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Binding of a T cell to an appropriate antigen-presenting cell (APC) induces the rapid reorientation of the T cell cytoskeleton and secretory apparatus towards the cell-cell contact site in a T cell antigen receptor (TCR) and peptide/major histocompatibility complex-dependent process. Such T cell polarization directs the delivery of cytokines and cytotoxic mediators towards the APC and contributes to the highly selective and specific action of effector T cells. To study the signaling pathways that regulate cytoskeletal rearrangements in T lymphocytes, we set up a conjugate formation assay using Jurkat T cells as effectors and cell-sized latex beads coated with various antibodies as artificial APCs. Here, we report that beads coated with antibodies specific for the TCR-CD3 complex were sufficient to induce T cell polarization towards the bead attachment site, as judged by reorientation of the microtubule-organizing center (MTOC) and localized actin polymerization. Thus, these cytoskeletal changes did not depend on activation of additional coreceptors. Moreover, single subunits of the TCR complex, namely TCR-zeta and CD3epsilon, were equally effective in inducing cytoskeletal polarization. However, mutagenesis of the immunoreceptor tyrosine-based activation motifs (ITAMs), present three times in TCR-zeta and once in CD3epsilon, revealed that the induction of cytoskeletal rearrangements required the presence of at least one intact ITAM. In agreement with this result, lack of functional Lck, the protein tyrosine kinase responsible for ITAM phosphorylation, abolished both MTOC reorientation and polarized actin polymerization. Both inhibitor and transient overexpression studies demonstrated that MTOC reorientation could occur in the absence of Ras activation. Our results suggest that APC-induced T cell polarization is a TCR-mediated event that is coupled to the TCR by the same signaling motif as TCR-induced gene activation, but diverges in its distal signaling requirements.

Glyphosate is controversially discussed because of its alleged harmful effects on human health and the environment. Although it is approved until December 2022 in the European Union, the Austrian government discusses a national ban. Research on farmers' intentions to deal with upcoming pesticide policy changes is limited and planned responses to a national glyphosate ban may inform accompanying measures and the development of weed management alternatives. Therefore, we have conducted 41 qualitative semi-structured interviews with farmers to explore their intended weed management if glyphosate-based herbicides were no longer available in Austria. The interviews were systematically analyzed, whereby the Theory of Planned Behavior (TPB) with its three social-psychological constructs served as guidance, i.e., attitude toward the planned behavior, subjective norm, and perceived behavioral control toward the planned behavior. We grouped farmers based on differences in their behavioral intentions toward glyphosate-free weed management, and identified four types of farmers by assigning group-specific attributes of the TPB constructs to the groups of farmers with similar behavioral intentions. Given a national glyphosate ban, the farmers intend to implement either mechanical or chemical alternatives, which would be solely applied or combined with changes in cultivation. Attitude toward the planned behavior, descriptive norms, and perceived behavioral control affect behavioral intentions, whereas injunctive norms do not differ much between the interviewed farmers. What unites the four types of farmers is that they would rather accept a glyphosate ban, if weed management alternatives with similar effectiveness and costs were available., (© 2022. The Author(s).)

Invasive species are considered a major threat for global agricultural production, biodiversity and ecosystem services. Their spread and establishment is mainly influenced by bio-physical factors, but also by people's activities such as tourism or farming. Understanding farmers' behavior is necessary to develop effective control measures. We conducted qualitative semi-structured interviews in south-east Austria to explore factors that facilitate or impede farmers' behavior to individually or collectively control the invasive Western Corn Rootworm (WCR, Diabrotica virgifera virgifera). We analyze the interview contents using the Capability-Opportunity-Motivation-Behavior model (COM-B model). Our results show that farmers' motivation and related behavior is influenced by intra- and interpersonal factors such as their knowledge about WCR control measures (capability psychological), perceived societal acceptance of WCR control measures or perceived normative obligations to participate in collective measures (opportunity social). Farmers' motivation (reflective and automatic) for implementing individual or collective WCR control measures is mainly determined by their perceived self-efficacy, their perceived efficacy of WCR control measures and the perceived severity of WCR damages. Contextual factors such as environmental conditions, legal regulations, the landscape composition, the farm type or financial impacts of WCR control measures (opportunity physical) are essential prerequisites for farmers' behavior. The results suggest that new modes of knowledge transfer are required to facilitate the proactive implementation of individual and collective WCR control measures prior to trigger events, such as severe WCR damages. The development of a trusting and communicative environment between farmers is key for collective WCR control. Exchange with residents about WCR and applied control measures may help to create a shared understanding and increase societal acceptance. Moreover, a long-term and proactive coordination which meets individual famers' needs is required to implement collective WCR control measures. Farmers who have successfully implemented individual and collective WCR measures may encourage non-applicants and sceptics by "learning from peers"., (Copyright © 2020. Published by Elsevier Ltd.)

