Spatiotemporal modulation of growth factors directs the generation of multilineage mouse embryonic stem cell-derived mammary organoids.

Ectodermal appendages, such as the mammary gland (MG), are thought to have evolved from hair-associated apocrine glands to serve the function of milk secretion. Through the directed differentiation of mouse embryonic stem cells (mESCs), here, we report the generation of multilineage ESC-derived mammary organoids (MEMOs). We adapted the skin organoid model, inducing the dermal mesenchyme to transform into mammary-specific mesenchyme via the sequential activation of Bone Morphogenetic Protein 4 (BMP4) and Parathyroid Hormone-related Protein (PTHrP) and inhibition of hedgehog (HH) signaling. Using single-cell RNA sequencing, we identified gene expression profiles that demonstrate the presence of mammary-specific epithelial cells, fibroblasts, and adipocytes. MEMOs undergo ductal morphogenesis in Matrigel and can reconstitute the MG in vivo. Further, we demonstrate that the loss of function in placode regulators LEF1 and TBX3 in mESCs results in impaired skin and MEMO generation. In summary, our MEMO model is a robust tool for studying the development of ectodermal appendages, and it provides a foundation for regenerative medicine and disease modeling.

Subgroup analyses of a randomized trial of DHA supplementation for infants born preterm with assessments of cognitive development up to 7-years of age: What happens in infants born <29 weeks' gestation?

A recent trial showed that high-dose docosahexaenoic acid (high-DHA) supplementation of infants born <29 weeks' gestation improves intelligence quotient (IQ) at five years' corrected age. However, this finding has not been detected by other trials of DHA, which either did not measure IQ or included more mature infants. We analyzed the subgroup of 204 infants born <29 weeks' from our earlier randomized trial of high-DHA (∼1 % total fatty acids) or standard-DHA (∼ 0.3 % total fatty acids). Participants were assessed for cognition at 18 months, and IQ and behavior at seven years' corrected age. No group differences were detected for mean cognitive, IQ or behavior scores. At 18 months, 18.8 % of children in the high-DHA group had a cognitive score <85, compared with 31.1 % of children in the standard-DHA group, but at seven years there was no difference. Although an underpowered post-hoc subgroup analysis, this study provides limited support to recommendations that infants born <29 weeks' gestation require supplemental DHA.

Effect of season on weaned piglet mortality during transport greater than 8 h under Canadian conditions.

North American swine producers commonly transport piglets away from sow farms at weaning. However, limited information on factors associated with piglet mortality during these transports is available. The objectives of this study were to identify transport characteristics that were associated with the occurrence of in-transit mortality (≥1 piglet found dead on arrival) and/or associated with increased rates of in-transit mortality using records of weaned piglet transport voluntarily provided by Canadian swine companies. Following cleaning and validation, records of 810 long duration (>8 h to <28 h) weaned piglet (min.: 4.2, max.: 7.7 kg) transports conducted between 2016 and 2017 by four companies were available to investigate risk factors for the occurrence of in-transit mortality. Transports originated in Ontario, Saskatchewan, or Alberta and 30% of transports had one or more mortality events recorded. Season of transport was significant in the final logistic regression model; the odds of a transport having mortality occur was greater during the winter (December, January, February) compared to the spring (March, April, May) (OR=1.8; 95% CI: 1.13-2.96; P = 0.013), summer (June, July, August) (OR= 2.1; 95% CI: 1.28-3.34; P = 0.003) and fall (September, October, November) (OR=2.2; 95% CI: 1.36-3.62; P = 0.001), with no differences observed between the other seasons. Additionally, records from 755 long duration weaned piglet (min.: 4.2, max.: 7.9 kg) transports conducted between 2014 and 2017 by a single company was used to identify risk factors for increased rates of in-transit mortality. Transports originated in Saskatchewan or Alberta and 44.9% of transports had one or more mortality events recorded with in-transit mortality rates ranging from 0.00% to 6.16%. Season of transport was again significant in the final negative binomial regression model. The rate of piglet death in-transit was increased in all seasons compared to the summer with winter transport having the highest predicted rate of in-transit mortality. Transports conducted during the winter had a rate of in-transit mortality nearly five times greater compared to transports conducted during the summer (IRR= 4.94; 95% CI: 3.11-7.87; P = 0.000) and approximately three times greater compared to transports conducted during the fall (IRR=2.7; 95% CI: 1.73-4.30; P = 0.000) and spring (IRR=3.1; 95% CI: 1.96-4.99; P = 0.000). These results suggest that winter transport in Western Canada is an area of opportunity to reduce in-transit mortality during long duration weaned piglet transport. Research investigating transport practices that may mitigate the effects of extreme cold (e.g., space allowance, bedding provision) would be useful for informing specific recommendations for this age group.

