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Background Overcoming immune suppression is a major barrier to eliciting potent CD8+ T cell responses against cancer. Treatment with anti-CD4 monoclonal antibody is an effective means for eliminating CD4+Foxp3+ regulatory (Treg) cells in preclinical models and has also demonstrated efficacy in early clinical trials. However, the underlying basis for treatment efficacy, more specifically the implications of codepleting other CD4-expressing cell compartments in tumor-bearing hosts, is not well understood.Methods Tumor-bearing mice were treated with anti-CD4 versus other therapies that preserve helper T cell function, and the priming, tissue distribution, and maintenance of tumor antigen-specific CD8 T cells were assessed. Antibody blockade and transgenic mouse models were used to determine the mechanisms of CD8 T cell priming. Single-cell RNA-sequencing (scRNAseq) was used to further characterize CD8 T cells that are primed by anti-CD4 therapy and to identify immunosuppressive CD4 T cell subsets in human melanoma following immune checkpoint blockade (ICB).Results Comparing anti-CD4 to dual ICB therapy, we show that anti-CD4 facilitates more robust priming of TCF-1+, IL-2-producing, tumor-specific CD8+ T cells that disseminate to tissues and form memory. By decoupling priming from homeostatic proliferation and associated cytokines, we find that anti-CD4 functions independently of creating homeostatic space for CD8+ T cells. We also show that depletion of CD4-expressing antigen-presenting cell subsets is not required for anti-CD4 efficacy. Instead, robust tumor-specific CD8+ T cell priming and memory generation required the removal of total antigen-specific CD4+ T cells, including both Tregs and CD4+ Foxp3-negative conventional (Tconv) cells. In particular, the elimination of CD4+ Tconv cells was necessary for the accumulation and maturation of conventional type-1 dendritic cells in tumor-draining LNs, which were required for CD8+ T cell priming. Accordingly, anti-CD4 treatment restored CD8+ T cell responses in mice cotreated with dual ICB. scRNAseq of melanoma tumors from patients who received ICB revealed the presence of Tr1 and Treg subsets, as well as CD4+ Tconv subsets that lacked clear transcriptional evidence of helper differentiation.Conclusions These findings underscore the underappreciated benefit of depleting CD4+ Tconv cells to promote systemic primary and memory CD8+ T cell responses against cancer.

Abstract Background Understanding current substance use practices is critical to reduce and prevent overdose deaths among individuals at increased risk including persons who use and inject drugs. Because individuals participating in harm reduction and syringe service programs are actively using drugs and vary in treatment participation, information on their current drug use and preferred drugs provides a unique window into the drug use ecology of communities that can inform future intervention services and treatment provision. Methods Between March and June 2023, 150 participants in a harm reduction program in Burlington, Vermont completed a survey examining sociodemographics; treatment and medication for opioid use disorder (MOUD) status; substance use; injection information; overdose information; and mental health, medical, and health information. Descriptive analyses assessed overall findings. Comparisons between primary drug subgroups (stimulants, opioids, stimulants-opioids) of past-three-month drug use and treatment participation were analyzed using chi-square and Fisher’s exact test. Results Most participants reported being unhoused or unstable housing (80.7%) and unemployed (64.0%) or on disability (21.3%). The drug with the greatest proportion of participants reporting past three-month use was crack cocaine (83.3%). Fentanyl use was reported by 69.3% of participants and xylazine by 38.0% of participants. High rates of stimulant use were reported across all participants independent of whether stimulants were a participant’s primary drug. Fentanyl, heroin, and xylazine use was less common in the stimulants subgroup compared to opioid-containing subgroups (p

This article introduces a model-based robust control framework for electrohydraulic soft robots. The methods presented herein exploit linear system control theory as it applies to a nonlinear soft robotic system. We employ dynamic mode decomposition with control (DMDc) to create appropriate linear models from real-world measurements. We build on the theory by developing linear models in various operational regions of the system to result in a collection of linear plants used in uncertainty analysis. To complement the uncertainty analyses, we utilize H∞ (“H Infinity”) synthesis techniques to determine an optimal controller to meet performance requirements for the nominal plant. Following this methodology, we demonstrate robust control over a multi-input multi-output (MIMO) hydraulically amplified self-healing electrostatic (HASEL)-actuated system. The simplifications in the proposed framework help address the inherent uncertainties and complexities of compliant robots, providing a flexible approach for real-time control of soft robotic systems in real-world applications.

Abstract The convergence of urban expansion, deteriorating infrastructure, and a changing climate will escalate the risks of stormwater pollution and urban flooding in the coming decades. Using outputs from an ensemble of global climate models to drive a high spatial resolution stormwater model, we analyzed climate change impacts on urban stormwater runoff and control measures for 23 cities across the United States. Runoff model outputs for two future emissions scenarios ending in 2055 were compared against a historical scenario to assess changes. All cities showed increases in average annual stormwater runoff, with changes up to 30% over the next 30 years due to a greater frequency of high intensity storm events. Runoff model outputs showed substantial variation across cities with untreated stormwater runoff increasing by as much as 48%. Patterns of future runoff impacts within cities will affect the performance of distributed treatment strategies such as Green Stormwater Infrastructure (GSI) to meet municipal water quality improvement and runoff reduction goals. Results indicate that adoption of adaptable design standards and decision support tools that readily accommodate projected precipitation changes are critical for supporting more resilient designs of stormwater control measures.

IntroductionTerrestrial species in riverine ecosystems face unique constraints leading to diverging patterns of population structure, connectivity, and disease dynamics. Desert bighorn sheep (Ovis canadensis nelsoni) in Grand Canyon National Park, a large native population in the southwestern USA, offer a unique opportunity to evaluate population patterns and processes in a remote riverine system with ongoing anthropogenic impacts. We integrated non-invasive, invasive, and citizen-science methods to address questions on abundance, distribution, disease status, genetic structure, and habitat fragmentation.MethodsWe compiled bighorn sightings collected during river trips by park staff, commercial guides, and private citizens from 2000–2018 and captured bighorn in 2010–2016 to deploy GPS collars and test for disease. From 2011–2015, we non-invasively collected fecal samples and genotyped them at 9–16 microsatellite loci for individual identification and genetic structure. We used assignment tests to evaluate genetic structure and identify subpopulations, then estimated gene flow and recent migration to evaluate fragmentation. We used spatial capture-recapture to estimate annual population size, distribution, and trends after accounting for spatial variation in detection with a resource selection function model.Results and discussionFrom 2010–2018, 3,176 sightings of bighorn were reported, with sightings of 56–145 bighorn annually on formal surveys. From 2012–2016, bighorn exhibiting signs of respiratory disease were observed along the river throughout the park. Of 25 captured individuals, 56% were infected by Mycoplasma ovipneumoniae, a key respiratory pathogen, and 81% were recently exposed. Pellet sampling for population estimation from 2011–2015 yielded 1,250 genotypes and 453 individuals. We detected 6 genetic clusters that exhibited mild to moderate genetic structure (FST 0.022–0.126). The river, distance, and likely topography restricted recent gene flow, but we detected cross-river movements in one section via genetic recaptures, no subpopulation appeared completely isolated, and genetic diversity was among the highest reported. Recolonization of one large stretch of currently empty habitat appears limited by the constrained topology of this system. Annual population estimates ranged 536–552 (95% CrI range 451–647), lamb:ewe ratios varied, and no significant population decline was detected. We provide a multi-method sampling framework useful for sampling other wildlife in remote riverine systems.

