OBJECTIVES/GOALS: The EWSR1-FLI1 gene fusion is implicated as a source of oncogenic activity in the majority of Ewing sarcoma (EwS) cases (>70%). Our studies will provide unparalleled insight into the transformative mechanics of EWS-FLI1 METHODS/STUDY POPULATION: EWS-FLI1 is an intrinsically disordered protein (IDP). IDPs do not form stable secondary or tertiary folds in biological conditions and ofter contain proline-rich regions and other multivalent binding regions. These regions interact with numerous partners resulting in highly dynamic and complex protein-protein interactions. Recent advances in proximity labelling techniques, such as the use of a biotin ligase fused to a protein of interest are exceptionally well suited to identifying IDP interactomes since they do not rely on the binding affinities but rather the distance between interacting proteins. Combining novel discovery proteomics with Nuclear Magnetic Resonance (NMR) approaches will provide unparalleled insight into the transformative mechanics of EWS-FLI1. RESULTS/ANTICIPATED RESULTS: Using the proximity labeling technique TurboID, which limits the number of false-positive interactions, we have shown that EWSR1 and EWS-FLI1 act within the spliceosome (responsible for mRNA splicing and processing). Using bioinformatics techniques, we show EWS-FLI1 interacts with peptidyl-prolyl isomerase (PPI) proteins, specifically PPIL1, through the EWS-FLI1 N-terminal region (EWS-LCD). We used NMR to provide insight into the interaction between PPIL1 and EWS-LCD, showing that EWS-LCD interacts with the catalytic region of PPIL1. We anticipate that PPIL1 isomerizes EWS-FLI1 to modulate its activity, and that EWS-FLI1 interacts with the spliceosome through formation of a biological condensate. However, future proximity labeling and NMR studies are needed to verify this activity. DISCUSSION/SIGNIFICANCE: Our studies will yield actionable insights regarding the protein-protein interfaces in EWS-FLI1 that can be targeted to attenuate the oncogenic activity of EWS-FLI1. More broadly, our results will be applicable toward understanding the etiology of other pediatric cancers and for guiding the development of novel targeted treatments.

The aquatic and terrestrial clades of species of Trypanosoma could provide insight into the evolutionary history of the genus, as well as complementary information for biomedical studies of medically and economically important species of Trypanosoma. The ecological interactions and phylogeny of aquatic trypanosomes are currently not well-understood, mostly due to their complex life cycles and a deficiency of data. The species of Trypanosoma from African anuran hosts are of the least understood taxa in the genus. Trypanosomes were collected from South African frogs and subjected to morphological and phylogenetic analyses. This study redescribes Trypanosoma (Trypanosoma) nelspruitense Laveran, 1904 and Trypanosoma (Haematomonas) grandicolor Pienaar, 1962, with morphological and molecular data. The present study aims to create a platform for further future research on African anuran trypanosomes.

Vertebral osteomyelitis (VO) represents 4–10% of bone and joint infections. In Western countries, its incidence seems to increase, simultaneously with an increasing number of comorbidities among an ageing population. This study aimed to assess the evolution of VO epidemiology in France over the 2010–2019 decade. A nationwide cross-sectional study was conducted using the French hospital discharge data collected through the French diagnosis-related groups ‘Programme de Médicalisation des Systèmes d'Information’. VOs were detected with a previously validated case definition using International Classification of Diseases 10 (ICD-10) codes, implemented with the French current procedural terminology codes. The study population included all patients hospitalised in France during the 2010–2019 decade, aged 15 years old and more. Patient and hospital stay characteristics and their evolutions were described. During the study period, 42 105 patients were hospitalised for VO in France involving 60 878 hospital stays. The mean VO incidence was 7.8/100 000 over the study period, increasing from 6.1/100 000 in 2010 to 11.3/100 000 in 2019. The mean age was 64.8 years old and the sex ratio was 1.56. There were 31 341 (74.4%) patients with at least one comorbidity and 3059 (7.3%) deceased during their hospital stay. Even if rare, device-associated VOs (4450 hospital stays, 7.3%) highly increased over the period. The reliability of the method, based upon an exhaustive database and a validated case definition, provided an effective tool to compare data over time in real-life conditions to regularly update the epidemiology of VO.