A descriptive study of weaned piglet transport practices in Alberta, Ontario, and Saskatchewan, Canada between 2014 and 2018.

Canadian transport practices for shipments of newly weaned piglets are not well-described despite documentation requirements for those conducting the movement of these animals. The objective of this study was to describe the characteristics of weaned piglet transport events that occurred between 2014 and 2018 using records provided by five Canadian swine companies. Following cleaning and validation, the dataset included records from 6203 transport events involving the transport of approximately 6.9 million piglets (5.7 kg, 4.1-7.9 kg) from 62 origin sites in Alberta, Ontario, or Saskatchewan, Canada. This represents approximately 4.7% of the piglets estimated to have been weaned in Canada between 2014 and 2018, and 1.7% of sow farms in Canada according to 2016 National census data. Most transport events ended at farms in Canada (71.3%), while the remaining delivered piglets to one of eight American states. The predominant trailer types used were Straightdeck (51.4%) and Potbelly (36.6%), but this did not reflect the number of piglets transported as Potbelly trailers have greater load capacity. Transport events most frequently involved loading piglets from one origin barn and delivering them to a single destination barn (78.1%). Only transport events involving export to the United States picked up piglets from, or delivered them to, more than one farm site. Most transport events had very short trip distances (median distance: 48.0 km; IQR: 497.0), but a marked range was observed (1.8-2931.2 km). Average daily temperature data matched to the transport records by origin and destination location demonstrated ambient environmental conditions during these transport events ranged from - 30.3-28.7 °C. Overall, less than 10% of transport events had mortality occur. Comparable with other observational studies documenting weaned piglet mortality, the average in-transit mortality rate observed over the multiple seasons, companies, trip distances, and other characteristics in this dataset was 0.027%. However, instances of mortality over 1% did sporadically occur and could translate to considerable losses given the large load sizes common for piglets of this age and size (median load size: 1105 piglets; IQR: 1036 piglets). These data provide a better understanding of the interconnectedness of the Canadian swine industry as well as common transport practices which may inform future research on disease transmission in swine transport networks, or piglet welfare during transport. Additionally, variables that were not present in this dataset that would further strengthen these types of investigations are highlighted (e.g., space allowance).

Immunotolerance of dairy heifers in response to repeated exposure to bacterial lipopolysaccharide endotoxin.

Dairy cattle face a variety of stressful events on a daily basis. More specifically, climate change has resulted in more frequent heat stress events that increase the incidence of chronic bacterial infections by inducing conditions like leaky gut syndrome, whereby the integrity of the intestinal epithelium is compromised allowing for luminal bacterial lipopolysaccharide (LPS) endotoxin to infiltrate the host's bloodstream resulting in acute or chronic systemic stimulation of the innate immune system. Repeated exposure to LPS over a short period of time is reported to induce immunotolerance within the host. This LPS tolerance is an essential immunohomeostatic response that can protect against over activation of the inflammatory response during subsequent exposure to LPS. In the present study, Holstein calves (n = 20) were initially stress challenged with either saline, or 100, 200 or 400 ng/kg of LPS administered intramuscular, and again re-challenged with 200 ng/kg of LPS 2-weeks later. Serum was collected every 2 hr for 6 hr to profile changes in circulatory stress biomarkers after the repeated LPS exposures. Heifers that were initially challenged with 100, 200 and 400 ng/kg of LPS demonstrated significantly attenuated cortisol responses in the second challenge (p < 0.01, 0.01 and 0.05, respectively), whereas control animals who previously received saline demonstrated a strong cortisol response at 2 hr after receiving 200 ng/kg of LPS (p < 0.05). The cytokine/chemokine (IL-6, CCL2, CCL3 and CCL4) responses were also attenuated during the LPS rechallenge (p < 0.05). Finally, microRNA expression profiles were determined to assess the epigenetic response to repeated LPS exposure. Interestingly, miR-31 and miR-223 were downregulated in response to the second LPS challenge. The present study demonstrates the dynamic nature of the stress response in dairy cattle as it relates to the development of LPS tolerance. Understanding the roles of various stress biomarkers in the context of innate immune cell tolerance is essential for evaluating their impact on immune system homeostasis.

