive protein (CRP), 43.6 mg/L. Sputum cultures were negative for bacteria, fungus, and Mycobacterium tuberculosis. Chest X-ray demonstrates a large anterior mediastinal mass and a left pleural effusion with a light contralateral shift of the trachea and mediastinum (Fig. 1). Chest computed tomography (CT) showed an anterior and middle mediastinal mass with a light contralateral shift of the trachea, pleural thickening of the left hemithorax, and left-sided pleural effusion (Fig. 2). Chest ultrasonography revealed massive left pleural effusion. Echocardiography showed little pericardial effusion. And ultrasonography of superficial lymph node showed lymphadenopathy in bilateral axillary region (left 21.1 × 11.4 mm; right 15.4 × 4.4 mm), from bilateral cervical region, (left, 18.7 × 17.1 mm; right 12 × 5.2 mm), and bilateral inguinal region (left 16 × 4.8 mm; right 11.3 × 9.3 mm), but not in retroperitoneal region. Thoracentesis were performed and revealed exudate with lactate dehydrogenase level of 721 U/L, ADA value of 25 U/L, and

TP 15.3 g/L. Pleural fluid were grossly bloody and the routine examination of pleural fluid showed leukocytes 5 × 109/L (55% percent multinucleated cells, 54% percent mononuclear cells). The cytologic examination of the effusion smears revealed massive lymphocytes, a small amount of mesothelial cells, and partly abnormal cells (tumor cell?). Pleural fluid cultures were negative for M. tuberculosis. Then the medical thoracoscopy was performed under local anesthesia, cardiovascular and respiratory monitoring, in the endoscopy suite by experienced operator. The inspection of the pleural by a direct vision optic revealed massive bloody pleural fluid in the pleural cavity, and widely membrane hyperemia with lots of small white apophysis involving the parietal pleura (Fig. 3).

1) The concurrent expressions of CD25 and FOXP3, following expan

1). The concurrent expressions of CD25 and FOXP3, following expansion of CD4+CD25+CD127lo/− and CD4+CD25− cultures, were analysed and compared to each other, as seen in Fig. 5a–c. The cut-offs for the gates were set after the fluorescence of a biologically FOXP3 negative and CD25 negative population. Data was analysed using the FlowJo software (Tree Star) and expressed as mean fluorescence intensity (MFI; geometrical and standard mean) and percentages of cells expressing each marker. Tregs were find more expanded according to a protocol adapted from Putnam et al.

[24]. Briefly, on day 0 sorted cells were resuspended in AIM-V medium (Gibco/Invitrogen) containing 10% HS and amphotericin B and plated according to Table 2. Dynabeads® Human Treg Expander anti-CD3/anti-CD28 coated microbeads (Invitrogen; catalogue number 111.61D) were added at a 1:1 bead to cell ratio. When Treg numbers were lower than 40.000, ALK inhibitor 96-well flat-bottomed plates were used. The cell culture volume was doubled at day 2 and IL-2 (Proleukin, Chiron Therapeutics) added at a final concentration of 300 U/ml.

On days 5 and 7, cells were counted, washed in AIM-V and resuspended as above, adding fresh IL-2. Restimulation with anti-CD3/anti-CD28 coated microbeads, was performed on day 9, as described for day 0. Cells were counted again on day 11 and 13, washed, resuspended according to Table 2, and supplemented with fresh IL-2. Cultures were terminated on day 15 and cells stained for FOXP3 analysis. CD4+CD25− cells were expanded according to a scheme similar to Tregs, with the following alterations. As anti-CD3/anti-CD28 coated microbeads caused overstimulation and activation-induced apoptosis, CD4+CD25− cells were expanded using anti-CD3 (OKT3, 10 μg/ml) coated culturing vessels (Table 3) and soluble anti-CD28 (1 μg/ml). IL-2 addition was added at a concentration of 30 U/ml. As expression of Treg-markers was not normally distributed, two-group comparisons were performed with the Mann–Whitney U-test, while three or more groups were compared using the Kruskal–Wallis

test for unpaired observations. For pair-wise comparisons, Wilcoxon signed rank test was Y-27632 mouse used. A probability level <0.05 was considered statistically significant. Calculations were performed using the statistical package GraphPad Prism version 5.01 for Windows (GraphPad Software, Inc.). The study was approved by the Research Ethics Committee of the Faculty of Health Sciences, Linköping University. Informed consent was obtained from all volunteers and/or their parents. Before sorting of Tregs, a small pre-study of healthy volunteering adults was performed assessing the stability of Treg-markers through cryopreservation and thawing. We did not find cryopreservation and thawing of PBMC to yield any differences in the percentage of FOXP3 expressing cells or the FOXP3 MFI, in the CD4+CD25hi cell population (Fig. 2a,b).