High-Throughput Transcriptomics Screen of ToxCast Chemicals in U-2 OS Cells.

New approach methodologies (NAMs) aim to accelerate the pace of chemical risk assessment while simultaneously reducing cost and dependency on animal studies. High Throughput Transcriptomics (HTTr) is an emerging NAM in the field of chemical hazard evaluation for establishing in vitro points-of-departure and providing mechanistic insight. In the current study, 1201 test chemicals were screened for bioactivity at eight concentrations using a 24-h exposure duration in the human- derived U-2 OS osteosarcoma cell line with HTTr. Assay reproducibility was assessed using three reference chemicals that were screened on every assay plate. The resulting transcriptomics data were analyzed by aggregating signal from genes into signature scores using gene set enrichment analysis, followed by concentration-response modeling of signatures scores. Signature scores were used to predict putative mechanisms of action, and to identify biological pathway altering concentrations (BPACs). BPACs were consistent across replicates for each reference chemical, with replicate BPAC standard deviations as low as 5.6 × 10-3 µM, demonstrating the internal reproducibility of HTTr-derived potency estimates. BPACs of test chemicals showed modest agreement (R2 = 0.55) with existing phenotype altering concentrations from high throughput phenotypic profiling using Cell Painting of the same chemicals in the same cell line. Altogether, this HTTr based chemical screen contributes to an accumulating pool of publicly available transcriptomic data relevant for chemical hazard evaluation and reinforces the utility of cell based molecular profiling methods in estimating chemical potency and predicting mechanism of action across a diverse set of chemicals.

Searching for LINCS to Stress: Using Text Mining to Automate Reference Chemical Curation.

Adaptive stress response pathways (SRPs) restore cellular homeostasis following perturbation but may activate terminal outcomes like apoptosis, autophagy, or cellular senescence if disruption exceeds critical thresholds. Because SRPs hold the key to vital cellular tipping points, they are targeted for therapeutic interventions and assessed as biomarkers of toxicity. Hence, we are developing a public database of chemicals that perturb SRPs to enable new data-driven tools to improve public health. Here, we report on the automated text-mining pipeline we used to build and curate the first version of this database. We started with 100 reference SRP chemicals gathered from published biomarker studies to bootstrap the database. Second, we used information retrieval to find co-occurrences of reference chemicals with SRP terms in PubMed abstracts and determined pairwise mutual information thresholds to filter biologically relevant relationships. Third, we applied these thresholds to find 1206 putative SRP perturbagens within thousands of substances in the Library of Integrated Network-Based Cellular Signatures (LINCS). To assign SRP activity to LINCS chemicals, domain experts had to manually review at least three publications for each of 1206 chemicals out of 181,805 total abstracts. To accomplish this efficiently, we implemented a machine learning approach to predict SRP classifications from texts to prioritize abstracts. In 5-fold cross-validation testing with a corpus derived from the 100 reference chemicals, artificial neural networks performed the best (F1-macro = 0.678) and prioritized 2479/181,805 abstracts for expert review, which resulted in 457 chemicals annotated with SRP activities. An independent analysis of enriched mechanisms of action and chemical use class supported the text-mined chemical associations (p < 0.05): heat shock inducers were linked with HSP90 and DNA damage inducers to topoisomerase inhibition. This database will enable novel applications of LINCS data to evaluate SRP activities and to further develop tools for biomedical information extraction from the literature.

Superior Ophthalmic Vein Thrombosis Associated With Asymptomatic COVID-19 Infection.

Superior ophthalmic vein thrombosis is a rare condition scarcely described in clinical literature with potentially severe consequences including permanent vision loss. This report details the case of a 70-year-old woman who presented with acute binocular horizontal diplopia, relative proptosis of the OD by 4 mm, and pain OD. On exam, visual acuity was 20/20 OD and 20/30 OS with full extraocular movements. CT revealed proptosis OD with a thrombosed superior ophthalmic varix. Evaluation for etiology of hypercoagulability was unremarkable, although the patient did have an asymptomatic COVID-19 infection 1 month prior. To the authors' knowledge, this is the first reported case of superior ophthalmic vein thrombosis secondary to an asymptomatic COVID-19 infection.