Genetic Covariation Between the Vertically Transmitted Endophyte Epichloë canadensis and Its Host Canada Wildrye.

Symbiotic mutualisms are thought to be stabilized by correlations between the interacting genotypes which may be strengthened via vertical transmission and/or reduced genetic variability within each species. Vertical transmission, however, may weaken interactions over time as the endosymbionts would acquire mutations that could not be purged. Additionally, temporal variation in a conditional mutualism could create genetic variation and increased variation in the interaction outcome. In this study, we assessed genetic variation in both members of a symbiosis, the endosymbiotic fungal endophyte Epichloë canadensis and its grass host Canada wildrye (Elymus canadensis). Both species exhibited comparable levels of diversity, mostly within populations rather than between. There were significant differences between populations, although not in the same pattern for the two species, and the differences were not correlated with geographic distance for either species. Interindividual genetic distance matrices for the two species were significantly correlated, although all combinations of discriminant analysis of principle components (DAPC) defined multilocus genotype groups were found suggesting that strict genotype matching is not necessary. Variation in interaction outcome is common in grass/endophyte interactions, and our results suggest that the accumulation of mutations overtime combined with temporal variation in selection pressures increasing genetic variation in the symbiosis may be the cause.

A National Referral Service for Paediatric Brachytherapy: An Evolving Practice and Outcomes Over 13 Years.

AIMS: Most children requiring radiotherapy receive external beam treatment and few have tumours suitable for brachytherapy. No paediatric radiotherapy centre will treat enough patients from its own normal catchment population for expertise in brachytherapy to be developed and sustained. Following discussion and agreement in the national paediatric radiotherapy group, a service for paediatric brachytherapy in the UK has been developed. We report the process that has evolved over more than 10 years, with survival and functional outcome results. MATERIALS AND METHODS: Since 2009, potential patients have been referred to the central paediatric oncology multidisciplinary team meeting, where imaging, pathology and treatment options are discussed. Since 2013, the National Soft Tissue Sarcoma Advisory Panel has also reviewed most patients, with the principal aim of advising on the most suitable primary tumour management for complex patients. Clinical assessment and examination under anaesthetic with biopsies may be undertaken to confirm the appropriateness of brachytherapy, either alone or following conservative surgery. Fractionated high dose rate brachytherapy was delivered to a computed tomography planned volume after implantation of catheters under ultrasound imaging guidance. Since 2019, follow-up has been in a dedicated multidisciplinary clinic. RESULTS: From 2009 to 2021 inclusive, 35 patients (16 female, 19 male, aged 8 months to 17 years 6 months) have been treated. Histology was soft-tissue sarcoma in 33 patients and carcinoma in two. The treated site was pelvic in 31 patients and head and neck in four. With a median follow-up of 5 years, the local control and overall survival rates are 100%. Complications have been few, and functional outcome is good. CONCLUSION: Brachytherapy is effective for selected paediatric patients, resulting in excellent tumour control and good functional results. It is feasible to deliver paediatric brachytherapy at a single centre within a national referral service.

Metals in urban soils of Europe: A systematic review.

