Allosteric inhibition of Aurora-A kinase by a synthetic vNAR domain

The vast majority of clinically-approved protein kinase inhibitors target the ATP binding pocket directly. Consequently, many inhibitors have broad selectivity profiles and most have significant off-target effects. Allosteric inhibitors are generally more selective, but are difficult to identify because allosteric binding sites are often unknown or poorly characterized. Aurora-A is activated through binding of TPX2 to an allosteric site on the kinase catalytic domain, and this knowledge could be exploited to generate an inhibitor. Here, we generated an allosteric inhibitor of Aurora-A kinase based on a synthetic, vNAR single domain scaffold, vNAR-D01. Biochemical studies and a crystal structure of the Aurora-A/vNAR-D01 complex show that the vNAR domain stabilizes an inactive conformation, in which the αC-helix is distorted, the canonical Lys-Glu salt bridge is broken, and the regulatory (R-) spine is disrupted by an additional hydrophobic side chain from the activation loop. These studies illustrate how single domain antibodies can be used to characterize the regulatory mechanisms of kinases and provide a rational basis for structure-guided design of allosteric Aurora-A kinase inhibitors

Playing in or out of character: User role differences in the experience of Interactive Storytelling

Interactive storytelling (IS) is a promising new entertainment technology synthesizing preauthored narrative with dynamic user interaction. Existing IS prototypes employ different modes to involve users in a story, ranging from individual avatar control to comprehensive control over the virtual environment. The current experiment tested whether different player modes (exerting local vs. global influence) yield different user experiences (e.g., senses of immersion vs. control). A within-subject design involved 34 participants playing the cinematic IS drama "Emo Emma

Transcriptional Regulator CNOT3 Defines an Aggressive Colorectal Cancer Subtype.

Cancer cells exhibit dramatic alterations of chromatin organization at cis-regulatory elements, but the molecular basis, extent, and impact of these alterations are still being unraveled. Here, we identify extensive genome-wide modification of sites bearing the active histone mark H3K4me2 in primary human colorectal cancers, as compared with corresponding benign precursor adenomas. Modification of certain colorectal cancer sites highlighted the activity of the transcription factor CNOT3, which is known to control self-renewal of embryonic stem cells (ESC). In primary colorectal cancer cells, we observed a scattered pattern of CNOT3 expression, as might be expected for a tumor-initiating cell marker. Colorectal cancer cells exhibited nuclear and cytoplasmic expression of CNOT3, suggesting possible roles in both transcription and mRNA stability. We found that CNOT3 was bound primarily to genes whose expression was affected by CNOT3 loss, and also at sites modulated in certain types of colorectal cancers. These target genes were implicated in ESC and cancer self-renewal and fell into two distinct groups: those dependent on CNOT3 and MYC for optimal transcription and those repressed by CNOT3 binding and promoter hypermethylation. Silencing CNOT3 in colorectal cancer cells resulted in replication arrest. In clinical specimens, early-stage tumors that included >5% CNOT3(+) cells exhibited a correlation to worse clinical outcomes compared with tumors with little to no CNOT3 expression. Together, our findings implicate CNOT3 in the coordination of colonic epithelial cell self-renewal, suggesting this factor as a new biomarker for molecular and prognostic classification of early-stage colorectal cancer. Cancer Res; 77(3); 766-79. ©2016 AACR

Ectopic expression of CXCL13, BAFF, APRIL and LT-ß is associated with artery tertiary lymphoid organs in giant cell arteritis

Objectives To investigate whether artery tertiary lymphoid organs (ATLOs) are present in giant cell arteritis (GCA) and that their formation is associated with the ectopic expression of constitutive lymphoid tissue-homing chemokines. Methods Reverse transcriptase PCR, immunohistochemical and immunofluorescence analysis were used to determine the presence of ectopic ATLOs in GCA and the expression of chemokines/chemokine receptors and cytokines involved in lymphoneogenesis in the temporal artery samples obtained from 50 patients with GCA and 30 controls. The presence of lymphatic conduits, of follicular dendritic cells (FDCs) precursors and lymphoid tissue inducer cells was also investigated. Finally, expression of CXCL13, B cell activating factor (BAFF), a proliferation-inducing ligand (APRIL) and CCL21 by isolated myofibroblasts was evaluated before and after stimulation with Toll-like receptors (TLRs) agonists and cytokines. Results ATLOs were observed in the media layer of 60% of patients with GCA in close proximity to high endothelial venules and independently by the age of patients and the presence of atherosclerosis. ATLO formation was also accompanied by the expression of CXCL13, BAFF, a proliferation-inducing ligand (APRIL), lymphotoxin (LT)-ß, interleukin (IL)-17 and IL-7, the presence of FDC precursors and of lymphoid conduits. Stimulation of myofibroblasts with TLR agonists and cytokines resulted in the upregulation of BAFF and CXCL13. Conclusions ATLOs occur in the inflamed arteries of patients with GCA possibly representing the immune sites where immune responses towards unknown arterial wall-derived antigens may be organised