The water-energy-land nexus requires long-sighted approaches that help avoid maladaptive pathways to ensure its promise to deliver insights and tools that improve policy-making. Climate services can form the foundation to avoid myopia in nexus studies by providing information about how climate change will alter the balance of nexus resources and the nature of their interactions. Nexus studies can help climate services by providing information about the implications of climate-informed decisions for other economic sectors across nexus resources. First-of-its-kind guidance is provided to combine nexus studies and climate services. The guidance consists of ten principles and a visual guide, which are discussed together with questions to compare diverse case studies and with examples to support the application of the principles., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: Determine whether ambient light affects the accuracy of pulse oximetry readings., Design: Prospective, repeated-measures study., Setting: A photographic darkroom., Subjects: Forty-five faculty and students at a university, none of whom had pale skin, dark skin, or evidence of cardiopulmonary disease. Any nail polish was removed., Methods: Five light sources were individually tested: incandescent, quartz-halogen, infrared, fluorescent, and bilirubin light. A pulse oximetry probe was placed on the subject's finger, and the finger and probe were placed sideways under each light source, on a predetermined mark., Results: The greatest difference in pulse oximetry reading between any of the light sources was 0.5%. Repeated-measures analysis of variance yielded a p value of 0.204., Conclusions: Ambient light has no statistically significant effect on pulse oximetry readings. Even had the differences been statistically significant, the magnitude of the differences was small and thus clinically unimportant.

NK cell function is regulated by a dual receptor system, which integrates signals from triggering receptors and MHC class I-specific inhibitory receptors. We show here that the src family kinase Fyn is required for efficient, NK cell-mediated lysis of target cells, which lack both self-MHC class I molecules and ligands for NKG2D, an activating NK cell receptor. In contrast, NK cell inhibition by the MHC class I-specific receptor Ly49A was independent of Fyn, suggesting that Fyn is specifically required for NK cell activation via non-MHC receptor(s). Compared to wild type, significantly fewer Fyn-deficient NK cells expressed the inhibitory Ly49A receptor. The presence of a transgenic Ly49A receptor together with its H-2(d) ligand strongly reduced the usage of endogenous Ly49 receptors in Fyn-deficient mice. These data suggest a model in which the repertoire of inhibitory Ly49 receptors is formed under the influenced of Fyn-dependent NK cell activation as well as the respective MHC class I environment. NK cells may acquire Ly49 receptors until they generate sufficient inhibitory signals to balance their activation levels. Such a process would ensure the induction of NK cell self-tolerance.

NK cell function is negatively regulated by MHC class I-specific inhibitory receptors. Transduction of the inhibitory signal involves protein tyrosine phosphatases such as SHP-1 (SH2-containing protein tyrosine phosphatase-1). To investigate the role of SHP-1 for NK cell development and function, we generated mice expressing a catalytically inactive, dominant-negative mutant of SHP-1 (dnSHP-1). In this paper we show that expression of dnSHP-1 does not affect the generation of NK cells even though MHC receptor-mediated inhibition is partially impaired. Despite this defect, these NK cells do not kill syngeneic, normal target cells. In fact dnSHP-1-expressing NK cells are hyporesponsive toward MHC-deficient target cells, suggesting that non-MHC-specific NK cell activation is significantly reduced. In contrast, these NK cells mediate Ab-dependent cell-mediated cytotoxicity and prevent the engraftment with beta2-microglobulin-deficient bone marrow cells. A similar NK cell phenotype is observed in viable motheaten (mev) mice, which show reduced SHP-1 activity due to a mutation in the Shp-1 gene. In addition, NK cells in both mouse strains show a tendency to express more inhibitory MHC-specific Ly49 receptors. Our results demonstrate the importance of SHP-1 for the generation of functional NK cells, which are able to react efficiently to the absence of MHC class I molecules from normal target cells. Therefore, SHP-1 may play an as-yet-unrecognized role in some NK cell activation pathways. Alternatively, a reduced capacity to transduce SHP-1-dependent inhibitory signals during NK cell development may be compensated by the down-modulation of NK cell triggering pathways.