Metal contamination of soils is widespread across Europe and is of great concern as it may impact food production, the supply of drinking water and human health (European Environment Agency, 2014; Panagos et al., 2013). Most research to date on soil metal contamination has focussed on agricultural soils (Tóth et al., 2016a). Current knowledge of the extent of urban soil metal contamination in Europe, however, is limited, especially for soils in recreational areas, which is particularly concerning as these areas may have a high footfall. Here, we conducted a systematic analysis of metal contamination in European urban soils based on 174 peer-reviewed studies spanning 143 urban sites and 29 European countries. The results show that reporting of data on urban soil metals is highly heterogeneous across the study area. Over half of all studies are from only five countries (Italy, Spain, UK, Poland and Serbia) and no data are available for 14 other European countries. The metals that most commonly exceed national safety thresholds are Pb, Zn, Cu, Cr and Ni. Elevated levels of these metals are usually attributed to anthropogenic sources, primarily traffic and industry. Some 22 % of urban sites studied show anthropogenic enrichment; this phenomenon is most common in Italy, Serbia and Finland. In contrast, 44 % of urban sites studied show geogenic metal enrichment; this is most common in Italy, the UK and Serbia. The dataset is subject to a sample size bias, whereby soil metal enrichment is identified more frequently in regions with more data. Future studies should focus on key knowledge gaps, such as urban soils in locations with current or historical heavy industrialisation and locations in central and eastern Europe. Study methods should be standardised to facilitate comparison of soil metal data from different studies and European safety thresholds should be identified for key elements.

Lumbar Level Significantly Influences Postoperative Global Sagittal Balance Following Pedicle Subtraction Osteotomy.

STUDY DESIGN: Retrospective cohort study. OBJECTIVES: The current evidence regarding how level of lumbar pedicle subtraction osteotomy (PSO) influences correction of sagittal alignment is limited. This study sought to investigate the relationship of lumbar level and segmental angular change (SAC) of PSO with the magnitude of global sagittal alignment correction. METHODS: This study retrospectively evaluated 53 consecutive patients with adult spinal deformity who underwent lumbar PSO at a single institution. Radiographs were evaluated to quantify the effect of PSO on lumbar lordosis (LL), thoracic kyphosis (TK), sacral slope (SS), pelvic tilt (PT), pelvic incidence (PI), T1-spinopelvic inclination (T1SPI), T1-pelvic alignment (TPA), and sagittal vertical axis (SVA). RESULTS: Significant correlations were found between PSO SAC and the postoperative increase in LL (r = 0.316, P = .021) and PT (r = 0.352, P = .010), and a decrease in TPA (r = -0.324, P = .018). PSO level significantly correlated with change in T1SPI (r = -0.305, P = .026) and SVA (r = -0.406, P = .002), with more caudal PSO corresponding to a greater correction in sagittal balance. On multivariate analysis, more caudal PSO level independently predicted a greater reduction in T1SPI (ß = -3.138, P = .009) and SVA (ß = -29.030, P = .001), while larger PSO SAC (ß = -0.375, P = .045) and a greater number of fusion levels (ß = -1.427, P = .036) predicted a greater reduction in TPA. CONCLUSION: This study identified a gain of approximately 3 degrees and 3 cm of correction for each level of PSO more caudal to L1. Additionally, a larger PSO SAC predicted greater improvement in TPA. While further investigation of these relationships is warranted, these findings may help guide preoperative PSO level selection.

Preoperative Multifidus Muscle Quality is Associated With Patient Reported Outcomes After Lateral Lumbar Interbody Fusion.

STUDY DESIGN: Retrospective cohort. OBJECTIVE: Lateral lumbar interbody fusion (LLIF) commonly involves a transpsoas approach. Despite the association between LLIF, postoperative iliopsoas weakness, and iatrogenic neuropraxia, no study has yet examined the effect of psoas or multifidus muscle quality on patient-reported outcomes (PROs). METHODS: This study retrospectively reviewed patients who underwent LLIF with 1-year minimum follow-up. Psoas and multifidus muscle qualities were graded on preoperative magnetic resonance imaging using two validated classification systems for muscle atrophy. Average muscle quality was calculated as the mean score from all levels (L1-2 through L5-S1). Univariate and multivariate statistics were utilized to investigate the relationship between psoas/multifidus muscle quality and preoperative, 6-weeks postoperative, and final postoperative PROs. RESULTS: 74 patients (110 levels) with a mean follow-up of 18.71 ± 8.02 months were included for analysis. Greater multifidus atrophy was associated with less improvement on ODI, SF12, and VR12 (P < .05) on univariate analysis. On multivariate analysis, worse multifidus atrophy predicted less improvement on SF12 and VR12 (P < .05). CONCLUSION: Despite the direct manipulation of the psoas muscle inherent to LLIF, preoperative psoas muscle quality did not affect postoperative outcomes. Rather, the extent of preoperative multifidus fatty infiltration and atrophy was more likely to predict postoperative pain and disability. These findings suggest that multifidus atrophy may be more pertinent than psoas atrophy in its association with patient-reported outcome measures after LLIF.

