An adverse in utero experience negatively impacts perinatal growth in livestock. Maternal heat stress (HS) during gestation reduces placental growth and function. This progressive placental insufficiency ultimately leads to fetal growth restriction (FGR). Studies in chronically catheterized fetal sheep have shown that FGR fetuses exhibit hypoxemia, hypoglycemia, and lower anabolic hormone concentrations. Under hypoxic stress and nutrient deficiency, fetuses prioritize basal metabolic requirements over tissue accretion to support survival. Skeletal muscle is particularly vulnerable to HS-induced placental insufficiency due to its high energy demands and large contribution to total body mass. In FGR fetuses, skeletal muscle growth is reduced, evidenced by smaller myofiber size and mass, reduced satellite cell proliferation, and slower rate of protein synthesis. Disruptions in skeletal muscle growth are associated with mitochondrial dysfunction, including reduced pyruvate flux into the mitochondrial matrix and lower complex I activity in the mitochondrial electron transport chain. This review summarizes current research on the mechanisms by which HS-induced placental insufficiency affects skeletal muscle growth in the fetus, with an emphasis on myogenesis, hypertrophy, protein synthesis, and energy metabolism. The evidence presented is primarily drawn from experiments using chronically catheterized fetal sheep exposed to maternal HS during mid-gestation. Additionally, we explore emerging nutritional strategies aimed at enhancing skeletal muscle growth in animals with FGR. These strategies hold promise not only for improving reproductive efficiency in livestock affected by prenatal stress but also for their translational relevance to human pregnancies complicated by placental insufficiency.

Through compositional inclusion or exclusion, the photograph can assert and communicate what belongs in a picture, in a landscape, in an ecosystem. It can illuminate what we deem conservation-worthy, or, on a larger scale, which extinctions are attention-worthy. Photographic practice helps to illuminate the active nature of extinction, and our choices as actors and witnesses within that process. Here, researchers from the University of Leeds’ Extinction Studies Doctoral Training Programme present individual reflections on interdisciplinary practice-led research in the Scottish Small Isles. We consider how photography, as a form of praxis, can generate new forms of knowledge surrounding extinction: its meanings, representations, and legacies, particularly through visual representation. We offer seven perspectives on contemporary image-making, from disciplines including philosophy, conservation biology, literature, sociology, geology, cultural anthropology, and palaeontology. Researchers gathered experiential, ethical, even biological meanings from considering what to include or exclude in images: from the micro to the macro, the visible to the invisible, the aesthetic to the ecological. We draw conclusions around meaning-making through the process of photography itself, and the tensions encountered through framing and decision-making in a time of mass ecological decline.

A method is presented for constructing a covariance matrix Σ*0 that is the sum of a matrix Σ(γ0) that satisfies a specified model and a perturbation matrix,E, such that Σ*0=Σ(γ0) +E. The perturbation matrix is chosen in such a manner that a class of discrepancy functions F(Σ*0, Σ(γ0)), which includes normal theory maximum likelihood as a special case, has the prespecified parameter value γ0 as minimizer and a prespecified minimum δ A matrix constructed in this way seems particularly valuable for Monte Carlo experiments as the covariance matrix for a population in which the model does not hold exactly. This may be a more realistic conceptualization in many instances. An example is presented in which this procedure is employed to generate a covariance matrix among nonnormal, ordered categorical variables which is then used to study the performance of a factor analysis estimator.

Lynchets, often the defining component of historic agricultural landscapes in northern Europe, are generally associated with soft-limestone geologies and are particularly well developed on loess-mantled landscapes. To understand their formation and chronology, the authors present their geoarchaeological analyses of lynchet soils and loess deposits at Blick Mead and Charlton Forest in southern England, and Sint Martens-Voeren in Belgium. The lynchets date from the late prehistoric to the medieval periods and were constructed by plough action at the English sites, and by both cut-and-fill and ploughing in Belgium. This has resulted in the preservation of highly fertile loessic soils across chalk slopes, lost elsewhere. Although each example is associated with local/regional agricultural histories, the lynchets’ effective soil-retention capacities allowed them to survive as important heritage features with environmental benefits over millennia.

