IL-17RA Signaling in Prx1+ Mesenchymal Cells Influences Fracture Healing in Mice.

Fracture healing is a complex series of events that requires a local inflammatory reaction to initiate the reparative process. This inflammatory reaction is important for stimulating the migration and proliferation of mesenchymal progenitor cells from the periosteum and surrounding tissues to form the cartilaginous and bony calluses. The proinflammatory cytokine interleukin (IL)-17 family has gained attention for its potential regenerative effects; however, the requirement of IL-17 signaling within mesenchymal progenitor cells for normal secondary fracture healing remains unknown. The conditional knockout of IL-17 receptor a (Il17ra) in mesenchymal progenitor cells was achieved by crossing Il17raF/F mice with Prx1-cre mice to generate Prx1-cre; Il17raF/F mice. At 3 months of age, mice underwent experimental unilateral mid-diaphyseal femoral fractures and healing was assessed by micro-computed tomography (µCT) and histomorphometric analyses. The effects of IL-17RA signaling on the osteogenic differentiation of fracture-activated periosteal cells was investigated in vitro. Examination of the intact skeleton revealed that the conditional knockout of Il17ra decreased the femoral cortical porosity but did not affect any femoral trabecular microarchitectural indices. After unilateral femoral fractures, Il17ra conditional knockout impacted the cartilage and bone composition of the fracture callus that was most evident early in the healing process (day 7 and 14 post-fracture). Furthermore, the in vitro treatment of fracture-activated periosteal cells with IL-17A inhibited osteogenesis. This study suggests that IL-17RA signaling within Prx1+ mesenchymal progenitor cells can influence the early stages of endochondral ossification during fracture healing.

Enhancing reniform nematode management in sweetpotato by complementing host-plant resistance with non-fumigant nematicides.

The reniform nematode (Rotylenchulus reniformis Linford and Oliveira) adversely impacts the quality and quantity of sweetpotato storage roots. Management of R. reniformis in sweetpotato remains a challenge because host plant resistance is not available, fumigants are detrimental to the environment and health, and crop rotation is not effective. We screened a core set of 24 sweetpotato plant introductions (PIs) against R. reniformis. Four PIs were resistant and 10 were moderately resistant to R. reniformis suggesting these PIs can serve as sources of resistance for sweetpotato resistance breeding programs. PI 595869, PI 153907, and PI 599386 suppressed 83% to 89% egg production relative to the susceptible control 'Beauregard', and these PIs were employed in subsequent experiments to determine if their efficacy against R. reniformis can be further increased by applying non-fumigant nematicides oxamyl, fluopyram and fluensulfone. A 34% to 93% suppression of nematode reproduction was achieved by the application of non-fumigant nematicides, with oxamyl providing the best suppression followed by fluopyram and fluensulfone. Although sweetpotato cultivars resistant to R. reniformis are currently not available and there is a need for the development of safer yet highly effective non-fumigant nematicides, results from the current study suggest that complementing host plant resistance with non-fumigant nematicides can serve as an important tool for effective and sustainable nematode management.

PFAS Electroanalysis in Low-Oxygen River Water Using Electrogenerated Dioxygen.

Per- and polyfluoroalkyl substances (PFAS), nicknamed "forever chemicals" due to the strength of their carbon-fluorine bonds, are a class of potent micropollutants that cause deleterious health effects in mammals. The current state-of-the-art detection method requires the collection and transport of water samples to a centralized facility where chromatography and mass spectrometry are performed for the separation, identification, and quantification of PFAS. However, for efficient remediation efforts to be properly informed, a more rapid in-field testing method is required. We previously demonstrated the development and use of dioxygen as the mediator molecule. The use of dioxygen is predicated on the assumption that there will be consistent ambient dioxygen levels in natural waters. This is not always the case in hypoxic groundwater and at high altitudes. To overcome this challenge and further advance the strategies that will enable in-field electroanalysis of PFAS, we demonstrate, as a proof of concept, that dioxygen can be generated in solution through the hydrolysis of water. The electrogenerated dioxygen can then be used as a mediator molecule for the indirect detection of PFOS via molecularly imprinted polymer (MIP)-based electroanalysis. We demonstrate that calibration curves can be constructed with high precision and sensitivity (LOD < 1 ppt or 1 ng/L). Our results provide a foundation for enabling in-field hypoxic PFAS electroanalysis.