Abstract Background Deaf and hard‐of‐hearing (DHH) patients are a priority population for emergency medicine health services research. DHH patients are at higher risk than non‐DHH patients of using the emergency department (ED), have longer lengths of stay in the ED and report poor patient–provider communication. This qualitative study aimed to describe ED care‐seeking and patient‐centred care perspectives among DHH patients. Methods This qualitative study is the second phase of a mixed‐methods study. The goal of this study was to further explain quantitative findings related to ED outcomes among DHH and non‐DHH patients. We conducted semistructured interviews with 4 DHH American Sign Language (ASL)‐users and 6 DHH English speakers from North Central Florida. Interviews were transcribed and analysed using a descriptive qualitative approach. Results Two themes were developed: (1) DHH patients engage in a complex decision‐making process to determine ED utilization and (2) patient‐centred ED care differs between DHH ASL‐users and DHH English speakers. The first theme describes the social‐behavioural processes through which DHH patients assess their need to use the ED. The second theme focuses on the social environment within the ED: patients feeling stereotyped, involvement in the care process, pain communication, receipt of accommodations and discharge processes. Conclusions This study underscores the importance of better understanding, and intervening in, DHH patient ED care‐seeking and care delivery to improve patient outcomes. Like other studies, this study also finds that DHH patients are not a monolithic group and language status is an equity‐relevant indicator. We also discuss recommendations for emergency medicine. Patient or Public Contribution This study convened a community advisory group made up of four DHH people to assist in developing research questions, data collection tools and validation of the analysis and interpretation of data. Community advisory group members who were interested in co‐authorship are listed in the byline, with others in the acknowledgements. In addition, several academic‐based co‐authors are also deaf or hard of hearing.

Piperonyl butoxide (PBO) is a popular insecticide synergist present in thousands of commercial, agricultural, and household products. PBO inhibits cytochrome P450 activity, impairing the ability of insects to detoxify insecticides. PBO was recently discovered to also inhibit Sonic hedgehog signaling, a pathway required for embryonic development, and rodent studies have demonstrated the potential for in utero PBO exposure to cause structural malformations of the brain, face, and limbs, or more subtle neurodevelopmental abnormalities. The current understanding of the pharmacokinetics of PBO in mice is limited, particularly with respect to dosing paradigms associated with developmental toxicity. To establish a pharmacokinetic (PK) model for oral exposure, PBO was administered to female C57BL/6J mice acutely by oral gavage (22–1800 mg/kg) or via diet (0.09 % PBO in chow). Serum and adipose samples were collected, and PBO concentrations were determined by HPLC-MS/MS. The serum concentrations of PBO were best fit by a linear one-compartment model. PBO concentrations in visceral adipose tissue greatly exceeded those in serum. PBO concentrations in both serum and adipose tissue decreased quickly after cessation of dietary exposure. The elimination half-life of PBO in the mouse after gavage dosing was 6.5 h (90 % CI 4.7–9.5 h), and systemic oral clearance was 83.3 ± 20.5 mL/h. The bioavailability of PBO in chow was 41 % that of PBO delivered in olive oil by gavage. Establishment of this PK model provides a foundation for relating PBO concentrations that cause developmental toxicity in the rodent models to Sonic hedgehog signaling pathway inhibition.

Abstract Although glaucoma is a disease modulated by eye pressure, the mechanisms of pressure sensing in the eye are not well understood. Here, we investigated associations between mechanosensitive ion channel gene variants and primary open-angle glaucoma (POAG). Common (minor allele frequency > 5%) single nucleotide polymorphisms located within the genomic regions of 20 mechanosensitive ion channel genes in the K2P, TMEM63, PIEZO and TRP channel families were assessed using genotype data from the NEIGHBORHOOD consortium of 3853 cases and 33,480 controls. Rare (minor allele frequency

Abstract Robust estimates of historical changes in aerosols are key for accurate constraints on climate sensitivity. Dry deposition is a primary sink of aerosols from the atmosphere. However, most global climate models do not accurately represent observed strong dependencies of dry deposition following turbulent transport on aerosol size. It is unclear whether there is a substantial impact of mischaracterized aerosol deposition velocities on historical aerosol changes. Here we describe improved mechanistic representation of aerosol dry deposition in the NASA Goddard Institute for Space Studies (GISS) global climate model, ModelE, and illustrate the impact on 1850–2000 changes in global aerosol burdens as well as aerosol direct and cloud albedo effects using a set of 1850 and 2000 time slice simulations. We employ two aerosol configurations of ModelE (a “bulk” mass‐based configuration and a configuration that more explicitly represents aerosol size distributions, internal mixing, and microphysics) to explore how model structural differences in aerosol representation alter the response to representation of dry deposition. Both configurations show larger historical increases in the global burdens of non‐dust aerosols with the new dry deposition scheme, by 11% in the simpler mass‐based configuration and 23% in the more complex microphysical configuration. Historical radiative forcing responses, which vary in magnitude from 5% to 12% as well as sign, depend on the aerosol configuration.

Introduction. Inflammatory sacroiliitis is common in rheumatology practice. Spondyloarthritis is often underdiagnosed due to the lack of proper evaluation of the sacroiliac joints (SIJs), clinically and radiographically. If SIJ is inflamed or arthritic, the arthritic said patient typically has spondyloarthritis, in the absence of infections or crystal arthritis. Sacroiliitis, in particular, when diagnosed between 12 and 45 years of age, is indicative of spondyloarthritis. People are often misdiagnosed and mislabeled as fibromyalgia because their serologies are negative. Our goal is to point out the importance of proper evaluation, diagnosis, and importance of inflammatory SIJ disease and conditions that involve SIJ inflammation. Cases. We present three rare conditions presenting with bilateral and symmetric SIJ disease, none of which is ankylosing spondylitis, Crohn’s colitis, ulcerative colitis, psoriatic arthritis, and reactive arthritis (Reiter syndrome); there are reports of concurrent SIJ disease in rheumatoid arthritis and SLE. Conclusion. The authors believe that SIJ disease is overlooked, is underdiagnosed, and can lead to incorrect treatment. We suggest a greater focus on SIJ imaging in the diagnosis and treatment of unexplained illnesses associated with low back pain, morning stiffness, or unexplained buttock pain. Providers should review their own SIJ films. The meaning of SIJ widening, cortical irregularity, spurs, and the significance of the anterior inferior SI joints, bone marrow edema, and fusion (namely, the natural history of sacroiliac pathophysiology).