Ambiguity, Familiarity and Learning Behavior in the Adoption of ICT for Irrigation Management

Subjective behavior of decision makers (DMs) is paramount when modeling information and communication technology (ICT) adoption choices in irrigated agriculture. Here, efficient ICT aided irrigation plans often involve a certain degree of uncertainty, and differential attitudes toward it can cause uncoordinated actions between actors. Some DMs will implement ICT information, while others will not because they do not trust ICT reliability. This risks undermining the achievement of ICT benefits in terms of water saving at the irrigation district level. By distinguishing between differ ent sources of uncertainty, taking the form of risk and ambiguity, in the present paper, we developed a new decision model to assess the impact that subjective behavior and learning processes have on the efficiency of ICT-aided irrigation plans. A case study was selected to implement the model in simplified settings. The results revealed the potential of ambiguity to limit ICT information implemen tation and to hinder water governance. Implications mainly concern the development of uncertainty management policies to favor DMs becoming familiar with the new ICT with lower ambiguit

Facing the Emotional Barriers to Colorectal Cancer Screening. The Roles of Reappraisal and Situation Selection

Background Disgust, embarrassment, and fear can hinder the attendance of colorectal cancer (CRC) screening. However, individuals can respond to these emotions differently. The present study tested whether reappraising a negative stimulus versus avoiding a negative stimulus is associated with age; whether these two emotion regulation strategies (reappraisal and situation selection) moderate the effects of disgust, embarrassment and fear on CRC screening intention; and the efficacy of a message based on participants' preferred emotion regulation strategy.Methods We recruited 483 Italian participants (aged 40-84 years) through snowball sampling. Participants were randomly assigned to one of four conditions differing for a message promoting CRC screening with an affective lever, a cognitive lever, both levers or none. Key variables included emotion regulation strategies, emotional barriers and intention to get screened.Results The preference for reappraisal over situation selection increased with age. Reappraisal neutralized the effect of disgust on CRC screening intention. The combined message with both affective and cognitive levers increased CRC screening intention (b = 0.27, beta = 0.11, SE = 0.13 p = .049), whereas reading the message based only on the affective (b = 0.16, beta = 0.06, SE = 0.14 p = .258) or the cognitive (b = 0.22, beta = 0.09, SE = 0.14 p = .107) lever was not effective.Conclusions Communication campaigns should support the activation of a reappraisal strategy of emotion control, and messages promoting CRC screening should highlight both the instrumental (i.e., early detection) and affective (i.e., peace of mind) benefits of attendance

Enhancing intentions to reduce meat consumption: An experiment comparing the role of self- and social pro-environmental identities

Research has consistently shown that a pro-environmental identity plays a critical role in motivating and sustaining pro-environmental actions. However, few studies have directly compared the effects of pro- environmental self- and social identities on pro-environmental behaviors. In the present study, we experimentally tested the effect of increasing self- and social identity salience on the intention to reduce meat consumption. A total of 678 young Italian adults were randomly assigned to one of four conditions: 1) past behavior recall plus personal feedback aimed at enhancing pro-environmental self-identity salience; 2) past behavior recall plus social feedback to reinforce pro-environmental social identity salience; 3) no feedback, where participants only recalled their past behavior; 4) control condition, i.e., recall of past behaviors unrelated to sustainability. In addition, we explored the mediating role of attitudes, subjective norms, and perceived behavioral control. The results revealed that recalling past eating-related pro-environmental behaviors enhances both the pro- environmental self-identity and the pro-environmental social identity, regardless of whether feedback was received. All experimental conditions indirectly fostered the intention to reduce meat consumption with respect to the control condition. In conclusion, recalling past pro-environmental behaviors, even without receiving specific feedback, can activate pro-environmental identities, thereby creating pathways toward stronger intentions to reduce meat consumption

Noninvasive ventilation for acute exacerbations of chronic obstructive pulmonary disease in the emergency department: early predictors of in-hospital outcome

The applications of early non-invasive ventilation (NIV) are nowadays expanding in the Emergency Department (ED) for the treatment of different types and etiologies of acute respiratory failure (ARF) Emergency physicians (EP) often face the challenge of predicting short-term outcomes for patients with ARF in the E