Mesenchymal Transglutaminase 2 Activates Epithelial ADAM17: Link to G-Protein-Coupled Receptor 56 (ADGRG1) Signalling.

A wound healing model was developed to elucidate the role of mesenchymal-matrix-associated transglutaminase 2 (TG2) in keratinocyte re-epithelialisation. TG2 drives keratinocyte migratory responses by activation of disintegrin and metalloproteinase 17 (ADAM17). We demonstrate that epidermal growth factor (EGF) receptor ligand shedding leads to EGFR-transactivation and subsequent rapid keratinocyte migration on TG2-positive ECM. In contrast, keratinocyte migration was impaired in TG2 null conditions. We show that keratinocytes express the adhesion G-protein-coupled receptor, ADGRG1 (GPR56), which has been proposed as a TG2 receptor. Using ADAM17 activation as a readout and luciferase reporter assays, we demonstrate that TG2 activates GPR56. GPR56 activation by TG2 reached the same level as observed with an agonistic N-GPR56 antibody. The N-terminal GPR56 domain is required for TG2-regulated signalling response, as the constitutively active C-GPR56 receptor was not activated by TG2. Signalling required the C-terminal TG2 ß-barrel domains and involved RhoA-associated protein kinase (ROCK) and ADAM17 activation, which was blocked by specific inhibitors. Cell surface binding of TG2 to the N-terminal GPR56 domain is rapid and is associated with TG2 and GPR56 endocytosis. TG2 and GPR56 represent a ligand receptor pair causing RhoA and EGFR transactivation. Furthermore, we determined a binding constant for the interaction of human TG2 with N-GPR56 and show for the first time that only the calcium-enabled "open" TG2 conformation associates with N-GPR56.

Metamotivational beliefs about intrinsic and extrinsic motivation.

Although intrinsic motivation is often viewed as preferable to more extrinsic forms of motivation, there is evidence that the adaptiveness of these motivational states depends on the nature of the task being completed (e.g., Cerasoli et al., 2014). Specifically, research suggests task-motivation fit such that intrinsic motivation tends to benefit performance on open-ended tasks (tasks that involve qualitative performance assessment; e.g., creative writing) and extrinsic motivation benefits performance on closed-ended tasks (tasks that involve quantitative performance assessment; e.g., multiple choice). We examined people's metamotivational beliefs about intrinsic and extrinsic motivation in the context of this task-motivation fit. Across 11 studies (seven primary, four supplemental; N = 3,544), participants provided beliefs about the utility of different types of motivation-regulation strategies: strategies that enhance one's interest and enjoyment in a task versus strategies that focus on the value associated with task outcomes (self-relevance strategies and reward strategies). Across all studies, participants recognized that the adaptiveness of these strategies depends on the nature of the task being completed. Consistent with an understanding of normative task-motivation fit, participants generally reported that interest-enhancing strategies were more useful for open-ended tasks and that reward strategies were more useful for closed-ended tasks; however, in some studies, participants reported that reward strategies were equally useful across task types (Studies 2, 3, and 5). More normatively accurate beliefs were associated with more normatively accurate consequential behavioral choices (Study 6) and better task performance (Study 7). We discuss the implications of these results for theories of motivation and self-regulation. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Attitudes and beliefs about Vermont's 2021 buprenorphine decriminalization law among residents who use illicit opioids.

BACKGROUND: In July 2021, Vermont removed all criminal penalties for possessing 224mg or less of buprenorphine. METHODS: Vermont residents (N=474) who used illicit opioid drugs or received treatment for opioid use disorder in the past 90 days were recruited for a mixed-methods survey on the health and criminal legal effects of decriminalization. Topics assessed included: motivations for using non-prescribed buprenorphine, awareness of and support for decriminalization, and criminal legal system experiences involving buprenorphine. We examined the frequencies of quantitative measures and qualitatively summarized themes from free-response questions. RESULTS: Three-quarters of respondents (76%) reported lifetime use of non-prescribed buprenorphine. 80% supported decriminalization, but only 28% were aware buprenorphine was decriminalized in Vermont. Respondents described using non-prescribed buprenorphine to alleviate withdrawal symptoms and avoid use of other illicit drugs. 18% had been arrested while in buprenorphine, with non-White respondents significantly more likely to report such arrests (15% v 33%, p<0.001). CONCLUSION: Decriminalization of buprenorphine may reduce unnecessary criminal legal system involvement, but its health impact was limited by low awareness at the time of our study.