This pilot study sought to evaluate the circulating levels of immune cells, particularly regulatory T-cell (Treg) subsets, before and after lung resection for non-small cell lung cancer. Twenty-five patients consented and had specimens collected. Initially, peripheral blood of 21 patients was collected for circulating immune cell studies. Two of these patients were excluded due to technical issues, leaving 19 patients for the analyses of circulating immune cells. Standard gating and high-dimensional unsupervised clustering flow cytometry analyses were performed. The blood, tumors and lymph nodes were analyzed via single-cell RNA and TCR sequencing for Treg analyses in a total of five patients (including four additional patients from the initial 21 patients). Standard gating flow cytometry revealed a transient increase in neutrophils immediately following surgery, with a variable neutrophil–lymphocyte ratio and a stable CD4–CD8 ratio. Unexpectedly, the total Treg and Treg subsets did not change with surgery with standard gating in short- or long-term follow-up. Similarly, unsupervised clustering of Tregs revealed a dominant cluster that was stable perioperatively and long-term. Two small FoxP3hi clusters slightly increased following surgery. In the longer-term follow-up, these small FoxP3hi Treg clusters were not identified, indicating that they were likely a response to surgery. Single-cell sequencing demonstrated six CD4+FoxP3+ clusters among the blood, tumors and lymph nodes. These clusters had a variable expression of FoxP3, and several were mainly, or only, present in tumor and lymph node tissue. As such, serial monitoring of circulating Tregs may be informative, but not completely reflective of the Tregs present in the tumor microenvironment.

Mixed methods research relies on the integration of quantitative and qualitative data to develop meta-inferences. Despite an increase in techniques to facilitate integration at the methods and reporting levels, there is less practical guidance on how to conduct mixed methods integrative data analysis. Qualitative data analysis software is rapidly advancing to facilitate integrative analysis in mixed methods research studies. However, examples on how these features support analysis are relatively limited. The purpose of this article is to promote the use of qualitative data analysis software for integrative analysis in mixed methods research by describing the value of the major features available in software (i.e., code queries, crosstabulations, and quote matrices). In addition to a brief overview of these features, this article demonstrates the use of MAXQDA's Interactive Quote Matrix with a real-life example of a completed mixed methods study and delineates the step-by-step process on how to use this feature to develop meta-inferences.

BackgroundPeople with physical disabilities often experience premature multimorbidity and adverse health events. A tailored primary care approach for this vulnerable population that also accounts for social and functional risk factors could promote healthier aging and more equitable health care. ObjectiveThis project will evaluate the implementation of a health program designed for people with physical disabilities. The proposed evaluation result is to generate the first best-practice protocol focused specifically on developing primary care to help reduce preventable causes of morbidity and improve functioning among people with physical disabilities. MethodsWe will design and implement a pilot health program for people with physical disabilities at a primary care clinic within Michigan Medicine. The health program for people with physical disabilities will be an integrated intervention involving a tailored best practice alert designed to prompt family medicine providers to screen and monitor for common, preventable health conditions. The program will also collect social and functional status information to determine the patient’s need for further care coordination and support. Adult participants from this clinic with identified physical disabilities will be targeted for potential enrollment. To create a quasi-experimental setting, a separate departmental clinic will serve as a control site for comparison purposes. A quantitative analysis to estimate the treatment effect of implementing this health program will be conducted using a difference-in-differences approach. Outcomes of interest will include the use of preventative services (eg, hemoglobin A1c for diabetes screening), social work assistance, and emergency and hospital services. These data will be extracted from electronic health records. Time-invariant covariates, particularly sociodemographic covariates, will be included in the models. A qualitative analysis of patient and health care provider interviews will also be completed to assess the effect of the health program. Patient Health Questionnaire-9 and Generalized Anxiety Disorder 7-item scores will be assessed to both screen for depression and anxiety as well as explore program impacts related to addressing health and functioning needs related to physical disabilities in a primary care setting. These will be summarized through descriptive analyses. ResultsThis study was funded in September 2018, data collection started in September 2021, and data collection is expected to be concluded in September 2023. ConclusionsThis study is a mixed methods evaluation of the effectiveness of an integrated health program designed for people with physical disabilities, based on a quasi-experimental comparison between an intervention and a control clinic site. The intervention will be considered successful if it leads to improvements in greater use of screening and monitoring for preventable health conditions, increased social worker referrals to assist with health and functioning needs, and improvements in emergency and hospital-based services. The findings will help inform best practices for people with physical disabilities in a primary care setting. International Registered Report Identifier (IRRID)DERR1-10.2196/50105

Abstract Purpose Triple-negative breast cancer continues to be one of the leading causes of death in women, making up 7% of all cancer deaths. Tumor-treating electric fields are low-energy, low-frequency oscillating electric fields that induce an anti-proliferative effect on mitotic cells in glioblastoma multiforme, non-small cell lung cancer, and ovarian cancer. Little is known about effects of tumor-treating fields on triple-negative breast cancer and known research for tumor-treating fields only utilizes low (

Abstract Background The genus Sphingobium within the class Alpha-proteobacteria contains a small number of plant-growth promoting rhizobacteria (PGPR), although it is mostly comprised of organisms that play an important role in biodegradation and bioremediation in sediments and sandy soils. A Sphingobium sp. isolate was obtained from the rhizosphere of the beachgrass Ammophila breviligulata with a variety of plant growth-promoting properties and designated as Sphingobium sp. strain AEW4. Results Analysis of the 16S rRNA gene as well as full genome nucleotide and amino acid identities revealed that this isolate is most similar to Sphingobium xenophagum and Sphingobium hydrophobicum. Comparative genomics analyses indicate that the genome of strain AEW4 contains unique features that explain its relationship with a plant host as a PGPR, including pathways involved in monosaccharide utilization, fermentation pathways, iron sequestration, and resistance to osmotic stress. Many of these unique features are not broadly distributed across the genus. In addition, pathways involved in the metabolism of salicylate and catechol, phenyl acetate degradation, and DNA repair were also identified in this organism but not in most closely related organisms. Conclusion The genome of Sphingobium sp. strain AEW4 contains a number of distinctive features that are crucial to explain its role as a plant-growth promoting rhizobacterium, and comparative genomics analyses support its classification as a relevant Sphingobium strain involved in plant growth promotion of beachgrass and other plants.

Abstract Depression, anxiety, and stress continue to be among the largest burdens of disease, globally. The Depression, Anxiety, and Stress Scale-21 Items (DASS-21) is a shortened version of DASS-41 developed to measure these mental health conditions. The DASS-41 has strong evidence of validity and reliability in multiple contexts. However, the DASS-21, and the resulting item properties, has been explored less in terms of modern test theories. One such theory is Item Response Theory (IRT), and we use IRT models to explore latent item and person traits of each DASS-21 sub-scale among people living in Malaysia. Specifically, we aimed to assess Classical Test Theory and IRT properties including dimensionality, internal consistency (reliability), and item-level properties. We conducted a web-based cross-sectional study and sent link-based questionnaires to people aged 18 and above in a private university and requested to roll out the link. Overall and individual sub-scales’ Cronbach’s alpha of the DASS-21 indicates an excellent internal consistency. The average inter-item correlation and corrected inter-item correlations for each of the sub-scales indicated acceptable discrimination. On average, DASS-21 total scores and sub-scale scores were significantly higher among female participants than males. The Graded Response Model had better empirical fit to sub-scale response data. Raw summated and latent (IRT estimated) scores of the Depression, Anxiety, and Stress sub-scales, and overall DASS-21 were strongly correlated. Thus, this study provides evidence of validity supporting the use of the DASS-21 as a mental health screening tool among Malaysians. Specifically, standard error of measurement was minimized to provide robust evidence of potential utility in identifying participants who are and are not experiencing these mental health issues. Additional research is warranted to ensure that test content culturally appropriate and accurately measuring cultural norms of depression, anxiety, and stress.