Variation of body weight supported treadmill training parameters during a single session can modulate muscle activity patterns in post-stroke gait.

Evidence supporting the benefits of locomotor training (LT) to improve walking ability following stroke are inconclusive and could likely be improved with a better understanding of the effects of individual parameters i.e., body weight support (BWS), speed, and therapist assistance and their interactions with walking ability and specific impairments. We evaluated changes in muscle activity of thirty-seven individuals with chronic stroke (> 6 months), in response to a single session of LT at their self-selected or fastest-comfortable speed (FS) with three levels of BWS (0%, 15%, and 30%), and at FS with 30% BWS and seven different combinations of therapist assistance at the paretic foot, non-paretic foot, and trunk. Altered Muscle Activation Pattern (AMAP), a previously developed tool in our lab was used to evaluate the effects of LT parameter variation on eight lower-extremity muscle patterns in individuals with stroke. Repeated-measures mixed-model ANOVA was used to determine the effects of speed, BWS, and their interaction on AMAP scores. The Wilcoxon-signed rank test was used to determine the effects of therapist-assisted conditions on AMAP scores. Increased BWS mostly improved lower-extremity muscle activity patterns, but increased speed resulted in worse plantar flexor activity. Abnormal early plantar flexor activity during stance decreased with assistance at trunk and both feet, exaggerated plantar flexor activity during late swing decreased with assistance to the non-paretic foot or trunk, and diminished gluteus medius activity during stance increased with assistance to paretic foot and/or trunk. Therefore, different sets of training parameters have different immediate effects on activation patterns of each muscle and gait subphases.

Walking-related knee contact forces and associations with knee pain across people with mild, moderate and severe radiographic knee osteoarthritis: a cross-sectional study.

OBJECTIVE: To investigate knee contact forces (KCFs), and their relationships with knee pain, across grades of radiographic knee osteoarthritis (OA) severity. DESIGN: Cross-sectional exploratory analysis of 164 participants with medial knee OA. Radiographic severity was classified as mild (grade 2), moderate (grade 3) or severe (grade 4) using the Kellgren & Lawrence (KL) scale. Walking knee pain was assessed using an 11-point numerical rating scale. External knee adduction moment (external KAM) and internal muscle forces were used to calculate medial, lateral and total KCFs using a musculoskeletal computational model. Force-time series across stance phase of gait were compared across KL grades using Statistical Parametric Mapping. Associations between KCFs and pain across KL grades were assessed using linear models. RESULTS: Medial KCFs during early and middle stance were higher in participants with KL3 and KL4 compared to those with KL2. In contrast, lateral KCFs were higher in those with KL2 compared to KL3 and KL4 in middle to late stance. The external loading component (i.e., KAM) of the medial KCF during middle to late stance was also greater in participants with KL3 and KL4 compared to those with KL2, whereas the internal (i.e., muscle) component was greater in those with KL3 and KL4 compared to KL3 during early stance. There were no associations between medial KCF and knee pain in any KL grade. CONCLUSIONS: Medial and lateral KCFs differ between mild, moderate and severe radiographic knee OA but are not associated with knee pain severity for any radiographic OA grade.

A qualitative study of fourteen African countries' nursing workforce and labour market.

AIM: The aim of this study was to describe factors affecting nursing education and labour markets in countries in East, Central, and Southern Africa, and critical areas for investment. BACKGROUND: An understanding about the relationship between the supply of nurses (determined by types of educational programmes, and the quantity and quality of nurse graduates), and workforce demand is critical to health policy development. METHODS: Six focus groups and 14 key informant interviews with nursing leaders and experts were conducted. Participants included government chief nursing officers, registrars of regulatory bodies, association leaders and heads of nursing education. The data were transcribed, coded and analysed using inductive techniques. FINDINGS: Participants discussed challenges and strengths of nursing education, school and regulatory infrastructure, financing mechanisms for the nursing workforce, the state of nursing jobs and scope of nursing practice. CONCLUSION: Strengthened regulations and leadership are needed to improve investment in nursing, the quality of nursing education, and working conditions and to promote the achievement of better health outcomes. IMPLICATIONS FOR NURSING POLICY: Clarifying scope of practice for nurses in the health sector and creating competency-based requirements is important. Governments should establish positions that align with updated competencies and provide fair and safe working conditions. The current and ongoing investment case for nursing requires improved data systems and a commitment to use labour market data for decision-making.