Impact of Lower Extremity Arthroplasty on Improvement of Quality-of-Life Outcomes Following Lumbar Fusion.

BACKGROUND: Both hip-spine and knee-spine syndromes can significantly impact a patient's quality of life; however, few studies have investigated their effect on postoperative outcomes following lumbar fusion. OBJECTIVE: Our study aimed to evaluate the impact of a prior lower extremity arthroplasty on the improvement of patient-reported outcome measures (PROMs) following lumbar fusion surgery. METHODS: Patients undergoing primary, single, or multilevel lumbar interbody fusion were retrospectively reviewed. Patients missing preoperative PROMs were excluded. PROMs were collected preoperatively and postoperatively and included the Oswestry Disability Index (ODI), 12-Item Short Form Physical Component Summary, Patient-Reported Outcomes Measurement Information System Physical Function, and visual analog scale (VAS). A minimum clinically important difference (MCID) was calculated. Patients were categorized based on a history of hip/knee arthroplasty and propensity score matched. Intragroup improvement of PROM scores and intergroup differences in mean scores were evaluated using a paired t test and linear regression. MCID achievement differences were evaluated using logistic regression. RESULTS: A total of 335 patients were included, with 25 having a history of hip/knee arthroplasty. Arthroplasty patients were significantly older (P = 0.001) and typically had a higher Charlson Comorbidity Index (P ≤ 0.003, both). Patients differed in spinal pathology of degenerative spondylolisthesis (P = 0.049). Nonarthroplasty patients demonstrated significant improvements in all PROMs by 2 years (P < 0.001, all). The arthroplasty group demonstrated significant improvements in all PROMs by 1 year (P < 0.031, all). Preoperative VAS back was significantly worse for nonarthroplasty patients (P = 0.035). MCID achievement did not significantly differ between groups except at 6 months for ODI (P = 0.035). CONCLUSION: Following lumbar fusion, patients with a past surgical history did not demonstrate differences in outcome measures or MCID from those without. These results suggest that comorbid orthopedic conditions requiring surgery do not negatively impact the ability of patients to improve following lumbar fusion. CLINICAL RELEVANCE: Prior surgical history of lower extremity arthroplasty should not discourage the use of lumbar fusion when properly indicated, as patients reported clinical improvement regardless of history of hip or knee arthroplasty.

How often does the preoperative plan for freehand pedicle screw placement match the actual surgical execution in adolescent idiopathic scoliosis?

PURPOSE: To provide a baseline for comparison with future advancements, this study determined the accuracy of preoperative planning of pedicle screw placement using standard radiographs for posterior fusion (PSF) for adolescent idiopathic scoliosis (AIS). METHODS: Ninety-five patients with AIS planned for PSF were prospectively enrolled. Preoperative planning was based upon standard upright posteroanterior and lateral radiographs. The planned number of screws to be placed at each level was recorded. Intraoperatively, all screws were placed by freehand technique. The number of successfully placed screws and the reasons for abandoning screw placement were documented. RESULTS: There were a total of 1783 pedicle screws planned preoperatively. The average planned implant density was 2.0 implants/vertebra. A total of 1723 (96.6%) of the planned screws were placed successfully. Fourteen (0.8%) screws were abandoned after attempted placement (range 0-2 screws/case). Of 241 screws planned in pedicles noted to be "hypoplastic," 13 resulted in the use of a hook or no instrumentation. The placement was not attempted for 49 (range 0-7/case) planned screws due to intraoperative decision-making and a sense that the curve was flexible enough not to require every screw. Three cases (3.2%) required instrumentation of an additional level. CONCLUSIONS: Standard spine radiographs allow for accurate preoperative planning for freehand pedicle screw placement in AIS. Ninety-seven percent of planned screws were placed successfully. The primary reason for deviation from the preoperative plan was intraoperative surgeon decision-making rather than difficulty with screw placement. This study will serve as a baseline when considering the utilization of navigation in PSF for AIS.

