Lessons learned from tolDC trials, relating particularly to biomarker identification, should assist the development and clinical translation of new tolerance-inducing strategies, e.g. strategies that directly target and enhance the tolerogenic function of DC in vivo, or strategies that combine tolDC therapy with other treatments. For example, it has been shown that the combination Buparlisib of tolDC treatment with CTLA-4Ig prolongs allograft survival significantly in an animal model [31]. The success of human tolDC trials will be enhanced by the definition of a robust set of biomarkers; without such a set it may prove difficult to establish if immune tolerance has been achieved.

Furthermore, defining and standardizing biomarker analyses will be important to compare the results from different therapeutic tolerance strategies and trials. The authors are supported by grants from Arthritis Research

UK, Medical Research Council (MRC), Biotechnology and Biological Sciences Research Council (BBSRC) and the J.G.W. Patterson Foundation. Research in the Musculoskeletal Research Group is supported by the National Institute for Health Research Newcastle Biomedical Research Centre based at Newcastle Hospitals Foundation Trust and Newcastle University. The views expressed FDA-approved Drug Library supplier are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. The authors have no competing interests. “
“Reperfusion injury remains one of the major problems in transplantation. Repair from ischaemic acute renal failure (ARF) involves stimulation of tubular epithelial cell proliferation. The aim of this very exploratory study was to evaluate the effects of preconditioning donor animals with rapamycin and tacrolimus to prevent ischaemia–reperfusion (I/R) injury. Twelve hours before nephrectomy, the donor animals received immunosuppressive drugs. The animals were divided into four groups, as follows: group 1 control: no treatment; group 2: rapamycin (2 mg/kg); group 3 FK506 (0, 3 mg/kg); and group 4: FK506 (0, 3 mg/kg) plus rapamycin (2 mg/kg). The left

kidney was removed and after 3 h of cold ischaemia, the graft was transplanted. Twenty-four hours after transplant, the kidney was recovered for histological analysis and cytokine expression. Preconditioning treatment with rapamycin or tacrolimus significantly reduced blood urea nitrogen and creatinine compared with control [blood urea nitrogen (BUN): P < 0·001 versus control and creatinine: P < 0·001 versus control]. A further decrease was observed when rapamycin was combined with tacrolimus. Acute tubular necrosis was decreased significantly in donors treated with immunosuppressants compared with the control group (P < 0·001 versus control). Moreover, the number of apoptotic nuclei in the control group was higher compared with the treated groups (P < 0·001 versus control). Surprisingly, only rapamycin preconditioning treatment increased anti-apoptotic Bcl2 levels (P < 0·001).

A potential caveat of the above results is that the CD3lo DP cells from Bcl11bdp−/− mice may not represent a pure population of immature,

unselected, DP cells, and might contain cells derived from more mature populations, possibly owing to the difficulty to resolve the mutant cell populations with the CD8, CD4, and CD3 markers. To address this issue, we analyzed the expression of several genes previously found to be induced in WT DP cells during positive selection, using transcriptome data from a published comparison of gene expression profiles of unselected DP cells (CD69− DP cells from Zap70-deficient mice) to selected, buy OSI-906 CD69hi cells from WT animals 41 (data accessible at Selleckchem GSI-IX NCBI GEO database accession GSE2262). Although some selection-induced genes were indeed overexpressed in the CD3lo DP cells from Bcl11bdp−/− mice (Zbtb7b, Id2, Klf2, CD53, IL7r, and Irf7), several others were expressed at similar low levels in WT and mutant cells (Itm2a, Nr4a1, Bcl2a1a, Slfn1, Mapk11, Nr4a3, Tnfrsf9,

Acvrl1, Ccr7, Ephx1, Ms4a4b, St6gal1, Tes, Nab2, and Ccl22), suggesting that the mutant CD3lo DP cells do not exhibit a general induction of the gene expression program associated with thymocyte maturation. We selected five of these genes (Ccr7, Slfn1, Ephx1, Ms4a4b, and Mapk11) for further analysis, as these genes displayed strong differences in gene expression levels between unselected and selected cells in the data from Sun et al.41 (>3 log induction), Interleukin-3 receptor and were thus likely to be informative with respect to the selection/purity status of the analyzed populations. We sorted CD3loDP, CD3+DP, CD3+CD4+ SP, and CD3+CD8+ SP cells from two WT and two Bcl11bdp−/− mice (see Supporting Information Fig. 6 for

