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Abstract Pericytes function to maintain tissue homeostasis by regulating capillary blood flow and maintaining endothelial barrier function. Pericyte dysfunction is associated with various pathologies and has recently been found to aid cancer progression. Despite having critical functions in health and disease, pericytes remain an understudied population due to a lack of model systems which accurately reflect in vivo biology. In this study we developed a protocol to isolate and culture murine lung, brain, bone, and liver pericytes, that maintains their known phenotypes and functions. We demonstrate that pericytes, being inherently plastic, benefit from controlled oxygen tension culture conditions, aiding their expansion ex vivo. Primary pericytes grown in physiologically relevant oxygen tensions (10% O2 for lung; 5% O2 for brain, bone, and liver) also better retain pericyte phenotypes indicated by stable expression of characteristic transcriptional and protein markers. In functional tube formation assays, pericytes were observed to significantly associate with endothelial junctions. Importantly, we identified growth conditions that limit expression of the plasticity factor Klf4 to prevent spontaneous phenotypic switching in vitro. Additionally, we were able to induce pathological pericyte phenotypic switching in response to metastatic stimuli to accurately recapitulate in vivo biology. Here, we present a robust method for studying pericyte biology in both physiology and disease.

Abstract Background This project aimed to design and evaluate the potential to integrate an exercise oncology service into clinical care in a local healthcare system. The goal was to inform the design of an implementation strategy to promote its sustainable use in standard care. Methods This two-phase, exploratory study used a mixed-methods approach. First, qualitative measures were used to understand the context for exercise integration into oncology care by clinicians and administrators in the healthcare system. Next, the Reach, Effectiveness, Adoption, Implementation, Maintenance (RE-AIM) framework guided a comprehensive evaluation of the exercise service. Reach and Adoption were measured via number of patients enrolled in the program relative to those treated. Adoption was measured via referrals from stakeholders. Implementation was described using the program structure and delivery. Maintenance was described using hospital funding decisions. Effectiveness was assessed using the Bellarmine Norton Assessment Tool (BNAT) and Functional Assessment of Cancer Therapy – Fatigue Scale. Results Phase 1 context analysis suggested critical elements to guide program delivery, including limiting participants to post-primary treatment (surgery, chemotherapy and radiation) and streamlining referral process. The Phase 2 evaluation demonstrated suboptimal program reach (2%); significant program effectiveness (improved physical function (BNAT; p = 0.05 ) and decreased fatigue (p

The paper integrates well-established methods from other spheres of flood risk assessment in a novel way to explore the currently poorly understood Safe Development Paradox (SDP) phenomenon in coastal settings. The study contributes to addressing this knowledge gap based on insights from contrasting UK case studies: Portsmouth, Weston-super-Mare, and Southport. Differential analysis of historic LiDAR Digital Surface Models (DSMs) was used to identify temporal changes in the urban landscape to create a DSM of Difference (DoD), representing elevation change between two locations over time. Geostatistical testing, specifically t-tests, were then used to infer statistical significance of changes in urban development. The findings reveal a consistent pattern: following completion or improvement of large-scale structural coastal flood defences, there is subsequent, and statistically significant, increases in urban development within/near flood-exposed areas across all three case studies, contrary to the limited flood-exposed development in neighbouring settlements, with no comparable defences constructed during the same period. On average, new urban development occurs approximately 2 years after the completion of coastal flood defence projects. The study emphasises the importance of context-specific and neighbourhood analyses, considering settlement size, economy, and extraneous factors influencing the nature of development to better understand the SDP’s long-term implications.

Non-invasive behavioural indicators of welfare can be particularly useful for managing captive breeding populations of endangered species; these allow individual welfare to be monitored and reproductive success maximised without the need for capture and restraint methods. However, most studies focus on the behaviours whose presence or frequency can predict welfare issues; the absence of a behaviour is less frequently considered an indicator of welfare. Here, we investigate potential behavioural correlates with welfare-related health states in captive Livingstone’s fruit bats (Pteropus livingstonii), a critically endangered species that can become obese due to restricted space and reduced activity rates compared with wild populations. In this study, behavioural data were collected on males (which are particularly prone to obesity). Hurdle models were used to separately determine the factors predicting the presence or absence of behaviour and the frequency of observed behaviours. Whilst significantly lower levels of vigilance were observed in males with a larger body mass, those with diagnosed health issues were significantly more likely to show an absence of locomotion and foraging behaviour. Males with a lower body mass were also more likely to show an absence of foraging behaviour. Our study demonstrates how the absence of a behaviour can be informative as to an individual’s welfare state. This study has identified behavioural profiles that can be used to flag at-risk individuals, reducing the need for potentially stressful handling and improving our ability to safeguard the welfare of individuals within a large captive group.

Proton therapy presents an interesting and important challenge to accelerator physics. Practical restrictions such as room size, gantry weight, and cost have led to a demand for more compact and affordable devices. Success would benefit a significant number of patients every year. Dielectric wall accelerators could provide this breakthrough, but there is limited work available regarding their feasibility or possible integration into medical devices, and no solutions exist for longitudinal beam control. In this work, we discuss the development of a linear beam dynamics model for dielectric wall accelerators. The model is implemented as part of the open-source code transoptr. It is tested against the rf LINAC subroutine in transoptr and is used to study bunch injection and transport. Building on recent work, the effect of time-varying fields on beam dynamics is also investigated. Conditions for limiting transverse beam size and maintaining longitudinal stability are discussed, and considerations are given to accelerator and beamline design.