A Comparison of the Incidence and Type of Ocular Motility Defects in Patients Presenting to Birmingham Midland Eye Centre Emergency Department during 2019 and 2020 to Assess the Impact of the Covid-19 Pandemic.

Aim: This audit aimed to investigate whether COVID-19 had any impact on the incidence and type of ocular motility defects in patients presenting to Birmingham Midland Eye Centre (BMEC) Emergency Department (ED) during the COVID-19 pandemic in 2020 compared to the previous year. Methods: Medical records were reviewed for all patients presenting to BMEC ED during 2019 and 2020. Patients were classified depending on their diagnosis. The incidence and classification of ocular motility defect were analysed. Factors considered during analysis were number of presentations by year and month; COVID-19 tests; and pre-existing conditions. Results: Two hundred and twenty-one patients presented in 2019, and 260 patients in 2020, an increase in incidence of 17.6% was observed. One hundred and eighty-five patients were classified with new-onset neurogenic conditions in 2019, and 222 patients in 2020, an increase of 20.0%. In 2020, most patients presented in July, November, and December. Overall, there was a 91.3% increase in new-onset fourth cranial nerve palsies in 2020. Fifty-seven patients in 2020 had a Polymerase Chain Reaction COVID-19 test, of these 5 were COVID-19 positive. Conclusion: There was a higher incidence of ocular motility defects in 2020 compared to 2019. The majority of ocular motility defects were classified as neurogenic. It is difficult to attribute this increase to COVID-19 due to lack of testing and results, and confounding variables such as pre-existing conditions and lockdown restrictions. Some potential explanations for the change in presentations across the year of 2020 are proposed.

Perceived barriers and rewards to sexual consent communication: A qualitative analysis.

Increasingly, affirmative consent - direct, unambiguous and voluntary agreement to engage in sexual activity (Craig & McKinley, 2015) - is the standard being adopted by educational institutions in North America (Bennett, 2016). Yet, studies show that most individuals continue to communicate consent through nonresistance (Jozkowski et al., 2014a). Given this discrepancy, it is critical to understand what factors prevent individuals from engaging in affirmative consent. Furthermore, a better understanding of the perceived rewards of consent communication could incentivize the adoption of affirmative consent. To understand the range of perceived barriers and rewards, we conducted an online, qualitative study where 231 participants answered two open-ended questions. We used inductive content analysis to categorize participants' perceptions of sexual consent barriers and rewards into four general content areas: (1) Communication Quality, (2) Relational and Emotional Experiences, (3) Sexual Quality and (4) Safety and Coercion. These perceived rewards and barriers were examined through the lens of the Information-Motivation-Behavioural Skills Model. Participants viewed consent communication not only as a means of ensuring safety but also as a way to enhance relational and sexual quality. However, they also perceived barriers in all three of these domains as well as barriers to ensuring that sexual consent communication is fluid and easily understood. These findings provide important avenues for future research investigating how individuals reconcile perceived rewards and costs of affirmative consent communication. We also suggest ways to enhance sexual education by discussing potential rewards and validating the normative nature of fears and anxieties around affirmative consent.

Zfhx3 modulates retinal sensitivity and circadian responses to light.

Mutations in transcription factors often exhibit pleiotropic effects related to their complex expression patterns and multiple regulatory targets. One such mutation in the zinc finger homeobox 3 (ZFHX3) transcription factor, short circuit (Sci, Zfhx3Sci/+ ), is associated with significant circadian deficits in mice. However, given evidence of its retinal expression, we set out to establish the effects of the mutation on retinal function using molecular, cellular, behavioral and electrophysiological measures. Immunohistochemistry confirms the expression of ZFHX3 in multiple retinal cell types, including GABAergic amacrine cells and retinal ganglion cells including intrinsically photosensitive retinal ganglion cells (ipRGCs). Zfhx3Sci/+ mutants display reduced light responsiveness in locomotor activity and circadian entrainment, relatively normal electroretinogram and optomotor responses but exhibit an unexpected pupillary reflex phenotype with markedly increased sensitivity. Furthermore, multiple electrode array recordings of Zfhx3Sci/+ retina show an increased sensitivity of ipRGC light responses.

