Etiology of lipid-laden macrophages in the lung.

Uniquely positioned as sentinel cells constantly exposed to the environment, pulmonary macrophages are vital for the maintenance of the lung lining. These cells are responsible for the clearance of xenobiotics, pathogen detection and clearance, and homeostatic functions such as surfactant recycling. Among the spectrum of phenotypes that may be expressed by macrophages in the lung, the pulmonary lipid-laden phenotype is less commonly studied in comparison to its circulatory counterpart, the atherosclerotic lesion-associated foam cell, or the acutely activated inflammatory macrophage. Herein, we propose that lipid-laden macrophage formation in the lung is governed by lipid acquisition, storage, metabolism, and export processes. The cellular balance of these four processes is critical to the maintenance of homeostasis and the prevention of aberrant signaling that may contribute to lung pathologies. This review aims to examine mechanisms and signaling pathways that are involved in lipid-laden macrophage formation and the potential consequences of this phenotype in the lung.

The CHALLENGE trial: the effects of a virtual reality-assisted exposure therapy for persistent auditory hallucinations versus supportive counselling in people with psychosis: study protocol for a randomised clinical trial.

BACKGROUND: Many patients suffering from schizophrenia spectrum disorders continue having distressing auditory hallucinations in spite of treatment with antipsychotic medication. The aim of this trial is to examine the effect of a targeted virtual reality therapy for persistent auditory hallucinations in individuals with psychosis. The trial explores whether this type of therapy can decrease the severity, frequency and distress of auditory hallucinations and, additionally, whether it can reduce clinical symptoms and enhance daily functioning in individuals with psychosis. METHODS: The study is a randomised, assessor-blinded parallel-group superiority clinical trial, allocating a total of 266 patients to either the experimental intervention or supportive counselling. The participants will be randomised to either (1) seven sessions of virtual reality therapy or (2) seven sessions of supportive counselling to be delivered within the first 12 weeks after inclusion in the study. All participants will be assessed at baseline and 12 and 24 weeks post-baseline. Independent assessors blinded to the treatment allocation will evaluate the outcome. The primary outcome is the level of auditory hallucinations measured with the Psychotic Symptoms Rating Scales (PSYRATS-AH) total score at the cessation of treatment at 12 weeks. Secondary outcomes are frequency of auditory hallucinations, the distress caused by auditory hallucinations, perceived voice power, patient acceptance of voices, patients' ability to respond to voices in an assertive way and social and daily function. DISCUSSION: Promising evidence of the efficacy of this immersive virtual reality-based therapy for auditory hallucinations exist, but evidence needs to be established in a large, methodological rigorous trial. If the therapy proves to be beneficial in reducing the severity of refractory auditory hallucinations, a large group of patients with schizophrenia and related disorders could be the target group of this short-term psychotherapeutic intervention.

Decreasing surface albedo signifies a growing importance of clouds for Greenland Ice Sheet meltwater production.

Clouds regulate the Greenland Ice Sheet's surface energy balance through the competing effects of shortwave radiation shading and longwave radiation trapping. However, the relative importance of these effects within Greenland's narrow ablation zone, where nearly all meltwater runoff is produced, remains poorly quantified. Here we use machine learning to merge MODIS, CloudSat, and CALIPSO satellite observations to produce a high-resolution cloud radiative effect product. For the period 2003-2020, we find that a 1% change in cloudiness has little effect (±0.16 W m-2) on summer net radiative fluxes in the ablation zone because the warming and cooling effects of clouds compensate. However, by 2100 (SSP5-8.5 scenario), radiative fluxes in the ablation zone will become more than twice as sensitive (±0.39 W m-2) to changes in cloudiness due to reduced surface albedo. Accurate representation of clouds will therefore become increasingly important for forecasting the Greenland Ice Sheet's contribution to global sea-level rise.

Seventy years of data from the world's longest grazed and irrigated pasture trials.

