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Case report This report documents an outbreak of acute renal failure in a flock of 9-month-old White Suffolk-cross ewe lambs. The mortality rate was moderate but the morbidity was high. The lambs had grazed a wheat stubble paddock in which there were several weed species, including Amaranthus spp. Clinical significance Renal failure in livestock has been reported internationally following the ingestion of Amaranthus spp. and although it has previously been suspected in Australia no cases have been documented. It is important for Australian veterinarians and farmers to realise the toxic potential of this group of plants. [ABSTRACT FROM AUTHOR]

Case report An outbreak of photosensitisation affecting approximately 25% of a flock of 120 meat lambs that was grazing a monoculture of the pasture legume Biserrula pelecinus var. Casbah (biserrula) was investigated. Blood samples were taken from sheep with moderate to severe clinical signs, and from apparently normal animals, for a complete blood count and biochemistry; 5 affected animals were subjected to a full necropsy. Histopathological investigation showed lesions consistent with photosensitisation of the exposed unpigmented skin of the face and ears. No histopathological or clinical pathological abnormalities suggestive of a hepatopathy were detected in any of the cases, indicating that the lesions observed in this flock were caused by a primary photosensitising agent present in B. pelecinus. Conclusion This is the first confirmation that photosensitisation caused by ingestion of biserrula is caused by a primary photosensitising agent. [ABSTRACT FROM AUTHOR]

Background: Embryonic stem (ES) cells can differentiate into all cell types and have been used extensively to study factors affecting neuronal differentiation. ES cells containing mutations in known genes have the potential to provide useful in vitro models for the study of gene function during neuronal differentiation. Recently, mouse ES cell lines lacking the neurogenic transcription factor Pax6 were reported; neurons derived from these Pax6-/- ES cells died rapidly after neuronal differentiation in vitro.Results: Here we report the derivation of new lines of Pax6-/- ES cells and the assessment of their ability to survive and differentiate both in vitro and in vivo. Neurons derived from our new Pax6-/- lines were viable and continued to elaborate processes in culture under conditions that resulted in the death of neurons derived from previously reported Pax6-/- ES cell lines. The new lines of Pax6-/-ES cells showed reduced neurogenic potential, mimicking the effects of loss of Pax6 in vivo. We used our new lines to generate Pax6-/- <--> Pax6+/+ chimeras in which the mutant cells survived and displayed the same phenotypes as Pax6-/- cells in Pax6-/- <--> Pax6+/+ chimeras made by embryo aggregation.Conclusions: We suggest that loss of Pax6 from ES cells reduces their neurogenic capacity but does not necessarily result in the death of derived neurons. We offer these new lines as additional tools for those interested in the generation of chimeras and the analysis of in vitro ES cell models of Pax6 function during neuronal differentiation, embryonic and postnatal development. [ABSTRACT FROM AUTHOR]

Waste-to-energy systems can provide a functional demonstration of the economic and environmental benefits of circularity, innovation, and reimagining existing systems. This study offers a robust quantification of the greenhouse gas (GHG) emission reduction potential of the adoption of anaerobic digestion (AD) technology on applicable large-scale dairy farms in the contiguous United States. GHG reduction estimates were developed through a robust life cycle modeling framework paired with sensitivity and uncertainty analyses. Twenty dairy configurations were modeled to capture important differences in housing and manure management practices, applicable AD technologies, regional climates, storage cleanout schedules, and methods of land application. Monte Carlo results for the 90% confidence interval illustrate the potential for AD adoption to reduce GHG emissions from the large-scale dairy industry by 2.45-3.52 MMT of CO 2 -eq per year considering biogas use only in renewable natural gas programs and as much as 4.53-6.46 MMT of CO 2 -eq per year with combined heat and power as an additional biogas use case. At the farm level, AD technology may reduce GHG emissions from manure management systems by 58.1-79.8% depending on the region. Discussion focuses on regional differences in GHG emissions from manure management strategies and the challenges and opportunities surrounding AD adoption.

The seamless adoption of electric vehicles (EVs) in the United States necessitates the development of extensive and effective charging infrastructure. Various charging systems have been proposed, including Direct Current Fast Charging, Battery Swapping, and Dynamic Wireless Power Transfer. While many studies have evaluated the charging costs and greenhouse gas (GHG) intensity of EVs, a comprehensive analysis comparing these systems and their implications across vehicle categories remains unexplored. This study compares the total cost of ownership (TCO) and GHG-intensity of EVs using these charging systems. Based on nationwide infrastructure deployment simulations, the change to TCO from adopting EVs varies by scenario, vehicle category, and location, with local fuel prices, electricity prices, and traffic volumes dramatically impacting results. Further, EV GHG-intensity depends on local electricity mixes and infrastructure utilizations. This research highlights the responsiveness of EV benefits resulting from technology advancements, deployment decisions, and policymaking., (© 2024. The Author(s).)