Abrogation of Rb Tumor Suppression Initiates GBM in Differentiated Astrocytes by Driving a Progenitor Cell Program.

Glioblastoma (GBM) remains lethal with no effective treatments. Despite the comprehensive identification of commonly perturbed molecular pathways, little is known about the disease's etiology, particularly in early stages. Several studies indicate that GBM is initiated in neural progenitor and/or stem cells. Here, we report that differentiated astrocytes are susceptible to GBM development when initiated by perturbation of the RB pathway, which induces a progenitor phenotype. In vitro and in vivo inactivation of Rb tumor suppression (TS) induces cortical astrocytes to proliferate rapidly, express progenitor markers, repress differentiation markers, and form self-renewing neurospheres that are susceptible to multi-lineage differentiation. This phenotype is sufficient to cause grade II astrocytomas which stochastically progress to GBM. Together with previous findings, these results demonstrate that cell susceptibility to GBM depends on the initiating driver.

Segmental range of motion after cervical total disc arthroplasty at long-term follow-up: a systematic review and meta-analysis.

OBJECTIVE: As an alternative procedure to anterior cervical discectomy and fusion, total disc arthroplasty (TDA) facilitates direct neural decompression and disc height restoration while also preserving cervical spine kinematics. To date, few studies have reported long-term functional outcomes after TDA. This paper reports the results of a systematic review and meta-analysis that investigated how segmental range of motion (ROM) at the operative level is maintained with long-term follow-up. METHODS: PubMed and MEDLINE were queried for all published studies pertaining to cervical TDA. The methodology for screening adhered strictly to the PRISMA guidelines. All English-language prospective studies that reported ROM preoperatively, 1 year postoperatively, and/or at long-term follow-up of 5 years or more were included. A meta-analysis was performed using Cochran's Q and I2 to test data for statistical heterogeneity, in which case a random-effects model was used. The mean differences (MDs) and associated 95% confidence intervals (CIs) were reported. RESULTS: Of the 12 studies that met the inclusion criteria, 8 reported the long-term outcomes of 944 patients with an average (range) follow-up of 99.86 (60-142) months and were included in the meta-analysis. There was no difference between preoperative segmental ROM and segmental ROM at 1-year follow-up (MD 0.91°, 95% CI -1.25° to 3.07°, p = 0.410). After the exclusion of 1 study from the comparison between preoperative and 1-year ROM owing to significant statistical heterogeneity according to the sensitivity analysis, ROM significantly improved at 1 year postoperatively (MD 1.92°, 95% CI 1.04°-2.79°, p < 0.001). However, at longer-term follow-up, the authors again found no difference with preoperative segmental ROM, and no study was excluded on the basis of the results of further sensitivity analysis (MD -0.22°, 95% CI -1.69° to -1.23°, p = 0.760). In contrast, there was a significant decrease in ROM from 1 year postoperatively to final long-term follow-up (MD -0.77°, 95% CI -1.29° to -0.24°, p = 0.004). CONCLUSIONS: Segmental ROM was found to initially improve beyond preoperative values for as long as 1 year postoperatively, but then ROM deteriorated back to values consistent with preoperative motion at long-term follow-up. Although additional studies with further longitudinal follow-up are needed, these findings further support the notion that cervical TDA may successfully maintain physiological spinal kinematics over the long term.

Dynamic changes in Holstein heifer circulatory stress biomarkers in response to lipopolysaccharide immune challenge.