ABSTRACT Haemophilus parainfluenzae (Hp) is a Gram-negative, highly prevalent, and abundant commensal in the human oral cavity, and an infrequent extraoral opportunistic pathogen. Hp occupies multiple niches in the oral cavity, including the supragingival plaque biofilm. Little is known about how Hp interacts with its neighbors in healthy biofilms nor its mechanisms of pathogenesis as an opportunistic pathogen. To address this, we identified the essential genome and conditionally essential genes in in vitro biofilms aerobically and anaerobically. Using transposon insertion sequencing (TnSeq) with a highly saturated mariner transposon library in two strains, the ATCC33392 type-strain (Hp 392) and oral isolate EL1 (Hp EL1), we show that the essential genomes of Hp 392 and Hp EL1 are composed of 395 (20%) and 384 (19%) genes, respectively. The core essential genome, consisting of 341 (17%) essential genes conserved between both strains, was composed of genes associated with genetic information processing, carbohydrate, protein, and energy metabolism. We also identified conditionally essential genes for aerobic and anaerobic biofilm growth, which were associated with carbohydrate and energy metabolism in both strains. RNAseq analysis determined that most genes upregulated during anaerobic growth are not essential for Hp 392 anaerobic survival. The completion of this library and analysis under these conditions gives us a foundational insight into the basic biology of H. parainfluenzae in differing oxygen conditions, similar to its in vivo habitat. This library presents a valuable tool for investigation into conditionally essential genes for an organism that lives in close contact with many microbial species in the human oral habitat.IMPORTANCEHaemophilus parainfluenzae is a highly abundant human commensal microbe, present in most healthy individuals where it colonizes the mouth. H. parainfluenzae correlates with good oral health and may play a role in preservation of healthy host status. Also, H. parainfluenzae can cause opportunistic infections outside of the oral cavity. To date, little is known about how H. parainfluenzae colonizes the human host, despite being such a frequent and abundant part of our human microbiome. Here, we demonstrate the creation and use of a powerful tool, a TnSeq library, used to identify genes necessary for both the outright growth of this organism and also genes conditionally essential for growth in varying oxygen status which it can encounter in the human host. This tool and these data serve as a foundation for further study of this relatively unknown organism that may play a role in preserving human health.

Abstract Analysis of single cancer cells is critical to obtain accurate patient diagnosis and prognosis. In this work, we report the selective dielectrophoretic capture and electrochemical analysis of single melanoma cells at an array of interlocked spiral bipolar electrodes (iBPEs). Following dielectrophoretic capture, individual melanoma cells are hydrodynamically transferred into picoliter‐scale chambers for subsequent analysis. The interlocked spiral end of the iBPE (the sensing pole) is utilized to read out an electrochemical enzyme‐linked immunosorbent assay (eELISA), which quantifies the expression of a cell surface antigen, melanoma cell adhesion marker (MCAM). The opposite pole of each BPE is located in a fluidically isolated compartment containing reagents for electrogenerated chemiluminescence (ECL), such that luminescence reports iBPE current. In a preliminary device design, the ECL intensity was insufficient to detect MCAM expression on single cells. To achieve single‐cell analysis, we decreased the gap size between the interlocked spirals tenfold (5.0 μm to 0.5 μm), thereby creating a more sensitive biosensor by enhanced redox cycling of the product of eELISA. This work is significant because it allows for the selective isolation and sensitive analysis of individual melanoma cells in a device amenable to point‐of‐care (POC) application by combining dielectrophoresis (DEP) with interdigitated bipolar electrodes (IDBPEs).

IntroductionChildhood undernutrition is associated with increased morbidity, mortality and a high socio-economic burden.MethodsSupporting Pediatric GRowth and Health OUTcomes (SPROUT) is a randomized, controlled trial evaluating the effects of an oral nutritional supplement (ONS) with dietary counseling (DC; n = 164) compared to a DC-only group who continued consuming their habitual milk (n = 166; NCT05239208). Children aged 24–60 months who were at risk or with undernutrition, as defined by weight-for-age [WAZ]

Arctic marine mammals face many challenges linked to climate change, including increasing anthropogenic noise from vessel traffic. The bowhead whale (Balaena mysticetus Linnaeus, 1758), an Arctic endemic cetacean, relies on acoustic communication, with documented overlapping frequencies between communication and vessel noise. Bering–Chukchi–Beaufort (BCB) bowhead whales migrate through areas with the highest levels of vessel traffic in the Pacific Arctic. Here, we document the spatial and temporal overlap between 25 satellite-tagged BCB bowhead whales and vessels during July–December, 2012–2018. We report 1332 occasions when a vessel was within 125 km of a tagged whale, and where possible, quantified changes in swim speed to investigate individual behavioural responses to vessel approaches within a 50 km radius (n = 18 encounters). In the quantitative analysis, bowhead whales were not observed to alter swim speed within 8–50 km of vessels (we could not assess distances

Summary: Tight regulation of macrophage immune gene expression is required to fight infection without risking harmful inflammation. The contribution of RNA-binding proteins (RBPs) to shaping the macrophage response to pathogens remains poorly understood. Transcriptomic analysis reveals that a member of the serine/arginine-rich (SR) family of mRNA processing factors, SRSF7, is required for optimal expression of a cohort of interferon-stimulated genes in macrophages. Using genetic and biochemical assays, we discover that in addition to its canonical role in regulating alternative splicing, SRSF7 drives transcription of interferon regulatory transcription factor 7 (IRF7) to promote antiviral immunity. At the Irf7 promoter, SRSF7 maximizes STAT1 transcription factor binding and RNA polymerase II elongation via cooperation with the H4K20me1 histone methyltransferase KMT5a (SET8). These studies define a role for an SR protein in activating transcription and reveal an RBP-chromatin network that orchestrates macrophage antiviral gene expression.

Abstract We report a method for dielectrophoretic capture of melanoma cells and their electrochemical analysis for tumor‐specific markers, using a microfluidic device composed of microwells overlying an array of interdigitated bipolar electrodes (IDBPEs). In this device, cells are captured on each IDBPE by dielectrophoresis, and microfabricated wells allow for retention of the cells to enable their subsequent electrochemical analysis. This advancement is significant because it addresses a need for cancer diagnostics with few‐ to single‐cell resolution amenable to use in low‐resource settings. Specifically, this approach combines the selectivity of dielectrophoresis for cell phenotype, the low cost of electrochemical methods, and the ability of BPEs to be arrayed, with the sensitivity afforded by interdigitation. The IDBPE detects a redox‐active species produced by an enzyme‐linked antibody targeted to a cell surface antigen, and reports the current by electrochemiluminescence. In this work, we first characterize cell capture by dielectrophoresis at the IDBPE array. Second, the retention of cells in the microwells and their viability is demonstrated. Then, the sensitivity of the IDBPE for the redox‐active species is quantified. Finally, we demonstrate capture and analysis of melanoma cells at the IDBPE array, which underscores the ability of the IDBPE to achieve biologically relevant detection limits.