Cardiac transplantation outcomes in patients with amyloid cardiomyopathy.

OBJECTIVE: Amyloid cardiomyopathy (ACM) is a progressive and life-threatening disease caused by abnormal protein deposits within cardiac tissue. The most common forms of ACM are caused by immunoglobulin derived light chains (AL) and transthyretin (TTR). Orthotopic heart transplantation (OHT) remains the definitive treatment for patients with end stage heart failure. In this study, we perform a contemporary multicenter analysis evaluating post OHT survival in patients with ACM. METHODS: We conducted a multicenter analysis of 40,044 adult OHT recipients captured in the United Network for Organ Sharing (UNOS) registry from 1987-2018. Patients were characterized as ACM or non-ACM. Baseline characteristics were obtained, and summary characteristics were calculated. Outcomes of interest included post-transplant survival, infection, treated rejection, and the ability to return to work. Racial differences in OHT survival were also analyzed. Unadjusted associations between ACM and non-ACM survival were determined using the Kaplan-Meier estimations and confounding was addressed using multivariable Cox proportional hazards models. RESULTS: Three hundred ninety-eight patients with a diagnosis of ACM were identified of which 313 underwent heart only OHT. ACM patients were older (61 vs 53; P < .0001) and had a higher proportion of African Americans (30.7% vs 17.6%; P < .0001). Median survival for ACM was 10.2 years vs 12.5 years in non-ACM (P = .01). After adjusting for confounding, ACM patients had a higher likelihood of death post-OHT (HR 1.39 CI: 1.14, 1.70; P = .001). African American ACM patients had a higher likelihood of survival compared to White ACM patients (HR 0.51 CI 0.31-0.85; P = .01). No difference was observed in episodes of treated rejection (OR 0.63 CI 0.23, 1.78; P = .39), hospitalizations for infections (OR 1.24 CI: 0.85, 1.81; P = .26), or likelihood of returning to work for income (OR 1.23 CI: 0.84, 1.80; P = .30). CONCLUSIONS: In this analysis of OHT in ACM, ACM was associated with a higher likelihood of post-OHT mortality. Racial differences in post-OHT were observed with African American patients with ACM having higher likelihood of survival compared to White patients with ACM. No differences were observed in episodes of treated rejection, hospitalization for infection, or likelihood to return to work for income.

Histological Analysis of the Effect of ATX-101 (Deoxycholic Acid Injection) on Subcutaneous Fat: Results From a Phase 1 Open-Label Study.

BACKGROUND: ATX-101 is approved for submental fat reduction. OBJECTIVE: To characterize the histological effect of ATX-101 injection into subcutaneous fat. METHODS: This Phase 1 open-label study enrolled 14 adults to receive injections of ATX-101 into abdominal fat at varying concentrations (0.5%, 1.0%, 2.0%, or 4.0%), volumes (0.2 or 0.4 mL), spacing (0.7, 1.0, or 1.5 cm), and time points before scheduled abdominoplasty (1, 3, 7, or 28 days). During abdominoplasty, tissue was excised and preserved for histology. RESULTS: All injection paradigms resulted in histological changes confined to the subcutaneous layer, which were more prominent at higher concentrations and independent of volume and spacing. Key features at Day 1 after injection were adipocytolysis, blood vessel injury, neutrophilic inflammation, and lysis of locally present neutrophils. At Day 3, inflammation was reduced versus Day 1, and hemorrhage and lipid lake formation (at higher concentrations) were observed. Day 7 samples exhibited prominent adipocytolysis, mild inflammation, lipid-laden macrophages in the septae, and repair of vascular injury. At Day 28, inflammation was largely resolved and prominent features were septal thickening, neovascularization, and atrophy of fat lobules. CONCLUSION: Subcutaneous injection of ATX-101 induces adipocytolysis and local inflammation with septal thickening and resolution of inflammation by 28 days after injection.