Pastures are the most widespread land use, globally. The Winchmore trials were established in 1948-1949 in Canterbury, New Zealand and examined either different rates of phosphorus (P) fertiliser on the same irrigation schedule (Fertiliser trial), or different irrigation scheduling at the same rate of P application (Irrigation trial). About 96,000 records of soil chemistry and physical data and pasture yield and botanical composition are available along with nearly 7000 soil samples. These data have been used in 475 publications that have explored topics as diverse as: improvements in sheep, dairy and deer production; the efficacy and scheduling of irrigation; improvements in pasture and crop production; agronomic and environmental soil and water research; and entomology. In addition to above topics, these data are invaluable for calibrating models to predict long-term issues like the accumulation of soil carbon or contaminants like cadmium and informing policy on climate change and agricultural practices. The data and soil samples are available for use and may yet yield discoveries, unforeseen 70 years ago.

Preferential transcription of the mutated allele in NPM1 mutated acute myeloid leukaemia.

Nucleophosmin is commonly both over-expressed and mutated in acute myeloid leukemia (AML). NPM1 mutations are always heterozygous. In addition, NPM1 has a number of different splice variants with the major variant encoded by exons 1-9 and 11-12 (NPM1.1). Further variants include NPM1.2 which lacks exons 8 and 10 and NPM1.3 which comprises exons 1-10 (and so lacks the region of sequence mutated in AML). In this study we quantified the expression of these three variants in 108 AML patient samples with and without NPM1 mutations and also assessed the level of expression from the wild-type and mutant alleles in variants NPM1.1 and NPM1.2. The results show that NPM1.1 is the most commonly expressed variant, however transcripts from wild-type and mutated alleles do not occur at equal levels, with a significant bias toward the mutated allele. Considering the involvement of mutant nucleophosmin in the progression and maintenance of AML, a bias towards mutated transcripts could have a significant impact on disease maintenance.

Collapse dynamics of spherical cavities in a solid under shock loading.

Extraordinary states of highly localised pressure and temperature can be generated upon the collapse of impulsively driven cavities. Direct observation of this phenomenon in solids has proved challenging, but recent advances in high-speed synchrotron radiography now permit the study of highly transient, subsurface events in real time. We present a study on the shock-induced collapse of spherical cavities in a solid polymethyl methacrylate medium, driven to shock states between 0.49 and 16.60 GPa. Utilising multi-MHz phase contrast radiography, extended sequences of the collapse process have been captured, revealing new details of interface motion, material failure and jet instability formation. Results reveal a rich array of collapse characteristics dominated by strength effects at low shock pressures and leading to a hydrodynamic response at the highest loading conditions.

Greenland Ice Sheet surface melt amplified by snowline migration and bare ice exposure.

Greenland Ice Sheet mass loss has recently increased because of enhanced surface melt and runoff. Since melt is critically modulated by surface albedo, understanding the processes and feedbacks that alter albedo is a prerequisite for accurately forecasting mass loss. Using satellite imagery, we demonstrate the importance of Greenland's seasonally fluctuating snowline, which reduces ice sheet albedo and enhances melt by exposing dark bare ice. From 2001 to 2017, this process drove 53% of net shortwave radiation variability in the ablation zone and amplified ice sheet melt five times more than hydrological and biological processes that darken bare ice itself. In a warmer climate, snowline fluctuations will exert an even greater control on melt due to flatter ice sheet topography at higher elevations. Current climate models, however, inaccurately predict snowline elevations during high melt years, portending an unforeseen uncertainty in forecasts of Greenland's runoff contribution to global sea level rise.

Development of the central nervous system in equine twin fetuses derived by somatic cell nuclear transfer.

Because of the growing importance of horses in leisure and several sports, somatic cell nuclear transfer (SCNT) is being used more frequently for cloning animals for performance and reproductive purposes. However, because of the need to perforate the zona pellucida during microsurgical reconstruction of the oocyte, it is possible that SCNT-derived embryos undergo premature hatching, resulting in embryo bisection and twinning. Therefore, because equine twin pregnancies often lead to abnormal embryo development and pregnancy failure, we performed a detailed comparative assessment of equine twin fetuses derived by SCNT with particular attention on the development of the central nervous system at 40 and 60 days gestation. The results of this study indicate that although cloned twin embryos show small differences in size, they do not exhibit apparent macro- or microscopic developmental discrepancies in the central nervous system, suggesting that the twining phenomenon resulting from SCNT does not affect fetal differentiation.

