Interleukin 31 mediates MAP kinase and STAT1/3 activation in intestinal epithelial cells and its expression is upregulated in inflammatory bowel disease

Background/aim: Interleukin 31 (IL31), primarily expressed in activated lymphocytes, signals through a heterodimeric receptor complex consisting of the IL31 receptor alpha (IL31R\textgreeka) and the oncostatin M receptor (OSMR). The aim of this study was to analyse IL31 receptor expression, signal transduction, and specific biological functions of this cytokine system in intestinal inflammation.Methods: Expression studies were performed by RT-PCR, quantitative PCR, western blotting, and immunohistochemistry. Signal transduction was analysed by western blotting. Cell proliferation was measured by MTS assays, cell migration by restitution assays.Results: Colorectal cancer derived intestinal epithelial cell (IEC) lines express both IL31 receptor subunits, while their expression in unstimulated primary murine IEC was low. LPS and the proinflammatory cytokines TNF-\textgreeka, IL1\textgreekb, IFN-\textgreekg, and sodium butyrate stimulation increased IL31, IL31R\textgreeka, and OSMR mRNA expression, while IL31 itself enhanced IL8 expression in IEC. IL31 mediates ERK-1/2, Akt, STAT1, and STAT3 activation in IEC resulting in enhanced IEC migration. However, at low cell density, IL31 had significant antiproliferative capacities (p<0.005). IL31 mRNA expression was not increased in the TNF\textgreekDARE mouse model of ileitis but in inflamed colonic lesions compared to non-inflamed tissue in patients with Crohn's disease (CD; average 2.4-fold increase) and in patients with ulcerative colitis (UC; average 2.6-fold increase) and correlated with the IL-8 expression in these lesions (r = 0.564 for CD; r = 0.650 for UC; total number of biopsies analysed: n = 88).Conclusion: IEC express the functional IL31 receptor complex. IL31 modulates cell proliferation and migration suggesting a role in the regulation of intestinal barrier function particularly in intestinal inflammation

Suv4-20h Histone Methyltransferases Promote Neuroectodermal Differentiation by Silencing the Pluripotency-Associated Oct-25 Gene

Post-translational modifications (PTMs) of histones exert fundamental roles in regulating gene expression. During development, groups of PTMs are constrained by unknown mechanisms into combinatorial patterns, which facilitate transitions from uncommitted embryonic cells into differentiated somatic cell lineages. Repressive histone modifications such as H3K9me3 or H3K27me3 have been investigated in detail, but the role of H4K20me3 in development is currently unknown. Here we show that Xenopus laevis Suv4-20h1 and h2 histone methyltransferases (HMTases) are essential for induction and differentiation of the neuroectoderm. Morpholino-mediated knockdown of the two HMTases leads to a selective and specific downregulation of genes controlling neural induction, thereby effectively blocking differentiation of the neuroectoderm. Global transcriptome analysis supports the notion that these effects arise from the transcriptional deregulation of specific genes rather than widespread, pleiotropic effects. Interestingly, morphant embryos fail to repress the Oct4-related Xenopus gene Oct-25. We validate Oct-25 as a direct target of xSu4-20h enzyme mediated gene repression, showing by chromatin immunoprecipitaton that it is decorated with the H4K20me3 mark downstream of the promoter in normal, but not in double-morphant, embryos. Since knockdown of Oct-25 protein significantly rescues the neural differentiation defect in xSuv4-20h double-morphant embryos, we conclude that the epistatic relationship between Suv4-20h enzymes and Oct-25 controls the transit from pluripotent to differentiation-competent neural cells. Consistent with these results in Xenopus, murine Suv4-20h1/h2 double-knockout embryonic stem (DKO ES) cells exhibit increased Oct4 protein levels before and during EB formation, and reveal a compromised and biased capacity for in vitro differentiation, when compared to normal ES cells. Together, these results suggest a regulatory mechanism, conserved between amphibians and mammals, in which H4K20me3-dependent restriction of specific POU-V genes directs cell fate decisions, when embryonic cells exit the pluripotent state

Comparative analysis of the lambda-interferons IL-28A and IL-29 regarding their transcriptome and their antiviral properties against hepatitis C virus.