Taking embodiment seriously in public policy and practice: adopting a procedural approach to health and welfare.

It is a common refrain amongst phenomenologists, disability theorists, and feminist legal theorists that medical practice pays insufficient attention to people's embodiment. The complaint that we take insufficient account of people's embodiment isn't limited to the clinical interaction. It has also been directed at healthcare regulation and welfare policy. In this paper, I examine the arguments for taking embodiment seriously in both medical practice and welfare policy, concluding we have good reasons to take better account of people's embodiment. I then set out two challenges to taking embodiment seriously in public policy. First, given the amount of variation in how people are embodied, there is strong possibility that adjusting policy to benefit particular individuals based on an appreciation of their embodied experiences could be detrimental towards other individuals. The second challenge concerns how to ensure that people's testimony about their first-person embodied experience is subject to adequate scrutiny without this resulting in epistemic injustice. I argue that the solution to both of these challenges is to devise a just procedure for soliciting people's testimony and taking it into account in the policy development process. As such, I also provide an outline of what a just procedure should look like.

Systemic inflammatory and gut microbiota responses to fracture in young and middle-aged mice.

Age is a patient-specific factor that can significantly delay fracture healing and exacerbate systemic sequelae during convalescence. The basis for this difference in healing rates is not well-understood, but heightened inflammation has been suggested to be a significant contributor. In this study, we investigated the systemic cytokine and intestinal microbiome response to closed femur fracture in 3-month-old (young adult) and 15-month-old (middle-aged) female wild-type mice. Middle-aged mice had a serum cytokine profile that was distinct from young mice at days 10, 14, and 18 post-fracture. This was characterized by increased concentrations of IL-17a, IL-10, IL-6, MCP-1, EPO, and TNFα. We also observed changes in the community structure of the gut microbiota in both young and middle-aged mice that was evident as early as day 3 post-fracture. This included an Enterobacteriaceae bloom at day 3 post-fracture in middle-aged mice and an increase in the relative abundance of the Muribaculum genus. Moreover, we observed an increase in the relative abundance of the health-promoting Bifidobacterium genus in young mice after fracture that did not occur in middle-aged mice. There were significant correlations between serum cytokines and specific genera, including a negative correlation between Bifidobacterium and the highly induced cytokine IL-17a. Our study demonstrates that aging exacerbates the inflammatory response to fracture leading to high levels of pro-inflammatory cytokines and disruption of the intestinal microbiota.

Impact of non-fumigant nematicides on reproduction and pathogenicity of Meloidogyne enterolobii and disease severity in tobacco.

Meloidogyne enterolobii is a highly aggressive quarantine pathogen which threatens the multibillion-dollar tobacco industry and is not manageable with the currently available management methods in tobacco. There is currently no known host plant resistance in tobacco and previous studies have shown that the lower level of the currently recommended rate of non-fumigant nematicides does not provide satisfactory management of M. enterolobii. The current study was conducted with the hypothesis that M. enterolobii can be better managed using a single soil application of the maximum allowed rate of non-fumigant nematicides. Treatments involved three non-fumigant chemical nematicides (oxamyl, fluopyram, and fluensulfone), a biological nematicide derived from Burkholderia, and a non-treated control. Fluensulfone significantly suppressed the nematode reproduction relative to the control, the suppression being 71% for eggs and 86% for the second stage juveniles (J2). Fluopyram also suppressed nematode reproduction, although this was statistically insignificant, with the suppression being 26% and 37% for eggs and J2, respectively. Oxamyl significantly suppressed J2 (80%), but not eggs (50%) in relation to the control. The most significant reduction of disease severity was achieved by the application of fluensulfone (64%), followed by oxamyl (54%) and fluopyram (48%). Except for fluensulfone, which significantly reduced the root biomass, none of the nematicides significantly impacted root and shoot biomass. The biological nematicide did not significantly affect nematode reproduction, pathogenicity, or disease severity. The results from the current study suggest that while the non-fumigant nematicides provided a good level of the nematode suppression, more research is needed to improve the efficacy of non-fumigant nematicides through employing better application methods or finding better chemistries.

Retinoic Acid-Related Orphan Receptor α Is Required for Generation of Th2 Cells in Type 2 Pulmonary Inflammation.