Introduction People with disabilities have a higher prevalence of cigarette smoking than people without disabilities. However, little information exists on smoking cessation interventions tailored to address the unique needs of people with disabilities. This paper describes a systematic review protocol to identify and evaluate tobacco smoking cessation interventions designed to improve outcomes for people with disabilities.Methods and analysis We will conduct a systematic review of the literature using the procedures outlined by Cochrane. We will search four electronic databases (CINAHL Plus (EBSCO), Embase (Ovid), Medline (Ovid) and PsycINFO (Ovid)) with no date restriction to identify tobacco cessation interventions tailored to meet the needs of people with disabilities. We will extract data and assess risk of bias using the RoB2 and ROBINS-I for included studies using Covidence systematic review software. Quantitative and qualitative syntheses will summarise key study characteristics and outcomes with text, tables and forest plots; a meta-analysis will be conducted, if appropriate.Ethics and dissemination Ethical approval is not required as there are no primary data associated with the study. Data will be disseminated through a peer-reviewed articles and conference presentations.PROSPERO registration number CRD42022337434.

People with disabilities represent the largest minority group in the United States and a priority population for health services research. Despite federal civil rights law, people with disabilities face inaccessible health care environments that fail to accommodate their disability. We present Michigan Medicine’s Disability and Accommodations Tab. This patient-facing questionnaire and shared data field in the electronic health record enables the collection and reporting of patient disability-related accommodations. The Disability Tab seeks to address provider- and clinic staff–reported barriers to providing accommodations and fosters an opportunity to redesign health care to meet the needs of people with disabilities.

Current methods of root sampling typically only obtain small or incomplete sections of root systems and do not capture their true complexity. To facilitate the visualization and analysis of full-sized plant root systems in 3-dimensions, we developed customized mesocosm growth containers. While highly scalable, the design presented here uses an internal volume of 45 ft3 (1.27 m3), suitable for large crop and bioenergy grass root systems to grow largely unconstrained. Furthermore, they allow for the excavation and preservation of 3-dimensional root system architecture (RSA), and facilitate the collection of time-resolved subterranean environmental data. Sensor arrays monitoring matric potential, temperature and CO2 levels are buried in a grid formation at various depths to assess environmental fluxes at regular intervals. Methods of 3D data visualization of fluxes were developed to allow for comparison with root system architectural traits. Following harvest, the recovered root system can be digitally reconstructed in 3D through photogrammetry, which is an inexpensive method requiring only an appropriate studio space and a digital camera. We developed a pipeline to extract features from the 3D point clouds, or from derived skeletons that include point cloud voxel number as a proxy for biomass, total root system length, volume, depth, convex hull volume and solidity as a function of depth. Ground-truthing these features with biomass measurements from manually dissected root systems showed a high correlation. We evaluated switchgrass, maize, and sorghum root systems to highlight the capability for species wide comparisons. We focused on two switchgrass ecotypes, upland (VS16) and lowland (WBC3), in identical environments to demonstrate widely different root system architectures that may be indicative of core differences in their rhizoeconomic foraging strategies. Finally, we imposed a strong physiological water stress and manipulated the growth medium to demonstrate whole root system plasticity in response to environmental stimuli. Hence, these new “3D Root Mesocosms” and accompanying computational analysis provides a new paradigm for study of mature crop systems and the environmental fluxes that shape them.

In recent decades, GNSS radio occultation (RO) soundings have proven to be an invaluable input to global weather forecasting. The success of government programs such as COSMIC is now complemented by commercial low-cost cubesats. The result is access to more than 10,000 soundings per day. We examine aerial platforms for commercial GNSS-RO, specifically high-altitude balloons and commercial aviation. Meteorological balloons (radiosondes) are deployed daily in over 900 locations globally. Adding GNSS-RO capability to radiosondes would expand capability and enable local area monitoring. Commercial aviation offers scale with more than 100,000 flights daily. A barrier to entry for the inclusion of GNSS-RO sensors is cost and complexity, as GNSS-RO traditionally requires highly specialized equipment. This paper describes a low-cost and scalable approach to aerial GNSS-RO based on commercial-off-the-shelf (COTS) GNSS receivers. We present hardware prototypes and data processing techniques that demonstrate technical feasibility through the results from several flight testing campaigns.

People who are deaf and hard-of-hearing (DHH) struggle with information marginalization and limited health literacy, challenging their ability to access information on preventing coronavirus disease 2019 (COVID-19). This study assessed the relationship between language preference, health literacy, and COVID-19 information barriers among parents who are DHH in the United States. Data were drawn from a larger study focused on individuals who are DHH who had given birth in the past 10 years. Respondents completed a web-based survey between March 2020 and July 2021. We segmented respondents by language preference [i.e., American Sign Language (ASL), English, or bilingual ASL/English] and used logistic regression models to test the hypothesis that language preference and health literacy were both associated with COVID-19 information marginalization. Of the total sample (N = 417), approximately 17% had limited health literacy, and 22% reported experiencing difficulty accessing information about COVID-19. In adjusted analyses, respondents with limited health literacy ([adjusted odds ratio] aOR = 2.245) and Hispanic ethnicity (aOR = 2.149) had higher risk of reporting information access barriers. There was no association between language preference and reporting COVID-19 information barriers. However, individuals who are DHH with limited health literacy were at higher risk of experiencing information marginalization during the ongoing COVID-19 pandemic, highlighting the need for tailored information based on access needs. [HLRP: Health Literacy Research and Practice. 2022;6(4):e310–e315.]

ABSTRACT Aspergillus fumigatus causes invasive aspergillosis (IA) in immunocompromised patients, resulting in high mortality rates. Currently, no vaccine formulations to promote immune protection in at-risk individuals have been developed. In this work, we deleted the sterylglucosidase-encoding gene, sglA, in Aspergillus fumigatus and investigated its role in fungal virulence and host vaccine protection. The ΔsglA mutant accumulated sterylglucosides (SGs), newly studied immunomodulatory glycolipids, and exhibited reduced hyphal growth and altered compositions of cell wall polysaccharides. Interestingly, the ΔsglA mutant was avirulent in two murine models of IA and was fully eliminated from the lungs. Both corticosteroid-induced immunosuppressed and cyclophosphamide-induced leukopenic mice vaccinated with live or heat-killed ΔsglA conidia were fully protected against a lethal wild-type A. fumigatus challenge. These results highlight the potential of SG-accumulating strains as safe and promising vaccine formulations against invasive fungal infections. IMPORTANCE Infections by Aspergillus fumigatus occur by the inhalation of environmental fungal spores called conidia. We found that live mutant conidia accumulating glycolipids named sterylglucosides are not able to cause disease when injected into the lung. Interestingly, these animals are now protected against a secondary challenge with live wild-type conidia. Remarkably, protection against a secondary challenge persists even with vaccination with heat-killed mutant conidia. These results will significantly advance the field of the research and development of a safe fungal vaccine for protection against the environmental fungus A. fumigatus.