Synovial sepsis is rare following intrasynovial medication in equine ambulatory practice.

BACKGROUND: There are few observational studies regarding the potentially serious complication of synovial sepsis following intrasynovial medication in general equine practice. Quantification of risk is fundamental to fully inform decision-making and owner consent prior to undertaking procedures. OBJECTIVES: To describe the occurrence of synovial sepsis following intrasynovial injections in a large ambulatory equine practice between 2006-2011. STUDY DESIGN: Retrospective cohort study. METHODS: Medication records were cross-referenced against synovial cytology submissions and hospital admissions for synovial sepsis. The occurrence of synovial sepsis within strata of interest, including horse age, sex and product used were described as proportions with corresponding 95% confidence intervals, and measures of attributable risks. RESULTS: In the study period, 9456 intrasynovial medications were performed in 4331 sessions. The most frequently used medications were: corticosteroids (3869/4331, 89.3% [95% CI 88.4, 90.3%]), hyaluronate (3617/4331, 83.5% [95% CI 82.4, 84.6%]) and amikacin sulphate (4044/4331, 93.4% [95% CI 92.6, 94.1%]). Overall four horses developed post medication synovial sepsis (0.04% [95% CI 0.0, 0.08%] of all medications), two of these cases were given polysulphated glycosaminoglycans and hyaluronate (2/14, 14.3% [95% CI 0.0, 32.6%]), two were given triamcinolone acetonide (2/3592, 0.1% [95% CI 0.0, 0.1%]), and hyaluronate (2/3617, 0.06% [95% CI 0.0, 0.1%]). One of these cases had received concurrent medication with amikacin sulphate (1/4044, 0.02% [95% CI 0.0, 0.1%]). All four cases returned to racing following joint lavage. MAIN LIMITATIONS: Due to the low frequency of cases multivariable statistical analysis was not performed. Although the study was conducted in an ambulatory setting, the population included a high number of racehorses, limiting application to the general horse population. CONCLUSIONS: The frequency of synovial sepsis in this population of horses was 0.04%. These data may be helpful in informing clients regarding the potential risks of adverse complications resulting from intrasynovial medication.

Early phosphoproteomic changes for adverse outcome pathway development in the fathead minnow (Pimephales promelas) brain.

Adverse outcome pathways (AOPs) are conceptual frameworks that organize and link contaminant-induced mechanistic molecular changes to adverse biological responses at the individual and population level. AOPs leverage molecular and high content mechanistic information for regulatory decision-making, but most current AOPs for hormonally active agents (HAAs) focus on nuclear receptor-mediated effects only despite the overwhelming evidence that HAAs also activate membrane receptors. Activation of membrane receptors triggers non-genomic signaling cascades often transduced by protein phosphorylation leading to phenotypic changes. We utilized label-free LC-MS/MS to identify proteins differentially phosphorylated in the brain of fathead minnows (Pimephales promelas) aqueously exposed for 30 minutes to two HAAs, 17α-ethinylestradiol (EE2), a strong estrogenic substance, and levonorgestrel (LNG), a progestin, both components of the birth control pill. EE2 promoted differential phosphorylation of proteins involved in neuronal processes such as nervous system development, synaptic transmission, and neuroprotection, while LNG induced differential phosphorylation of proteins involved in axon cargo transport and calcium ion homeostasis. EE2 and LNG caused similar enrichment of synaptic plasticity and neurogenesis. This study is the first to identify molecular changes in vivo in fish after short-term exposure and highlights transduction of rapid signaling mechanisms as targets of HAAs, in addition to nuclear receptor-mediated pathways.

A longitudinal study of fractures in 1488 Thoroughbred racehorses receiving intrasynovial medication: 2006-2011.