Abstract Introduction In the PINETREE study, early remdesivir treatment reduced risk of coronavirus disease 2019 (COVID-19)-related hospitalizations or all-cause death versus placebo by 87% by day 28 in high-risk, non-hospitalized patients. Here we report results of assessment of heterogeneity of treatment effect (HTE) of early outpatient remdesivir, focusing on time from symptom onset and number of baseline risk factors (RFs). Methods PINETREE was a double-blind, placebo-controlled trial of non-hospitalized patients with COVID-19 who were randomized within 7 days of symptom onset and had ≥ 1 RF for disease progression (age ≥ 60 years, obesity [body mass index ≥ 30], or certain coexisting medical conditions). Patients received remdesivir intravenously (200 mg on day 1 and 100 mg on days 2 and 3) or placebo. Results In this subgroup analysis, HTE of remdesivir by time from symptom onset at treatment initiation and number of baseline RFs was not detected. Treatment with remdesivir reduced COVID-19-related hospitalizations independent of stratification by time from symptom onset to randomization. Of patients enrolled ≤ 5 days from symptom onset, 1/201 (0.5%) receiving remdesivir and 9/194 (4.6%) receiving placebo were hospitalized (hazard ratio [HR] 0.10; 95% confidence interval [CI] 0.01–0.82). Of those enrolled at > 5 days from symptom onset, 1/78 (1.3%) receiving remdesivir and 6/89 (6.7%) receiving placebo were hospitalized (HR 0.19; 95% CI 0.02–1.61). Remdesivir was also effective in reducing COVID-19-related hospitalizations when stratified by number of baseline RFs for severe disease. Of patients with ≤ 2 RFs, 0/159 (0.0%) receiving remdesivir and 4/164 (2.4%) receiving placebo were hospitalized; of those with ≥ 3 RFs, 2/120 (1.7%) receiving remdesivir and 11/119 (9.2%) receiving placebo were hospitalized (HR 0.16; 95% CI 0.04–0.73). Conclusions In the outpatient setting, benefit of remdesivir initiated within 7 days of symptoms appeared to be consistent across patients with RFs. Therefore, it may be reasonable to broadly treat patients with remdesivir regardless of comorbidities. Trial Registration ClinicalTrials.gov number NCT04501952.

During their quest for growth, adaptation, and survival, cancer cells create a favorable environment through the manipulation of normal cellular mechanisms. They increase anabolic processes, including protein synthesis, to facilitate uncontrolled proliferation and deplete the tumor microenvironment of resources. As a dynamic adaptation to the self-imposed oncogenic stress, cancer cells promptly hijack translational control to alter gene expression. Rewiring the cellular proteome shifts the phenotypic balance between growth and adaptation to promote therapeutic resistance and cancer cell survival. The integrated stress response (ISR) is a key translational program activated by oncogenic stress that is utilized to fine-tune protein synthesis and adjust to environmental barriers. Here, we focus on the role of ISR signaling for driving cancer progression. We highlight mechanisms of regulation for distinct mRNA translation downstream of the ISR, expand on oncogenic signaling utilizing the ISR in response to environmental stresses, and pinpoint the impact this has for cancer cell plasticity during resistance to therapy. There is an ongoing need for innovative drug targets in cancer treatment, and modulating ISR activity may provide a unique avenue for clinical benefit.

Abstract Background Systemic drug delivery to the central nervous system is limited by presence of the blood–brain barrier (BBB). Low intensity focused ultrasound (LiFUS) is a non-invasive technique to disrupt the BBB, though there is a lack of understanding of the relationship between LiFUS parameters, such as cavitation dose, time of sonication, microbubble dose, and the time course and magnitude of BBB disruption. Discrepancies in these data arise from experimentation with modified, clinically untranslatable transducers and inconsistent parameters for sonication. In this report, we characterize microbubble and cavitation doses as LiFUS variables as they pertain to the time course and size of BBB opening with a clinical Insightec FUS system. Methods Female Nu/Nu athymic mice were exposed to LiFUS using the ExAblate Neuro system (v7.4, Insightec, Haifa, Israel) following target verification with magnetic resonance imaging (MRI). Microbubble and cavitation doses ranged from 4–400 μL/kg, and 0.1–1.5 cavitation dose, respectively. The time course and magnitude of BBB opening was evaluated using fluorescent tracers, ranging in size from 105–10,000 Da, administered intravenously at different times pre- or post-LiFUS. Quantitative autoradiography and fluorescence microscopy were used to quantify tracer accumulation in brain. Results We observed a microbubble and cavitation dose dependent increase in tracer uptake within brain after LiFUS. Tracer accumulation was size dependent, with 14C-AIB (100 Da) accumulating to a greater degree than larger markers (~ 625 Da–10 kDa). Our data suggest opening of the BBB via LiFUS is time dependent and biphasic. Accumulation of solutes was highest when administered prior to LiFUS mediated disruption (2–fivefold increases), but was also significantly elevated at 6 h post treatment for both 14C-AIB and Texas Red. Conclusion The magnitude of LiFUS mediated BBB opening correlates with concentration of microbubbles, cavitation dose as well as time of tracer administration post-sonication. These data help define the window of maximal BBB opening and applicable sonication parameters on a clinically translatable and commercially available FUS system that can be used to improve passive permeability and accumulation of therapeutics targeting the brain.

Abstract Human visual systems have evolved to extract ecologically relevant information from complex scenery. In some cases, the face in the crowd visual search task demonstrates an anger superiority effect, where anger is allocated preferential attention. Across three studies (N = 419), we tested whether facial hair guides attention in visual search and influences the speed of detecting angry and happy facial expressions in large arrays of faces. In Study 1, participants were faster to search through clean-shaven crowds and detect bearded targets than to search through bearded crowds and detect clean-shaven targets. In Study 2, targets were angry and happy faces presented in neutral backgrounds. Facial hair of the target faces was also manipulated. An anger superiority effect emerged that was augmented by the presence of facial hair, which was due to the slower detection of happiness on bearded faces. In Study 3, targets were happy and angry faces presented in either bearded or clean-shaven backgrounds. Facial hair of the background faces was also systematically manipulated. A significant anger superiority effect was revealed, although this was not moderated by the target’s facial hair. Rather, the anger superiority effect was larger in clean-shaven than bearded face backgrounds. Together, results suggest that facial hair does influence detection of emotional expressions in visual search, however, rather than facilitating an anger superiority effect as a potential threat detection system, facial hair may reduce detection of happy faces within the face in the crowd paradigm.

