Network approach reveals preferential T-cell and macrophage association with α-linked ß-cells in early stage of insulitis in NOD mice.

One of the challenges in studying islet inflammation - insulitis - is that it is a transient phenomenon. Traditional reporting of the insulitis progression is based on cumulative, donor-averaged values of leucocyte density in the vicinity of pancreatic islets, that hinders intra- and inter-islet heterogeneity of disease progression. Here, we aimed to understand why insulitis is non-uniform, often with peri-insulitis lesions formed on one side of an islet. To achieve this, we demonstrated applicability of network theory in detangling intra-islet multi-cellular interactions during insulitis. Specifically, we asked the question "what is unique about regions of the islet which interact with immune cells first". This study utilized the non-obese diabetic mouse model of type one diabetes and examined the interplay among α-, ß-, T-cells, myeloid cells, and macrophages in pancreatic islets during the progression of insulitis. Disease evolution was tracked based on T/ß cell ratio in individual islets. In the early stage, we found that immune cells are preferentially interacting with α-cell-rich regions of an islet. At the islet periphery α-linked ß-cells were found to be targeted significantly more compared to those without α-cell neighbors. Additionally, network analysis revealed increased T-myeloid, and T-macrophage interactions with all ß-cells.

FishShapes v1: Functionally relevant measurements of teleost shape and size on three dimensions.

Teleost fishes account for 96% of all fish species and exhibit a spectacular variety of body forms. Teleost lineages range from deep bodied to elongate (e.g., eels, needlefish), laterally compressed (e.g., ribbonfish) to globular (e.g., pufferfish), and include uniquely shaped lineages such as seahorses, flatfishes, and ocean sunfishes. Adaptive body shape convergence within fishes has long been hypothesized but the nature of the relationships between fish form and ecological and environmental variables remain largely unknown at the macroevolutionary scale. To facilitate the investigation of the interacting factors influencing teleost body shape evolution we measured eight functionally relevant linear traits on adult-sized specimens along with specimen mass. Linear measurements of standard length, maximum body depth, maximum fish width, lower jaw length, mouth width, head depth, minimum caudal peduncle depth, and minimum caudal peduncle width were taken in millimeters with calipers, or tape measures for oversized specimens. We measured these traits on a total of 16,523 specimens (1-3 specimens per species) at the Smithsonian National Museum of Natural History and took approximately 7000 person hours of data collection to complete. The data went through a three-step error-checking process to clean and validate the data and then species averages were calculated. We present the complete specimen data set, which encompasses approximately one-fifth of extant teleost species diversity, spanning ~90% of teleost families and ~96% of orders. The species and family names are compatible with the taxonomy used by FishBase and the order information with the phylogenetically informed taxonomy of Betancur-R and colleagues published in 2014. This dataset is licensed under Creative Commons CC0 1.0 Universal (CC0 1.0) but please cite this paper when using the data or a subset of it.

Ena/VASP Protein-Mediated Actin Polymerization Contributes to Naïve CD8+ T Cell Activation and Expansion by Promoting T Cell-APC Interactions In Vivo.

Naïve T cell activation in secondary lymphoid organs such as lymph nodes (LNs) occurs upon recognition of cognate antigen presented by antigen presenting cells (APCs). T cell activation requires cytoskeleton rearrangement and sustained interactions with APCs. Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) proteins are a family of cytoskeletal effector proteins responsible for actin polymerization and are frequently found at the leading edge of motile cells. Ena/VASP proteins have been implicated in motility and adhesion in various cell types, but their role in primary T cell interstitial motility and activation has not been explored. Our goal was to determine the contribution of Ena/VASP proteins to T cell-APC interactions, T cell activation, and T cell expansion in vivo. Our results showed that naïve T cells from Ena/VASP-deficient mice have a significant reduction in antigen-specific T cell accumulation following Listeria monocytogenes infection. The kinetics of T cell expansion impairment were further confirmed in Ena/VASP-deficient T cells stimulated via dendritic cell immunization. To investigate the cause of this T cell expansion defect, we analyzed T cell-APC interactions in vivo by two-photon microscopy and observed fewer Ena/VASP-deficient naïve T cells interacting with APCs in LNs during priming. We also determined that Ena/VASP-deficient T cells formed conjugates with significantly less actin polymerization at the T cell-APC synapse, and that these conjugates were less stable than their WT counterparts. Finally, we found that Ena/VASP-deficient T cells have less LFA-1 polarized to the T cell-APC synapse. Thus, we conclude that Ena/VASP proteins contribute to T cell actin remodeling during T cell-APC interactions, which promotes the initiation of stable T cell conjugates during APC scanning. Therefore, Ena/VASP proteins are required for efficient activation and expansion of T cells in vivo.