The effect of weight gain during mid- and late gestation in dairy heifers on performance at the start of first lactation was studied. In this experiment, 47 Holstein heifers with first calving at 36 months of age were used. The plane of nutrition aimed to have a high (900 g/d, H; n = 23) and low (500, L; n = 24) average daily gain (ADG) from the 4th month of gestation until 3 weeks before the expected day of calving, achieved by ad libitum intake of high quality pasture (H) or controlled intake of a total mixed ration (L). Body weight (BW), body condition score (BCS), milking, and reproductive performances were recorded. Concentrations of plasma non-esterified fatty acids (NEFA), glucose, beta-hydroxybutyric acid (BHBA), and urea were characterised at weeks 2, 4, 6 and 8 of lactation. Milk fatty acid composition was determined at weeks 3 and 6. A total of 39 heifers successfully calved and completed first lactation. During feeding treatment the required ADG were achieved. BW and BCS were higher in H heifers at calving compared to L heifers: 707 vs. 640 kg, and 3.91 vs. 3.01 respectively. H heifers lost more weight, BCS and had lower feed intake during the beginning of first lactation (−0.8 kg DM/d/heifer over the first 4 weeks of lactation). Per day of lactation, H heifers produced significantly more milk (29.2 vs. 26.2 kg), fat (1.27 vs. 1.07 kg) and protein (0.84 vs. 0.477 kg) from 0 to 8 weeks of lactation. Concentrations of NEFA, glucose and BHBA were higher in H heifers compared to L heifers, but urea concentration was not affected. Concentration of preformed fatty acids in the milk (C16 and more) was higher. As a result, the calculated daily net energy balance during the first 8 weeks of lactation was −1.53 and −5.95 MJ for L and H heifers, respectively.

Weeds are often spatially aggregated in maize fields, and the level of aggregation varies across and within fields. Several annual weed species are present in maize fields before postemergence herbicide application, and herbicides applied will control several species at a time. The goal of this study was to assess the spatial distribution of multispecies weed infestation in maize fields. Ground-based imagery was used to map weed infestations in rain-fed maize fields. Image segmentation was used to extract weed cover information from geocoded images, and an expert-based threshold of 0.102% weed cover was used to generate maps of weed presence/absence. From 19 site-years, 13 (68%) demonstrated a random spatial distribution, whereas six site-years demonstrated an aggregated spatial pattern of either monocotyledons, dicotyledons, or both groups. The results of this study indicated that monocotyledonous and dicotyledonous weed groups were not spatially segregated, but discriminating these weed groups slightly increased the chances of detecting an aggregated pattern. It was concluded that weeds were not always spatially aggregated in maize fields. These findings emphasize the need for techniques allowing the assessment of weed aggregation prior to conducting site-specific weed management.

Weed management decisions based on weed threshold models offer the opportunity to reduce herbicide use by allowing the possibility of forgoing treatment or lowering rates. Weed thresholds based on a relative leaf-cover model were tested during a 4-yr period at two locations. Two 1.62-ha fields, planted to conventional and glyphosate-resistant corn (2004, 2005, 2007) or soybean (2006), were divided in 900 m2 sections. Herbicides were applied postemergence to each of these sections with either variable rates based on weed thresholds, or constant full rates. Variable herbicide rates included: no application, half rate, or full rate. Relative weed cover values of 0.2 and 0.4 (corn) or 0.1 and 0.3 (soybean) served as thresholds for incremental rates. Digital images were used to evaluate the relative weed cover. Weed density was assessed before and after herbicide application. Weed seed production was estimated for two species in 2004 and 2005. No difference in crop yield, relative weed cover, weed density, or weed seed production was observed between conventional and glyphosate-resistant cropping systems. During the first year, herbicide use reduction was obtained (−85.4%) with marginal crop yield loss (5 to 15%). In the subsequent 3 yr, preherbicide weed densities increased and concomitant increases in relative weed cover values did not allow more than a 10% overall reduction in herbicide use. This threshold model designed to maintain crop yields within a given year did not allow significant reduction in herbicide use during the following 3 yr. Residual weed populations most likely replenished the seed bank to levels that allowed weed densities to increase afterward. Increased weed density over time in plots treated with full rates of herbicide every year also indicated that a single postemergence herbicide treatment was not sufficient to contain weed populations at low levels every year in this corn–soybean rotation.

The implementation of site-specific weed management requires information about weed cover and decision support systems to determine weed cover thresholds and concomitant herbicide rates. Although it is possible to create accurate weed cover maps over large areas, weed cover thresholds have generally been evaluated using tedious weed density counts. To bridge this gap between weed cover obtained by machine vision and the concept of economic threshold, crop advisers specializing in weed scouting were asked to evaluate over 2,500 weed cover images (2 m by 3 m) and determine if a given image would require herbicide application or not. Using the area under the “receiver operating characteristic” curve method, an optimal weed cover threshold was established. The derived economic thresholds ranged from 0.06 to 0.31% weed cover contingent on the level of tolerance of the expert adviser. Although this threshold seems low, it is comparable with economic threshold values based on weed density.