Introduction: Transient ischaemic attack (TIA) is associated with increased risk of cognitive decline and dementia as early as one-year post-event. Regional brain atrophy measurements may predict future cognitive decline. Aims: 1) To determine whether Medial Temporal Atrophy (MTA) scores and interseptal distance (ISD) measurements are greater in patients with TIA compared to controls; and 2) To determine whether MTA and ISD predicts cognitive change one year after TIA. Methods: Baseline demographic, vascular risk factors, structural imaging and cognitive tests scores were compared between 103 Patients with TIA and 103 age-and-sex-matched controls from the Predementia Neuroimaging of Transient Ischaemic Attack (PREVENT) Study. MTA was assessed using the Schelten's Scale, and ISD was calculated as the distance between the septal nucleus of each hemisphere. Multiple linear regression models were used to evaluate how MTA and ISD related to cognitive change after adjusting for covariates. Results: Patients with TIA had larger ISD measurements (1.4 mm [SD=1.2] vs. 0.9 mm [SD=1.0]); p

Abstract Chimeric antigen receptor (CAR) T cells are a promising form of cancer immunotherapy, although they are often associated with severe toxicities. Here, we present a split-CAR design incorporating separate antigen recognition and intracellular signaling domains. These exploit the binding between the tetracycline repressor protein and a small peptide sequence (TIP) to spontaneously assemble as a functional CAR. Addition of the FDA-approved, small molecule antibiotic minocycline, acts as an “off-switch” by displacing the signaling domain and down-tuning CAR T activity. Here we describe the optimization of this split-CAR approach to generate a CAR in which cytotoxicity, cytokine secretion and proliferation can be inhibited in a dose-dependent and reversible manner. Inhibition is effective during on-going CAR T cell activation and inhibits activation and tumor control in vivo. This work shows how optimization of split-CAR structure affects function and adds a novel design allowing easy CAR inhibition through an FDA-approved small molecule.

The volcanic Intersalar Range of western Bolivia provides a unique opportunity to examine geochemical variations spanning the arc and reararc regions of the Central Andean Plateau. In this study we report 23 new 40Ar/39Ar ages, 15 whole-rock Sr-Nd-Pb isotope analyses, and 50 whole-rock major and trace element analyses from samples collected across ∼115 km of the Intersalar Range. Most samples are classified as trachyandesites and trachydacites, with the most mafic lavas (slightly alkaline, basaltic trachyandesites) erupting from the Pliocene Coracora volcano in the central Altiplano. We identify two distinct pulses of reararc magmatism: a Miocene phase between 20 Ma and 14 Ma that corresponds with local compressional shortening, and a Plio-Pleistocene phase between 5 and 1 Ma that postdates observed structural deformation in the region. 87Sr/86Sr values (0.70512–0.70600) and 143Nd/144Nd values (0.51226–0.51255) are generally higher, and lower, respectively, in the younger phase, whereas Pb isotopes (206Pb/204Pb = 17.7315–18.5095; 207Pb/204Pb = 15.5714–15.6279; 208Pb/204Pb = 37.7862–38.6156) show little variation with age. Isotope values are only loosely correlated with distance from the modern Central Volcanic Zone. Higher Sr/Y, Dy/Yb, and [La/Yb]N values in the Plio-Pleistocene samples are consistent with homogenization at the base of a thicker continental crust compared to the Miocene samples. Nb concentrations show the strongest correlation with distance into the reararc compared to all other trace elements (arc Nb = 6–16 ppm; reararc Nb = 12–26 ppm). Nb/Nb* values (a measurement of the depth of the negative Nb anomaly) correspondingly increase into the reararc (indicating smaller anomalies), reaching a maximum at Coracora volcano before decreasing in the far rear arc region. Compiled data across the Central Andean Plateau reveal a strong correlation between Nb/Nb* and the presence of intact mantle lithosphere beneath the central Altiplano. We interpret this distinct Nb signal to reflect melting triggered by the breakdown of Nb-rich hydrous minerals within foundering (delaminating) mantle lithosphere. In conjunction with spatiotemporal data, Nb systematics provide the clearest indication of mantle lithosphere in regions where mafic samples are not present.

Trophic factors are secreted proteins that can modulate neuronal integrity, structure, and function. Previous preclinical studies have shown synergistic effects on decreasing apoptosis and improving behavioral performance after stroke when combining two such trophic factors, erythropoietin (EPO) and insulin-like growth factor-1 (IGF-1). However, EPO can elevate the hematocrit level, which can be life-threatening for non-anemic individuals. A chemically engineered derivative of EPO, carbamoylated EPO (CEPO), does not impact hematological parameters but retains neurotrophic effects similar to EPO. To obtain insight into CEPO and IGF-1 combination signaling, we examined immediate early gene (IEG) expression after treatment with CEPO, IGF-1, or CEPO + IGF-1 in rat pheochromocytoma (PC-12) cells and found that combining CEPO and IGF-1 produced a synergistic increase in IEG expression. An in vivo increase in the protein expression of Npas4 and Nptx2 was also observed in the rat hippocampus. We also examined which kinase signaling pathways might be mediating these effects and found that while AKT inhibition did not alter the pattern of IEG expression, both ERK and JAK2 inhibition significantly decreased IEG expression. These results begin to define the molecular effects of combining CEPO and IGF-1 and indicate the potential for these trophic factors to produce positive, synergistic effects.

Purpose: This study assessed the biological reliability of peripheral human cytokines and adipokines, and the influence of participant characteristics on total error. This has essential application to interventional cytokine measurement to ensure that reported results are interpreted with confidence.Methods: Participants (49% female, 18–85 years, n = 84) completed two consecutive-day testing sessions. Participants provided a venous blood sample at the same time of day across two consecutive days, under standardized participant presentation, including 24-h rested and 12-h fasted conditions. Multiplex immunoassay was used to assess inflammatory analytes from samples (predominantly plasma). Repeat measurements were conducted between-day for total precision quantification, and technical (technique) error was negated from the total to provide an estimate of biological (attributed to participant presentation) error.Results: Whilst there was no evidence of statistically significant biological error, a small amount of biological error was consistently present across most analytes (∼3.3%/0.07 pg/ml), which was largest for measurement of leptin (7.3%/210 pg/ml). There was also an influence of sex on reliability of leptin and adiponectin (total model explained 6–7% of error variation), where females demonstrated the greatest error.Conclusion: Biological error reported in this study should be applied to any future study or individual with a repeated measurement of cytokine concentrations over time that maintain best practice procedures (12-h fasted, 24-h rested). In most cases, raw error should be used, with exceptions for women for measurement of leptin and adiponectin. This approach will ensure that results are reported with certainty for improved reporting of intervention efficacy.