Diversity, Equity, and Inclusion Milestones: Creation of a Tool to Evaluate Graduate Medical Education Programs.

Background: As the Accreditation Council for Graduate Medical Education (ACGME) began to ask programs to report their efforts surrounding diversity, equity, and inclusion (DEI), program directors felt ill prepared to evaluate their programs and measure change. Objective: To develop a tool that would allow graduate medical education (GME) programs to evaluate the current state of DEI within their residencies, identify areas of need, and track progress; to evaluate feasibility of using this assessment method within family medicine training programs; and to analyze and report pilot data from implementation of these milestones within family medicine residency programs. Methods: The Association of Family Medicine Residency Directors (AFMRD) Diversity and Health Equity (DHE) Task Force developed a tool for program DEI evaluation modeled after the ACGME Milestones. These milestones focus on DEI assessment in 5 key domains: Institution, Curriculum, Evaluation, Resident Personnel, and Faculty Personnel. After finalizing a draft, a pilot implementation of the milestones was conducted by a convenience sample of 10 AFMRD DHE Task Force members for their own programs. Results: Scores varied widely across surveyed programs for all milestones. Highest average scores were seen for the Curriculum milestone (2.65) and the lowest for the Faculty Personnel milestone (2.0). Milestone assessments were completed within 10 to 40 minutes using various methods. Conclusions: The AFMRD DEI Milestones were developed for program assessment, goal setting, and tracking of progress related to DEI within residency programs. The pilot implementation showed these milestones were easily used by family medicine faculty members in diverse settings.

Food Systems Development: The Necessary Paradigm Shift for a Healthy and Sustainable Agrarian Transition, a Case Study from Bougainville, Papua New Guinea.

Classical agricultural development paradigms prioritise basic requirements such as agronomic, caloric and economic needs for the target environment and for beneficiaries. As challenges associated with climate change, globalisation, and population growth compound and amplify one another, project scope must be broadened to take a holistic food systems approach that includes sociocultural and historical contexts, as well as climate impacts as underpinning project design. In this paper, we illustrate the importance of adopting a food systems development paradigm rather than a classical agricultural development paradigm through a case study in Bougainville, Papua New Guinea. The case uses Rich Picturing, targeted and focus-group interviews, and garden visits in remote Bougainville; it provides a poignant illustration of the importance of this more holistic perspective given the historical inefficacy of food systems development, as well as Papua New Guinea's exposure to a plethora of compounding environmental, social, economic, and political stresses and shocks that demonstrate the important linkages between ecosystem services and health. The study aims to demonstrate how including localised gender dynamics, climate vulnerability, rapidly morphing social norms, and climate analogue environments is critical in building food systems resilience and is key to designing policies, programs, and development projects that more effectively address environmental, sociocultural, and health considerations. Building on the inadequacies in agricultural development efforts previously documented for Papua New Guinea, we propose an improved framing for food systems development and identify areas for future research.

Telemedicine Familiarity and Post-Disaster Utilization of Emergency and Hospital Services for Ambulatory Care Sensitive Conditions.