Vaccines are one of the greatest medical accomplishments to date, yet no fungal vaccines are currently available in humans mainly because opportunistic mycoses generally occur during immunodeficiencies necessary for vaccine protection. In previous studies, a live, attenuated Cryptococcus neoformans Δsgl1 mutant accumulating sterylglucosides was found to be avirulent and protected mice from a subsequent lethal infection even in absence of CD4+ T cells, a condition most associated with cryptococcosis (e.g., HIV). Here, we tested three strategies of vaccination against cryptococcosis. First, in our preventative model, protection was achieved even after a 3-fold increase of the vaccination window. Second, because live C. neoformans Δsgl1-vaccinated mice challenged more than once with WT strain had a significant decrease in lung fungal burden, we tested C. neoformans Δsgl1 as an immunotherapeutic. We found that therapeutic administrations of HK C. neoformans Δsgl1 post WT challenge significantly improves the lung fungal burden. Similarly, therapeutic administration of HK C. neoformans Δsgl1 post WT challenge resulted in 100% or 70% survival depending on the time of vaccine administration, suggesting that HK Δsgl1 is a robust immunotherapeutic option. Third, we investigated a novel model of vaccination in preventing reactivation from lung granuloma using C. neoformans Δgcs1. Remarkably, we show that administration of HK Δsgl1 prevents mice from reactivating Δgcs1 upon inducing immunosuppression with corticosteroids or by depleting CD4+ T cells. Our results suggest that HK Δsgl1 represents a clinically relevant, efficacious vaccine that confers robust host protection in three models of vaccination against cryptococcosis even during CD4-deficiency.

Abstract Background Age-related macular degeneration (AMD) is a progressive retinal disease contributing to blindness worldwide. Multiple estimates for AMD heritability (h 2 ) exist; however, a substantial proportion of h 2 is not attributable to known genomic loci. The International AMD Genomics Consortium (IAMDGC) gathered the largest dataset of advanced AMD (ADV) cases and controls available and identified 34 loci containing 52 independent risk variants defining known AMD h 2 . To better define AMD heterogeneity, we used Pathway Analysis by Randomization Incorporating Structure (PARIS) on the IAMDGC data and identified 8 statistical driver genes (SDGs), including 2 novel SDGs not discovered by the IAMDGC. We chose to further investigate these pathway-based risk genes and determine their contribution to ADV h 2 , as well as the differential ADV subtype h 2 . Methods We performed genomic-relatedness-based restricted maximum-likelihood (GREML) analyses on ADV, geographic atrophy (GA), and choroidal neovascularization (CNV) subtypes to investigate the h 2 of genotyped variants on the full DNA array chip, 34 risk loci (n = 2758 common variants), 52 variants from the IAMDGC 2016 GWAS, and the 8 SDGs, specifically the novel 2 SDGs, PPARA and PLCG2. Results Via GREML, full chip h 2 was 44.05% for ADV, 46.37% for GA, and 62.03% for CNV. The lead 52 variants’ h 2 (ADV: 14.52%, GA: 8.02%, CNV: 13.62%) and 34 loci h 2 (ADV: 13.73%, GA: 8.81%, CNV: 12.89%) indicate that known variants contribute ~ 14% to ADV h 2 . SDG variants account for a small percentage of ADV, GA, and CNV heritability, but estimates based on the combination of SDGs and the 34 known loci are similar to those calculated for known loci alone. We identified modest epistatic interactions among variants in the 2 SDGs and the 52 IAMDGC variants, including modest interactions between variants in PPARA and PLCG2. Conclusions Pathway analyses, which leverage biological relationships among genes in a pathway, may be useful in identifying additional loci that contribute to the heritability of complex disorders in a non-additive manner. Heritability analyses of these loci, especially amongst disease subtypes, may provide clues to the importance of specific genes to the genetic architecture of AMD.

Abstract Background Laminin-α2-related congenital muscular dystrophy (LAMA2-CMD) is a devastating genetic disease caused by mutations in the LAMA2 gene. These mutations result in progressive muscle wasting and inflammation leading to delayed milestones, and reduced lifespan in affected patients. There is currently no cure or treatment for LAMA2-CMD. Preclinical studies have demonstrated that mouse laminin-111 can serve as an effective protein replacement therapy in a mouse model of LAMA2-CMD. Methods In this study, we generated a novel immunocompromised dyW mouse model of LAMA2-CMD to study the role the immune system plays in muscle disease progression. We used this immune-deficient dyW mouse model to test the therapeutic benefits of recombinant human laminin-111 and laminin-211 protein therapy on laminin-α2-deficient muscle disease progression. Results We show that immunodeficient laminin-α2 null mice demonstrate subtle differences in muscle regeneration compared to immunocompetent animals during early disease stages but overall exhibit a comparable muscle disease progression. We found human laminin-111 and laminin-211 could serve as effective protein replacement strategies with mice showing improvements in muscle pathology and function. We observed that human laminin-111 and laminin-211 exhibit differences on satellite and myoblast cell populations and differentially affect muscle repair. Conclusions This study describes the generation of a novel immunodeficient mouse model that allows investigation of the role the immune system plays in LAMA2-CMD. This model can be used to assess the therapeutic potential of heterologous therapies that would elicit an immune response. Using this model, we show that recombinant human laminin-111 can serve as effective protein replacement therapy for the treatment of LAMA2-CMD.

Abstract Background Temperature exerts a strong influence on protein evolution: species living in thermally distinct environments often exhibit adaptive differences in protein structure and function. However, previous research on protein temperature adaptation has focused on small numbers of proteins and on proteins adapted to extreme temperatures. Consequently, less is known about the types and quantity of evolutionary change that occurs to proteins when organisms adapt to small shifts in environmental temperature. In this study, these uncertainties were addressed by developing software that enabled comparison of structural changes associated with temperature adaptation (hydrogen bonding, salt bridge formation, and amino acid use) among large numbers of proteins from warm- and cold-adapted species of marine mussels, Mytilus galloprovincialis and Mytilus trossulus, respectively. Results Small differences in habitat temperature that characterize the evolutionary history of Mytilus mussels were sufficient to cause protein structural changes consistent with temperature adaptation. Hydrogen bonds and salt bridges that increase stability and protect against heat-induced denaturation were more abundant in proteins from warm-adapted M. galloprovincialis compared with proteins from cold-adapted M. trossulus. These structural changes were related to deviations in the use of polar and charged amino acids that facilitate formation of hydrogen bonds and salt bridges within proteins, respectively. Enzymes, in particular those within antioxidant and cell death pathways, were over-represented among proteins with the most hydrogen bonds and salt bridges in warm-adapted M. galloprovincialis. Unlike extremophile proteins, temperature adaptation in Mytilus proteins did not involve substantial changes in the number of hydrophobic or large volume amino acids, nor in the content of glycine or proline. Conclusions Small shifts in organism temperature tolerance, such as that needed to cope with climate warming, may result from structural and functional changes to a small percentage of the proteome. Proteins in which function is dependent on large conformational change, notably enzymes, may be particularly sensitive to temperature perturbation and represent foci for natural selection. Protein temperature adaptation can occur through different types and frequencies of structural change, and adaptive mechanisms used to cope with small shifts in habitat temperature appear different from mechanisms used to retain protein function at temperature extremes.