Sustainable fuel initiatives in the United States such as the Environmental Protection Agency's Renewable Fuel Standard and the Department of Energy's Sustainable Aviation Fuel Grand Challenge have increased the production of corn ethanol and soybean biodiesel. However, the lack of precise information regarding biomass sourcing at a localized level has hindered accurate understanding of both biofuel costs and environmental impact of these production pathways. By harnessing the power of geospatial analysis and leveraging United States Department of Agriculture (USDA) crop census data, this dataset fills this critical knowledge gap. This dataset offers a novel estimation of geospatial biomass sourcing for biofuel production in the United States by synthesizing 2017 USDA crop census data, biorefinery data from the United States Energy Information Administration, and publicly available information about biomass sourcing for biofuel production. This dataset provides a detailed understanding of biomass use for first generation biofuel production, enabling stakeholders to make informed decisions about resource allocation, investment strategies, and infrastructure development. Furthermore, the county-level granularity of the dataset allows for increased fidelity in the techno-economic assessments and life-cycle analyses of first-generation biofuels in the United States., (© 2024 The Authors. Published by Elsevier Inc.)

Objective: Reimbursement trends for common procedures have persistently declined over the past 2 decades. Spinal instrumentational and fusion procedures are increasingly utilized and have increased in clinical complexity, yet longitudinal inflation-adjusted data for Medicare reimbursements of these procedures have not been evaluated., Methods: The Centers for Medicare and Medicaid Services (CMS) Physician Fee Schedule Look-Up Tool was used to extract Medicare reimbursements for the 5 most common spinal procedures and associated instrumentations from 2000-2020. Current Procedural Terminology (CPT) codes include 22551, 22600, 22633, 63030, and 63047 as well as instrumentation CPT codes 22840 and 22842-6. The nominal values were adjusted for inflation according to the latest consumer price index (U.S. Bureau of Labor Statistics; reported as 2020 USD) and used to calculate average annual percent changes and compound annual growth rates (CAGRs) in reimbursements., Results: After inflation adjustment, the physician fee reimbursement decreased by 11.05% ± 8.46% (mean ± s.d., from $2,009.89 in 2011 to $1,787.85 in 2020) for anterior cervical discectomy and fusion (ACDF), 28.38% ± 8.42% (from $1,889.38 in 2000 to $1,353.14 in 2020) for posterior cervical fusion, 7.85% ± 8.20% (from $2,111.20 in 2012 to $1,945.49 in 2020) for transforaminal lumbar interbody fusion (TLIF), 28.17% ± 13.88% (from $1,421.78 in 2000 to $1,021.22 in 2020) for lower back disc surgery, and 31.88% ± 8.22% (from $1,700.38 in 2000 to $1,158.25 in 2020) for lumbar laminectomy. Instrumentation reimbursements showed an average decrease of 33.43% ± 8.4% over this period. Average CAGR was -1.7% ± .41% for procedures and -2.02% ± .14% for instrumentation., Conclusion: Our analysis reveals a persistent decline in reimbursement rates of the most common spine procedures and instrumentation since the year 2000. If unaddressed, this trend can serve as a substantial disincentive for physicians to perform these procedures and can significantly limit access to spinal care at the population level., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objective: The current Roussouly classification identifies four groups of "normal" sagittal spine morphology, which has greatly expanded the understanding of normal heterogeneity of the spine. While there has been extensive characterization of the influence of spinopelvic parameters on outcomes after degenerative spine surgery, the influence of spinopelvic parameters on thoracolumbar trauma has yet to be described. The goal of this study was to determine if spinopelvic parameters and global spine morphology influence fracture location, fracture morphology, and rate of neurological deficit in the setting of thoracolumbar trauma., Methods: Of 2896 patients reviewed in the authors' institutional spine database between January 2014 and April 2020 with an ICD-9/10 diagnosis of thoracolumbar trauma, 514 met the inclusion criteria of acute thoracolumbar fracture on CT and visible femoral heads on sagittal CT. Pelvic incidence (PI) was calculated on sagittal CT. Demographic and clinical data including age, sex, BMI, smoking status, concomitant cervical fracture, mechanism of injury, major fracture location, neurological deficit, AO Spine thoracolumbar injury classification, and management type (operative vs nonoperative) were collected. Patients were stratified into high-PI (≥ 50°) and low-PI (< 50°) groups., Results: Patients with high PI had a lower incidence of fractures in the lower lumbar spine (below L2) compared with patients with low PI (16% vs 8%, p < 0.01). The last lordotic vertebrae were observed between T10 and L4, and of fractures that occurred at these levels, 75% were at the last lordotic vertebrae. Fall from height was the most common cause of neurological deficit, accounting for 47%. Of the patients presenting with a fall from height, AO Spine type B distraction injuries were more common in the high-PI group (41% vs 18%, p = 0.01). Similarly, within the same subgroup, AO Spine type A compression injuries were more common in the low-PI group (73% vs 53%, p = 0.01)., Conclusions: Spinopelvic parameters and sagittal balance influence the location and morphology of thoracolumbar fractures. Fractures of the thoracolumbar junction are strongly associated with the inflection point, which is defined by sagittal alignment. While the importance of considering sagittal balance is known for decision-making in degenerative spinal pathology, further studies are required to determine if spinopelvic parameters and sagittal balance should play a role in the decision-making for management of thoracolumbar fractures.