INTRODUCTION: In this study, we examined the association between telemedicine use before a disaster and utilization of emergency or hospital services for ambulatory care sensitive conditions post-disaster. METHODS: Difference-in-differences analyses were conducted in 2020‒2021 to assess pre- to post-fire changes in emergency or hospital utilization for 5 ambulatory care sensitive conditions: asthma, diabetes, hypertension, coronary artery disease, and heart failure across all Kaiser Permanente Santa Rosa patients (N=108,113) based on telemedicine utilization before the 2017 Tubbs wildfire. Inverse probability of treatment weighting was employed for cohort balancing across telemedicine familiar status. RESULTS: Utilization for any ambulatory care sensitive condition increased from 9.03% pre-fire to 9.45% post-fire across the full cohort. Telemedicine familiarity (ref: not familiar) was associated with decreased absolute risk in pre- to post-fire inpatient and emergency department utilization for 4 conditions: asthma (absolute risk= -1.59%, 95% CI= -2.02%, -1.16%), diabetes (absolute risk= -0.68%, 95% CI= -0.89%, -0.47%), hypertension (absolute risk= -2.07%, 95% CI= -2.44%, -1.71%), and coronary artery disease (absolute risk= -0.43%, 95% CI= -0.61%, -0.24%). Telemedicine familiarity was associated with decreased relative change in pre- to post-fire utilization for 5 conditions: asthma (RRR=0.70, 95% CI=0.64, 0.75), diabetes (RRR=0.54, 95% CI=0.47, 0.63), hypertension (RRR=0.57, 95% CI=0.52, 0.62), heart failure (RRR=0.64, 95% CI=0.50, 0.82), and coronary artery disease (RRR=0.56, 95% CI=0.47, 0.67). Similar results were seen among patients residing in evacuation zones. CONCLUSIONS: Telemedicine familiarity pre-fire was associated with decreased inpatient and emergency department utilization for certain ambulatory care sensitive conditions for 1-year post-fire. These results suggest a role for telemedicine in preventing unnecessary emergency and hospital utilization following disasters.

Islet Lymphocytes Maintain a Stable Regulatory Phenotype Under Homeostatic Conditions and Metabolic Stress.

T cells and B cells have been identified in human and murine islets, but the phenotype and role of islet lymphocytes is unknown. Resident immune populations set the stage for responses to inflammation in the islets during homeostasis and diabetes. Thus, we sought to identify the phenotype and effector function of islet lymphocytes to better understand their role in normal islets and in islets under metabolic stress. Lymphocytes were located in the islet parenchyma, and were comprised of a mix of naïve, activated, and memory T cell and B cell subsets, with an enrichment for regulatory B cell subsets. Use of a Nur77 reporter indicated that CD8 T cells and B cells both received local antigen stimulus, indicating that they responded to antigens present in the islets. Analysis of effector function showed that islet T cells and B cells produced the regulatory cytokine IL-10. The regulatory phenotype of islet T cells and B cells and their response to local antigenic stimuli remained stable under conditions of metabolic stress in the diet induced obesity (DIO) model. T cells present in human islets retained a similar activated and memory phenotype in non-diabetic and T2D donors. Under steady-state conditions, islet T cells and B cells have a regulatory phenotype, and thus may play a protective role in maintaining tissue homeostasis.

Conceptualizing ecosystem services using social-ecological networks.

Social-ecological networks (SENs) represent the complex relationships between ecological and social systems and are a useful tool for analyzing and managing ecosystem services. However, mainstreaming the application of SENs in ecosystem service research has been hindered by a lack of clarity about how to match research questions to ecosystem service conceptualizations in SEN (i.e., as nodes, links, attributes, or emergent properties). Building from different disciplines, we propose a typology to represent ecosystem service in SENs and identify opportunities and challenges of using SENs in ecosystem service research. Our typology provides guidance for this growing field to improve research design and increase the breadth of questions that can be addressed with SEN to understand human-nature interdependencies in a changing world.

Autoimmunity in motion: Mechanisms of immune regulation and destruction revealed by in vivo imaging.