The quenching of cluster satellite galaxies is inextricably linked to the suppression of their cold interstellar medium (ISM) by environmental mechanisms. While the removal of neutral atomic hydrogen (H i) at large radii is well studied, how the environment impacts the remaining gas in the centres of galaxies, which are dominated by molecular gas, is less clear. Using new observations from the Virgo Environment traced in CO survey (VERTICO) and archival H i data, we study the H i and molecular gas within the optical discs of Virgo cluster galaxies on 1.2-kpc scales with spatially resolved scaling relations between stellar ($\Sigma_{\star}$), H i ($\Sigma_{\text{H}\,{\small\text{I}}}$), and molecular gas ($\Sigma_{\text{mol}}$) surface densities. Adopting H i deficiency as a measure of environmental impact, we find evidence that, in addition to removing the H i at large radii, the cluster processes also lower the average $\Sigma_{\text{H}\,{\small\text{I}}}$ of the remaining gas even in the central $1.2\,$kpc. The impact on molecular gas is comparatively weaker than on the H i, and we show that the lower $\Sigma_{\text{mol}}$ gas is removed first. In the most H i-deficient galaxies, however, we find evidence that environmental processes reduce the typical $\Sigma_{\text{mol}}$ of the remaining gas by nearly a factor of 3. We find no evidence for environment-driven elevation of $\Sigma_{\text{H}\,{\small\text{I}}}$ or $\Sigma_{\text{mol}}$ in H i-deficient galaxies. Using the ratio of $\Sigma_{\text{mol}}$-to-$\Sigma_{\text{H}\,{\small\text{I}}}$ in individual regions, we show that changes in the ISM physical conditions, estimated using the total gas surface density and midplane hydrostatic pressure, cannot explain the observed reduction in molecular gas content. Instead, we suggest that direct stripping of the molecular gas is required to explain our results.

OBJECTIVES/GOALS: In this study, the ability of a pathologist to detect malignancy on digital pseudo-H&E slides obtained via structured illumination microscopy (SIM) imaging of fresh diagnostic breast biopsies was assessed. The speed of imaging and processing was also assessed for potential clinical implementation. METHODS/STUDY POPULATION: This study was conducted in accordance with an Ochsner Medical Center of New Orleans IRB. 200 patients undergoing either stereotactic or ultrasound-guided diagnostic breast biopsies were consented and an additional core from the suspicious lesion was collected for research use. Research biopsies were transported to the lab and stained with DRAQ5 and Eosin-Y and imaged with SIM before being submitted for histology processing. Imaging and digital processing times were recorded. The resulting SIM images and histology slides were given to a pathologist for blind review to assess accuracy. RESULTS/ANTICIPATED RESULTS: The ex-vivo structured illumination microscopy images and subsequent histology slides from 79 research cores have been assessed to date. Some samples were excluded from the total data set and not included in the final assessment due to technical failures of the imaging protocol. Of the current set, the pathologist has a specificity of 88% and a sensitivity of 65%, as well as an NPV of 88% and a PPV of 65%. Staining time for each biopsy was completed within 3 and a half minutes and imaging at 20x magnification took between 4 and 12 minutes, depending on size and implementation of autofocus to the imaging system. Image processing took approximately 5 minutes per biopsy and is a direct function of biopsy size. DISCUSSION/SIGNIFICANCE: Decreased time between cancer suspicion and treatment will improve the prognosis of breast cancer patients. SIM imaging of fresh breast biopsies could ultimately allow primary and secondary histology to be performed simultaneously and minimize histopathology time, thus allowing clinicians and patients to implement treatment course more quickly.