Techno-economic analyses (TEAs) and life cycle assessments (LCAs) of algal biofuels often focus on locations in suboptimal latitudes for algal cultivation, which can under-represent the sustainability potential of the technology. This study identifies the optimal global productivity potential, environmental impacts, and economic viability of algal biofuels by using validated biophysical and sustainability modeling. The biophysical model simulates growth rates of Scenedesmus obliquus based on temperature, photoinhibition, and respiration effects at 6685 global locations. Region-specific labor costs, construction factors, and tax rates allow for spatially resolved TEA, while the LCA includes regional impacts of electricity, hydrogen, and nutrient markets across ten environmental categories. The analysis identifies optimal locations for algal biofuel production in terms of environmental impacts and economic viability which are shown to follow biomass yields. Modeling results highlight the global variability of productivity with maximum yields ranging between 24.8 and 27.5 g m -2 d -1 in equatorial regions. Environmental impact results show favorable locations tracked with low-carbon electricity grids, with the well-to-wheels global warming potential (GWP) ranging from 31 to 45 g CO 2eq MJ -1 in South America and Central Africa. When including direct land use change impacts, the GWP ranged between 44 and 55 g CO 2eq MJ -1 in these high-productivity regions. Low-carbon electricity also favors air quality and eutrophication impacts. The TEA shows that minimum algal fuel prices of $1.89-$2.15 per liter of gasoline-equivalent are possible in southeast Asia and Venezuela. This discussion focuses on the challenges and opportunities to reduce fuel prices and the environmental impacts of algal biofuels in various global regions.

Study Design: Case report., Purpose: The purpose of this case report is to illustrate the utility of ECMO as possible treatment in the management of perioperative massive pulmonary embolism (PE) during spine surgery. We present a case report of an intra-operative massive PE managed with ECMO and review relevant literature of perioperative massive PE. Treatment poses substantial risk given the need for anticoagulation. ECMO has not previously been described in the management of this condition in spine surgery., Methods: The patient is a 75-year-old male with hypertension, diabetes mellitus type II, chronic obstructive pulmonary disease, and prior cerebral infarction with minimal residual deficits who presented for Lumbar 2-Sacral 1 (L2-S1) revision posterior spinal decompression and fusion with L3-4 TLIF (Transforaminal Lumbar Interbody Fusion). The operation was routine until removing him from the operative table when he suffered a massive pulmonary embolism., Results: The patient became extremely unstable hemodynamically requiring massive doses of three vasopressors. He was too unstable for additional imaging or to transport for mechanical thrombectomy so with the guidance of our cardiologist we initiated ECMO. The patient ultimately recovered well and was discharged from the hospital at his neurological baseline and preserved cardiac function., Conclusion: ECMO is a viable option for management of acute perioperative massive pulmonary embolism when less invasive treatments are not sufficient., (© 2023. The Author(s), under exclusive licence to Scoliosis Research Society.)

While algal biofuels have the potential to reduce the national reliance on fossil fuels, high water consumption associated with algal biomass cultivation represents a major concern potentially compromising the sustainable commercialization of this technology. This study focuses on quantifying the water footprint (WF) and water scarcity footprint (WSF) of renewable diesel derived from algal biomass and provides insights into where algal cultivation is less water-intensive than traditional ethanol and biodiesel feedstocks. Results are generated with an engineering process model developed to predict the life-cycle water consumption, considering green, blue, and gray water, of algae facilities across the United States at a high spatiotemporal resolution. The total WFs for Florida and Arizona are determined to be 13.1 and 17.6 m 3 GJ -1 , respectively. The blue WF in Arizona is shown to be 8.5 times larger than in Florida, while the green WF is 4.5 times smaller, but when combined into a total WF, there is just a 26% difference between the two locations. The analysis reveals that the total life-cycle WFs of algal renewable diesel are smaller than the optimal WFs of corn ethanol and soybean biodiesel. Algal systems benefit from higher growth rates and offer the opportunity to manage wastewater streams, therefore generating smaller green and gray WFs than those of conventional biofuels. The WSF analysis identifies the Gulf Coast as the most suitable region for algal cultivation, with cultivation in the western US shown to exacerbate local water stress levels.