Assessments of the potential responses of animal species to climate change often rely on correlations between long-term average temperature or precipitation and species’ occurrence or abundance. Such assessments do not account for the potential predictive capacity of either climate extremes and variability or the indirect effects of climate as mediated by plant phenology. By contrast, we projected responses of wildlife in desert grasslands of the southwestern United States to future climate means, extremes, and variability and changes in the timing and magnitude of primary productivity. We used historical climate data and remotely sensed phenology metrics to develop predictive models of climate-phenology relations and to project phenology given anticipated future climate. We used wildlife survey data to develop models of wildlife-climate and wildlife-phenology relations. Then, on the basis of the modeled relations between climate and phenology variables, and expectations of future climate change, we projected the occurrence or density of four species of management interest associated with these grasslands: Gambel’s Quail ( Callipepla gambelii ), Scaled Quail ( Callipepla squamat ), Gunnison’s prairie dog ( Cynomys gunnisoni ), and American pronghorn ( Antilocapra americana ). Our results illustrated that climate extremes and plant phenology may contribute more to projecting wildlife responses to climate change than climate means. Monthly climate extremes and phenology variables were influential predictors of population measures of all four species. For three species, models that included climate extremes as predictors outperformed models that did not include extremes. The most important predictors, and months in which the predictors were most relevant to wildlife occurrence or density, varied among species. Our results highlighted that spatial and temporal variability in climate, phenology, and population measures may limit the utility of climate averages-based bioclimatic niche models for informing wildlife management actions, and may suggest priorities for sustained data collection and continued analysis.

Cryptococcus neoformans is a fungal pathogen causing life-threatening meningoencephalitis in susceptible individuals. Fungal vaccine development has been hampered by the fact that cryptococcosis occurs during immunodeficiency. We previously reported that a C. neoformans mutant (Δsgl1) accumulating sterylglucosides (SGs) is avirulent and provides complete protection to WT challenge, even under CD4+ T cell depletion, an immunodeficient condition commonly associated with cryptococcosis. We found high levels of SGs in the lungs post-immunization with Δsgl1 that decreased upon fungal clearance. Th1 cytokines increased whereas Th2 cytokines concurrently decreased, coinciding with a large recruitment of leukocytes to the lungs. Depletion of B or CD8+ T cells did not affect either Δsgl1 clearance or protection from WT challenge. Although CD4+ T cell depletion affected clearance, mice were still protected indicating that clearance of the mutant was not necessary for host protection. Protection was lost only when both CD4+ and CD8+ T cells were depleted, highlighting a previously unexplored role of fungal-derived SGs as an immunoadjuvant for host protection against cryptococcosis.

Qualitative thematic analysis is a commonly used and widely applicable form of qualitative analysis, though it can be challenging to implement. Due to its use across research questions, qualitative traditions, and fields, thematic analysis is also prevalent in mixed methods studies. Despite its widespread use, the term "thematic analysis" is often confused and not always well described in empirical studies. In addition, there is limited guidance on how to use thematic analysis within the context of mixed methods research to ensure rigorous study design and mixed methods integration. Therefore, the purpose of this paper is to describe the definitions, use, and variations of thematic analysis and explore how integration and mixed methods validity may be impacted in mixed methods studies.

The molecular mechanisms of mitochondrial dysfunction in the premature ageing Werner syndrome were elusive. Here the authors show that NAD+ depletion-induced impaired mitophagy contributes to this phenomenon, shedding light on potential therapeutics.

We propose a set of Bell-type nonlocal games that can be used to prove an unconditional quantum advantage in an objective and hardware-agnostic manner. In these games, the circuit depth needed to prepare a cyclic cluster state and measure a subset of its Pauli stabilizers on a quantum computer is compared to that of classical Boolean circuits with the same, nearest-neighboring gate connectivity. Using a circuit-based trapped-ion quantum computer, we prepare and measure a six-qubit cyclic cluster state with an overall fidelity of 60.6% and 66.4%, before and after correcting for measurement-readout errors, respectively. Our experimental results indicate that while this fidelity readily passes conventional (or depth-0) Bell bounds for local hidden-variable models, it is on the cusp of demonstrating a higher probability of success than what is possible by depth-1 classical circuits. Our games offer a practical and scalable set of quantitative benchmarks for quantum computers in the pre-fault-tolerant regime as the number of qubits available increases.

Background: Lateral interbody fusion (LIF) is traditionally performed with the patient in lateral decubitus, requiring repositioning to prone for adjunctive posterior procedures, or modifying traditional posterior techniques to be done while positioned lateral. The benefits of lateral anterior column access may be achievable with the patient prone, allowing for concomitant posterior techniques in a more familiar single-position setting. Methods: Prone transpsoas (PTP) access was outlined and vetted by a group of LIF-experienced spine surgeons. Early clinical experience included prospectively capturing procedural details and perioperative outcomes across a multi-centre cohort of clinicians to assess feasibility and to identify efficiencies and/or challenges. Results: Perioperative data was prospectively collected from 120 consecutive cases (176 levels) from 22 surgeons. Lateral exposure was achieved in an average 18 min/level, guided by triggered EMG; and retraction time averaged 25 min/level, with continued plexus monitoring via saphenous SSEP. Fixation was via percutaneous pedicle screws (65%), open pedicle screws (24%), other (11%). No re-positioning was required. Concomitant procedures facilitated by prone position included direct decompression (37%), treatment at L5-S1 (18%), posterior instrumentation revision (7%), and osteotomy/bony releases (9%). PTP procedure time, blood loss, and length of stay were consistent with established LIF experience. Challenges included patient movement with lateral instrument forces, retractor sag, stability of access relative to the patient, and surgeon ergonomics of the working channel. These challenges were overcome later in the experience through development of a specialized positioner and retractor system specific to this approach and a prescribed workflow developed by consensus of the surgeons. Conclusion: Initial multi-centre clinical experience suggests that PTP is not only feasible but creates efficiencies by allowing for single-position surgery maximizing both anterior and posterior column access and corrective techniques, with perioperative outcomes consistent with lateral decubitus experience. Learnings included the need for development of procedure-specific technologies and technique refinement.