Dairy cattle routinely face a variety of stressors. For example, climate change has resulted in more frequent heat stress events that increase the incidence of bacterial infections by inducing conditions like leaky gut syndrome, whereby the integrity of the intestinal epithelium is compromised allowing for luminal bacteria and their membrane components to infiltrate the host's bloodstream resulting in systemic inflammation. Lipopolysaccharide (LPS) is a well-characterized and biologically relevant microbe-associated molecular pattern (MAMP) that makes up the outer membrane of pathogenic and commensal Gram-negative bacteria and is known to contribute to inflammatory disorders including mastitis, acidosis and septicemia. In the present study, Holstein heifers (n = 20) were randomly allocated into different treatment groups receiving saline, 100, 200 or 400 ng/kg of LPS intramuscularly to create an experimentally induced endotoxemic state. Serum was collected hourly for 8 hr and then again at 24 hr to profile changes in circulatory stress biomarkers. All LPS -challenged animals demonstrated distinct cortisol responses 2 hr post-LPS challenge, and in the 200 ng/kg and 400 ng/kg of LPS treatments cortisol concentrations remained significantly induced for up to 4 hr. Rectal temperature was significantly increased for heifers challenged with 100 and 200 ng/kg of LPS at 2 and 4 hr as compared to their pre-challenge temperature. All LPS-challenged animals demonstrated marked leukopenia and thrombocytopenia as compared to the saline control animals. A total of 8 cytokines, including TNFα, and IL-10, were found to be induced between 2 and 4 hr. Finally, we report that miR-1246, miR-223, miR-29 and miR-31 were significantly induced in animals challenged with LPS as compared to the saline controls. The present study demonstrated that the stress response in dairy heifers is dynamic and there are peak windows of time when cortisol, cytokines and also miRNA are induced, and blood cells are sequestered as part of the systemic inflammatory response. Variability in the response to LPS warrants further investigation in dairy cattle to better understand the contribution of genetics and associations between LPS-induced stress and health and performance.

The diabetes community exercise programme plus usual care versus usual care in patients with type 2 diabetes: A randomised, two-arm, parallel, open-label trial.

Background: Exercise is important in type 2 diabetes (T2D) management. Focussing on Maori and Pacific people and those from deprived circumstances, the Diabetes Community Exercise Programme (DCEP) was developed to engage people with T2D in exercise. We report the evaluation of whether being offered DCEP (plus usual care) was more effective than usual care in improving glycaemic control at 1-year. Methods: A randomised, two-arm, parallel, open-label trial with blinding of outcome assessor and data analyst. Adults (age ≥35 years) with T2D recruited from two New Zealand (NZ) communities were randomised, using opaque sealed envelopes and stratified by centre with random block lengths, to DCEP or usual care. DCEP comprises twice-weekly, two-hour sessions of exercise and education over 12-weeks, followed by a twice-weekly maintenance exercise class. The primary outcome was between-group differences in mean changes of glycated haemoglobin (HbA1c) from baseline to 1-year follow-up with intention-to treat analysis. This trial is registered with the Australian NZ Clinical Trials Registry (ANZCTR): ACTRN12617001624370p and is closed to new participants. Findings: From 2018 - 2019, of 294 people screened, 165 (mean age 63·8, SD16·2 years, 56% female, 78·5% European, 14% Maori, 6% Pacific, 27% most deprived) were baseline evaluated, randomised, and analysed at study end (DCEP = 83, control = 82). Multimorbidity (≥2) and polypharmacy (>5 medications) were high (82%, 69%). We found no statistically significant between-groups differences in HbA1c (mmol/mol) change at 15 months (mean 3% higher in DCEP, 95% CI 2% lower to 8% higher, p = 0·23). Twelve-week intervention adherence was good (41% attended >80% available sessions). No adverse events were reported. Interpretation: DCEP was not effective in improving glycaemic control, possibly due to insufficient exercise intensity. Our attendance demonstrated DCEP's cultural accessibility. DCEP might be good to engage in exercise marginalised people with high Hb1Ac levels, multimorbidity, and high polypharmacy. Funding: Health Research Council of New Zealand.

Modeled Vegetation Community Trajectories: Effects from Climate Change, Atmospheric Nitrogen Deposition, and Soil Acidification Recovery.