Currently, there is no standardized rearing method or production guidelines for non-replacement male dairy calves that maximizes their economic viability. Producers have highlighted the need to match consumer expectations, but even with broadscale welfare improvement across the dairy industry, challenges remain at providing reliable and valuable pathways for non-replacement male dairy calves for beef production. A key consumer concern has been the use of on-farm euthanasia. Euthanasia has been a catalyst for change in the industry from a human and animal welfare perspective. The practice of euthanasia can lead to a decline in personnel wellbeing. To investigate the relationship between on-farm management practices of non-replacement male dairy calves and producer perceptions of their value proposition, an online questionnaire was provided to Australian dairy producers between June and October 2021. The aim was to identify supply-chain profitability of non-replacement male calves and investigate the attitudes and effects of euthanasia on producer wellbeing as part of managing these calves. A total of 127 useable responses were obtained, and a Bayesian network (BN) was utilized to model the interdependencies between management practices and wellbeing among participants. The results indicated that in general, dairy producers desired high welfare standards in their enterprises with regard to non-replacement male calves as well as expressed a desire to meet industry and consumers' expectations. In line with anecdotal reports of a reduction in practice, euthanasia was not identified as common practice in this group; however, producers were still accessing early-life markets for non-replacement male calves with operational requirements and environmental factors influencing their decisions. Producers expressed dissatisfaction with market access for their calves, as well as the lack of suitability of Australian beef grading standards for dairy-bred carcasses. Australian dairy managers and owners identified that euthanasia influenced employee wellbeing; however, they did not acknowledge euthanasia had an effect on their own wellbeing. Overall, the findings of this study indicate that all non-replacement male calf breeds had the potential to access profitable markets, and avoidance of euthanasia is a strong driver of change among dairy beef production systems in Australia., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Vicic, Saliba, Campbell, Xie and Quinn.)

Qualitative and quantitative PCR-based tests are widely used in both diagnostics and research to assess the prevalence of disease-causing pathogens in veterinary medicine. The efficacy of these tests, usually measured in terms of sensitivity and specificity, is critical in confirming or excluding a clinical diagnosis. We undertook a meta-analysis to assess the inherent value of published PCR diagnostic approaches used to confirm and quantify bacteria and viruses associated with bovine respiratory disease (BRD) in cattle. This review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A thorough search of nine electronic databases (Web of Science, EBSCOhost, Cambridge journals online, ProQuest, PubMed, Sage journals online, ScienceDirect, Wiley online library and MEDLINE) was undertaken to find studies that had reported on the use of PCR and/or qPCR for the detection and/or quantification of BRD associated organisms. All studies meeting the inclusion criteria for reporting quantitative PCR for identification of BRD associated microorganisms were included in the analysis. Studies were then assessed on the applications of the Minimum Information for Publication of Quantitative Real-Time PCR Experiment (MIQE) and PCR primer/probe sequences were extracted and tested for in silico specificity using a high level of stringency. Fourteen full-text articles were included in this study. Of these, 79% of the analysed articles did not report the application of the MIQE guidelines in their study. High stringency in silico testing of 144 previously published PCR primer/probe sequences found many to have questionable specificity. This review identified a high occurrence of primer/probe sequences with a variable in silico specificity such that this may have implications for the accuracy of reporting. Although this analysis was only applied to one specific disease state, identification of animals suspected to be suffering from bovine respiratory disease, there appears to be more broadly a need for veterinary diagnostic studies to adopt international best practice for reporting of quantitative PCR diagnostic data to be both accurate and comparable between studies and methodologies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Barnewall, Marsh and Quinn.)

Sexual dysfunction is a common toxicity and detrimental for the quality of life of women treated with chemoradiotherapy for anal cancer. Sexual dysfunction occurs because the vagina is closely approximated to the anal canal and typically receives substantial doses of radiation. Strategies for mitigation have largely been focused on posttreatment therapy and symptom management. The use of daily vaginal dilator placement during radiotherapy to mitigate dose to the vagina has been previously explored with modest gains, while proton therapy is under active investigation for the treatment of anal cancer. Use of proton therapy for anal cancer reduces dose to some organs at risk but may inadvertently increase vaginal toxicity if the proton beam terminates in the vaginal tissue. Herein, we present the case histories of 2 women treated for squamous cell carcinoma of the anal canal with the novel combination of intensity-modulated proton therapy and daily vaginal dilator placement to maximally reduce dose to the vagina and protect it from areas of increased energy deposition at the end of the proton range., Competing Interests: Conflicts of Interest: The authors have no relevant conflicts of interest to disclose., (©Copyright 2022 The Author(s).)

Male non-replacement calves in dairy systems represent an underutilized economic resource for dairy producers worldwide. Despite this, increasing the practice of rearing non-replacement male calves has significant barriers both in on-farm adoption and practice. Poor neonatal rearing practices, higher levels of morbidity and mortality, and disaggregated production pathways with multiple points of handling, have all been described as barriers to adoption of surplus calf production. To identify the critical decision-determining challenges associated with broader adoption of raising non-replacement stock, and to investigate the whole-of-value chain issues faced by dairy producers to rear non-replacement male calves, we undertook a series of semi-structured interviews with Australian dairy producers to interrogate their key challenges. To achieve this, a constructivist grounded theory approach was used to inform the process of analysis of in-depth interviews with Australian dairy producers regarding their current practices and perceptions. Five major themes emerged from these conversations that were key barriers to on-farm non-replacement calf rearing in the producer group participants. These were: impacts of drought on cost and availability of feed for these calves and the whole herd; the management requirements of non-replacement male calves as an additional workload to that of their current operation; their attitudes and current practices to and surrounding euthanasia; perceived ease of supply-chain access for these calves, and their perceptions of the economic value of dairy-beef product as a return on investment. Understanding the barriers to adoption of non-replacement calf rearing, and addressing the value proposition for dairy beef, can assist increased uptake of non-replacement calf rearing. These findings will allow development of strategies to address these barriers, and extension of viable management strategies to increase adoption of profitable business practices surrounding non-replacement male calf production., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Vicic, Saliba, Campbell and Quinn.)