Spatio-temporal insights into microbiology of the freshwater-to-hypersaline, oxic-hypoxic-euxinic waters of Ursu Lake.

Ursu Lake is located in the Middle Miocene salt deposit of Central Romania. It is stratified, and the water column has three distinct water masses: an upper freshwater-to-moderately saline stratum (0-3 m), an intermediate stratum exhibiting a steep halocline (3-3.5 m), and a lower hypersaline stratum (4 m and below) that is euxinic (i.e. anoxic and sulphidic). Recent studies have characterized the lake's microbial taxonomy and given rise to intriguing ecological questions. Here, we explore whether the communities are dynamic or stable in relation to taxonomic composition, geochemistry, biophysics, and ecophysiological functions during the annual cycle. We found: (i) seasonally fluctuating, light-dependent communities in the upper layer (≥0.987-0.990 water-activity), a stable but phylogenetically diverse population of heterotrophs in the hypersaline stratum (water activities down to 0.762) and a persistent plate of green sulphur bacteria that connects these two (0.958-0.956 water activity) at 3-3.5 to 4 m; (ii) communities that might be involved in carbon- and sulphur-cycling between and within the lake's three main water masses; (iii) uncultured lineages including Acetothermia (OP1), Cloacimonetes (WWE1), Marinimicrobia (SAR406), Omnitrophicaeota (OP3), Parcubacteria (OD1) and other Candidate Phyla Radiation bacteria, and SR1 in the hypersaline stratum (likely involved in the anaerobic steps of carbon- and sulphur-cycling); and (iv) that species richness and habitat stability are associated with high redox-potentials. Ursu Lake has a unique and complex ecology, at the same time exhibiting dynamic fluctuations and stability, and can be used as a modern analogue for ancient euxinic water bodies and comparator system for other stratified hypersaline systems.

Osmotolerance as a determinant of microbial ecology: A study of phylogenetically diverse fungi.

Osmotic stress induced by high solute concentration can prevent fungal metabolism and growth due to alterations in properties of the cytosol, changes in turgor, and the energy required to synthesize and retain compatible solutes. We used germination to quantify tolerance/sensitivity to the osmolyte KCl (0.1-4.5 M, in 0.1 M increments) for 71 strains (40 species) of ecologically diverse fungi. These include 11 saprotrophic species (17 strains, including two xerophilic species), five mycoparasitic species (five strains), six plant-pathogenic species (13 strains), and 19 entomopathogenic species (36 strains). A dendrogram obtained from cluster analyses, based on KCl inhibitory concentrations 50 % and 90 % calculated by Probit Analysis, revealed three groups of fungal isolates accordingly to their osmotolerance. The most-osmotolerant group (Group 3) contained the majority of saprotrophic fungi, and Aspergillus niger (F19) was the most tolerant. The highly xerophilic Aspergillus montevidense and Aspergillus pseudoglaucus were the second- and third-most tolerant species, respectively. All Aspergillus and Cladosporium species belonged to Group 3, followed by the entomopathogens Colletotrichum fioriniae, Simplicillium lanosoniveum, and Trichothecium roseum. Group 2 exhibited a moderate osmotolerance, and included plant-pathogens such as Colletotrichum and Fusarium, mycoparasites such as Clonostachys spp, some saprotrophs such as Mucor and Penicillium spp., and some entomopathogens such as Isaria, Lecanicillium, Mariannaea, Simplicillium, and Torrubiella. Group 1 contained the osmo-sensitive strains: the rest of the entomopathogens and the mycoparasitic Gliocladium and Trichoderma. Although stress tolerance did not correlate with their primary ecological niche, classification of these 71 fungal strains was more closely aligned with their ecology than with their phylogenetic relatedness. We discuss the implications for both microbial ecology and fungal taxonomy.

Addressing unmet needs in understanding asthma mechanisms: From the European Asthma Research and Innovation Partnership (EARIP) Work Package (WP)2 collaborators.