Paracrine signaling in the tissue microenvironment is a central mediator of morphogenesis, and modeling this dynamic intercellular activity in vitro is critical to understanding normal and abnormal development. For example, Sonic Hedgehog (Shh) signaling is a conserved mechanism involved in multiple developmental processes and strongly linked to human birth defects including orofacial clefts of the lip and palate. SHH ligand produced, processed, and secreted from the epithelial ectoderm is shuttled through the extracellular matrix where it binds mesenchymal receptors, establishing a gradient of transcriptional response that drives orofacial morphogenesis. In humans, complex interactions of genetic predispositions and environmental insults acting on diverse molecular targets are thought to underlie orofacial cleft etiology. Consequently, there is a need for tractable in vitro approaches that model this complex cellular and environmental interplay and are sensitive to disruption across the multistep signaling cascade. We developed a microplate-based device that supports an epithelium directly overlaid onto an extracellular matrix-embedded mesenchyme, mimicking the basic tissue architecture of developing orofacial tissues. SHH ligand produced from the epithelium generated a gradient of SHH-driven transcription in the adjacent mesenchyme, recapitulating the gradient of pathway activity observed in vivo. Shh pathway activation was antagonized by small molecule inhibitors of epithelial secretory, extracellular matrix transport, and mesenchymal sensing targets, supporting the use of this approach in high-content chemical screening of the complete Shh pathway. Together, these findings demonstrate a novel and practical microphysiological model with broad utility for investigating epithelial-mesenchymal interactions and environmental signaling disruptions in development.

Abstract The intersection of wildlife and people on roads raises two critical issues: the barrier and mortality effects of roads on wildlife and risks to people from animal‐vehicle collisions (AVCs). Road mitigation decisions are typically made at the discretion of transportation departments that are mandated to primarily address motorist safety. Therefore, prioritization of road sections for mitigation currently focuses on identification of spatial clusters of AVCs. We sought to understand if AVC clusters align with multispecies connectivity across roads to accurately identify multipurpose mitigation hotspots. We developed a decision‐support tool based on weighted priorities for mitigation planning across 7,900 km of roads over an 84,000‐km2 area of southern Alberta, Canada. To assess alignment, we built functional connectivity models for four focal species (prairie rattlesnake, grizzly bear, mule deer, and pronghorn) and a species‐neutral structural connectivity model. We integrated AVC risk and wildlife connectivity indices into Mitigation Priority Indices that varied the weighting of individual indices. Our results demonstrated poor spatial alignment between road sections of high motorist safety risk and those of high value for wildlife connectivity. Transportation planning would benefit from integrating motorist safety risk and wildlife management needs to prioritize mitigation neighborhoods along roadways.

Type I lissencephaly is a neuronal migration disorder caused by haploinsuffiency of the PAFAH1B1 (mouse: Pafah1b1) gene and is characterized by brain malformation, developmental delays, and epilepsy. Here, we investigate the impact of Pafah1b1 mutation on the cellular migration, morphophysiology, microcircuitry, and transcriptomics of mouse hippocampal CA1 parvalbumin-containing inhibitory interneurons (PV+INTs). We find that WT PV+INTs consist of two physiological subtypes (80% fast-spiking (FS), 20% non-fast-spiking (NFS)) and four morphological subtypes. We find that cell-autonomous mutations within interneurons disrupts morphophysiological development of PV+INTs and results in the emergence of a non-canonical ‘intermediate spiking (IS)’ subset of PV+INTs. We also find that now dominant IS/NFS cells are prone to entering depolarization block, causing them to temporarily lose the ability to initiate action potentials and control network excitation, potentially promoting seizures. Finally, single-cell nuclear RNAsequencing of PV+INTs revealed several misregulated genes related to morphogenesis, cellular excitability, and synapse formation.

Cryptococcus species are environmental fungal pathogens and the causative agents of cryptococcosis. Infection occurs upon inhalation of infectious particles, which proliferate in the lung causing a primary infection. From this primary lung infection, fungal cells can eventually disseminate to other organs, particularly the brain, causing lethal meningoencephalitis. However, in most cases, the primary infection resolves with the formation of a lung granuloma. Upon severe immunodeficiency, dormant cryptococcal cells will start proliferating in the lung granuloma and eventually will disseminate to the brain. Many investigators have sought to study the protective host immune response to this pathogen in search of host parameters that keep the proliferation of cryptococcal cells under control. The majority of the work assimilates research carried out using the primary infection animal model, mainly because a reactivation model has been available only very recently. This review will focus on anti-cryptococcal immunity in both the primary and reactivation models. An understanding of the differences in host immunity between the primary and reactivation models will help to define the key host parameters that control the infections and are important for the research and development of new therapeutic and vaccine strategies against cryptococcosis.

Assessments of organisms’ vulnerability to potential climatic shifts are increasingly common. Such assessments are often conducted at the species level and focused primarily on the magnitude of anticipated climate change (i.e., climate exposure). However, wildlife management would benefit from population-level assessments that also incorporate measures of local or regional potential for organismal adaptation to change. Estimates of genetic diversity, gene flow, and landscape connectivity can address this need and complement climate exposure estimates to establish management priorities at broad to local scales. We provide an example of this holistic approach for desert bighorn sheep (Ovis canadensis nelsoni) within and surrounding lands administered by the U.S. National Park Service. We used genetic and environmental data from 62 populations across the southwestern U.S. to delineate genetic structure, evaluate relationships between genetic diversity and isolation, and estimate relative climate vulnerability for populations as a function of five variables associated with species’ responses to climate change: genetic diversity, genetic isolation, geographic isolation, forward climate velocity within a population’s habitat patch (a measure of geographic movement rate required for an organism to maintain constant climate conditions), and maximum elevation within the habitat patch (a measure of current climate stress, as lower maximum elevation is associated with higher temperature, lower precipitation, and lower population persistence). Genetic structure analyses revealed a high-level division between populations in southeastern Utah and populations in the remainder of the study area, which were further differentiated into four lower-level genetic clusters. Genetic diversity decreased with population isolation, whereas genetic differentiation increased, but these patterns were stronger for native populations than for translocated populations. Populations exhibited large variation in predicted vulnerability across the study area with respect to all variables, but native populations occupying relatively intact landscapes, such as Death Valley and Grand Canyon national parks, had the lowest overall vulnerability. These results provide local and regional context for conservation and management decisions regarding bighorn populations in a changing climate. Our study further demonstrates how assessments combining multiple factors could allow a more integrated response, such as increasing efforts to maintain connectivity and thus potential for adaptation in areas experiencing rapid climate change.

Premise High‐resolution cameras are very helpful for plant phenotyping as their images enable tasks such as target vs. background discrimination and the measurement and analysis of fine above‐ground plant attributes. However, the acquisition of high‐resolution images of plant roots is more challenging than above‐ground data collection. An effective super‐resolution (SR) algorithm is therefore needed for overcoming the resolution limitations of sensors, reducing storage space requirements, and boosting the performance of subsequent analyses. Methods We propose an SR framework for enhancing images of plant roots using convolutional neural networks. We compare three alternatives for training the SR model: (i) training with non‐plant‐root images, (ii) training with plant‐root images, and (iii) pretraining the model with non‐plant‐root images and fine‐tuning with plant‐root images. The architectures of the SR models were based on two state‐of‐the‐art deep learning approaches: a fast SR convolutional neural network and an SR generative adversarial network. Results In our experiments, we observed that the SR models improved the quality of low‐resolution images of plant roots in an unseen data set in terms of the signal‐to‐noise ratio. We used a collection of publicly available data sets to demonstrate that the SR models outperform the basic bicubic interpolation, even when trained with non‐root data sets. Discussion The incorporation of a deep learning–based SR model in the imaging process enhances the quality of low‐resolution images of plant roots. We demonstrate that SR preprocessing boosts the performance of a machine learning system trained to separate plant roots from their background. Our segmentation experiments also show that high performance on this task can be achieved independently of the signal‐to‐noise ratio. We therefore conclude that the quality of the image enhancement depends on the desired application.