Water scarcity footprint (WSF) is a recent addition to life cycle assessment methodology that has advanced the understanding of freshwater environmental impact. The Available Water Remaining (AWARE) method is one approach that has gained significant traction in WSF applications. While an effective method for determining WSF, the methodology has limitations that constrain capabilities for determining freshwater environmental impact in arid regions. The primary limitation is the inability to compare regions when more water demand exists than what is available which typically occurs in arid regions. This limitation reduces resolution and therefore decision-making capabilities. This work proposes a novel method for determining WSF in arid regions by capturing and quantifying scarcity when water demand is greater than availability. The approach presented here, called the demand to availability (DTA) method, is intended to be used for small-scale, or subregion analyses in areas where truncation occurs using standard AWARE methods. With the regional specificity, unique characterization factors can be developed to enhance deterministic resolution and ultimately improve decision-making abilities. The DTA methods are presented universally, allowing for application and implementation to any region. A case study was developed to demonstrate the effectiveness of the DTA method by analyzing characterization factors (CFs) and alfalfa WSFs in the arid Southwestern United States. Using the standard AWARE methods, this region originally truncated 38% of counties resulting in zero resolution or decision-making abilities. Results of the case study that used the proposed DTA method show an improved resolution in 100% of these counties, both within CF and alfalfa WSF. Although the proposed method is an improvement for understanding WSFs in arid regions, limitations and constraints still exist and are discussed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Nitrogen and sulfur emissions from oil sands operations in northern Alberta, Canada have resulted in increasing deposition of N and S to the region's ecosystems. To assess whether a changing N and S deposition regime affects bog porewater chemistry, we sampled bog porewater at sites at different distances from the oil sands industrial center from 2009 to 2012 (10-cm intervals to a depth of 1 m) and from 2009 to 2019 (top of the bog water table only). We hypothesized that: (1) as atmospheric N and S deposition increases with increasing proximity to the oil sands industrial center, surface porewater concentrations of NH 4 + , NO 3 - , DON, and SO 4 2- would increase and (2) with increasing N and S deposition, elevated porewater concentrations of NH 4 + , NO 3 - , DON, and SO 4 2- would be manifested increasingly deeper into the peat profile. We found weak evidence that oil sands N and S emissions affect bog porewater NH 4 + -N, NO 3 - -N, or DON concentrations. We found mixed evidence that increasing SO 4 2- deposition results in increasing porewater SO 4 2- concentrations. Current SO 4 2- deposition, especially at bogs closest to the oil sands industrial center, likely exceeds the ability of the Sphagnum moss layer to retain S through net primary production, such that atmospherically deposited SO 4 2- infiltrates downward into the peat column. Increasing porewater SO 4 2- availability may stimulate dissimilatory sulfate reduction and/or inhibit CH 4 production, potentially affecting carbon cycling and gaseous fluxes in these bogs., (© 2021. The Author(s).)

Australia has over 30 Panicum spp. (panic grass) including several non-native species that cause crop and pasture loss and hepatogenous photosensitisation in livestock. It is critical to correctly identify them at the species level to facilitate the development of appropriate management strategies for efficacious control of Panicum grasses in crops, fallows and pastures. Currently, identification of Panicum spp. relies on morphological examination of the reproductive structures, but this approach is only useful for flowering specimens and requires significant taxonomic expertise. To overcome this limitation, we used multi-locus DNA barcoding for the identification of ten selected Panicum spp. found in Australia. With the exception of P. buncei, other native Australian Panicum were genetically separated at the species level and distinguished from non-native species. One nuclear (ITS) and two chloroplast regions (matK and trnL intron-trnF) were identified with varying facility for DNA barcode separation of the Panicum species. Concatenation of sequences from ITS, matK and trnL intron-trnF regions provided clear separation of eight regionally collected species, with a maximum intraspecific distance of 0.22% and minimum interspecific distance of 0.33%. Two of three non-native Panicum species exhibited a smaller genome size compared to native species evaluated, and we speculate that this may be associated with biological advantages impacting invasion of non-native Panicum species in novel locations. We conclude that multi-locus DNA barcoding, in combination with traditional taxonomic identification, provides an accurate and cost-effective adjunctive tool for further distinguishing Panicum spp. at the species level., (© 2021. The Author(s).)