At the field scale, weeds generally appear aggregated rather than randomly distributed, and this aggregation is linked to the spatial heterogeneity of biotic and abiotic factors. Crop management practices shape the spatial pattern of weed infestations by modifying certain factors having an impact on weed emergence and growth. Although crop seeding is often the last in-field disturbance before crop and weed emergence, its effect on the distribution of weeds has received little attention in the literature. The purpose of this study was to assess the influence of the planting operation on weed cover and presence in corn fields using digital images to investigate the possibility of sensing the interrow to infer the presence or absence of weeds on the corn row. A total of 18 site-years under conventional tillage treated with a single POST application of herbicide were selected across seven locations. Image analysis, at the V2 to V4 growth stage of corn, was used to compare the weed cover in three zones: the undisturbed interrows, the corn rows, and the interrows compacted by tractor wheel traffic. For 61% of site-years, there was no significant difference among the zones. When there was a significant difference compared with the other two zones, the undisturbed interrow was usually less infested. Point-to-point comparisons of weed presence or absence (based on a threshold of five pixels) between the interrow and the corn row revealed 70 or 73% correspondence, depending on the type of interrow (undisturbed or tracked). However the error of inference of the corn row weed cover generated by sensing only adjacent interrows may be too high for efficient commercial weed control.

The morphologic radiation of Early Jurassic ammonites following the near extinction at the end of the Triassic is analyzed from 436 species of 156 genera that form a representative sample of morphs occurring worldwide in the first three stages of the Jurassic (Hettangian, Sinemurian, Pliensbachian: 36 subzones, 24 m.y.). Morphologic diversity is analyzed independently of taxonomy by processing 18 shape parameters using multivariate analysis and clustering techniques. The morphospace thus defined indicates that morphs fall readily into two groups made up of four and five adjacent morpho-subsets. The temporal pattern of morphospace occupation in the 36 Lower Jurassic subzones displays diversification, depletion (sometimes total), and displacement of successive parts of the morphospace, reflecting a complex history in which morphologic radiation appears to be more than a process of diffusion. The history of the morphologic evolution is tentatively related to sea-level changes and there is a suggestion that morphologic diversity increases during second-order transgressive periods.

New sedimentological, biostratigraphical and geochemical data recording the Toarcian Oceanic Anoxic Event (T-OAE) are reported from a marginal marine succession in southern Beaujolais, France. The serpentinum and bifrons ammonite zones record black shales with high (1–10 wt%) total organic carbon contents (TOC) and dysoxia-tolerant benthic fauna typical of the ‘Schistes Carton’ facies well documented in contemporaneous nearby basins. The base of the serpentinum ammonite zone, however, differs from coeval strata of most adjacent basinal series in that it presents several massive storm beds particularly enriched in juvenile ammonites and the dysoxia-tolerant, miniaturized gastropod Coelodiscus. This storm-dominated interval records a marked negative 5‰ carbonate and organic carbon isotope excursion being time-equivalent with that recording storm- and mass flow-deposits in sections of the Lusitanian Basin, Portugal, pointing to the existence of a major tempestite/turbidite event over tropical areas during the T-OAE. Although several explanations remain possible at present, we favour climatically induced changes in platform morphology and storm activity as the main drivers of these sedimentological features. In addition, we show that recent weathering, most probably due to infiltration of O2-rich meteoric water, resulted in the preferential removal of 12C-enriched organic carbon, dramatic TOC loss and total destruction of the lamination of the black shale sequence over most of the studied exposure. These latter observations imply that extreme caution should be applied when interpreting the palaeoenvironmental significance of sediments lacking TOC enrichment and lamination from outcrops with limited surface exposures.

Care-related infections are a major public health concern. Their transmission can be associated with environmental factors. This study looks at air contamination around 45 patients colonized with multidrug-resistant organisms (MDROs). We found that 30 hospital rooms (67%) were contaminated with MDRO species and 10 rooms (22%) were contaminated with at least 1 MDRO.

Le présent article traite des interactions fluide/structure dans le domaine des moyennes fréquences. En effet, dans ce domaine fréquentiel, il n'est pas possible de modéliser un système fluide/structure en FEM car la densité modale du milieu fluide est trop importante, ni de le modéliser en SEA car cette modélisation ne serait pas valide. Une méthode hybride FEM/SEA est donc développée : la méthode SEA-like. La méthode SEA-like est une méthode énergétique dont la matrice est calculée avec des méthodes de type "basses fréquences". Les coefficients SEA-like sont calculés avec une condensation dynamique afin de prendre en compte la densité modale importante dans le système. Dans cet article, la matrice SEA-like d'un système couplé plaque/cavité est calculé avec une analyse modale. On obtient alors une expression simplifiée des coefficients SEA-like. La matrice SEA-like est alors validée dans le domaine des basses fréquences avec modélisation FEM du système couplé et dans le domaine des hautes fréquences avec une modélisation SEA du système couplé.