The New York Bight is undergoing rapid anthropogenic change amidst an apparent increase in baleen whale sightings. Though survey efforts have increased in recent years, the lack of published knowledge on baleen whale occurrence prior to these efforts impedes effective assessments of distributional or behavioural shifts due to increasing human activities. Here we synthesize opportunistic sightings of baleen whales from 1998–2017, which represent the majority of sightings data prior to recent survey efforts, and which are largely unpublished. Humpback and fin whales were the most commonly sighted species, followed by North Atlantic right whales and North Atlantic minke whales. Important behaviours such as feeding and nursing were observed, and most species (including North Atlantic right whales) were seen during all seasons. Baleen whales overlapped with multiple anthropogenic use areas, and all species, but of particular importance North Atlantic right whales, were sighted outside the spatial and temporal bounds of the Seasonal Management Areas for North Atlantic right whales. These opportunistic data are vital for providing a baseline and context of baleen whales in the New York Bight prior to broad-scale efforts and facilitate interpretation of current and future observations and trends, which can more accurately inform effective management and mitigation efforts.

Amiodarone may be considered for patients with junctional ectopic tachycardia refractory to treatment with sedation, analgesia, cooling, and electrolyte replacements. There are currently no published pediatric data regarding the hemodynamic effects of the newer amiodarone formulation, PM101, devoid of hypotensive agents used in the original amiodarone formulation. We performed a single-center, retrospective, descriptive study from January 2012 to December 2020 in a pediatric ICU. Thirty-three patients were included (22 male and 11 female) between the ages of 1.1 and 1,460 days who developed post-operative junctional ectopic tachycardia or other tachyarrhythmias requiring PM101. Data analysis was performed on hemodynamic parameters (mean arterial pressures and heart rate) and total PM101 (mg/kg) from hour 0 of amiodarone administration to hour 72. Adverse outcomes were defined as Vasoactive-Inotropic Score >20, patients requiring ECMO or CPR, or patient death. There was no statistically significant decrease in mean arterial pressures within the 6 hours of PM101 administration (p > 0.05), but there was a statistically significant therapeutic decrease in heart rate for resolution of tachyarrhythmia (p < 0.05). Patients received up to 25 mg/kg in an 8-hour time for rate control. Average rate control was achieved within 11.91 hours and average rhythm control within 62 hours. There were four adverse events around the time of PM101 administration, with three determined to not be associated with the medication. PM101 is safe and effective in the pediatric cardiac surgical population. Our study demonstrated that PM101 can be used in a more aggressive dosing regimen than previously reported in pediatric literature with the prior formulation.

Remotely sensed land surface temperature (LST) data, such as the Moderate Resolution Imaging Spectroradiometer (MODIS) LST thermal infrared products, are useful for monitoring surface processes on the Greenland Ice Sheet in remote areas but must be validated to ensure accuracy. Using data from the Programme for Monitoring the Greenland Ice Sheet (PROMICE), we conducted a MODIS LST validation (MOD/MYD11 C6 swath level product) using radiometric in-situ skin temperature records from 2014 to 2017 over 17 PROMICE sites mostly in the ice sheet's ablation zone. There is a significant cold bias in MODIS LST when compared to PROMICE skin temperature, particularly when PROMICE records temperatures below 0°C (mean bias: 2.4 ± 0.01°C mean ± standard error, RMSE = 3.2°C). Multiple linear regression analysis reveals the difference between MODIS LST and PROMICE skin temperature is larger at lower temperatures, lower latent heat fluxes and higher specific humidity. Our results confirm the presence of a progressive cold bias in the MODIS LST that should be considered in use of this product, and we identify and corroborate areas for ongoing algorithm development.