Abstract C. difficile infection (CDI) is a worldwide healthcare problem with ~30% of cases failing primary therapy, placing a burden on healthcare systems and increasing patient morbidity. We have little understanding of why these therapies fail. Here, we use a clinically validated in vitro gut model to assess the contribution of biofilms towards recurrent disease and to investigate biofilm microbiota-C. difficile interactions. Initial experiments show that C. difficile cells became associated with the colonic biofilm microbiota and are not depleted by vancomycin or faecal microbiota transplant therapies. We observe that transferring biofilm encased C. difficile cells into a C. difficile naïve but CDI susceptible model induces CDI. Members of the biofilm community can impact C. difficile biofilm formation by acting either antagonistically or synergistically. We highlight the importance of biofilms as a reservoir for C. difficile, which can be a cause for recurrent infections.

Rock debris covers ~30% of glacier ablation areas in the Central Himalaya and modifies the impact of atmospheric conditions on mass balance. The thermal properties of supraglacial debris are diurnally variable but remain poorly constrained for monsoon-influenced glaciers over the timescale of the ablation season. We measured vertical debris profile temperatures at 12 sites on four glaciers in the Everest region with debris thickness ranging from 0.08 to 2.8 m. Typically, the length of the ice ablation season beneath supraglacial debris was 160 days (15 May to 22 October)—a month longer than the monsoon season. Debris temperature gradients were approximately linear (r2 > 0.83), measured as −40°C m–1 where debris was up to 0.1 m thick, −20°C m–1 for debris 0.1–0.5 m thick, and −4°C m–1 for debris greater than 0.5 m thick. Our results demonstrate that the influence of supraglacial debris on the temperature of the underlying ice surface, and therefore melt, is stable at a seasonal timescale and can be estimated from near-surface temperature. These results have the potential to greatly improve the representation of ablation in calculations of debris-covered glacier mass balance and projections of their response to climate change.

Salt-sensitive (SS) hypertension is accompanied with severe cardiorenal complications. In this condition, elevated blood pressure (BP) resulting from salt retention is associated with counterintuitively lower levels of atrial natriuretic peptide (ANP). In plasma, ANP is degraded by the neprilysin; therefore, pharmacological inhibition of this metalloprotease (i.e., with sacubitril) can be employed to increase ANP level. We have shown earlier that sacubitril in combination with valsartan (75 μg/day each) had beneficial effects on renal function in Dahl SS rats. The goal of this study was to evaluate the effects of a higher dose of sacubitril on renal damage in this model. To induce hypertension, male Dahl SS rats were fed a 4% NaCl diet (HS) for 21 days, and were administered sacubitril (125 μg/day) or vehicle via s.c. osmotic pumps. At the end of the HS challenge, both groups exhibited similar outcomes for GFR, heart weight, plasma electrolytes, BUN, and creatinine. Sacubitril exacerbated kidney hypertrophy, but did not affect levels of renal fibrosis. We also observed aggravated glomerular lesions and increased formation of protein casts in the sacubitril-treated animals compared to controls. Thus, in Dahl SS rats, administration of sacubitril without renin-angiotensin-system blockage had adverse effects on renal disease progression, particularly in regards to glomerular damage and protein cast formation. We can speculate that while ANP levels are increased because of neprilysin inhibition, there are off-target effects of sacubitril, which are detrimental to renal function in the SS hypertensive state.

Clearance phagocytosis is a documented function of Müller glia in the retina. However, the molecular mechanisms of Müller glia phagocytosis remain largely undefined. Here, we show that extracellular galectin-3 and protein S promote clearance phagocytosis by immortalized human MIO-M1 Müller cells in an additive, saturable manner. Galectin-3 promotes phagocytosis by primary Müller glia from wild-type (WT) mice but not from mice that lack the engulfment receptor MERTK and therefore develop postnatal photoreceptor degeneration. Probing a possible functional link between Müller galectin-3 and MERTK, we discovered that mertk−/− Müller glia in situ show excess galectin-3 at postnatal day 20 (P20), an age prior to detectable photoreceptor degeneration. Moreover, double knockout (DKO) mice lacking both galectin-3 and MERTK show increased activation of Müller cells (but not of microglia) at P20 and more pronounced photoreceptor loss at P35 compared to mice lacking MERTK alone. Exploring the well-established sodium iodate injury model, we also found more severe activation specifically of Müller glia, and worse retinal damage in mice lacking galectin-3 compared to WT mice. Indeed, galectin-3 deficiency significantly increased sensitivity to injury, yielding Müller activation and retinal damage at a sodium iodate concentration that had no effect on the WT retina. Altogether, our results from both inherited and acutely induced models of retinal degeneration agree that eliminating galectin-3 exacerbates Müller cell activation and retinal degeneration. These data identify an important protective role for the MERTK ligand galectin-3 in the retina in restraining Müller glia activation.

The unmet needs of people living with dementia have been shown to be multidimensional and impact well-being. However, there are a lack of studies examining variability of unmet needs and need-related distress from the person living with dementia’s perspective. The current study ( n = 12) examined the self-reported unmet needs and need-related distress of people with mild to moderate dementia. Seventy-five percent of participants ( n = 9) identified at least one unmet need and 50% ( n = 6) reported 10 or more unmet needs. “Finding and Arranging Services” and “Health Information” subscales had the highest reported average unmet needs. The most frequently reported unmet need-item was “ getting information about your memory problems? ” Participants reported variability in distress for both unmet and met needs. Continued research can provide beneficial information on the relationship between unmet needs, need-related distress, and outcomes of well-being for future interventions.