BACKGROUND: There is a need for investigation of associations between intrasynovial medication and fracture risk in racehorses. OBJECTIVES: To document the proportion and type of fracture occurring following intrasynovial medication. STUDY DESIGN: Retrospective longitudinal study. METHODS: Review of clinical records from all Thoroughbred flat racehorses receiving intrasynovial medication under the care of a first-opinion veterinary practice between 2006 and 2011. Fractures pre- and post-medication were categorised by location, type, and severity; analysis of fractures sustained within 56-days of medication was undertaken. Survival analyses using Cox proportional hazards models were conducted for associations between medication used (corticosteroid vs. non-corticosteroid), age at first medication, sex and total number of medications and fracture. RESULTS: A total of 1488 horses received intrasynovial medication during the period of study; 8692 synovial spaces were medicated in 3925 sessions. There were 96 fractures that occurred within 56-days of medication, of which 44 were classified as 'serious' (44/1488; 3.0% total population, 95% CI 2.1, 3.8%) and 11 (11/1488, 0.7%, 95% CI 0.3, 1.2%) were euthanased due to severity of injury. Fifty-four of 96 cases (56.4%, 95% CI 46.3, 66.2%) returned to racing. Targeted imaging of the subsequent injury site was undertaken prior to injury in only 7 (7/96, 7.3%) horses that injured post-medication. Horses that had received ≥3 previous intrasynovial medication sessions had an increased hazard of sustaining a fracture within 56 days (HR 2.31, 95% CI 1.51, 3.54, P<0.001) compared with those receiving ≤3 medication sessions, adjusted for each increasing year of age (HR 0.67, 95% CI 0.53, 0.86, P = 0.001). MAIN LIMITATIONS: Absence of an unexposed cohort of nonmedicated racehorses. CONCLUSIONS: Serious musculoskeletal injury following intrasynovial medication occurred in 3% (44/1488) horses. Reduction in injury rates may be possible through greater use of premedication diagnostic imaging.

Mesenchymal-like stem cells in canine ovary show high differentiation potential.

OBJECTIVES: Recent studies have reported the existence of stem cells in ovarian tissue that show enhanced proliferative and differentiation potential compared to other adult tissues. Based on this evidence, we hypothesized that ovarian tissue contained mesenchymal-like stem cells (MSC) that could be isolated using a novel rapid plastic adhesion technique. MATERIALS AND METHODS: We established MSC lines derived from ovarian and adipose tissue based on their ability to rapidly adhere to plastic culture dishes in the first 3 hours after plating and studied their potentiality in terms of molecular markers and differentiation capacity. RESULTS: Morphological and kinetic properties of in vitro cultured ovarian MSC were similar to adipose-derived MSC, and both reached senescence after similar passage numbers. Ovarian-derived MSC expressed mesenchymal (CD90 and CD44) but not haematopoietic markers (CD34 and CD45), indicating similarity to adipose-derived MSC. Moreover, ovarian-derived MSC expressed NANOG, TERT, SOX2, OCT4 and showed extensive capacity to differentiate not only into adipogenic, osteogenic and chondrogenic tissue but also towards neurogenic and endodermal lineages and even precursors of primordial germ cells. CONCLUSION: These results show for the first time the derivation of ovarian cells with the molecular properties of MSC as well as wide differentiation potential. Canine ovarian tissue is accessible, expandable, multipotent and has high plasticity, holding promise for applications in regenerative medicine.

Visualisation of Kiss1 Neurone Distribution Using a Kiss1-CRE Transgenic Mouse.