OBJECTIVES/GOALS: In a familial case where 10 of 17 members inherited EA/LVNC in an autosomal dominant pattern, we discovered a novel, damaging missense variant in the gene KLHL26 that segregates with disease and comprises an altered electrostatic surface profile, likely decoupling the CUL3-interactome. We hypothesize that this KLHL26 variant is etiologic of EA/LVNC. METHODS/STUDY POPULATION: We differentiated a family trio (a heart-healthy daughter and EA/LVNC-affected mother and daughter) of induced pluripotent stem cells into cardiomyocytes (iPSC-CMs) in a blinded manner on three iPSC clones per subject. Using flow cytometry, immunofluorescence, and biomechanical, electrophysiological, and automated contraction methods, we investigated iPSC-CM differentiation efficiency between D10-20, contractility analysis and cell cycle regulation at D20, and sarcomere organization at D60. We further conducted differential analyses following label-free protein and RNA-Seq quantification at D20. Via CRISPR-Cas9 gene editing, we plan to characterize KLHL26 variant-specific iPSC-CM alterations and connect findings to discoveries from patient-specific studies. RESULTS/ANTICIPATED RESULTS: All iPSC lines differentiated into CMs with an increased percentage of cTnT+ cells in the affected daughter line. In comparison to the unaffected, affected iPSC-CMs had fewer contractions per minute and altered calcium transients, mainly a higher amount of total calcium release, faster rate of rise and faster rate of fall. The affected daughter line further had shorter shortening and relaxation times, higher proliferation, lower apoptosis, and a smaller cell surface area per cardiac nucleus. The affected mother line trended in a similar direction to the affected daughter line. There were no gross differences in sarcomere organization between the lines. We also discovered differential expression of candidate proteins such as kinase VRK1 and collagen COL5A1 from proteomic profiling. DISCUSSION/SIGNIFICANCE: These discoveries suggest that EA/LVNC characteristics or pathogenesis may result from decreased contractile ability, altered calcium transients, and cell cycle dysregulation. Through the KLHL26 variant correction and introduction in the daughter lines, we will build upon this understanding to inform exploration of critical clinical targets.

Introduction. It has been estimated that smokers tend to fail to report unsuccessful quit attempts that lasted a short time and occurred a longer time ago. However, it is unclear whether the failure to report unsuccessful quit attempts varies by the type of cessation aid used. Methods. A total of 5,892 smokers aged 16+ years who had made 1+ quit attempts in the past year were surveyed between January 2014 and December 2020 as part of the Smoking Toolkit Study. Respondents indicated when their most recent quit attempt started, how long it lasted, and which cessation aid(s) were used (e.g., unaided, varenicline, and behavioural support). The percentage failure to report for each cessation aid and 95% bootstrap confidence intervals (CIs) were estimated with an established method. Test for equality of proportions was performed to examine whether quit attempts lasting between one day and one week and that started >6 months ago failed to be reported at a different rate depending on the cessation aid used. Results. We estimated that after three months, 97% (95% CI = 96%-98%) of unaided quit attempts lasting less than one day, 80% (95% CI = 79%-81%) of those lasting between one day and one week, and 60% (95% CI = 59%-61%) of those lasting between one week and one month fail to be reported. Compared with unaided attempts, the estimated percentage failure to report quit attempts that lasted between one day and one week and that started >6 months ago was significantly lower for attempts involving behavioural support (92% of unaided attempts vs. 75% of attempts involving behavioural support, ${\chi }^2\left(1\right)=9.29$, p = 0.002). No other significant differences were detected. Conclusions. Smokers in England appear to fail to report a substantial proportion of unsuccessful quit attempts. This failure appears particularly prominent for attempts that last a short time or occurred longer ago and appears lower for attempts involving behavioural support compared with unaided attempts.

The Quaternary Isotope Laboratory (QIL) at the University of Washington was launched in 1969 and directed by Minze Stuiver until his retirement in 1998. Here we review some of the scientific work undertaken in the QIL and the memories of some of Minze’s former students and colleagues.

Anelaphus villosus (Fabricius) and A. parallelus (Newman) are longhorned beetles (Coleoptera: Cerambycidae: Cerambycinae: Elaphidiini) that closely resemble each other in appearance. In practice, if antennomere 3 is distinctly longer than antennomere 4, the specimen is considered to be A. villosus, but the accuracy of this methodology is unknown. Authorities disagree about the ease of separating these two species based on morphological traits, and recent work hypothesises that they should be synonymised due to the difficulty of distinguishing them by either morphology or natural history. Assuming correct initial species determinations of curated specimens, as well as of those determined with DNA barcoding and by the primary author, data from 23 body measurements were collected from 50 A. villosus and 60 A. parallelus specimens. Stepwise discriminant analyses and discriminant functions were used to evaluate the ability to distinguish these species based on morphology. Species assignments from discriminant functions were very accurate and were supported by assignments determined with DNA barcoding. The ability to distinguish A. villosus and A. parallelus based on morphological and molecular differences provides evidence against the taxonomic hypothesis of synonymy under one species. A need for greater ecological understanding of these species remains.