sorting gates and purity of the sorted populations) and analyzed the expression of the selected genes in these populations by RT-qPCR (Fig. 7). In WT samples, all five genes were expressed at low levels in CD3lo DP cells and strongly induced in the CD3+ DP and SP populations, thus validating previous microarray results 41. In agreement with our transcriptome data, all five genes were also expressed at very low levels in mutant CD3lo DP cells. Two genes (Ephx1 and Ms4a4b) were strongly induced in the mutant CD3+DP and SP-like populations. This observation reveals that the phenotypically more mature cells from Bcl11bdp−/− mice have retained the capacity to induce a subset of the genes normally upregulated during positive selection.

These previous studies in BALB/c mice suggested that the initial constraints regulating CDR-H3 content reflected germline sequence content; i.e. the product of natural selection. Superimposed upon these germline restrictions in diversity

were a series of somatic, presumably Ku-0059436 mw clonal, selective events that sequentially produced a CDR-H3 repertoire that had undergone “trimming” of apparently “disfavored” sequence content. This process included a reduction in the use of a specific VH gene segment, VH81X; a reduction in the use of very short CDR-H3s; enhanced use of reading frame 1; enhanced use of tyrosine and glycine in the CDR-H3 loop; and a sequential elimination of highly charged or heavily hydrophobic CDR-H3s with development. The present analysis of immunoglobulin repertoire development in the bone marrow of C57BL/6 mice again demonstrates the effects of germline-imposed restrictions on the range of initial diversity in the H-chain repertoire; but would point to significant differences in either the efficiency, the ability, or the direction of the late-stage somatic, clonal selective events in the bone marrow and the periphery. The end result is a mature, recirculating B-cell repertoire characterized by including IgM BCRs that bear antigen-binding sites that seemed to be not just “disfavored”,

but commonly “discarded” by the mature, recirculating Selleckchem PD0332991 B cells in BALB/c mice. At the progenitor B-cell stage, the influence of the germline on the C57BL/6 repertoire is obvious. VH, DH, and JH usage in C57BL/6 H-chain transcripts appears to differ from BALB/c H-chain transcripts both due to changes in number as well as the sequence of homologous gene segments. Germline variation appeared to be associated with changes in VDJ rearrangement frequency, although this latter point needs to be confirmed through the analysis of nonfunctional sequences. Among the features

of the C57BL/6 repertoire that most closely matched the BALB/c repertoire were similarities in the initial distribution OSBPL9 of N addition, lengths, charge, and the usage of 18 of the 20 different potential amino acids. One of the features that varied between the two strains reflected the diminished number of functional VH gene segments, including the absence of the most commonly used VH in the BALB/c genome, VH7183.10. Others included the enhanced use of serine-enriched DFL16.1; the presence of a DSP2.11 homologue, DSP2.x, that encodes serine in RF1; and an increased use of JH1 in place of JH4. Of these changes, the most apparent effect in early B-cell progenitors was on VH content, again due to the absence of many of the VH7183 variant sequences available to BALB/c mice.

Importantly, how commensals contribute to the expression

of these enzymes and metabolism of vitamin A remains unknown. Another important question is the timing necessary for DCs migrating in the GALT to acquire RA from epithelial cells and how these processes can be modified during infection. How RA contributes to oral tolerance, and at the same time protective immunity in the GI tract, also remains to be addressed. One possibility is that RA favours the induction of Tregs in the absence of secondary signals but enhances effector responses following exposure to inflammatory mediators. Treg populations require not only appropriate conditions for their induction, but also for their upkeep, particularly when confronted with an inflammatory environment. Very recently it has been shown that, in the gut, myeloid cell-derived IL-10 plays a crucial role in maintaining functional Treg activity by stimulating Caspase activity assay IL-10R directly on FoxP3+ Tregs and allowing them to play a fully protective role in the prevention of colitis [45]. Thus, in the absence of either innate IL-10 production, or IL-10R on Tregs, these cells lose