NK cells can kill MHC-different or MHC-deficient but not syngeneic MHC-expressing target cells. This MHC class I-specific tolerance is acquired during NK cell development. MHC recognition by murine NK cells largely depends on clonally distributed Ly49 family receptors, which inhibit NK cell function upon ligand engagement. We investigated whether these receptors play a role for the development of NK cells and provide evidence that the expression of a Ly49 receptor transgene on developing NK cells endowed these cells with a significant developmental advantage over NK cells lacking such a receptor, but only if the relevant MHC ligand was present in the environment. The data suggest that the transgenic Ly49 receptor accelerates and/or rescues the development of NK cells which would otherwise fail to acquire sufficient numbers of self-MHC-specific receptors. Interestingly, the positive effect on NK cell development is most prominent when the MHC ligand is simultaneously present on both hemopoietic and nonhemopoietic cells. These findings correlate with functional data showing that MHC class I ligand on all cells is required to generate functionally mature NK cells capable of reacting to cells lacking the respective MHC ligand. We conclude that the engagement of inhibitory MHC receptors during NK cell development provides signals that are important for further NK cell differentiation and/or maturation.

Most NK1.1+ T (NKT) cells express a biased TCRalphabeta repertoire that is positively selected by the monomorphic MHC class I-like molecule CD1d. The development of CD1d-dependent NKT cells is thymus dependent but, in contrast to conventional T cells, requires positive selection by cells of hemopoietic origin. Here, we show that the Src protein tyrosine kinase Fyn is required for development of CD1d-dependent NKT cells but not for the development of conventional T cells. In contrast, another Src kinase, Lck, is required for the development of both NKT and T cells. Impaired NKT cell development in Fyn-deficient mice cannot be rescued by transgenic expression of CD8, which is believed to increase the avidity of CD1d recognition by NKT cells. Taken together, our data reveal a selective and nonredundant role for Fyn in NKT cell development.

Families of clonally expressed major histocompatibility complex (MHC) class I-specific receptors provide specificity to and regulate the function of natural killer (NK) cells. One of these receptors, mouse Ly49A, is expressed by 20% of NK cells and inhibits the killing of H-2D(d) but not D(b)-expressing target cells. Here, we show that the trans-acting factor TCF-1 binds to two sites in the Ly49A promoter and regulates its activity. Moreover, we find that TCF-1 determines the size of the Ly49A NK cell subset in vivo in a dosage-dependent manner. We propose that clonal Ly49A acquisition during NK cell development is regulated by TCF-1.

Each cell is equipped with two copies (alleles) of each autosomal gene. While the vast majority use both alleles, occasional genes are expressed from a single allele. The reason for mono-allelic expression is not always evident and can serve distinct purposes. First, it may facilitate the tight control over the dosage of certain gene products such as some growth factors and their receptors or X-linked genes. Second, the differential usage of the two parental alleles may reflect the mechanisms that ensure mono-specificity, e.g. olfactory receptors, T and B cell receptors. The context of allele-specific expression of the murine Ly49 natural killer (NK) cell receptor genes suggests that their allele-specific expression reflects a process that generates clonal variability., (Copyright 1999 Academic Press.)

Two families of protein tyrosine kinases (PTKs), the Src and Syk/ZAP-70 families, are required for T cell development. Lck is the major Src family member required for thymopoiesis, since there is a severe deficit of CD4+CD8+ thymocytes and mature T cells in its absence. However, some peripheral T cells are evident in these mice, suggesting that additional PTKs may contribute to T cell development. Here we show that the combined disruption of Lck and Fyn (lck(-/-)fyn(-/-)) completely arrests alpha beta T cell development at the CD4-CD8- stage. The development of V gamma 3+ dendritic epidermal T cells is also severely impaired, but natural killer cell development and cytolytic activity is unaffected in lck(-/-)fyn(-/-) mice. These findings reveal the potential for redundant functions mediated by Src family PTKs while emphasizing crucial roles for Lck and Fyn in T cell development.