The Effect of Frailty on Discharge Location for Medicare Beneficiaries After Acute Stroke.

OBJECTIVE: To examine the effect of frailty on poststroke discharge location with respect to stroke severity and create a risk-adjusted model for understanding the effects of frailty on discharge to an inpatient rehabilitation facility. DESIGN: Retrospective cohort. SETTING: A 2014 5% Medicare sample. PARTICIPANTS: Patients hospitalized for a first-time acute ischemic stroke (N=7258). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: A prehospitalization 6-month baseline was used to calculate a frailty score. Logistic regression to predict odds of discharge to inpatient rehabilitation was used to calculate for 3 levels of baseline frailty, controlling for patient demographics, stroke severity, and comorbidities. RESULTS: About 1603 patients were discharged to inpatient rehabilitation. Patients who were nonfrail (odds ratio [OR] 1.716; 95% confidence interval [95% CI], 1.463-2.013) or prefrail (OR 1.519; 95% CI, 1.296-1.779) were more likely to be discharged to inpatient rehabilitation. The final logistic regression model had a C-statistic of 0.63. Most of the patients discharged to inpatient rehabilitation were nonfrail (44.2%) and had moderate strokes (38.9%). Individuals who were frail and suffered a moderate (OR 0.78; 95% CI, 0.558-1.091) or severe stroke (OR 0.509; 95% CI, 0.358-0.721) were less likely to be discharged to an inpatient rehabilitation facility. CONCLUSIONS: A lack of a claims-based measure for prestroke functional ability makes it difficult to understand discharge decision-making patterns for individuals' poststroke. Prestroke frailty was found to have a significant effect on predicating inpatient rehabilitation discharge after an acute stroke when controlling for stroke severity, comorbidities, and age. Further investigation is warranted to examine differences in rehabilitation utilization based on frailty and to quantify the effect of rehabilitation on frailty status in individuals poststroke.

Dysphagia Requiring Nasogastric Feeding Following Orthognathic Surgery: An Unusual Complication, Case Report, Literature Review, and Recommendations.

PURPOSE: To report on dysphagia following orthognathic surgery, to review the literature on this topic, and to make recommendations for the management of this highly unusual problem. PATIENTS AND METHODS: Two cases of dysphagia requiring nasogastric feeding tubes following two-jaw surgery are presented adding to the two already reported in the English language. Similarities and possible causes are discussed and the management of this entity is proposed. RESULTS: Both of our patients required nasogastric feeding tubes be placed to facilitate enteral nutrition. Similar to cases reported in the literature, none of the patients had neurological deficits other than to the second and third division of the trigeminal nerve. All patients recovered the ability to swallow. CONCLUSIONS: While less than 2% of patients undergoing orthognathic surgery may report difficulty with swallowing following jaw surgery, most will recover in a short period of time. For those with prolonged dysphagia, a neurologic examination, a swallow study, and a nasogastric feeding may be required until normal swallowing returns.

A quantitative case study assessment of biophysical and economic effects from altering season of calving in temperate pasture-based dairy systems.