Specific differences in signaling and antiviral properties between the different Lambda-interferons, a novel group of interferons composed of IL-28A, IL-28B and IL-29, are currently unknown. This is the first study comparatively investigating the transcriptome and the antiviral properties of the Lambda-interferons IL-28A and IL-29. Expression studies were performed by microarray analysis, quantitative PCR (qPCR), reporter gene assays and immunoluminometric assays. Signaling was analyzed by Western blot. HCV replication was measured in Huh-7 cells expressing subgenomic HCV replicon. All hepatic cell lines investigated as well as primary hepatocytes expressed both IFN-λ receptor subunits IL-10R2 and IFN-λR1. Both, IL-28A and IL-29 activated STAT1 signaling. As revealed by microarray analysis, similar genes were induced by both cytokines in Huh-7 cells (IL-28A: 117 genes; IL-29: 111 genes), many of them playing a role in antiviral immunity. However, only IL-28A was able to significantly down-regulate gene expression (n = 272 down-regulated genes). Both cytokines significantly decreased HCV replication in Huh-7 cells. In comparison to liver biopsies of patients with non-viral liver disease, liver biopsies of patients with HCV showed significantly increased mRNA expression of IL-28A and IL-29. Moreover, IL-28A serum protein levels were elevated in HCV patients. In a murine model of viral hepatitis, IL-28 expression was significantly increased. IL-28A and IL-29 are up-regulated in HCV patients and are similarly effective in inducing antiviral genes and inhibiting HCV replication. In contrast to IL-29, IL-28A is a potent gene repressor. Both IFN-λs may have therapeutic potential in the treatment of chronic HCV

rs1004819 Is the Main Disease-Associated IL23R Variant in German Crohn's Disease Patients: Combined Analysis of IL23R, CARD15, and OCTN1/2 Variants

The IL23R gene has been identified as a susceptibility gene for inflammatory bowel disease (IBD) in the North American population. The aim of our study was to test this association in a large German IBD cohort and to elucidate potential interactions with other IBD genes as well as phenotypic consequences of IL23R variants. Genomic DNA from 2670 Caucasian individuals including 833 patients with Crohn's disease (CD), 456 patients with ulcerative colitis (UC), and 1381 healthy unrelated controls was analyzed for 10 IL23R SNPs. Genotyping included the NOD2 variants p.Arg702Trp, p.Gly908Arg, and p.Leu1007fsX1008 and polymorphisms in SLC22A4/OCTN1 (1672 C-->T) and SLC22A5/OCTN2 (-207 G-->C). All IL23R gene variants analyzed displayed highly significant associations with CD. The strongest association was found for the SNP rs1004819 [P = 1.92x10(-11); OR 1.56; 95 % CI (1.37-1.78)]. 93.2% of the rs1004819 TT homozygous carriers as compared to 78% of CC wildtype carriers had ileal involvement [P = 0.004; OR 4.24; CI (1.46-12.34)]. The coding SNP rs11209026 (p.Arg381Gln) was protective for CD [P = 8.04x10(-8); OR 0.43; CI (0.31-0.59)]. Similar, but weaker associations were found in UC. There was no evidence for epistasis between the IL23R gene and the CD susceptibility genes CARD15 and SLC22A4/5. IL23R is an IBD susceptibility gene, but has no epistatic interaction with CARD15 and SLC22A4/5. rs1004819 is the major IL23R variant associated with CD in the German population, while the p.Arg381Gln IL23R variant is a protective marker for CD and UC

Analysis of IL12B Gene Variants in Inflammatory Bowel Disease

IL12B encodes the p40 subunit of IL-12, which is also part of IL-23. Recent genome-wide association studies identified IL12B and IL23R as susceptibility genes for inflammatory bowel disease (IBD). However, the phenotypic effects and potential gene-gene interactions of IL12B variants are largely unknow

Transient and sustained incentive effects on electrophysiological indices of cognitive control in younger and older adults

Preparing for upcoming events, separating task-relevant from task-irrelevant information and efficiently responding to stimuli all require cognitive control. The adaptive recruitment of cognitive control depends on activity in the dopaminergic reward system as well as the frontoparietal control network. In healthy aging, dopaminergic neuromodulation is reduced, resulting in altered incentive-based recruitment of control mechanisms. In the present study, younger adults (18–28 years) and healthy older adults (66–89 years) completed an incentivized flanker task that included gain, loss, and neutral trials. Event-related potentials (ERPs) were recorded at the time of incentive cue and target presentation. We examined the contingent negative variation (CNV), implicated in stimulus anticipation and response preparation, as well as the P3, which is involved in the evaluation of visual stimuli. Both younger and older adults showed transient incentive-based modulation of CNV. Critically, cue-locked and target-locked P3s were influenced by transient and sustained effects of incentives in younger adults, while such modulation was limited to a sustained effect of gain incentives on cue-P3 in older adults. Overall, these findings are in line with an age-related reduction in the flexible recruitment of preparatory and target-related cognitive control processes in the presence of motivational incentives

SUMO modification of a novel MAR-binding protein, SATB2, modulates immunoglobulin μ gene expression

Nuclear matrix attachment regions (MARs) are regulatory DNA sequences that are important for higher-order chromatin organization, long-range enhancer function, and extension of chromatin modifications. Here we characterize a novel cell type-specific MAR-binding protein, SATB2, which binds to the MARs of the endogenous immunoglobulin μ locus in pre-B cells and enhances gene expression. We found that SATB2 differs from the closely related thymocyte-specific protein SATB1 by modifications of two lysines with the small ubiquitive related modifier (SUMO), which are augmented specifically by the SUMO E3 ligase PIAS1. Mutations of the SUMO conjugation sites of SATB2 enhance its activation potential and association with endogenous MARs in vivo, whereas N-terminal fusions with SUMO1 or SUMO3 decrease SATB2-mediated gene activation. Sumoylation is also involved in targeting SATB2 to the nuclear periphery, raising the possibility that this reversible modification of a MAR-binding protein may contribute to the modulation of subnuclear DNA localization