The transcription factor retinoic acid-related orphan receptor α (RORα) is important in regulating several physiological functions, such as cellular development, circadian rhythm, metabolism, and immunity. In two in vivo animal models of type 2 lung inflammation, Nippostrongylus brasiliensis infection and house dust mite (HDM) sensitization, we show a role for Rora in Th2 cellular development during pulmonary inflammation. N. brasiliensis infection and HDM challenge induced an increase in frequency of Rora-expressing GATA3+CD4 T cells in the lung. Using staggerer mice, which have a ubiquitous deletion of functional RORα, we generated bone marrow chimera mice, and we observed a delayed worm expulsion and reduced frequency in the expansion of Th2 cells and innate lymphoid type 2 cells (ILC2s) in the lungs after N. brasiliensis infection. ILC2-deficient mouse (Rorafl/flIl7raCre) also had delayed worm expulsion with associated reduced frequency of Th2 cells and ILC2s in the lungs after N. brasiliensis infection. To further define the role for Rora-expressing Th2 cells, we used a CD4-specific Rora-deficient mouse (Rorafl/flCD4Cre), with significantly reduced frequency of lung Th2 cells, but not ILC2, after N. brasiliensis infection and HDM challenge. Interestingly, despite the reduction in pulmonary Th2 cells in Rorafl/flCD4Cre mice, this did not impact the expulsion of N. brasiliensis after primary and secondary infection, or the generation of lung inflammation after HDM challenge. This study demonstrates a role for RORα in Th2 cellular development during pulmonary inflammation that could be relevant to the range of inflammatory diseases in which RORα is implicated.

Dietary phosphorus consumption alters T cell populations, cytokine production, and bone volume in mice.

The intake of dietary phosphate far exceeds recommended levels; however, the long-term health consequences remain relatively unknown. Here, the chronic physiological response to sustained elevated and reduced dietary phosphate consumption was investigated in mice. Although serum phosphate levels were brought into homeostatic balance, the prolonged intake of a high-phosphate diet dramatically and negatively impacted bone volume; generated a sustained increase in the phosphate responsive circulating factors FGF23, PTH, osteopontin and osteocalcin; and produced a chronic low-grade inflammatory state in the BM, marked by increased numbers of T cells expressing IL-17a, RANKL, and TNF-α. In contrast, a low-phosphate diet preserved trabecular bone while increasing cortical bone volume over time, and it reduced inflammatory T cell populations. Cell-based studies identified a direct response of T cells to elevated extracellular phosphate. Neutralizing antibodies against proosteoclastic cytokines RANKL, TNF-α, and IL-17a blunted the high-phosphate diet-induced bone loss identifying bone resorption as a regulatory mechanism. Collectively, this study illuminates that habitual consumption of a high-phosphate diet in mice induces chronic inflammation in bone, even in the absence of elevated serum phosphate. Furthermore, the study supports the concept that a reduced phosphate diet may be a simple yet effective strategy to reduce inflammation and improve bone health during aging.

Bifidobacterium longum supplementation improves age-related delays in fracture repair.

Age-related delays in bone repair remains an important clinical issue that can prolong pain and suffering. It is now well established that inflammation increases with aging and that this exacerbated inflammatory response can influence skeletal regeneration. Recently, simple dietary supplementation with beneficial probiotic bacteria has been shown to influence fracture repair in young mice. However, the contribution of the gut microbiota to age-related impairments in fracture healing remains unknown. Here, we sought to determine whether supplementation with a single beneficial probiotic species, Bifidobacterium longum (B. longum), would promote fracture repair in aged (18-month-old) female mice. We found that B. longum supplementation accelerated bony callus formation which improved mechanical properties of the fractured limb. We attribute these pro-regenerative effects of B. longum to preservation of intestinal barrier, dampened systemic inflammation, and maintenance of the microbiota community structure. Moreover, B. longum attenuated many of the fracture-induced systemic pathologies. Our study provides evidence that targeting the gut microbiota using simple dietary approaches can improve fracture healing outcomes and minimize systemic pathologies in the context of aging.

Topdressing Biochar Compost Mixtures and Biological Control Organism Applications Suppress Foliar Pathogens in Creeping Bentgrass Fairway Turf.