In theory, a late winter-early spring calving date in temperate grazing systems best matches pasture supply and herd demand, thereby minimizing the need for nonpasture feeds and maximizing profitability. We used a quantitative case study approach to define the effects of season of calving on biophysical and financial performance in a grazing system without the confounding effects of imported feeds (i.e., milk production directly from grazed pasture). A 2-yr production system experiment was established to quantify the effects of changing onset of seasonal calving (i.e., planned start of calving; PSC) from winter (July in the Southern Hemisphere) to spring (October), summer, (January), or autumn (April) on pasture and animal production and profitability. Eighty Holstein-Friesian cows were randomly allocated to 1 of 4 PSC treatments, each of which had a different PSC [mean calving date of January 10 (JAN), April 10 (APR), July 10 (JUL), or October 10 (OCT)]. Data were analyzed for consistency of treatment response over years using ANOVA procedures with year, PSC treatment, and year × PSC treatment interactions as fixed effects. Collated biological data and financial data extracted from a national economic database were used as fixed variables to model the financial performance for the different treatments. A stochastic risk analysis was undertaken, where historical pasture growth and milk price data were used to estimate the probability distributions for stochastic input variables. Gross farm revenue and operating profit per hectare were modeled under 2 scenarios: (A) milk price did not include a premium for milk supplied during the winter, and (B) milk price included a realistic premium for milk supplied in winter. Annual and seasonal pasture growth did not differ between treatments, but the pasture growth (kg of dry matter/ha) and profile of the JUL treatment best matched the lactation nutrient demand profile. In comparison, profiles for JAN, APR, and OCT calving treatments had periods of greater surplus and deficit due to the time of calving and herd demand relative to the pasture growth profile. As a result, the JAN and OCT treatments conserved more pasture as silage and cows consumed a larger proportion of their annual diet as silage. Although the amount of silage conserved and consumed did not differ between the JUL and APR calving treatments, the timing of the silage consumption was different, with silage making up a greater proportion of the diets in the APR treatment 1 to 90 and 91 to 180 d postcalving and being offered to the JUL calving treatment only 271 to 365 d postcalving. As a result of differences in the quantity and proportion of pasture and pasture silage in the lactating diet, the JUL treatment herd tended to produce greater milk, 4% fat-corrected milk, fat, protein, and lactose yields (kg/cow) than the other PSC treatments, which did not differ from each other. Operating expenses per hectare did not differ materially between calving date scenarios, but operating expenses per kilogram of fat-corrected milk and kilogram of fat and protein were 15 to 20% less in the JUL treatment. With or without a realistic winter milk premium, gross farm revenue and operating profit per hectare were greater in the JUL treatment than in the APR treatment, which had greater revenue and profitability than the remaining 2 calving date treatments. In summary, our results indicate that a PSC in late winter is most profitable in a grazing system not importing feed, with or without a realistic price incentive scheme.

First Results from CUORE: A Search for Lepton Number Violation via 0νßß Decay of

The CUORE experiment, a ton-scale cryogenic bolometer array, recently began operation at the Laboratori Nazionali del Gran Sasso in Italy. The array represents a significant advancement in this technology, and in this work we apply it for the first time to a high-sensitivity search for a lepton-number-violating process: ^{130}Te neutrinoless double-beta decay. Examining a total TeO_{2} exposure of 86.3 kg yr, characterized by an effective energy resolution of (7.7±0.5) keV FWHM and a background in the region of interest of (0.014±0.002) counts/(keV kg yr), we find no evidence for neutrinoless double-beta decay. Including systematic uncertainties, we place a lower limit on the decay half-life of T_{1/2}^{0ν}(^{130}Te)>1.3×10^{25} yr (90% C.L.); the median statistical sensitivity of this search is 7.0×10^{24} yr. Combining this result with those of two earlier experiments, Cuoricino and CUORE-0, we find T_{1/2}^{0ν}(^{130}Te)>1.5×10^{25} yr (90% C.L.), which is the most stringent limit to date on this decay. Interpreting this result as a limit on the effective Majorana neutrino mass, we find m_{ßß}<(110-520) meV, where the range reflects the nuclear matrix element estimates employed.

Dairy cow breed interacts with stocking rate in temperate pasture-based dairy production systems.