Design and implementation of management strategy evaluation for the Great Barrier Reef inshore (MSE-GBR)

[Extract] Stakeholder engagement is important for successful management, both to make effective decisions and to obtain support. However, in the context of coastal management, question s rem in on h ow to effectively link decisions made in the catchment with objectives for marine biodiversity and fisheries productivity. Moreover, there is much uncertainty on how to best in form and elicit community input in a rigorous manner. A decision support process is described that elicits management objectives, priorities and management options using two case studies. The case studies show that demand for local input and regional management is high, but local conditions influence the relative success. Differences between case study outcomes high light the importance of discussing objectives prior to suggesting management actions. In that regard, eliciting the broader community’s objectives can now be undertaken cost effectively through new survey methods. Governance arrangements can be developed that link managers and community members, but continuity is essential. A big contributor to success is providing local information to the community group and embedding managers and influencers within the group. Of great value to positive outcomes were that the scientists, managers and community members were prepared to work together and offer enormous volunteer time to work towards a common solution. Two case studies were selected to develop an overall method of using a regional management process with local community groups to develop local management options – Mackay and Bowen-Burdekin. These two case studies were chosen for what they have in common and also what separates them. Both ca se studies have in common that the rural area is mostly farming for which accelerated management activity has been directed to reduce the a mount of sediment and nutrient runoff to the GBR. However, the two regions’ ports are distinct in that, during the study period, a major proposed port upgrade with associated dredging in the Abbott Point are(just south of the Burdekin) was a source of conflict in the region and great controversy within Australia. Whereas the Mackay port s were well established and are presently not as controversial. The population size is also very different with Mackay having a far larger urban footprint with a growing city although this may have slowed down in recent years due to the general downturn in mining activity. A hierarchical system of engagement was attempted in both region s. At the highest level, a community group, the Local Marine Advisory Committee (LM AC) run by GBRMPA was already established in the region. Its charter is to advise GBRMPA on local management issues (http://www.gbrmpa.gov.au/about-us/local-marine-advisory-committees). Since the LMACs met every quarter with a full agenda, a sub-committee was formed and called the LMAC Reference Group (RG). This was made up of LMAC members who volunteered for the group and additional members that would cover a broader skill set from people who were previously on the LMAC. The project lead facilitated the RG meetings, with a member elected as the RG chair. The project team included “managers” (defined as people that either directly or indirectly influence management decisions) from QDAFF and GBRMPA, and social, economic, mathematical and environmental scientists from both State and Commonwealth agencies. Within a few months of project engagement in the Bowen-Burdekin area, the Abbott point port expansion and associated dredging controversy meant that participation was minimal. An alternative approach was undertaken, but generally meant engaging with individuals directly and separately. Interactions between the different RG and LMAC members were minimal. In Mackay, the RG was very successful and was used throughout the process. At various stages in the process community and senior level managers’ input was sought. All documentation was kept in a traceable format, i.e. iterations of all steps could be backtracked through the various meetings to its original source. A local Mackay GBRMPA person devoted an enormous amount of time on support and engagement in between meetings. This support was essential and provided local continuity. A sequence of broad steps were undertaken: 1. Qualitative modelling of the Mackay coastal system; 2. A review of existing objectives from government organisations, NGO s and NRM bodies that were directly or indirectly relevant to the region was undertaken (both case studies). These were collated by the RGs into an objective hierarchy –one for each case study; 3. A survey of the RG, LM AC and Mackay public was undertaken to ascertain the relative importance of different objectives. A new method was developed during this process; 4. An issues register, direct and indirect management options, and responsible agencies for each topic relevant to managing the coastal zone fisheries and biodiversity were developed through a series of workshop with experts and RG. These were combined into management strategies and is a separate printed product for use by Mackay residents and NGOs; 5. An impact assessment was undertaken to determine the relative importance of the different management strategies. These were then turned into a series of management-orientated products for use by relevant management agencies. The project has uncovered a conundrum that does challenge the effectiveness of management because there can be a significant gap between the perception of managers with regard s to their action s and outcomes and the perception of the community as to the effectiveness (and wisdom) of the management action(s). A review of the successes and failures of the two case studies by the project tea m were undertaken through questionnaires to the Mackay RG and managers. The final and main product of the project is a semi-quantitative generic elicitation framework that ultimately provides a prioritised list of management option s in the context of clearly articulated management objectives that has broader application to coastal communities in Australia and beyond. It comes with detailed instructions, and generic objectives and management strategies