Kisspeptin neuropeptides are encoded by the Kiss1 gene and play a critical role in the regulation of the mammalian reproductive axis. Kiss1 neurones are found in two locations in the rodent hypothalamus: one in the arcuate nucleus (ARC) and another in the RP3V region, which includes the anteroventral periventricular nucleus (AVPV). Detailed mapping of the fibre distribution of Kiss1 neurones will help with our understanding of the action of these neurones in other regions of the brain. We have generated a transgenic mouse in which the Kiss1 coding region is disrupted by a CRE-GFP transgene so that expression of the CRE recombinase protein is driven from the Kiss1 promoter. As expected, mutant mice of both sexes are sterile with hypogonadotrophic hypogonadism and do not show the normal rise in luteinising hormone after gonadectomy. Mutant female mice do not develop mature Graafian follicles or form corpora lutea consistent with ovulatory failure. Mutant male mice have low blood testosterone levels and impaired spermatogenesis beyond the meiosis stage. Breeding Kiss-CRE heterozygous mice with CRE-activated tdTomato reporter mice allows fluorescence visualisation of Kiss1 neurones in brain slices. Approximately 80-90% of tdTomato positive neurones in the ARC were co-labelled with kisspeptin and expression of tdTomato in the AVPV region was sexually dimorphic, with higher expression in females than males. A small number of tdTomato-labelled neurones was also found in other locations, including the lateral septum, the anterodorsal preoptic nucleus, the amygdala, the dorsomedial and ventromedial hypothalamic nuclei, the periaquaductal grey, and the mammillary nucleus. Three dimensional visualisation of Kiss1 neurones and fibres by CLARITY processing of whole brains showed an increase in ARC expression during puberty and higher numbers of Kiss1 neurones in the caudal region of the ARC compared to the rostral region. ARC Kiss1 neurones sent fibre projections to several hypothalamic regions, including rostrally to the periventricular and pre-optic areas and to the lateral hypothalamus.

Differentiation of equine induced pluripotent stem cells into a keratinocyte lineage.

REASONS FOR PERFORMING STUDY: Skin trauma in horses often leads to the development of chronic nonhealing wounds that lack a keratinocyte cover, vital for healing. Reports in mouse and man confirm the possibility of generating functional keratinocytes from induced pluripotent stem cells (iPSC), thus presenting myriad potential applications for wound management or treatment of skin disease. Similarly, differentiation of equine iPSC (eiPSC) into a keratinocyte lineage should provide opportunities for the advancement of veterinary regenerative medicine. OBJECTIVES: The purpose of this study was to develop an efficient method for the differentiation of eiPSC into a keratinocyte lineage. It was hypothesised that eiPSC can form differentiated keratinocytes (eiPSC-KC) comparable with primary equine keratinocytes (PEK) in their morphological and functional characteristics. STUDY DESIGN: Experimental in vitro study. METHODS: Equine iPSC established using a nonviral system were treated for 30 days with retinoic acid and bone morphogenetic protein-4 to induce directed differentiation into iPSC-KC. Temporospatial gene and protein expression by eiPSC-KC was measured at weekly intervals of differentiation and in response to calcium switch. Proliferative and migratory capacities of eiPSC-KC were compared with those of PEK. RESULTS: Equine iPSC, upon directed differentiation, showed loss of pluripotency genes and progressive increase in pancytokeratin expression indicating ectodermal specification into keratinocytes. High differentiation efficiency was achieved, with 82.5% of eiPSC expressing keratin 14, a marker of epidermal-specific basal stem cells, after 30 days of directed differentiation. Moreover, the proliferative capacity of eiPSC-KC was superior, while the migratory capacity (measured as the ability to epithelise in vitro wounds) was comparable with that of PEK. CONCLUSIONS: This proof of concept study suggests that eiPSC can successfully be differentiated into equine keratinocytes (eiPSC-KC) with features that are promising to the development of a stem cell-based skin construct, with the potential to regenerate lost or damaged skin.

Mitochondrial DNA dynamics during in vitro culture and pluripotency induction of a bovine Rho0 cell line.

Large number of cellular changes and diseases are related to mutations in the mitochondrial DNA copy number. Cell culture in the presence of ethidium bromide is a known way of depleting mitochondrial DNA and is a useful model for studying such conditions. Interestingly, the morphology of these depleted cells resembles that of pluripotent cells, as they present larger and fragmented mitochondria with poorly developed cristae. Herein, we aimed to study the mechanisms responsible for the control of mitochondrial DNA replication during mitochondrial DNA depletion mediated by ethidium bromide and during the in vitro induction of cellular pluripotency with exogenous transcription factor expression in a bovine model. This article reports the generation of a bovine Rho0 mesenchymal cell line and describes the analysis of mitochondrial DNA copy number in a time-dependent manner. The expression of apoptosis and mitochondrial-related genes in the cells during mitochondrial DNA repletion were also analyzed. The dynamics of mitochondrial DNA during both the depletion process and in vitro reprogramming are discussed. It was possible to obtain bovine mesenchymal cells almost completely depleted of their mitochondrial DNA content (over 90%). However, the production of induced pluripotent stem cells from the transduction of both control and Rho0 bovine mesenchymal cells with human reprograming factors was not successful.