Autoimmunity arises when mechanisms of immune tolerance fail. Here we discuss mechanisms of T cell activation and tolerance and the dynamics of the autoimmune response at the site of disease. Live imaging of autoimmunity provides the ability to analyze immune cell dynamics at the single-cell level within the complex intact environment where disease occurs. These analyses have revealed mechanisms of T cell activation and tolerance in the lymph nodes, mechanisms of T cell entry into sites of autoimmune disease, and mechanisms leading to pathogenesis or protection in the autoimmune lesions. The overarching conclusions point to stable versus transient T cell antigen presenting cell interactions dictating the balance between T cell activation and tolerance, and T cell restimulation as a driver of pathogenesis at the site of autoimmunity. Findings from models of multiple sclerosis and type 1 diabetes are highlighted, however, the results have implications for basic mechanisms of T cell regulation during immune responses, tumor immunity, and autoimmunity.

MERTK on mononuclear phagocytes regulates T cell antigen recognition at autoimmune and tumor sites.

Understanding mechanisms of immune regulation is key to developing immunotherapies for autoimmunity and cancer. We examined the role of mononuclear phagocytes during peripheral T cell regulation in type 1 diabetes and melanoma. MERTK expression and activity in mononuclear phagocytes in the pancreatic islets promoted islet T cell regulation, resulting in reduced sensitivity of T cell scanning for cognate antigen in prediabetic islets. MERTK-dependent regulation led to reduced T cell activation and effector function at the disease site in islets and prevented rapid progression of type 1 diabetes. In human islets, MERTK-expressing cells were increased in remaining insulin-containing islets of type 1 diabetic patients, suggesting that MERTK protects islets from autoimmune destruction. MERTK also regulated T cell arrest in melanoma tumors. These data indicate that MERTK signaling in mononuclear phagocytes drives T cell regulation at inflammatory disease sites in peripheral tissues through a mechanism that reduces the sensitivity of scanning for antigen leading to reduced responsiveness to antigen.

Formin-like 1 mediates effector T cell trafficking to inflammatory sites to enable T cell-mediated autoimmunity.

Lymphocyte migration is essential for the function of the adaptive immune system, and regulation of T cell entry into tissues is an effective therapy in autoimmune diseases. Little is known about the specific role of cytoskeletal effectors that mediate mechanical forces and morphological changes essential for migration in complex environments. We developed a new Formin-like-1 (FMNL1) knock-out mouse model and determined that the cytoskeletal effector FMNL1 is selectively required for effector T cell trafficking to inflamed tissues, without affecting naïve T cell entry into secondary lymphoid organs. Here, we identify a FMNL1-dependent mechanism of actin polymerization at the back of the cell that enables migration of the rigid lymphocyte nucleus through restrictive barriers. Furthermore, FMNL1-deficiency impairs the ability of self-reactive effector T cells to induce autoimmune disease. Overall, our data suggest that FMNL1 may be a potential therapeutic target to specifically modulate T cell trafficking to inflammatory sites.

Resident mesenchymal vascular progenitors modulate adaptive angiogenesis and pulmonary remodeling via regulation of canonical Wnt signaling.

Adaptive angiogenesis is necessary for tissue repair, however, it may also be associated with the exacerbation of injury and development of chronic disease. In these studies, we demonstrate that lung mesenchymal vascular progenitor cells (MVPC) modulate adaptive angiogenesis via lineage trace, depletion of MVPC, and modulation of ß-catenin expression. Single cell sequencing confirmed MVPC as multipotential vascular progenitors, thus, genetic depletion resulted in alveolar simplification with reduced adaptive angiogenesis. Following vascular endothelial injury, Wnt activation in MVPC was sufficient to elicit an emphysema-like phenotype characterized by increased MLI, fibrosis, and MVPC driven adaptive angiogenesis. Lastly, activation of Wnt/ß-catenin signaling skewed the profile of human and murine MVPC toward an adaptive phenotype. These data suggest that lung MVPC drive angiogenesis in response to injury and regulate the microvascular niche as well as subsequent distal lung tissue architecture via Wnt signaling.

CD11c+ Cells Are Gatekeepers for Lymphocyte Trafficking to Infiltrated Islets During Type 1 Diabetes.