Economic optimum stocking rates for grazing dairy systems have been defined by accounting for the pasture production potential of the farm [t of dry matter (DM)/ha], the amount of feed imported from outside the farm (t of DM/ha), and the size of the cow (kg). These variables were combined into the comparative stocking rate [CSR; kg of body weight (BW)/t of feed DM available] measure. However, CSR assumes no effect of cow genetics beyond BW, and there is increasing evidence of within-breed differences in residual feed intake and between-breed differences in the gross efficiency with which cows use metabolizable energy for milk production. A multiyear production system experiment was established to determine whether Jersey (J) and Holstein-Friesian (HF) breeds performed similarly at the same CSR. Fifty-nine J cows and 51 HF cows were randomly allocated to 1 of 2 CSR in a 2 × 2 factorial arrangement; systems were designed to have a CSR of either 80 or 100 kg of BW/t of feed DM (J-CSR80, J-CSR100, HF-CSR80, and HF-CSR100 treatment groups). Data were analyzed for consistency of farmlet response over years using ANOVA procedures, with year and farmlet as fixed effects and the interaction of farmlet with year as a random effect. The collated biological data and financial data extracted from a national economic database were used to model the financial performance for the different breed and CSR treatments. On average, annual and individual season pasture DM production was greater for the J farmlets and was less in the CSR100 treatment; however, the effect of CSR was primarily driven by a large decline in pasture DM production in the HF-CSR100 treatment (breed × CSR interaction). This interaction in feed availability resulted in a breed × CSR interaction for the per-cow and per-hectare milk production variables, with HF cows producing more milk and milk components per cow in the CSR80 treatment but the same amount as the J cows in the CSR100 treatment. On a per-hectare basis, HF cows produced the same amount of 4% fat-corrected milk and lactose as J cows in the CSR80 treatment, but less fat; at CSR100, J cows produced more 4% fat-corrected milk, fat, and protein per hectare than HF cows. Our results support a greater gross efficiency for use of metabolizable energy by the J cow; 11% less total metabolizable energy was required to produce 1 kg of fat and protein at a system level. Economic modeling indicated that profitability of both breeds was less at CSR100, but the decline in profitability with increasing stocking rate was much greater in the HF breed. Holstein-Friesian cows were more profitable at CSR80 but were less profitable at CSR100.

Human gestation-associated tissues express functional cytosolic nucleic acid sensing pattern recognition receptors.

The role of viral infections in adverse pregnancy outcomes has gained interest in recent years. Innate immune pattern recognition receptors (PRRs) and their signalling pathways, that yield a cytokine output in response to pathogenic stimuli, have been postulated to link infection at the maternal-fetal interface and adverse pregnancy outcomes. The objective of this study was to investigate the expression and functional response of nucleic acid ligand responsive Toll-like receptors (TLR-3, -7, -8 and -9), and retinoic acid-inducible gene 1 (RIG-I)-like receptors [RIG-I, melanoma differentiation-associated protein 5 (MDA5) and Laboratory of Genetics and Physiology 2(LGP2)] in human term gestation-associated tissues (placenta, choriodecidua and amnion) using an explant model. Immunohistochemistry revealed that these PRRs were expressed by the term placenta, choriodecidua and amnion. A statistically significant increase in interleukin (IL)-6 and/or IL-8 production in response to specific agonists for TLR-3 (Poly(I:C); low and high molecular weight), TLR-7 (imiquimod), TLR-8 (ssRNA40) and RIG-I/MDA5 (Poly(I:C)LyoVec) was observed; there was no response to a TLR-9 (ODN21798) agonist. A hierarchical clustering approach was used to compare the response of each tissue type to the ligands studied and revealed that the placenta and choriodecidua generate a more similar IL-8 response, while the choriodecidua and amnion generate a more similar IL-6 response to nucleic acid ligands. These findings demonstrate that responsiveness via TLR-3, TLR-7, TLR-8 and RIG-1/MDA5 is a broad feature of human term gestation-associated tissues with differential responses by tissue that might underpin adverse obstetric outcomes.

Predicting the functional states of human iPSC-derived neurons with single-cell RNA-seq and electrophysiology.