Food additives are common components of processed foods consumed in a Western diet. In inflammatory bowel disease patients, some diets that exclude food additives improved clinical disease parameters, suggesting a link between food additives and disease pathogenesis. Food additives also enhanced disease severity in mouse colitis models through incompletely described mechanisms. This study examined the mechanisms by which the food additive maltodextrin (MDX) alters the development of colitis in a murine model. Interleukin-10 knockout (IL10KO) mice were fed diets supplemented with MDX or carboxymethyl cellulose (CMC) to determine their impact on colitis onset and severity; microbiome composition, function, and location; colonic immune cell infiltrates; and mucus layer integrity. Primary IL10KO colonic epithelial monolayers were used to dissect the impact of MDX directly on epithelial differentiation and mucus production. MDX or CMC consumption increased the incidence and severity of colitis, as well as decreased microbiome diversity, altered microbial composition, and decreased fecal acetic acid levels. The number of mucus producing cells were decreased in food additive fed mice and resulted in increased microbial proximity to the intestinal epithelium. Additionally, MDX supplementation resulted in crypt hyperplasia and expansion of the HopX+ injury renewal stem cell niche. In primary intestinal epithelial-derived monolayers devoid of microbes and immune cells, MDX exposure decreased goblet cell number and mucus production in association with downregulated expression of the transcription factor Klf4, a marker of terminally differentiated goblet cells. These results suggest MDX disrupts the balance of epithelial cell differentiation and proliferation to contribute to disease pathogenesis through direct and indirect actions on the intestinal epithelial barrier.

Abstract Island biogeography theory is a foundational ecological concept that describes how island or habitat patch size and isolation predict biodiversity, but few have studied how the effects of island biogeography vary with life history characteristics of temperate mammal communities. From 2014 to 2020, we surveyed volant and nonvolant mammals within the Apostle Islands archipelago (Wisconsin, USA) using camera trapping, live trapping, and acoustic monitoring. We used linear regression and information‐theoretic model selection to evaluate the effects of island size and isolation on indices of biodiversity. We also examined the effects of life history characteristics (i.e., body mass, winter inactivity) on the relative abundance and distribution of each species on the islands and the mainland. For volant mammals, island size and isolation were not associated with any metrics of diversity. We found island size was positively associated with nonvolant mammal species richness and diversity, but not species evenness. Island isolation was not a significant predictor for small nonvolant mammal diversity, although both size and isolation appeared to influence island biogeography of medium‐large nonvolant mammals. We also found that the interaction between winter inactivity and body mass influenced the vagility of nonvolant mammals in the archipelago. Our results suggest that life history characteristics are important in mediating species responses to biogeography, supporting the dispersal‐limitation nonequilibrium view of island biogeography theory, particularly for nonvolant mammals.

The profound impact of the gut microbiome on host health has led to a revolution in biomedical research, motivating researchers from disparate fields to define the specific molecular mechanisms that mediate host-beneficial effects. The advent of genomic technologies allied to the use of model microbiomes in gnotobiotic mouse models has transformed our understanding of intestinal microbial ecology and the impact of the microbiome on the host. However, despite incredible advances, our understanding of the host-microbiome dialogue that shapes host physiology is still in its infancy. Progress has been limited by challenges associated with developing model systems that are both tractable enough to provide key mechanistic insights while also reflecting the enormous complexity of the gut ecosystem. Simplified model microbiomes have facilitated detailed interrogation of transcriptional and metabolic functions of the microbiome but do not recapitulate the interactions seen in complex communities. Conversely, intact complex communities from mice or humans provide a more physiologically relevant community type, but can limit our ability to uncover high-resolution insights into microbiome function. Moreover, complex microbiomes from lab-derived mice or humans often do not readily imprint human-like phenotypes. Therefore, improved model microbiomes that are highly defined and tractable, but that more accurately recapitulate human microbiome-induced phenotypic variation are required to improve understanding of fundamental processes governing host-microbiome mutualism. This improved understanding will enhance the translational relevance of studies that address how the microbiome promotes host health and influences disease states. Microbial exposures in wild mice, both symbiotic and infectious in nature, have recently been established to more readily recapitulate human-like phenotypes. The development of synthetic model communities from such “wild mice” therefore represents an attractive strategy to overcome the limitations of current approaches. Advances in microbial culturing approaches that allow for the generation of large and diverse libraries of isolates, coupled to ever more affordable large-scale genomic sequencing, mean that we are now ideally positioned to develop such systems. Furthermore, the development of sophisticated in vitro systems is allowing for detailed insights into host-microbiome interactions to be obtained. Here we discuss the need to leverage such approaches and highlight key challenges that remain to be addressed.

D-amphetamine induces emergence from sevoflurane and propofol anesthesia in rats. Dexmedetomidine is an α2-adrenoreceptor agonist that is commonly used for procedural sedation, whereas ketamine is an anesthetic that acts primarily by inhibiting NMDA-type glutamate receptors. These drugs have different molecular mechanisms of action from propofol and volatile anesthetics that enhance inhibitory neurotransmission mediated by GABAA receptors. In this study, we tested the hypothesis that d-amphetamine accelerates recovery of consciousness after dexmedetomidine and ketamine. Sixteen rats (Eight males, eight females) were used in a randomized, blinded, crossover experimental design and all drugs were administered intravenously. Six additional rats with pre-implanted electrodes in the prefrontal cortex (PFC) were used to analyze changes in neurophysiology. After dexmedetomidine, d-amphetamine dramatically decreased mean time to emergence compared to saline (saline:112.8 ± 37.2 min; d-amphetamine:1.8 ± 0.6 min, p < 0.0001). This arousal effect was abolished by pre-administration of the D1/D5 dopamine receptor antagonist, SCH-23390. After ketamine, d-amphetamine did not significantly accelerate time to emergence compared to saline (saline:19.7 ± 18.0 min; d-amphetamine:20.3 ± 16.5 min, p = 1.00). Prefrontal cortex local field potential recordings revealed that d-amphetamine broadly decreased spectral power at frequencies

Multimodality ophthalmic imaging systems aim to enhance the contrast, resolution, and functionality of existing technologies to improve disease diagnostics and therapeutic guidance. These systems include advanced acquisition and post-processing methods using optical coherence tomography (OCT), combined scanning laser ophthalmoscopy and OCT systems, adaptive optics, surgical guidance, and photoacoustic technologies. Here, we provide an overview of these ophthalmic imaging systems and their clinical and basic science applications.