Biochar, compost, and biological control agents can suppress pathogens on their own; however, their reliability and efficacy are not as acceptable as synthetic fungicides commonly used to suppress pathogens. A multiyear field study was initiated to evaluate combinations of monthly applications of a biochar compost mixture and weekly or biweekly Bacillus subtilis QST713 applications for their ability to suppress foliar pathogens on a creeping bentgrass (Agrostis stolonifera L.) fairway and to measure their impact on strain QST713 establishment. Disease severity and turfgrass quality were measured every 14 days throughout the growing season. Populations of strain QST713 were quantified by quantitative PCR analysis on DNA extracted from foliage samples collected throughout the trial. Biochar compost mixture applications increased turfgrass quality in both years of the study and reduced dollar spot (Clarireedia jacksonii Salgado-Salazar) severity in 2021. Weekly strain QST713 applications reduced copper spot (Gloeocercospora sorghi D. C. Bain & Edgerton) severity compared with biweekly applications and the nontreated control in 2020, yet monthly biochar compost mixture with weekly strain QST713 applications completely suppressed copper spot in 2021. Populations of strain QST713 were highest in weekly treated plots, and monthly biochar compost mixture applications did not affect strain QST713 establishment. Although there was not an interaction between biochar compost mixture and strain QST713 applications, implementing both in a season-long program will benefit turfgrass health and reduce disease severity.

Sexually Dimorphic Increases in Bone Mass Following Tissue-specific Overexpression of Runx1 in Osteoclast Precursors.

Many metabolic bone diseases arise as a result excessive osteoclastic bone resorption, which has motivated efforts to identify new molecular targets that can inhibit the formation or activity of these bone-resorbing cells. Mounting evidence indicates that the transcription factor Runx1 acts as a transcriptional repressor of osteoclast formation. Prior studies using a conditional knockout approach suggested that Runx1 in osteoclast precursors acts as an inhibitor of osteoclastogenesis; however, the effects of upregulation of Runx1 on osteoclast formation remain unknown. In this study, we investigated the skeletal effects of conditional overexpression of Runx1 in preosteoclasts by crossing novel Runx1 gain-of-function mice (Rosa26-LSL-Runx1) with LysM-Cre transgenic mice. We observed a sex-dependent effect whereby overexpression of Runx1 in female mice increased trabecular bone microarchitectural indices and improved torsion biomechanical properties. These effects were likely mediated by delayed osteoclastogenesis and decreased bone resorption. Transcriptomics analyses during osteoclastogenesis revealed a distinct transcriptomic profile in the Runx1-overexpressing cells, with enrichment of genes related to redox signaling, apoptosis, osteoclast differentiation, and bone remodeling. These data further confirm the antiosteoclastogenic activities of Runx1 and provide new insight into the molecular targets that may mediate these effects.

The Utility of the Trauma Symptom Inventory as a Primary and Secondary Assessment Instrument for Forensic Practice in Legal Settings.

This paper examines the utility of the Trauma Symptom Inventory-2 (TSI-2) and its predecessor, the Trauma Symptom Inventory (TSI) in forensic psychology practice. The instrument's psychometric properties, use with special populations, legal case review and admissibility considerations are discussed. Recommendations regarding the strengths and limitations of the TSI/TSI-2 are suggested for forensic practitioners and lawyers. Considerations related to potential expert witness cross-examination are also presented. Psychological research and legal review suggest that the TSI/TSI-2 is admissible as an instrument under the Daubert Standard, especially as related to civil court disability claims. Still, lingering issues with the ATR validity scale remain and there is limited independent research establishing the predictive and discriminant validity of the TSI-2 across diverse forensic samples. In summary, this suggests the instrument is most effective as part of a comprehensive assessment battery for identifying PTSD symptomology within legal proceedings where a trauma diagnosis is relevant.

Deletion of IL-17ra in osteoclast precursors increases bone mass by decreasing osteoclast precursor abundance.

Metabolic bone diseases, such as osteoporosis, typically reflect an increase in the number and activity of bone-resorbing osteoclasts that result in a loss of bone mass. Inflammatory mediators have been identified as drivers of both osteoclast formation and activity. The IL-17 family of inflammatory cytokines has gained attention as important contributors to both bone formation and resorption. The majority of IL-17 cytokines signal through receptor complexes containing IL-17a receptor (IL-17ra); however, the role of IL-17ra signaling in osteoclasts remains elusive. In this study, we conditionally deleted Il17ra in osteoclast precursors using LysM-Cre and evaluated the phenotypes of skeletally mature male and female conditional knockout and control mice. The conditional knockout mice displayed an increase in trabecular bone microarchitecture in both the appendicular and axial skeleton. Assessment of osteoclast formation in vitro revealed that deletion of Il17ra decreased osteoclast number, which was confirmed in vivo using histomorphometry. This phenotype was likely driven by a lower abundance of osteoclast precursors in IL-17ra conditional knockout mice. This study suggests that IL-17ra signaling in preosteoclasts can contribute to osteoclast formation and subsequent bone loss.