Background: Quantitative PCR (qPCR) aims to measure the DNA or RNA concentration in diagnostic and biological samples based on the quantification cycle (Cq) value observed in the amplification curves. Results of qPCR experiments are regularly calculated as if all assays are 100% efficient or reported as just Cq, ΔCq, or ΔΔCq values., Contents: When the reaction shows specific amplification, it should be deemed to be positive, regardless of the observed Cq. Because the Cq is highly dependent on amplification efficiency that can vary among targets and samples, accurate calculation of the target quantity and relative gene expression requires that the actual amplification efficiency be taken into account in the analysis and reports. PCR efficiency is frequently derived from standard curves, but this approach is affected by dilution errors and hampered by properties of the standard and the diluent. These factors affect accurate quantification of clinical and biological samples used in diagnostic applications and collected in challenging conditions. PCR efficiencies determined from individual amplification curves avoid these confounders. To obtain unbiased efficiency-corrected results, we recommend absolute quantification with a single undiluted calibrator with a known target concentration and efficiency values derived from the amplification curves of the calibrator and the unknown samples., Summary: For meaningful diagnostics or biological interpretation, the reported results of qPCR experiments should be efficiency corrected. To avoid ambiguity, the Minimal Information for Publications on Quantitative Real-Time PCR Experiments (MIQE) guidelines checklist should be extended to require the methods that were used (1) to determine the PCR efficiency and (2) to calculate the reported target quantity and relative gene expression value., (© American Association for Clinical Chemistry 2021.)

Increasing gaseous emissions of nitrogen (N) and sulfur (S) associated with oil sands development in northern Alberta (Canada) has led to changing regional wet and dry N and S deposition regimes. We assessed the potential for using bog plant/lichen tissue chemistry (N and S concentrations, C:N and C:S ratios, in 10 plant/lichen species) to monitor changing atmospheric N and S deposition through sampling at five bog sites, 3-6 times per growing season from 2009 to 2016. During this 8-year period, oil sands N emissions steadily increased, while S emissions steadily decreased. We examined the following: (1) whether each species showed changes in tissue chemistry with increasing distance from the Syncrude and Suncor upgrader stacks (the two largest point sources of N and S emissions); (2) whether tissue chemistry changed over the 8 year period in ways that were consistent with increasing N and decreasing S emissions from oil sands facilities; and (3) whether tissue chemistry was correlated with growing season wet deposition of NH 4 + -N, NO 3 - -N, or SO 4 2- -S. Based on these criteria, the best biomonitors of a changing N deposition regime were Evernia mesomorpha, Sphagnum fuscum, and Vaccinium oxycoccos. The best biomonitors of a changing S deposition regime were Evernia mesomorpha, Cladonia mitis, Sphagnum fuscum, Sphagnum capillifolium, Vaccinium oxycoccos, and Picea mariana. Changing N and S deposition regimes in the oil sands region appear to be influencing N and S cycling in what once were pristine ombrotrophic bogs, to the extent that these bogs may effectively monitor future spatial and temporal patterns of deposition.

The United States' large-scale poultry meat industry is energy and water intensive, and opportunities may exist to improve sustainability during the broiler chilling process. By USDA regulation, after harvest the internal temperature of the chicken must be reduced to 40°F or less within 16 h to inhibit bacterial growth that would otherwise compromise the safety of the product. This step is accomplished most commonly by water immersion chilling in the United States, while air chilling methods dominate other global markets. A comprehensive understanding of the differences between these chilling methods is lacking. Therefore, we assessed the meat quality, shelf-life, microbial ecology, and techno-economic impacts of chilling methods on chicken broilers in a university meat laboratory setting. We discovered that air chilling methods resulted in superior chicken odor and shelf-life, especially prior to 14 days of dark storage. Moreover, we demonstrated that air chilling resulted in a more diverse microbiome that we hypothesize may delay the dominance of the spoilage organism Pseudomonas Finally, a techno-economic analysis highlighted potential economic advantages to air chilling compared to water chilling in facility locations where water costs are a more significant factor than energy costs. IMPORTANCE As the poultry industry works to become more sustainable and to reduce the volume of food waste, it is critical to consider points in the processing system that can be altered to make the process more efficient. In this study, we demonstrate that the method used during chilling (air versus water chilling) influences the final product microbial community, quality, and physiochemistry. Notably, the use of air chilling appears to delay the bloom of Pseudomonas spp. that are the primary spoilers in packaged meat products. By using air chilling to reduce carcass temperatures instead of water chilling, producers may extend the time until spoilage of the products and, depending on the cost of water in the area, may have economic and sustainability advantages. As a next step, a similar experiment should be done in an industrial setting to confirm these results generated in a small-scale university lab facility., (Copyright © 2021 Belk et al.)

The ongoing construction of natural gas combined cycle (NGCC) power plants is incompatible with a transition to global net-zero greenhouse gas emissions. This work evaluates the emission pricing and technology costs required to convert an existing NGCC power plant to a biomethane-based bioenergy with carbon capture and storage (BECCS) system. The conversion was evaluated using techno-economic analysis and time-resolved life cycle assessment. At current technology costs, carbon dioxide equivalent emission prices of $142 and $215 per metric ton are required to allow a BECCS conversion to compete with normal operation or shutdown, respectively, of an existing NGCC power plant. These results show further technological development must occur in parallel with emission pricing to make BECCS viable. If mid-range emission pricing estimates are implemented ($25-$105 per metric ton), BECCS capital cost targets range from $1434 to $2098 per kW of capacity, while operational costs range from $32 to $51 per MWh of electricity produced to enable conversion. These findings indicate that operational costs associated with fuel consumption and production must be significantly reduced to make a BECCS conversion viable, even with emission pricing. All data and methods of this work have been made publicly available in an open-source model.