Purpose. This study aims to identify clinical and imaging prognosticators associated with the successful bridging or downstaging to liver transplantation (LT) in patients undergoing Yttrium-90 radioembolization (Y90-RE) for hepatocellular carcinoma (HCC). Methods. Retrospectively, patients with Y90-RE naïve HCC who were candidates or potential candidates for LT and underwent Y90-RE were included. Patients were then divided into favorable (maintained or achieved Milan criteria (MC) eligibility) or unfavorable (lost eligibility or unchanged MC ineligibility) cohorts based on changes to their MC eligibility after Y90-RE. Penalized logistic regression analysis was performed to identify the significant baseline prognosticators. Results. Between 2013 and 2018, 135 patients underwent Y90-RE treatment. Among the 59 (42%) patients within MC, LT eligibility was maintained in 49 (83%) and lost in 10 (17%) patients. Within the 76 (56%) patients outside MC, eligibility was achieved in 32 (42%) and unchanged in 44 (58%). Among the 81 (60%) patients with a favorable response, 16 (20%) went on to receive LT. Analysis of the baseline characteristics revealed that lower Albumin-Bilirubin score, lower Child–Pugh class, lower Barcelona Clinic Liver Cancer stage, HCC diagnosis using dynamic contrast-enhanced imaging on CT or MRI, normal/higher albumin levels, decreased severity of tumor burden, left lobe HCC disease, and absence of HBV-associated cirrhosis, baseline abdominal pain, or fatigue were all associated with a higher likelihood of bridging or downstaging to LT eligibility (p’s

Environmental variability can be an important factor in the population dynamics of many species. In marine systems, for instance, whether environmental conditions facilitate or impede the movements of juvenile animals to nursery habitat can have a large influence on subsequent population abundance. Both subtle differences in the position of oceanographic features (such as meandering currents) and major disturbances (such as hurricanes) can greatly alter dispersal outcomes. Here, we use an ocean circulation model to explore seasonal and annual variation in the dispersal of post-hatchling Kemp’s ridley sea turtles (Lepidochelys kempii). We simulated the transport of 24 cohorts of young-of-the-year Kemp’s ridley sea turtles dispersing from the three primary nesting areas in the western Gulf of Mexico to describe variability in transport during the main hatching season and across years. We examined whether differences in transport distance among Kemp’s ridley cohorts could be explained by hurricane events. We found that years with high numbers of hurricanes corresponded to shorter dispersal distances and less variance within the first 6 months. Our findings suggest that differences in dispersal among sites and the impact of hurricane frequency and intensity could influence the survivorship and somatic growth rates of turtles from different nesting sites and hatching cohorts, either improving survival by encouraging retention in optimal pelagic habitat or decreasing survival by pushing hatchlings into dangerous shallow habitats. Considering such factors in future population assessments may aid in predicting how the potential for increasing tropical storms, a phenomenon linked to climate change, could affect Kemp’s ridley and other populations of sea turtles in the Atlantic Ocean.

The effectiveness of annual mailed fecal immunochemical testing (FIT) outreach is highest when return rates are optimized, which is aided by patient reminders. In a pilot patient-randomized controlled trial in two western Washington clinics of the Sea Mar Community Health Centers, we compared the effectiveness of two phone-based approaches to mailed FIT outreach reminders. In fall 2016, patients ages 50–75, due for colorectal cancer screening, and with a visit in the previous year at either of two clinics, were mailed an introductory letter and FIT. Those who did not return the FIT within 3 weeks (N = 427) were randomized to receive either: a) a series of up to 6 automated phone reminders; or b) the combination of automated and live phone reminders (up to 6 in total). The sole outcome was FIT return within 6 months after the FIT mailing. FIT completion rates were similar in the groups assigned to receive automated calls vs automated plus live calls (40% vs 39%; p = 0.89). The effectiveness of FIT reminder mode differed by language preference (p for interaction = 0.03): among Spanish-preferring patients (n = 106), FIT return rates were higher in the automated-only group than to the auto- plus live-call group (62% vs 39%, p = 0.02). Among English-preferring patients, no difference in modes was observed (n = 279, 32% vs 34%, p = 0.74). We observed no added benefit of live reminder calls in a mailed FIT plus automated call reminder program; our findings may inform efforts to efficiently optimize mailed-FIT outreach programs.ClinicalTrials.gov identifier NCT01742065 Keywords: Colorectal neoplasms, Early detection of cancer, Community health centers, Language

Guided by developmental models examining the legacy of childhood caregiving environments, we examined the longitudinal pattern of associations between harsh parenting and children's internalizing and externalizing symptoms across late childhood to late adolescence. Participants included 199 youth (48.7% female, 65.3% White, 32.2% Black, 2.5% biracial) and their mothers and fathers from a diverse range of socioeconomic backgrounds. The study utilized a multi-informant, longitudinal design including five waves of data (youths' mean ages were 9, 10, 11, 17, and 18 across waves). Harsh parenting at Age 9 predicted higher levels of (a) externalizing symptoms at Ages 11, 17, and 18 and (b) internalizing symptoms at Ages 17 and 18. Developmental sensitivity analyses revealed that the magnitude of the more distal association between early harsh parenting and later internalizing and externalizing symptoms was statistically stronger as compared to more proximal associations. Bidirectional analyses revealed that externalizing symptoms at Age 9 predicted harsh parenting at Ages 9, 10, 11, 17, and 18. Whereas links between harsh parenting and internalizing symptoms were consistent with a sleeper effects model, links between harsh parenting and externalizing symptoms provided some support for both enduring and sleeper effects models. Findings inform an understanding of youth developmental sensitivity to harsh parenting and the downstream consequences of harsh parenting. Results have important translational implications, including testing the long-term efficacy of therapeutic programs.

Abstract Prolonged tomato consumption can mitigate ultraviolet (UV) light induced sunburn via unknown mechanisms. Dietary carotenoids distributed to skin are hypothesized to protect skin against UV-induced damage, although other phytochemicals may play a role. We hypothesize that tomato consumption would protect against skin cancer. SKH-1 hairless and immunocompetent mice (n = 180) were fed AIN-93G or AIN-93G + 10% tangerine or red tomato powder for 35 weeks. From weeks 11–20, mice (n = 120) were exposed to 2240 J/m2 UV-B light, 3x/week, and tumors were tracked weekly. Control mice were fed the same diets but not exposed to UV. Tumor number was significantly lower in male mice consuming red tomato diets (1.73 ± 0.50, P = 0.015) or pooled tomato diets (2.03 ± 0.45, P = 0.017) compared to controls (4.04 ± 0.65). Carotenoid levels in plasma and skin were quantitated, with total lycopene higher in skin of tangerine fed animals despite a lower dose. Metabolomic analyses elucidated compounds derived from tomato glycoalkaloids (including tomatidine and hydroxylated-tomatidine) as significantly different metabolites in skin after tomato exposure. Here, we describe that tomato consumption can modulate risk for keratinocyte carcinomas; however, the role of the newly identified specific phytochemicals possibly responsible for this action require further investigation.