How to Regulate the Right to Self-Medicate.

In Pharmaceutical Freedom Professor Flanigan argues we ought to grant people self-medication rights for the same reasons we respect people's right to give (or refuse to give) informed consent to treatment. Despite being the most comprehensive argument in favour of self-medication written to date, Flanigan's Pharmaceutical Freedom leaves a number of questions unanswered, making it unclear how the safe-guards Flanigan incorporates to protect people from harming themselves would work in practice. In this paper, I extend Professor Flanigan's account by discussing a hypothetical case to illustrate how these safe-guards could work together to protect people from harms caused by their own ignorance or incompetence.

User and Healthcare Professional Perspectives on Do-It-Yourself Artificial Pancreas Systems: A Need for Guidelines.

A growing number of individuals with type 1 diabetes are choosing to use "do-it-yourself" artificial pancreas systems (DIY APS) to support their diabetes self-management. Observational and self-report data of glycemic benefits of DIY APS are promising; however, without rigorous clinical trials or regulation from governing bodies, liability and user safety continue to be central concerns for stakeholders. Despite DIY APS having been used for several years now, there are no guidelines to assist users and healthcare professionals in addressing DIY APS use in routine clinical care. This commentary reports key stakeholders' perspectives presented at the annual Advanced Technologies and Treatments in Diabetes conference in February 2020. Important considerations to inform the development of clinical care guidelines are also presented to generate further debate.

Investigating Chemical and Biological Control Applications for Pythium Root Rot Prevention and Impacts on Creeping Bentgrass Putting Green Rhizosphere Bacterial Communities.

Pythium root rot (PRR) is a disease that can rapidly devastate large swaths of golf course putting greens, with little recourse once symptoms appear. Golf courses routinely apply preventive fungicides for root diseases, which may alter the rhizosphere microbiome, leading to unintended effects on plant health. A multiyear field trial was initiated on a 'T-1' creeping bentgrass (Agrostis stolonifera L. cultivar T-1) putting green in College Park, Maryland to evaluate preventive PRR management for disease suppression and effects on rhizosphere bacterial communities. Fungicides commonly used to prevent PRR and a biological fungicide were repeatedly applied to experimental plots throughout the growing season. Rhizosphere samples were collected twice annually from each plot for evaluation of rhizosphere bacterial communities through amplicon sequencing and monitoring of biological control organism populations via quantitative PCR. Cyazofamid was the only treatment to suppress PRR in both years compared with the control. Fosetyl-Al on a 14-day interval and Bacillus subtilis QST713 also reduced PRR severity in 2019 compared with the nontreated control. Treatments did not significantly affect bacterial diversity or relative abundances of bacterial classes; however, seasonal environmental changes did. Repeated rhizosphere-targeted applications of B. subtilis QST713 appear to have established the bacterium into the rhizosphere, as populations increased between samples, even after applications stopped. These findings suggest that QST713 may reduce pathogen pressure when repeatedly applied and can reduce fungicide usage during periods of low PRR pressure.

Prescribing unapproved medical devices? The case of DIY artificial pancreas systems.

In response to slow progress regarding technological innovations to manage type 1 diabetes, some patients have created unregulated do-it-yourself artificial pancreas systems (DIY APS). Yet both in the United Kingdom (UK) and internationally, there is an almost complete lack of specific guidance - legal, regulatory, or ethical - for clinicians caring for DIY APS users. Uncertainty regarding their professional obligations has led to them being cautious about discussing DIY APS with patients, let alone recommending or prescribing them. In this article, we argue that this approach threatens to undermine trust and transparency. Analysing the professional guidance from the UK regulator - the General Medical Council - we demonstrate that nothing within it ought to be interpreted as precluding clinicians from initiating discussions about DIY APS. Moreover, in some circumstances, it may require that clinicians do so. We also argue that the guidance does not preclude clinicians from prescribing such unapproved medical devices.