Bogs and fens cover 6 and 21%, respectively, of the 140,329 km 2 Oil Sands Administrative Area in northern Alberta. Regional background atmospheric N deposition is low (<2 kg N ha -1  yr -1 ), but oil sands development has led to increasing N deposition (as high as 17 kg N ha -1  yr -1 ). To examine responses to N deposition, over five years, we experimentally applied N (as NH 4 NO 3 ) to a poor fen near Mariana Lake, Alberta, unaffected by oil sands activities, at rates of 0, 5, 10, 15, 20, and 25 kg N ha -1  yr -1 , plus controls (no water or N addition). At Mariana Lake Poor Fen (MLPF), increasing N addition: 1) progressively inhibited N 2 -fixation; 2) had no effect on net primary production (NPP) of Sphagnum fuscum or S. angustifolium, while stimulating S. magellanicum NPP; 3) led to decreased abundance of S. fuscum and increased abundance of S. angustifolium, S. magellanicum, Andromeda polifolia, Vaccinium oxycoccos, and of vascular plants in general; 4) led to an increase in stem N concentrations in S. angustifolium and S. magellanicum, and an increase in leaf N concentrations in Chamaedaphne calyculata, Andromeda polifolia, and Vaccinium oxycoccos; 5) stimulated root biomass and production; 6) stimulated decomposition of cellulose, but not of Sphagnum or vascular plant litter; and 7) had no or minimal effects on net N mineralization in surface peat, NH 4 + -N, NO 3 - -N or DON concentrations in surface porewater, or peat microbial composition. Increasing N addition led to a switch from new N inputs being taken up primarily by Sphagnum to being taken up primarily by shrubs. MLPF responses to increasing N addition did not exhibit threshold triggers, but rather began as soon as N additions increased. Considering all responses to N addition, we recommend a critical load for poor fens in Alberta of 3 kg N ha -1  yr -1 ., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

The use of noninvasive techniques to evaluate stress responses in animals has become an increasingly popular method of animal welfare assessment in both production animals and wildlife. In particular, using fecal samples to measure fecal cortisol metabolites (FCMs) as a quantitative measure has proven ideal as samples can be collected remote to the animal after defecation without the need for invasive procedures. Colorimetric enzyme immunoassays (EIAs) have been shown to have a high level of selectivity and sensitivity for FCM concentration analysis, equivalent to the traditionally used radioimmunoassay. Regardless of the assay system used, species- and sample-specific validation must be undertaken to ensure the reliability of results, particularly where sampling is undertaken in a novel species or where environmental conditions might impact FCM stability in the fecal sample. To determine the limit of environmental exposure acceptable for analysis of FCM concentrations in ovine scat samples collected from a paddock under conditions of stable heat and humidity, this study quantified FCMs in ovine feces shortly after defecation (2-7 h) and after timed environmental exposure (1-9 d). Samples were determined to show stable FCM concentrations for up to 5 d by this analysis. Understanding the impacts of environmental exposure, and therefore the viability of remote fecal collection methods for quantitative analysis of FCM by EIA, is important to assess the utility of noninvasive measures of endocrine status in animals where the exact timing of defecation may not be known., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Progesterone receptor membrane component 1 (PGRMC1) is often elevated in cancers, and exists in alternative states of phosphorylation. A motif centered on PGRMC1 Y180 was evolutionarily acquired concurrently with the embryological gastrulation organizer that orchestrates vertebrate tissue differentiation., Results: Here, we show that mutagenic manipulation of PGRMC1 phosphorylation alters cell metabolism, genomic stability, and CpG methylation. Each of several mutants elicited distinct patterns of genomic CpG methylation. Mutation of S57A/Y180/S181A led to increased net hypermethylation, reminiscent of embryonic stem cells. Pathways enrichment analysis suggested modulation of processes related to animal cell differentiation status and tissue identity, as well as cell cycle control and ATM/ATR DNA damage repair regulation. We detected different genomic mutation rates in culture., Conclusions: A companion manuscript shows that these cell states dramatically affect protein abundances, cell and mitochondrial morphology, and glycolytic metabolism. We propose that PGRMC1 phosphorylation status modulates cellular plasticity mechanisms relevant to early embryological tissue differentiation.