Forest understory plant communities in the United States harbor most of the vegetation diversity of forests and are often sensitive to changes in climate and atmospheric deposition of nitrogen (N). As temperature increases from human-caused climate change and soils recover from long term atmospheric deposition of N and sulfur (S), it is unclear how these important ecosystem components will respond. We used the newly developed US-PROPS model - based on species response functions for over 1,500 species - to evaluate the potential impacts of atmospheric N deposition and climate change on species occurrence probability for a case study in the forested ecosystems of the Great Smoky Mountains National Park (GRSM), an iconic park in the southeastern United States. We evaluated six future scenarios from various combinations of two potential recoveries of soil pH (no change, +0.5 pH units) and three climate futures (no change, +1.5, +3.0 deg C). Species critical loads (CLs) of N deposition and projected responses for each scenario were determined. Critical loads were estimated to be low (< 2 kg N/ha/yr) to protect all species under current and expected future conditions across broad regions of GRSM and these CLs were exceeded at large spatial extents among scenarios. Northern hardwood, yellow pine, and chestnut oak forests were among the most N-sensitive vegetation map classes found within GRSM. Potential future air temperature conditions generally led to decreases in the maximum occurrence probability for species. Therefore, CLs were considered "unattainable" in these situations because the specified level of protection used for CL determination (i.e., maximum occurrence probability under ambient conditions) was not attainable. Although some species showed decreases in maximum occurrence probability with simulated increases in soil pH, most species were favored by increased pH. The importance of our study is rooted in the methodology described here for establishing regional CLs and for evaluating future conditions, which is transferable to other national parks in the U.S. and in Europe where the original PROPS model was developed.

Characteristics of wastewater originating from dental practices using predominantly mercury-free dental materials.

Dental materials are currently undergoing a revolution. Mercury use, including traditional amalgam (mercury-containing) material used in dental fillings, is now being widely regulated under the Minamata convention, and dental amalgam is currently being replaced by resin formulations in dentistry. These resin-based materials can be tuned to offer varying material properties by incorporation of a range of nano- and micro-particle based 'fillers' for different dental properties and applications. However, these innovations may have a concomitant effect on the waste streams associated with common dental applications, in particular the potential for higher concentrations of novel micro- and nanomaterials within wastewater streams, and a potential route for novel nanomaterials into the wider Environment. These new materials may also mean that wastewater filtering apparatus commonly deployed at present, such as amalgam separators, may be less efficient or insufficient to capture these new filler materials in dental facility wastewater. In this work, we analyse dental wastewater streams from three dental facilities in Ireland with differing amalgam separators in place. The potential overall toxicity, particulate load and physicochemical properties are analysed. The overall risk posed by these new materials is also discussed.

Novel Humanized Mesothelin-Expressing Genetically Engineered Mouse Models Underscore Challenges in Delivery of Complex Therapeutics to Pancreatic Cancers.

Antibody-based therapies designed for human use frequently fail to cross-react with the murine isoform of their target. Because of this problem, preclinical studies of antibody-based mesothelin (Msl)-targeted therapeutics in immunocompetent systems have been limited by the lack of suitable mouse models. Here, we describe two immunocompetent humanized mesothelin transgenic mouse lines that can act as tolerant hosts for C57Bl/6-syngeneic cell lines expressing the human isoform of mesothelin. Thyroid peroxidase (TPO) mice have thyroid-restricted human mesothelin expression. Mesothelin (Msl) mice express human mesothelin in the typical serosal membrane distribution and can additionally be utilized to assess on-target, off-tumor toxicity of human mesothelin-targeted therapeutics. Both transgenic strains shed human mesothelin into the serum like human mesothelioma and patients with ovarian cancer, and serum human mesothelin can be used as a blood-based surrogate of tumor burden. Using these models, we examined the on-target toxicity and antitumor activity of human mesothelin-targeted recombinant immunotoxins. We found that immunotoxin treatment causes acute and chronic histologic changes to serosal membranes in Msl mice, while human mesothelin-expressing thyroid follicular cells in TPO mice are resistant to immunotoxin despite excellent drug delivery. Furthermore, poor delivery of immunotoxin to syngeneic orthotopic human mesothelin-expressing pancreatic adenocarcinoma limits antitumor activity both alone and in combination with immune checkpoint inhibition. In summary, we have developed two high-fidelity, immunocompetent murine models for human cancer that allow for rigorous preclinical evaluation of human mesothelin-targeted therapeutics.