Type 1 diabetes (T1D) is a T cell mediated autoimmune disease that affects more than 19 million people with incidence increasing rapidly worldwide. For T cells to effectively drive T1D, they must first traffic to the islets and extravasate through the islet vasculature. Understanding the cues that lead to T cell entry into inflamed islets is important because diagnosed T1D patients already have established immune infiltration of their islets. Here we show that CD11c+ cells are a key mediator of T cell trafficking to infiltrated islets in non-obese diabetic (NOD) mice. Using intravital 2-photon islet imaging we show that T cell extravasation into the islets is an extended process, with T cells arresting in the islet vasculature in close proximity to perivascular CD11c+ cells. Antigen is not required for T cell trafficking to infiltrated islets, but T cell chemokine receptor signaling is necessary. Using RNAseq, we show that islet CD11c+ cells express over 20 different chemokines that bind chemokine receptors expressed on islet T cells. One highly expressed chemokine-receptor pair is CXCL16-CXCR6. However, NOD. CXCR6-/- mice progressed normally to T1D and CXCR6 deficient T cells trafficked normally to the islets. Even with CXCR3 and CXCR6 dual deficiency, T cells trafficked to infiltrated islets. These data reinforce that chemokine receptor signaling is highly redundant for T cell trafficking to inflamed islets. Importantly, depletion of CD11c+ cells strongly inhibited T cell trafficking to infiltrated islets of NOD mice. We suggest that targeted depletion of CD11c+ cells associated with the islet vasculature may yield a therapeutic target to inhibit T cell trafficking to inflamed islets to prevent progression of T1D.

CD4 T Cells Reactive to Hybrid Insulin Peptides Are Indicators of Disease Activity in the NOD Mouse.

We recently established that hybrid insulin peptides (HIPs), formed in islet ß-cells by fusion of insulin C-peptide fragments to peptides of chromogranin A or islet amyloid polypeptide, are ligands for diabetogenic CD4 T-cell clones. The goal of this study was to investigate whether HIP-reactive T cells were indicative of ongoing autoimmunity. MHC class II tetramers were used to investigate the presence, phenotype, and function of HIP-reactive and insulin-reactive T cells in NOD mice. Insulin-reactive T cells encounter their antigen early in disease, but they express FoxP3 and therefore may contribute to immune regulation. In contrast, HIP-reactive T cells are proinflammatory and highly diabetogenic in an adoptive transfer model. Because the frequency of antigen-experienced HIP-reactive T cells increases over progression of disease, they may serve as biomarkers of autoimmune diabetes.

Live Imaging of IL-4-Expressing T Follicular Helper Cells in Explanted Lymph Nodes.

The generation of class-switched, high-affinity, antibody-producing B cells plays a critical role in the establishment of type 2 immunity to intestinal helminths as well as in the pathogenesis of allergy and asthma. The generation of these high-affinity, antibody-producing B cells occurs in germinal centers (GC) and relies on interactions with follicular dendritic cells (FDCs) and T follicular helper (Tfh) cells. One critical mediator produced by Tfh cells in GCs is interleukin-4 (IL-4). Tfh-derived IL-4 drives class switching to type 2 antibody isotypes IgE and IgG1 and is required for high-affinity IgG1 production. In vivo detection of IL-4-expressing Tfh cells is required to better understand the role of these cells during the GC response. Detection of IL-4-expressing cells has been greatly improved by the generation of the IL-44get reporter mice, which read out IL-4 expression as green fluorescent protein (GFP). Much has been learned from these mice with regard to type 2 immunity using flow cytometry and immunohistochemistry. However, these methods do not allow the study of cellular behavior and interactions in real time. In contrast, multi-photon microscopy allows for deep tissue imaging and tracking of multiple cell types in intact tissues over time. Here, we describe a protocol for in vivo detection of IL-4-expressing Tfh cells in an explanted popliteal lymph node by multi-photon microscopy. The dynamics of Tfh cell motility and their interactions with FDC networks in the GCs were analyzed.

Crowdfunding biodiversity conservation.