Objective: The experience of an illness such as breast cancer is not a static event. Just as physiological needs change as a patient transitions through diagnosis, treatment, to long-term survivorship, so too will their needs for social support. We applied a transitions theory framework to explore how African American women with breast cancer conceptualized and experienced support along their breast cancer journey. Design: We recruited 16 African American women with breast cancer from a regional cancer center in South Carolina to complete qualitative, semi-structured interviews. We iteratively examined verbatim transcripts using thematic analysis. Results: Three core themes emerged: 'I guess she was supposed to': When support meets patient expectations; 'I wasn't expecting that and that just made me feel so good': When reality exceeds expectations; and 'Don't try to make an invalid out of me': When support given wasn't what was desired. Survivors shared how their family, friends and clergy met their needs for emotional (e.g. prayer, sharing affirmations about God) and instrumental support (e.g. cooking meals, house cleaning). They emphasized how receiving emotional support from their healthcare providers was a pleasant surprise. However, survivors also described unexpected disappointments when family members offered support that was un-needed or un-desired. Conclusions : Applying transitions theory, we found that social support is a process of bidirectional negotiation where African American women with breast cancer perceive support as helpful and acceptable depending on who offers support, what type of support is offered, and when it is offered. Members of their social support network (e.g. family, friends, providers) should periodically assess the survivor's evolving needs to ensure the social support harmonizes with the needs and expectations of the survivor.

The indole diterpenoid toxin lolitrem B is a tremorgenic agent found in the common grass species, perennial ryegrass (Lolium perenne). The toxin is produced by a symbiotic fungus Epichloë festucae (var. lolii) and ingestion of infested grass with sufficient toxin levels causes a movement disorder in grazing herbivores known as 'ryegrass staggers'. Beside ataxia, lolitrem B intoxicated animals frequently show indicators of cognitive dysfunction or exhibition of erratic and unpredictable behaviours during handling. Evidence from field cases in livestock and controlled feeding studies in horses have indicated that intoxication with lolitrem B may affect higher cortical or subcortical functioning. In order to define the role of lolitrem B in voluntary motor control, spatial learning and memory under controlled conditions, mice were exposed to a known dose of purified lolitrem B toxin and tremor, coordination, voluntary motor activity and spatial learning and memory assessed. Motor activity, coordination and spatial memory were compared to tremor intensity using a novel quantitative piezo-electronic tremor analysis. Peak tremor was observed as frequencies between 15 and 25Hz compared to normal movement at approximately 1.4-10Hz. A single exposure to a known tremorgenic dose of lolitrem B (2 mg/kg IP) induced measureable tremor for up to 72 h in some animals. Initially, intoxication with lolitrem B significantly decreased voluntary movement. By 25 h post exposure a return to normal voluntary movement was observed in this group, despite continuing evidence of tremor. This effect was not observed in animals exposed to the short-acting tremorgenic toxin paxilline. Lolitrem B intoxicated mice demonstrated a random search pattern and delayed latency to escape a 3 h post intoxication, however by 27 h post exposure latency to escape matched controls and mice had returned to normal searching behavior indicating normal spatial learning and memory. Together these data indicate that the tremor exhibited by lolitrem B intoxicated mice does not directly impair spatial learning and memory but that exposure does reduce voluntary motor activity in intoxicated animals. Management of acutely affected livestock suffering toxicosis should be considered in the context of their ability to spatially orientate with severe toxicity., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Distiller's grains are a byproduct of corn ethanol production and provide an opportunity for increasing the economic viability and sustainability of the overall grain-to-fuels process. Typically, these grains are dried and sold as a ruminant feed adjunct. This study considers utilization of the residuals in a novel supplementary fermentation process to produce two products, enriched protein and fusel alcohols. The value-added proposition and environmental impact of this second fermentation step for distiller's grains are evaluated by considering three different processing scenarios. Techno-economic results show the minimum protein selling price, assuming fusel alcohol products are valued at $0.79 per liter gasoline equivalent, ranges between $1.65-$2.48 kg protein -1 for the different cases. Environmental impacts of the systems were evaluated through life cycle assessment. Results show a baseline emission results of 17 g CO 2-eq (MJ fuel) -1 for the fuel product and 10.3 kg CO 2-eq kg protein -1 for the protein product. Sensitivity to allocation methods show a dramatic impact with results ranging between -8 to 140 g CO 2-eq (MJ fuel) -1 for the fuel product and -0.3 to 6.4 kg CO 2-eq kg protein -1 for the protein product. The discussion is focused on the potential impact of the technology on corn ethanol production economics and sustainability.

Life cycle assessment is a fundamental tool used to evaluate the environmental impact of products. Standard life cycle assessment methodology ignores the impact of greenhouse gases relative to when they are emitted. In this paper, we present a method for leveraging the social cost of greenhouse gases to account for the temporal impacts of emissions in life cycle assessment and techno-economics. To demonstrate, we use this method to analyze the present value of the monetized impacts of emissions across multiple electricity generation technologies. Results show that accounting for time increases the present value across all but one of the technologies considered. Carbon intensive technologies show the highest increase, with coal rising between 26% and 62% depending on social cost scenario. Additionally, we demonstrate a second method that combines temporally resolved greenhouse gas emissions with techno-economic analysis. Considering temporal impacts of emissions within techno-economic analysis increases the levelized cost of electricity (LCOE) across all technologies considered. Carbon intensive technologies increase significantly, with the LCOE from coal rising between 37% and 263% depending on the social cost scenario. The proposed methods show that temporal resolution in life cycle assessment is critical for comparing the monetized impacts of greenhouse gas emissions across technologies.