the ability to block colitogenic effector T cells from causing inflammatory disease, and indeed succumb themselves to the inflammatory process by switching to the production of IFN-γ[45]. Hence, IL-10 is important for the maintenance of Treg activity and can be Phospholipase D1 pivotal at the tipping-point between regulation check details and inflammation. The regulation of Treg activity between the gut and the periphery is also of special interest, as IBD in humans may affect extraintestinal organs in up to 36% of cases [46]. IBD-related extraintestinal disorders are not specific to IBD. They can be classified into reactive manifestations dependent directly upon intestinal disease. The often co-existing presentation in the same patient points towards common underlying pathomechanisms that may involve enteric flora activating the immune system to turn against bacterial antigens and, based

on cross-reactivity, against intestinal antigens and antigens in extraintestinal organs (‘molecular mimicry’). A separate subset of IBD patients shows an increased frequency of other common autoimmune diseases that manifest mainly independently of the bowel disease. This may thus reflect susceptibility to autoimmunity in general. The complex relationship between intestinal and extraintestinal manifestations in IBD is also reflected by the complex multi-genetic control reported in animal models of IBD; genetic loci regulating intestinal and extra-intestinal manifestations are largely but not exclusively different [47]. The appearance of GI parasites is a major challenge to the discriminatory powers of the immune system, and one which in evolutionary time has been played out countless times.

Proteomic studies of Toxoplasma have revealed that many proteins exhibit multiple isoforms, indicating that post-translational modifications (PTMs) are fairly common

(69). Multiple studies ABT-263 mouse have been performed to examine the PTMs of α- and β-tubulin in Toxoplasma, as the microtubule cytoskeleton of the parasite plays an important role in host cell invasion (70). Initially, it was believed that α- and β-tubulin were only encoded by single genes in the parasite’s genome (71), such that PTMs would be the only way to supply tubulin diversity. However, the availability of genomic data from http://www.toxodb.org implies that there might be two additional genes for both α- and β-tubulin (7). Initial studies by Plessmann et al. utilized antibodies specific to various tubulin PTMs to show that Toxoplasmaα-tubulin can be acetylated and detyrosinated (removal of the last C-terminal residue, tyrosine 453). Additionally, mass spectrometry analysis revealed that the C-terminus of α-tubulin can be truncated by five amino acids and that glutamate 445 can be subjected to polyglutamylation (72). These findings were expanded upon by Xiao et al. (73), where cytoskeleton fractions were prepared from purified RH strain tachyzoites and subjected to 2DE followed by either immunoblotting Cilomilast ic50 with PTM-specific antibodies or identification of relevant bands with mass

spectrometry. Two β-tubulin isotypes and one α-tubulin isotype were detected from approximately 16 spots on a 2D gel. Between α- and β-tubulin, α-tubulin can undergo a wider spectrum of PTMs. The PTMs observed in the α-tubulin isotype included acetylation at lysine 40, detyrosination, polyglutamylation, methylation and C-terminal truncation of the last

two and last five amino acids. Of these modifications, only polyglutamylation Buspirone HCl and methylation were observed in β-tubulin. Methylation as a PTM has not been documented in tubulin previously, although Xiao et al. (73) mass spectrometry studies identified it on C-terminal α-tubulin peptides and peptides from one of the two β-tubulin isotypes. This tubulin methylation was not found in the human foreskin fibroblast host cells and may represent a specific modification for apicomplexa. As microtubules in Toxoplasma exhibit several functions that are specific to apicomplexans (gliding motility and invasion), garnering a greater understanding of the PTMs of tubulin could help to provide new therapeutic targets. The availability of a Toxoplasma reference genome sequence has been a great incentive for genomics studies, which have significantly shaped our understanding of unique cellular processes that drive Toxoplasma infection. Major progress has been made in the areas of host–parasite interaction, parasite cell division, intercellular transmission and stage differentiation.

The difference of plasma sRAGE between patients with normal Pirfenidone (>90 ml/min per 1.73 m2) and lower eGFR was not statistical significant (887.7 ± 82.5 pg/ml versus 949.5±155.1 pg/ml, P = 0.733). The positive rates for ANA, anti-dsDNA, AnuA, anti-Sm were 92.2% (95/103), 53.9% (55/102), 55.7% (54/97), 37.1% (30/89), respectively, in patients with SLE. There was no significant difference between sRAGE levels in patients

with negative ANA and those with different levels of ANA (Fig. 4A). In addition, there was no significant difference between the sRAGE levels in autoantibody-positive patients and those in autoantibody-negative patients (Fig. 4B,C,D). In patients