Human neural progenitors derived from pluripotent stem cells develop into electrophysiologically active neurons at heterogeneous rates, which can confound disease-relevant discoveries in neurology and psychiatry. By combining patch clamping, morphological and transcriptome analysis on single-human neurons in vitro, we defined a continuum of poor to highly functional electrophysiological states of differentiated neurons. The strong correlations between action potentials, synaptic activity, dendritic complexity and gene expression highlight the importance of methods for isolating functionally comparable neurons for in vitro investigations of brain disorders. Although whole-cell electrophysiology is the gold standard for functional evaluation, it often lacks the scalability required for disease modeling studies. Here, we demonstrate a multimodal machine-learning strategy to identify new molecular features that predict the physiological states of single neurons, independently of the time spent in vitro. As further proof of concept, we selected one of the potential neurophysiological biomarkers identified in this study-GDAP1L1-to isolate highly functional live human neurons in vitro.

Production and economic responses to intensification of pasture-based dairy production systems.

Production from pasture-based dairy farms can be increased through using N fertilizer to increase pasture grown, increasing stocking rate, importing feeds from off farm (i.e., supplementary feeds, such as cereal silages, grains, or co-product feeds), or through a combination of these strategies. Increased production can improve profitability, provided the marginal cost of the additional milk produced is less than the milk price received. A multiyear production system experiment was established to investigate the biological and economic responses to intensification on pasture-based dairy farms; 7 experimental farmlets were established and managed independently for 3 yr. Paddocks and cows were randomly allocated to farmlet, such that 3 farmlets had stocking rates of 3.35 cows/ha (LSR) and 4 farmlets had stocking rates of 4.41 cows/ha (HSR). Of the LSR farmlets, 1 treatment received no N fertilizer, whereas the other 2 received either 200 or 400 kg of N/ha per year (200N and 400N, respectively). No feed was imported from off-farm for the LSR farmlets. Of the 4 HSR farmlets, 3 treatments received 200N and the fourth treatment received 400N; cows on 2 of the HSR-200N farmlet treatments also received 1.3 or 1.1 t of DM/cow per year of either cracked corn grain or corn silage, respectively. Data were analyzed for consistency of farmlet response over years using mixed models, with year and farmlet as fixed effects and the interaction of farmlet with year as a random effect. The biological data and financial data extracted from a national economic database were used to model the statement of financial performance for the farmlets and determine the economic implications of increasing milk production/cow and per ha (i.e., farm intensification). Applying 200N or 400N increased pasture grown per hectare and milk production per cow and per hectare, whereas increasing stocking rate did not affect pasture grown or milk production per hectare, but reduced milk production per cow. Importing feed in the HSR farmlets increased milk production per cow and per hectare. Marginal milk production responses to additional feed (i.e., either pasture or imported supplementary feed) were between 0.8 and 1.2 kg of milk/kg of DM offered (73 to 97 g of fat and protein/kg of feed DM) and marginal response differences between feeds were explained by metabolizable energy content differences (0.08 kg of milk/MJ of metabolizable energy offered). The marginal milk production response to additional feed was quadratic, with the greatest milk production generated from the initial investment in feed; 119, 99, and 55 g of fat and protein were produced per kilogram of feed DM by reducing the annual feed deficit from 1.6 to 1.0, 1.0 to 0.5, and 0.5 to 0 t of DM, respectively. Economic modeling indicated that the marginal cost of milk produced from pasture resulting from applied N fertilizer was less than the milk price; therefore, strategic use of N fertilizer to increase pasture grown increased farm operating profit per hectare. In comparison, operating profit declined with purchased feed, despite high marginal milk production responses. The results have implications for the strategic direction of grazing dairy farms, particularly in export-oriented industries, where the prices of milk and feed inputs are subject to the considerable volatility of commodity markets.

A 100-Year Review: A century of change in temperate grazing dairy systems.