Precision Measurement of the p(e,e

New results are reported from a measurement of π^{0} electroproduction near threshold using the p(e,e^{'}p)π^{0} reaction. The experiment was designed to determine precisely the energy dependence of s- and p-wave electromagnetic multipoles as a stringent test of the predictions of chiral perturbation theory (ChPT). The data were taken with an electron beam energy of 1192 MeV using a two-spectrometer setup in Hall A at Jefferson Lab. For the first time, complete coverage of the ϕ_{π}^{*} and θ_{π}^{*} angles in the pπ^{0} center of mass was obtained for invariant energies above threshold from 0.5 up to 15 MeV. The 4-momentum transfer Q^{2} coverage ranges from 0.05 to 0.155 (GeV/c)^{2} in fine steps. A simple phenomenological analysis of our data shows strong disagreement with p-wave predictions from ChPT for Q^{2}>0.07 (GeV/c)^{2}, while the s-wave predictions are in reasonable agreement.

Bacterial and fungal pattern recognition receptors in homologous innate signaling pathways of insects and mammals.

In response to bacterial and fungal infections in insects and mammals, distinct families of innate immune pattern recognition receptors (PRRs) initiate highly complex intracellular signaling cascades. Those cascades induce a variety of immune functions that restrain the spread of microbes in the host. Insect and mammalian innate immune receptors include molecules that recognize conserved microbial molecular patterns. Innate immune recognition leads to the recruitment of adaptor molecules forming multi-protein complexes that include kinases, transcription factors, and other regulatory molecules. Innate immune signaling cascades induce the expression of genes encoding antimicrobial peptides and other key factors that mount and regulate the immune response against microbial challenge. In this review, we summarize our current understanding of the bacterial and fungal PRRs for homologous innate signaling pathways of insects and mammals in an effort to provide a framework for future studies.

Immune potential of allogeneic equine induced pluripotent stem cells.

REASONS FOR PERFORMING STUDY: Induced pluripotent stem cells (iPSC) have brought immense hope to cellular therapy and regenerative medicine. However, the antigenicity of iPSC has not been well documented and remains a hurdle for clinical applications. Expression of major histocompatibility complex (MHC) molecules by human and murine iPSC is downregulated, making these cells potentially safe for transplantation. No such data are available for any large animal model. OBJECTIVES: To measure expression of MHC molecules on equine iPSC (eiPSC) and describe their antigenicity using intradermal testing. The hypothesis was that allogeneic eiPSC weakly express MHC molecules and would not elicit a rejection response when injected intradermally. STUDY DESIGN: Experimental study involving both in vitro and in vivo components. METHODS: Two green fluorescent protein-expressing eiPSC lines were analysed by flow cytometry for MHC expression. One line was then transplanted intradermally, along with appropriate controls, into 2 unrelated experimental horses. Blood was collected pre- and 7 days post transplantation. The wheals formed at the sites of injection were measured at regular intervals beginning at 0.25 h until 4 weeks. Tissue samples of the injected sites were obtained at 2, 3, 7 and 30 days post transplantation and analysed by histopathology and immunofluorescence. RESULTS: Both eiPSC lines weakly expressed MHC molecules. eiPSC were detectable up to 7 days following allogeneic transplantation and elicited no apparent systemic response. Injection of eiPSC caused small wheal formation at the skin surface. Skin sections revealed CD4(+) and CD8(+) mononuclear cells up to 30 days post transplantation. CONCLUSIONS: These data suggest that while transplantation of allogeneic eiPSC elicits a moderate cellular response, it does not cause acute rejection. The feasibility of banking allogeneic iPSC for regenerative medicine applications should be explored.