with SLE, plasma sRAGE levels was negatively correlated with the leucocyte count (n = 95, r = −0.326, P = 0.001, Fig. 5A), absolute values of lymphocytes (n = 95, r = −0.357, P = 0.000, Fig. 5B), neutrophils (n = 95, r = −0.272, P = 0.008, Fig. 5C) and monocytes (n = 95, r = −0.286, P = 0.005, Fig. 5D) in peripheral blood. In this study, we found that plasma sRAGE level in patients with SLE was lower than that in HC, while there was no significant difference of sRAGE level between active and inactive patients. Decreased sRAGE levels in patients with SLE may be explained by the consumption of this soluble receptor. Renard et al. [36] postulated that sRAGE-ligand complexes were eliminated from the blood via spleen and/or liver. selleck compound It has been demonstrated that the level of HMGB1, one important RAGE ligand, is increased in the Pregnenolone circulation of SLE [19, 20], leading to the binding and consumption of sRAGE during the inflammatory process. It is also possible that sRAGE levels in patients with SLE may be regulated by alternative splicing and proteinases and this possibility needs to be clarified in the

future research. sRAGE might not only function as a decoy to exert their inhibitory effects on RAGE, but also act in a more direct way, e.g. binding to cell surface RAGE to block the formation of homodimers [28]. Therefore, decreased levels of sRAGE, which may contribute to enhanced RAGE-mediated pro-inflammatory signalling [27], support the essential role of RAGE in SLE pathology. Our results were different from the recent report showing that blood sRAGE levels in patients with SLE were higher than those in HC and compared with quiescent SLE, blood sRAGE levels are significantly increased during active disease [34]. One explanation for this discrepancy is that use of medication might influence the results. The discrepancy may also be caused by the low number of cases included in that study (only 10 cases of patients with SLE).

For determination of in vivo IL-4 production, total splenocytes were isolated on days 7 and 4 following the primary and secondary immunizations. In total, 106 splenocytes were cultured in cRPMI in the presence or absence of 2.5 μg/mL ConA for 24 h. Brefeldin A was added after 19 h of stimulation, 5 h prior to analysis, and cells were collected and analyzed using flow cytometry. Western blot analysis was performed as described previously 43. Briefly, protein

samples (5–20 μg) were isolated and resolved by electrophoresis on a 4–20% gradient Tris-HCl gel, transferred to Immobilon-P polyvinylidene Epigenetics Compound Library purchase difluoride membrances (Millipore), probed with either anti-CRAMP (Santa Cruz) at a 1:200 dilution or anti-actin at a 1:10 000 dilution, detected with HRP-labeled secondary Ab at a 1:1000–1:10 000 dilution, and developed with the SuperSignal West Pico kit (Thermo Scientific). Data with three or more groups were analyzed by a one-way ANOVA followed by post hoc analysis, while data with two groups were analyzed by a two-tailed unpaired t test. Statistically significant results were determined buy FK506 by a p value of *<0.05, **<0.01, ***<0.001. This research is part of the dissertation research conducted by Yao Chen who is a pre-doctoral student in the Microbiology Graduate Program, University

of Alabama at Birmingham, Birmingham, AL 35294, USA. The authors gratefully acknowledge Dr. Virginia M. Sanders oxyclozanide (The Ohio State University) for generously sharing the Sf-9/CD40L cells, Dr. Mark Lisanby (University of Alabama at Birmingham) for backcrossing the Camp−/− mice to C57BL/6, and Dr. Tamer Mahmoud (University of Alabama at Birmingham) for critical reading of the manuscript. This work was supported by research funds from the National Institutes of Health (NIH) Grant AI14782 (J. F. K.), AR052728 (R. L. G.), and AI052453 (R. L. G.). J. F. K. is a recipient of a Senior

Investigator Award from the American Asthma Foundation. N. W. K. is a recipient of an F32 NRSA Postdoctoral Fellowship Grant AI078662. Conflict of interest: The authors declare no financial or commercial conflict of interest. See accompanying Commentary: http://dx.doi.org/10.1002/eji.201142055 “
“The decoding of the Tritryp reference genomes nearly 7 years ago provided a first peek into the biology of pathogenic trypanosomatids and a blueprint that has paved the way for genome-wide studies. Although 60–70% of the predicted protein coding genes in Trypanosoma brucei, Trypanosoma cruzi and Leishmania major remain unannotated, the functional genomics landscape is rapidly changing. Facilitated by the advent of next-generation sequencing technologies, improved structural and functional annotation and genes and their products are emerging. Information is also growing for the interactions between cellular components as transcriptomes, regulatory networks and metabolomes are characterized, ushering in a new era of systems biology.