When one examines life in various countries of the world regardless of type of government, health system, or even culture, one sees the great diversity of the world but also large areas of commonality among all people and all places. The twentieth century’s history is one of almost constant war, two of those wars having affected the majority of the global population, but only one event parallels today’s pandemic in impact on almost every individual on planet Earth: the great pandemic of 1918, which by various accounts took between 20 to 100 million lives over a three-year period.

Abstract The United States Forest Service promulgated new planning regulations under the National Forest Management Act in 2012 (i.e., the Planning Rule). These new regulations include the first requirements in U.S. public land management history for National Forests to evaluate, protect, and/or restore ecological connectivity as they revise their land management plans. Data and resource limitations make single‐species, functional connectivity analyses for the myriad species that occur within the 78 million ha the Forest Service manages implausible. We describe an approach that relies on freely available data and generic species, virtual species whose profile consists of ecological requirements designed to reflect the needs of a group of real species, to address the new Planning Rule requirements. We present high‐resolution connectivity estimates for 10 different generic species across a 379,000 ha study area centered on the Custer Gallatin National Forest (CGNF) in Montana and South Dakota under two different movement models. We identify locations important for connectivity for multiple species and characterize the role of the CGNF for regional connectivity. Our results informed the Plan Revision process on the CGNF and could be readily exported to other National Forests currently or planning to revise their land management plans under the new Planning Rule.

We introduce a data-driven fractional modeling framework for complex materials, and particularly bio-tissues. From multi-step relaxation experiments of distinct anatomical locations of porcine urinary bladder, we identify an anomalous relaxation character, with two power-law-like behaviors for short/long long times, and nonlinearity for strains greater than 25%. The first component of our framework is an existence study, to determine admissible fractional viscoelastic models that qualitatively describe linear relaxation. After the linear viscoelastic model is selected, the second stage adds large-strain effects to the framework through a fractional quasi-linear viscoelastic approach for the nonlinear elastic response of the bio-tissue of interest. From single-step relaxation data of the urinary bladder, a fractional Maxwell model captures both short/long-term behaviors with two fractional orders, being the most suitable model for small strains at the first stage. For the second stage, multi-step relaxation data under large strains were employed to calibrate a four-parameter fractional quasi-linear viscoelastic model, that combines a Scott-Blair relaxation function and an exponential instantaneous stress response, to describe the elastin/collagen phases of bladder rheology. Our obtained results demonstrate that the employed fractional quasi-linear model, with a single fractional order in the range α = 0.25–0.30, is suitable for the porcine urinary bladder, producing errors below 2% without need for recalibration over subsequent applied strains. We conclude that fractional models are attractive tools to capture the bladder tissue behavior under small-to-large strains and multiple time scales, therefore being potential alternatives to describe multiple stages of bladder functionality.

Abstract Corneal diseases are an extensive cause of blindness worldwide and continue to persist as a challenging public health concern. Recently, various lipid-based therapies have been advocated for the modulation of corneal diseases; however, the number of studies is still very limited. Here we focus on developments and challenges on lipid-based therapies for dry eye disease, diabetic neuropathy, and Fuchs’ endothelial corneal dystrophy. All three diseases are highly prevalent conditions and involve corneal stress and inflammation. Lipid-based therapeutics discussed includes cyclooxygenase inhibitors, essential fatty acids, and resolvin analogs. Lipids also show increasing promise as biomarkers of disease and are explored in this review.

Ca2+/calmodulin-dependent protein kinase II (CaMKII) accounts for up to 2 percent of all brain protein and is essential to memory function. CaMKII activity is known to regulate dynamic shifts in the size and signaling strength of neuronal connections, a process known as synaptic plasticity. Increasingly, computational models are used to explore synaptic plasticity and the mechanisms regulating CaMKII activity. Conventional modeling approaches may exclude biophysical detail due to the impractical number of state combinations that arise when explicitly monitoring the conformational changes, ligand binding, and phosphorylation events that occur on each of the CaMKII holoenzyme's subunits. To manage the combinatorial explosion without necessitating bias or loss in biological accuracy, we use a specialized syntax in the software MCell to create a rule-based model of a twelve-subunit CaMKII holoenzyme. Here we validate the rule-based model against previous experimental measures of CaMKII activity and investigate molecular mechanisms of CaMKII regulation. Specifically, we explore how Ca2+/CaM-binding may both stabilize CaMKII subunit activation and regulate maintenance of CaMKII autophosphorylation. Noting that Ca2+/CaM and protein phosphatases bind CaMKII at nearby or overlapping sites, we compare model scenarios in which Ca2+/CaM and protein phosphatase do or do not structurally exclude each other's binding to CaMKII. Our results suggest a functional mechanism for the so-called "CaM trapping" phenomenon, wherein Ca2+/CaM may structurally exclude phosphatase binding and thereby prolong CaMKII autophosphorylation. We conclude that structural protection of autophosphorylated CaMKII by Ca2+/CaM may be an important mechanism for regulation of synaptic plasticity.

Protein ubiquitination is a highly orchestrated process that controls diverse aspects of human biology. Dysregulation of this process can lead to various disease states including cancer, neurodegeneration, and autoimmunity. It is the correction of these dysregulated pathways, as well as the enticing ability to manipulate protein stability, that have instigated intense research into the therapeutic control of protein ubiquitination. A major bottleneck in the development and validation of small molecule modulators is the availability of a suitable high-throughput assay for enzyme activity. Herein, we present a new assay, which we term UbiReal, that uses fluorescence polarization to monitor all stages of Ub conjugation and deconjugation in real time. We use the assay to validate a chemical inhibitor of the E1 ubiquitin-activating enzyme, as well as to assess the activities and specificities of E2s, E3s, and deubiquitinases. The sensitivity and accessibility of this approach make it an excellent candidate for high-throughput screens of activity modulators, as well as a valuable tool for basic research into the mechanisms of ubiquitin regulation.

Stroke is one of the leading causes of death and long-term disability worldwide. However, effective therapeutic approaches are still limited. The disruption of blood supply triggers complicated temporal and spatial events involving hemodynamic, biochemical, and neurophysiologic changes, eventually leading to pathological disturbance and diverse clinical symptoms. Ginseng (Panax ginseng), a popular herb distributed in East Asia, has been extensively used as medicinal and nutritional supplements for a variety of disorders worldwide. In recent years, ginseng has displayed attractive beneficial effects in distinct neurological disorders including stroke, involving multiple protective mechanisms. In this article, we reviewed the literature on ginseng studies in the experimental stroke field, particularly focusing on the in vivo evidence on the preventive or therapeutic efficacy and mechanisms of ginseng and ginsenosides in various stroke models of mice and rats. We also summarized the efficacy and underlying mechanisms of ginseng and ginsenosides on short- and long-term stroke outcomes.