Raising funds is critical for conserving biodiversity and hence so is scrutinizing emerging financial mechanisms that may help achieve this goal. Anecdotal evidence indicates crowdfunding is being used to support activities needed for biodiversity conservation, yet its magnitude and allocation remain largely unknown. To help address this knowledge gap, we conducted a global analysis based on conservation-focused projects extracted from crowdfunding platforms. For each project, we determined the funds raised, date, country of implementation, proponent characteristics, activity type, biodiversity realm, and target taxa. We identified 72 relevant platforms and 577 conservation-focused projects that raised $4,790,634 since 2009. Although proponents were based in 38 countries, projects were delivered across 80 countries, indicating a potential mechanism of resource mobilization. Proponents were affiliated with nongovernmental organizations (35%) or universities (30%) or were freelancers (26%). Most projects were for research (40%), persuasion (31%), and on-the-ground actions (21%). Projects were more focused on species (57.7%) and terrestrial ecosystems (20.3%), and less focused on marine (8.8%) and freshwater ecosystems (3.6%). Projects focused on 208 species, including a disproportionate number of threatened birds and mammals. Crowdfunding for biodiversity conservation is a global phenomenon and there is potential for expansion, despite possible pitfalls (e.g., uncertainty about effectiveness). Opportunities to advance conservation through crowdfunding arise from its capacity to mobilize funds spatially and increase steadily over time, inclusion of overlooked species, adoption by multiple actors, and funding of activities beyond research. Our findings pave the way for further research on key questions, such as campaign success rates, effectiveness of conservation actions, and drivers of crowdfunding adoption. Even though crowdfunding capital raised has been modest relative to other conservation-finance mechanisms, its contribution goes beyond funding research and providing capital. Embraced with due care, crowdfunding could become an important financial mechanism for biodiversity conservation.

Equity trade-offs in conservation decision making.

Conservation decisions increasingly involve multiple environmental and social objectives, which result in complex decision contexts with high potential for trade-offs. Improving social equity is one such objective that is often considered an enabler of successful outcomes and a virtuous ideal in itself. Despite its idealized importance in conservation policy, social equity is often highly simplified or ill-defined and is applied uncritically. What constitutes equitable outcomes and processes is highly normative and subject to ethical deliberation. Different ethical frameworks may lead to different conceptions of equity through alternative perspectives of what is good or right. This can lead to different and potentially conflicting equity objectives in practice. We promote a more transparent, nuanced, and pluralistic conceptualization of equity in conservation decision making that particularly recognizes where multidimensional equity objectives may conflict. To help identify and mitigate ethical conflicts and avoid cases of good intentions producing bad outcomes, we encourage a more analytical incorporation of equity into conservation decision making particularly during mechanistic integration of equity objectives. We recommend that in conservation planning motivations and objectives for equity be made explicit within the problem context, methods used to incorporate equity objectives be applied with respect to stated objectives, and, should objectives dictate, evaluation of equity outcomes and adaptation of strategies be employed during policy implementation.

Cognitive reappraisal and acceptance: Effects on emotion, physiology, and perceived cognitive costs.

Two emotion regulation strategies-cognitive reappraisal and acceptance-are both associated with beneficial psychological health outcomes over time. However, it remains unclear whether these 2 strategies are associated with differential consequences for emotion, physiology, or perceived cognitive costs in the short-term. The present study used a within-subjects design to examine the effects of reappraisal (reframing one's thoughts) and acceptance (accepting feelings without trying to control or judge them) on the subjective experience of negative emotions, positive emotions, and physiological responses during and following recovery from sad film clips shown in the laboratory. Participants also reported on perceived regulatory effort, difficulty, and success after deploying each emotion regulation strategy. In 2 samples of participants (N = 142), reappraisal (vs. acceptance) was associated with larger decreases in negative and larger increases in positive emotions, both during the film clips and recovery period. However, acceptance was perceived as less difficult to deploy than reappraisal, and was associated with a smaller dampening of skin conductance level (indicating more successful regulation) during the film clips in 1 sample. These results suggest that reappraisal and acceptance may exert differential short-term effects: Whereas reappraisal is more effective for changing subjective experiences in the short term, acceptance may be less difficult to deploy and be more effective at changing one's physiological response. Thus, these 2 strategies may both be considered "effective" for different reasons. (PsycINFO Database Record