Low birthweight as an index of IUGR reflects

the congenital defects of organs, which are associated with CKD through their direct influence on nephron number and function, also through related metabolic disease-induced kidney damage (Fig. 2). However, the role of LBW in the pathogenesis of CKD is not completely explicit and results of former studies are often inconsistent. Although a recent meta-analysis confirmed that LBW increases the risk of CKD, the authors still suggested additional well-designed population-based studies.51 In addition, it is worth looking for an alternative index to birthweight to better reflect the influence of IUGR on human health. The Authors state that there is no conflict of interest regarding the material discussed in the manuscript. “
“Aim:  Catheter-related infection is a major cause of catheter loss in peritoneal dialysis (PD). We KU-60019 order evaluated the effect of catheter revision on the treatment

of intractable exit site infection (ESI)/tunnel infection (TI) in PD patients who required catheter removal. Methods:  We reviewed Selleckchem SCH 900776 the medical records of 764 continuous ambulatory peritoneal dialysis (CAPD) patients from May 1995 to April 2011 at our hospital. One hundred and twenty six patients had more than one occurrence of ESI. Catheter revision was performed to treat intractable ESI/TI. Incidence of ESI, causative organisms and the outcomes of catheter revision were analyzed. Results:  The total PD duration of all patients was 32 581 months. Three hundred and twelve ESI episodes occurred in 126 patients and the incidence of ESI was 1/104 patient-months (0.12/patient-year). The most common causative organism was methicillin-sensitive Staphylococcus aureus (MSSA) (98 episodes), followed by Pseudomonas aeruginosa (63 episodes) and methicillin-resistant S. aureus (MRSA) (28 episodes). Among these, catheter revision was required due to intractable ESI/TI in 36 patients. The most common causative organism was MSSA (14 episodes) followed by P. aeruginosa (10 episodes) and MRSA (six episodes) in catheter revision cases. The outcomes of catheter

revision were as follows: ESI relapsed in 11 patients (30.6%) after catheter revision. Among them, five patients were treated with antibiotic treatment, two patients required secondary catheter revision, Fossariinae four patients required catheter removal due to ESI/TI accompanying peritonitis. The catheter survival rate after catheter revision was 89.7% in one year. There were no statistical differences in the rates of ESI relapse after catheter revision between ESI caused by P. aeruginosa (5/10, 50%) and ESI caused by S. aureus (6/21, 28.6%). Conclusion:  Catheter revision may be an alternative treatment option to treat intractable ESI/TI before catheter removal is considered in PD patients. “
“Aim:  Glomerular infiltration of macrophages is a characteristic alteration of renal pathology in hyperlipidaemic renal injury.

258 + 2T > C mutation [20]. Recently, there has been another report of a novel heterozygous mutation in the SBDS gene (exon 1, 98 A > C) in a 4-year-old girl with virtual absence of B cells but normal immunoglobulin levels [21]. Following our finding of the SBDS mutation in one patient, Selleck Everolimus we

subsequently checked for SBDS mutation in two other patients. One patient was a 77-year-old woman with CVID, chronic anaemia due possibly to underlying myelodysplasia (proved on bone marrow biopsy) and thrombocytopenia. The other patient was in his early 40s, with CVID and on IVIG for 8 years with a 2-year history of enteropathy (chronic diarrhoea, ongoing weight loss, coeliac-like disease with no response to gluten-free diet). No mutations Selleckchem GPCR Compound Library in the SBDS gene were found in either of these patients. SDS and CVID share common features, such as recurrent infections, malabsorption, cytopenias (neutropenia, thrombocytopenia, anaemia), low immunoglobulins ± absent vaccine responses in some cases [10], abnormal liver function tests,