From 1917 to 2017, dairy grazing systems have evolved from uncontrolled grazing of unimproved pastures by dual-purpose dairy-beef breeds to an intensive system with a high output per unit of land from a fit-for-purpose cow. The end of World War I signaled significant government investments in agricultural research institutes around the world, which coincided with technological breakthroughs in milk harvesting and a recognition that important traits in both plants and animals could be improved upon relatively rapidly through genetic selection. Uptake of milk recording and herd testing increased rapidly through the 1920s, as did the recognition that pastures that were rested in between grazing events yielded more in a year than those continuously grazed. This, and the invention and refinement of the electric fence, led to the development of "controlled" rotational grazing. This, in itself, facilitated greater stocking rates and a 5 to 10% increase in milk output per hectare but, perhaps more importantly, it allowed a more efficient use of nitrogen fertilizer, further increasing milk output/land area by 20%. Farmer inventions led to the development of the herringbone and rotary milking parlors, which, along with the "unshortable" electric fence and technological breakthroughs in sperm dilution rates, allowed further dairy farm expansion. Simple but effective technological breakthroughs in reproduction ensured that cows were identified in estrus early (a key factor in maintaining the seasonality of milk production) and enabled researchers to quantify the anestrus problem in grazing herds. Genetic improvement of pasture species has lagged its bovine counterpart, but recent developments in multi-trait indices as well as investment in genetic technologies should significantly increase potential milk production per hectare. Decades of research on the use of feeds other than pasture (i.e., supplementary feeds) have provided consistent milk production responses when the reduction in pasture intake associated with the provision of supplementary feed (i.e., substitution rate) is accounted for. A unique feature of grazing systems research over the last 70 yr has been the use of multi-year farm systems experimentation. These studies have allowed the evaluation of strategic changes to a component of the system on all the interacting features of the system. This technique has allowed excellent component research to be "systemized" and is an essential part of the development of the intensive grazing production system that exists today. Future challenges include the provision of skilled labor or specifically designed automation to optimize farm management and both environmental sustainability and animal welfare concerns, particularly relating to the concentration of nitrogen in each urine patch and the associated risk of nitrate leaching, as well as concerns regarding exposure of animals to harsh climatic conditions. These combined challenges could affect farmers' "social license" to farm in the future.

Evidence-Based Medicine and Regulatory Oversight: Opioid Prescribing for Treatment of Pain.

It is no secret that our country is in the midst of an opioid crisis. With hundreds of Americans dying everyday due to opioid use, it is hard to ignore. Closely related, though not as often talked about, is another crisis providers are facing: chronic pain and its treatment. The treatment approach to this problem is multifaceted. Evidence-based studies are the hallmark of appropriate treatment; however, increasing patient concerns and regulatory oversight have added an additional layer that physicians and other prescribers have to incorporate. Navigating a balance between evidence-based treatment, regulatory pressures, and patient concerns is now at the forefront of pain management and patient care.

Knee kinematics and kinetics are associated with early patellofemoral osteoarthritis following anterior cruciate ligament reconstruction.

OBJECTIVE: Patellofemoral osteoarthritis (PFOA) commonly occurs following anterior cruciate ligament reconstruction (ACLR). Our study aimed to compare knee kinematics and kinetics during a hop-landing task between individuals with and without early PFOA post-ACLR. DESIGN: Forty-five individuals (mean ± SD 26 ± 5 years) 1-2 years post-ACLR underwent 3T isotropic MRI scans and 3D biomechanical assessment of a standardised forward hop task. Knee kinematics (initial contact, peak, excursion) in all three planes and sagittal plane kinetics (peak) were compared between 15 participants with early PFOA (MRI-defined patellofemoral cartilage lesion) and 30 participants with no PFOA (absence of patellofemoral cartilage lesion on MRI) using analysis of covariance (ANCOVA), adjusted for age, BMI, sex and the presence of early tibiofemoral OA. RESULTS: Compared to participants without PFOA, those with early PFOA exhibited smaller peak knee flexion angles (mean difference, 95% confidence interval [CI]: -5.2°, -9.9 to -0.4; P = 0.035) and moments (-4.2 Nm/kg.m, -7.8 to -0.6; P = 0.024), and greater knee internal rotation excursion (5.3°, 2.0 to 8.6; P = 0.002). CONCLUSIONS: Individuals with early PFOA within the first 2-years following ACLR exhibit distinct kinematic and kinetic features during a high-load landing task. These findings provide new information regarding common post-ACLR biomechanical patterns and PFOA. Since management strategies, such as altering knee load, are more effective during the early stages of disease, this knowledge will help to inform clinical management of early PFOA post-ACLR.