The strength and direction of sexual selection via female choice on masculine facial traits in men is a paradox in human mate choice research. While masculinity may communicate benefits to women and offspring directly (i.e. resources) or indirectly (i.e. health), masculine men may be costly as long-term partners owing to lower paternal investment. Mating strategy theory suggests women's preferences for masculine traits are strongest when the costs associated with masculinity are reduced. This study takes a multivariate approach to testing whether women's mate preferences are context-dependent. Women (n = 919) rated attractiveness when considering long-term and short-term relationships for male faces varying in beardedness (clean-shaven and full beards) and facial masculinity (30% and 60% feminized, unmanipulated, 30% and 60% masculinized). Participants then completed scales measuring pathogen, sexual and moral disgust, disgust towards ectoparasites, reproductive ambition, self-perceived mate value and the facial hair in partners and fathers. In contrast to past research, we found no associations between pathogen disgust, self-perceived mate value or reproductive ambition and facial masculinity preferences. However, we found a significant positive association between moral disgust and preferences for masculine faces and bearded faces. Preferences for beards were lower among women with higher ectoparasite disgust, providing evidence for ectoparasite avoidance hypothesis. However, women reporting higher pathogen disgust gave higher attractiveness ratings for bearded faces than women reporting lower pathogen disgust, providing support for parasite-stress theories of sexual selection and mate choice. Preferences for beards were also highest among single and married women with the strongest reproductive ambition. Overall, our results reflect mixed associations between individual differences in mating strategies and women's mate preferences for masculine facial traits.

Background: Colorectal cancer screening (CRC) rates are low, particularly among individuals with low socioeconomic status. Organized CRC screening programs have demonstrated success in increasing screening rates. Little is known about provider attitudes, beliefs, and practices related to CRC screening or how they are influenced by an organized CRC screening program. Methods: In 2014 and 2016, providers from 26 safety net clinics in Oregon and Northern California were invited to complete baseline and follow-up online surveys for the Strategies and Opportunities to Stop Colon Cancer in Priority Populations (STOP CRC) study. The provider survey link was sent electronically to primary care providers serving adult patients. Providers were sent reminders every 2 weeks via email to complete the survey, up to 3 reminders total. In this article, we describe learnings about provider attitudes, beliefs, and practices related to CRC screening after implementation of the STOP CRC program. Results: A total of 166 unique providers completed baseline and/or follow-up surveys, representing 228 responses. Main themes included (1) favorable shifts in attitude toward fecal immunochemical test (FIT) and direct-mail cancer screening programs, (2) changes in provider perception of key barriers, and (3) growing interest in centralized automated systems for identifying patients due for CRC screening and eligible for population-based outreach. Discussion: Providers are interested in improved information systems for identifying patients due for CRC screening and delivering population-based outreach (ie, to distribute FIT kits outside of the clinic visit) to help reduce health system- and patient-level barriers to screening. Trial Registration: National Clinical Trial (NCT) Identifier NCT01742065.

Abstract Biotic interactions can shape species’ distributions through their impact on species’ realized niches, potentially constraining or expanding the range of conditions under which species occur. We examine whether fine‐scale plant–plant interactions scale up to shape broad‐scale species’ distributions, using Azorella selago, a widespread cushion plant that facilitates other species, and the rest of the vascular flora of sub‐Antarctic Marion Island as a model system. We compared the upper elevational distributional limit of each species when growing on vs. away from A. selago to test how the interaction with this cushion plant species affects species’ ranges. Three out of 19 vascular plant species occurred at higher altitudes in the presence of A. selago than in the absence of A. selago: Acaena magellanica (+26 m higher), Colobanthus kerguelensis (+37 m higher), and Lycopodium saururus (+19 m higher). Therefore, A. selago's fine‐scale impacts scaled up to shape the distribution of a subset of the vascular flora of Marion Island. Plant–plant interactions thus have the potential to expand species upper distributional limits by increasing the niche space that a species can occupy, although the influence of these interactions may be strongly species‐specific.

Background: HIV infection and antiretroviral therapy (ART) have both been linked to dyslipidemia and increased cardiovascular disease (CVD) risk. Alterations in the composition of saturated (SaFA), monounsaturated (MUFA), and polyunsaturated (PUFA) fatty acids are related to inflammation and CVD progression in HIV-uninfected (HIV–) populations. The relationships among the lipidome and markers of monocyte and immune activation in HIV-infected (HIV+) individuals are not well understood.Methods: Concentrations of serum lipids and their fatty acid composition were measured by direct infusion-tandem mass spectrometry in samples from 20 ART-treated HIV+ individuals and 20 HIV– individuals.Results: HIV+ individuals had increased levels of free fatty acids (FFAs) with enrichment of SaFAs, including palmitic acid (16:0) and stearic acid (18:0), and these levels were directly associated with markers of monocyte (CD40, HLA-DR, TLR4, CD36) and serum inflammation (LBP, CRP). PUFA levels were reduced significantly in HIV+ individuals, and many individual PUFA species levels were inversely related to markers of monocyte activation, such as tissue factor, TLR4, CD69, and SR-A. Also in HIV+ individuals, the composition of lysophosphatidylcholine (LPC) was enriched for SaFAs; LPC species containing SaFAs were directly associated with IL-6 levels and monocyte activation. We similarly observed direct relationships between levels of SaFAs and inflammation in HIV uninfected individuals. Further, SaFA exposure altered monocyte subset phenotypes and inflammatory cytokine production in vitro.Conclusions: The lipidome is altered in ART-treated HIV infection, and may contribute to inflammation and CVD progression. Detailed lipidomic analyses may better assess CVD risk in both HIV+ and HIV– individuals than does traditional lipid profiling.