autoimmunity and malignancy [myelodysplastic syndrome (MDS), leukaemia], and testing for mutations in the SBDS gene in CVID patients with most of the above features would be worthwhile. More importantly, testing for SBDS mutations would be important in children with persistent neutropenia, recurrent infections, growth and skeletal abnormalities where the immunodeficiency disorder may have been described as CVID. A scoring

system may prove useful in the future when more patients are described. Ribosomopathies and bone marrow failure syndromes have variable and overlapping clinical presentations, yet most have subtle immune defects and a strong tendency to develop leukaemic transformation. The role of p53 in ribosomal dysfunction is beginning to be understood, such as up-regulation of p53 in haploinsufficiency of certain ribosomal proteins and consequent apoptosis and cell-cycle arrest, offer interesting mechanisms of cellular effects in ribosomopathies [8]. Deciphering subtle defects in the immune system in these patients may help to unravel the complex interaction of ribosomal proteins in the development selleck chemicals llc of specific parts of the immune system. Table 2 lists the syndromes with known mutations in ribosomal genes and the immunological abnormalities. Future studies will determine whether our observations of polymorphisms in specific ribosomal genes associated with DBA and the association of symptomatic or asymptomatic hypogammaglobulinaemia. With expanding knowledge and detection of newer ribosomal proteins, sequencing of specific ribosomal genes and/or use of ‘functional’ assays that provide evidence of aberrant pre-ribosomal RNA precursor accumulation would provide more tools to detect newer ribosomopathies that currently do not have a genetic basis [8,57].

, 2004). Our observation suggests that this effect becomes more evident when the basal levels of EpoR expression are low and ARA290 is applied in nanomolar concentrations. Based on these initial results,

we chose an incubation time of 6 h and 10 nM or 100 nM ARA290 as an appropriate condition to prestimulate cells in further experiments. Epo and its analogues have been described to enhance Wnt inhibitor proliferation in healthy tissue, tumors and cell lines (Kumar et al., 2005; Hardee et al., 2007). Such activity would clearly constitute a strong adverse effect for the usage of ARA290 in the urinary tract. In addition to its clinical relevance, pronounced differences in cell growth would also skew the results from in vitro assays. Therefore, we investigated the cell proliferation and viability of cells cultured in the presence of ARA290 for 24 h and performed an XTT assay. On applying the assay, Selleck Quizartinib we could not detect any significant difference in cell proliferation and viability between treated and control cells in concentrations used for further experiments (T24: 102.7±5.8% for 100 nM ARA290; 5637: 97.1±3.2% for 100 nM ARA290) nor at higher concentrations (for 1 μM ARA290, T24: 90.33±7.6%; 5637: 98.3±0.7%). No changes

were observed when cells were costimulated with inactivated bacteria (data not shown). The neutrophil-attractant chemokine IL-8 serves a crucial function during UTI in mediating the elimination Etomidate of bacteria (Hedges et al., 1994; Agace, 1996). The treatment with recombinant Epo has repeatedly been demonstrated to reduce lipopolysaccharide-induced cytokine induction in leukocytes (Schultz et al., 2008; Strunk et al., 2008; Yazihan et al., 2008). To test whether ARA290 modulated this immune response, we costimulated bladder epithelial cell lines with E. coli NU14 and ARA290 in different concentrations. During the period corresponding to basal levels of EpoR expression, the additional presence of ARA290 enhanced IL-8 mRNA expression. At 3 h, an increase in the IL-8 mRNA levels was observed in T24 cells after costimulation with 100 nM ARA290, compared with

stimulation with bacteria alone (127% of 0 nM ARA290, P<0.05; Fig. 3a). This early proinflammatory effect was even stronger with 10 nM ARA290 (155% of 0 nM ARA290, P<0.05). Consequently, IL-8 protein levels were higher in cell culture supernatants 12 h after costimulation with 100 nM ARA290 (115% of 0 nM ARA290, Fig. 3b) or 10 nM ARA290 (125% of 0 nM ARA290, P<0.05). At later time points, when EpoR expression was upregulated, ARA290 costimulation did not further promote immune induction. In contrast, IL-8 levels were reduced on mRNA (61% of 0 nM ARA290, P<0.05; Fig. 3a) and protein levels (78% of 0 nM ARA290, P<0.05; Fig. 3b). This downregulation was also observed at 10 nM ARA290, even though not as pronounced (91% for mRNA and 81% of 0 nM ARA290 for protein, P<0.05).