Asthma is a heterogeneous, complex disease with clinical phenotypes that incorporate persistent symptoms and acute exacerbations. It affects many millions of Europeans throughout their education and working lives and puts a heavy cost on European productivity. There is a wide spectrum of disease severity and control. Therapeutic advances have been slow despite greater understanding of basic mechanisms and the lack of satisfactory preventative and disease modifying management for asthma constitutes a significant unmet clinical need. Preventing, treating and ultimately curing asthma requires co-ordinated research and innovation across Europe. The European Asthma Research and Innovation Partnership (EARIP) is an FP7-funded programme which has taken a co-ordinated and integrated approach to analysing the future of asthma research and development. This report aims to identify the mechanistic areas in which investment is required to bring about significant improvements in asthma outcomes.

Tissue Culture as a Source of Replicates in Nonmodel Plants: Variation in Cold Response in Arabidopsis lyrata ssp. petraea.

While genotype-environment interaction is increasingly receiving attention by ecologists and evolutionary biologists, such studies need genetically homogeneous replicates-a challenging hurdle in outcrossing plants. This could be potentially overcome by using tissue culture techniques. However, plants regenerated from tissue culture may show aberrant phenotypes and "somaclonal" variation. Here, we examined somaclonal variation due to tissue culturing using the response to cold treatment of photosynthetic efficiency (chlorophyll fluorescence measurements for Fv/Fm, Fv'/Fm', and ΦPSII, representing maximum efficiency of photosynthesis for dark- and light-adapted leaves, and the actual electron transport operating efficiency, respectively, which are reliable indicators of photoinhibition and damage to the photosynthetic electron transport system). We compared this to variation among half-sibling seedlings from three different families of Arabidopsis lyrata ssp. petraea Somaclonal variation was limited, and we could detect within-family variation in change in chlorophyll fluorescence due to cold shock successfully with the help of tissue-culture derived replicates. Icelandic and Norwegian families exhibited higher chlorophyll fluorescence, suggesting higher performance after cold shock, than a Swedish family. Although the main effect of tissue culture on Fv/Fm, Fv'/Fm', and ΦPSII was small, there were significant interactions between tissue culture and family, suggesting that the effect of tissue culture is genotype-specific. Tissue-cultured plantlets were less affected by cold treatment than seedlings, but to a different extent in each family. These interactive effects, however, were comparable to, or much smaller than the single effect of family. These results suggest that tissue culture is a useful method for obtaining genetically homogenous replicates for studying genotype-environment interaction related to adaptively-relevant phenotypes, such as cold response, in nonmodel outcrossing plants.

Fibrinogen Induces RUNX2 Activity and Osteogenic Development from Human Pluripotent Stem Cells.

Pluripotent stem cells, both human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC), provide an important resource to produce specialized cells such as osteogenic cells for therapeutic applications such as repair or replacement of injured, diseased or damaged bone. hESCs and iPSCs can also be used to better define basic cellular and genetic mechanisms that regulate the earliest stages of human bone development. However, current strategies to mediate osteogenic differentiation of hESC and iPSC are typically limited by the use of xenogeneic components such as fetal bovine serum (FBS) that make defining specific agents that mediate human osteogenesis difficult. Runt-related transcription factor 2 (RUNX2) is a key regulator required for osteogenic differentiation. Here, we used a RUNX2-YFP reporter system to characterize the novel ability of fibrinogen to mediate human osteogenic development from hESC and iPSC in defined (serum-free) conditions. These studies demonstrate that fibrinogen mediates significant osteo-induction potential. Specifically, fibrinogen binds to the surface integrin (α9ß1) to mediate RUNX2 gene expression through the SMAD1/5/8 signaling pathway. Additional studies characterize the fibrinogen-induced hESC/iPSC-derived osteogenic cells to demonstrate these osteogenic cells retain the capacity to express typical mature osteoblastic markers. Together, these studies define a novel fibrinogen-α9ß1-SMAD1/5/8-RUNX2 signaling axis can efficiently induce osteogenic differentiation from hESCs and iPSCs. Stem Cells 2016;34:2079-2089.