Background: Household activities are responsible for up to 80% of direct and indirect greenhouse gas emissions in the United States. These greenhouse gas emissions come from activities including actions taken in relation to food, energy, and water (FEW) resource consumption. Therefore, actions taken at a household level have the potential to significantly reduce greenhouse gas emissions. A game-based learning approach can be used to educate youth on what actions they can take around their household to reduce their carbon footprint. <br/>Aim: FEWS for change is a first player role-playing game developed to educate high school students on how their actions impact the FEW resources and carbon emissions. The game also aims to measure how player’s beliefs and worldview effect their game play regarding sustainability and the environment. <br/>Methods: We developed the FEWS (Food, Energy, and Water Systems) for Change role-playing game based on transdisciplinary research of the food, energy, and water nexus, social, economic, and environmental factors. We piloted the game with a few students for initial results and will have a high school classroom pilot the game in mid-May.<br/>Preliminary Results: Results from the 4 participants demonstrated achievement of the learning goal of the pilot testing. This is objective was met by measuring the players improvement on the postsurvey compared to the presurvey. Due to limitations of time and virtual facilitation of this game, the other two learning objectives could not be measured in this initial pilot because not all post-game activities were included which are needed to measure the other learning objectives. When the game is piloted in mid-May, the other two learning objectives will be tested and measured.

Background: Recurrent glioblastoma (GBM) is resistant to available treatments and continued growth of the tumor is inevitable; this process is facilitated by the expression of genes regulated by the Signal Transducer and Activator of Transcription (STAT) family of transcription factors, namely STAT5, active in the invasive rim of GBM tumors. Currently, there are no targeted therapies for recurrent GBM that increase the overall patient survival rate. This study aims to analyze the differential expression of genes regulated by STAT5 between primary and recurrent GBM.<br/>Methods: Analysis of whole exome and RNA sequencing were performed on matched bulk primary and multiple recurrent tumor samples from GBM patients who received the current standard care to determine significant changes in gene expression of STAT3/5 targets. <br/>Results: Statistical analysis reveals a decrease in Synaptotagmin 2 (SYT2) and Pleckstrin Homology Domain Containing A3 (PLEKHA3) at recurrence, previously identified as potential STAT5 targets. <br/>Conclusions: To get a better understanding of the roles of STAT5 in GBM recurrence, their downstream effects need to be better understood. The transcriptomic program initiated by STAT5 activation is distinct from that of STAT3 activation. The roles of STAT5 target genes in GBM are poorly characterized, so further research should focus on understanding the effects of altered expression of these genes as they relate to STAT3/5 in GBM recurrence.

Since nitrogen (N) is often limiting in permafrost soils, we investigated the N[subscript 2]-fixing genetic potential and the inferred taxa harboring those genes by sequencing nifH gene fragments in samples taken along a permafrost thaw gradient in an Alaskan boreal soil. Samples from minimally, moderately and extensively thawed sites were taken to a depth of 79 cm to encompass zones above and below the depth of the water table. NifH reads were translated with frameshift correction and 112,476 sequences were clustered at 5% amino acid dissimilarity resulting in 1,631 OTUs. Sample depth in relation to water table depth was correlated to differences in the NifH sequence classes with those most closely related to group I nifH-harboring Alpha- and Beta-Proteobacteria in higher abundance above water table depth while those related to group III nifH-harboring Delta Proteobacteria more abundant below. The most dominant below water table depth NifH sequences, comprising 1/3 of the total, were distantly related to Verrucomicrobia-Opitutaceae. Overall, these results suggest that permafrost thaw alters the class-level composition of N[subscript 2]-fixing communities in the thawed soil layers and that this distinction corresponds to the depth of the water table. These nifH data were also compared to nifH sequences obtained from a study at an Alaskan taiga site, and to those of other geographically distant, non-permafrost sites. The two Alaska sites were differentiated largely by changes in relative abundances of the same OTUs, whereas the non-Alaska sites were differentiated by the lack of many Alaskan OTUs, and the presence of unique halophilic, sulfate- and iron-reducing taxa in the Alaska sites.

Insulin-like growth factor 1 (IGF1) is an important biomarker for the management of growth hormone disorders. Recently there has been rising interest in deploying mass spectrometric (MS) methods of detection for measuring IGF1. However, widespread clinical adoption of any MS-based IGF1 assay will require increased throughput and speed to justify the costs of analyses, and robust industrial platforms that are reproducible across laboratories. Presented here is an MS-based quantitative IGF1 assay with performance rating of >1,000 samples/day, and a capability of quantifying IGF1 point mutations and posttranslational modifications. The throughput of the IGF1 mass spectrometric immunoassay (MSIA) benefited from a simplified sample preparation step, IGF1 immunocapture in a tip format, and high-throughput MALDI-TOF MS analysis. The Limit of Detection and Limit of Quantification of the resulting assay were 1.5 μg/L and 5 μg/L, respectively, with intra- and inter-assay precision CVs of less than 10%, and good linearity and recovery characteristics. The IGF1 MSIA was benchmarked against commercially available IGF1 ELISA via Bland-Altman method comparison test, resulting in a slight positive bias of 16%. The IGF1 MSIA was employed in an optimized parallel workflow utilizing two pipetting robots and MALDI-TOF-MS instruments synced into one-hour phases of sample preparation, extraction and MSIA pipette tip elution, MS data collection, and data processing. Using this workflow, high-throughput IGF1 quantification of 1,054 human samples was achieved in approximately 9 hours. This rate of assaying is a significant improvement over existing MS-based IGF1 assays, and is on par with that of the enzyme-based immunoassays. Furthermore, a mutation was detected in ∼1% of the samples (SNP: rs17884626, creating an A→T substitution at position 67 of the IGF1), demonstrating the capability of IGF1 MSIA to detect point mutations and posttranslational modifications.

Many studies link the compositions of microbial communities to their environments, but the energetics of organism-specific biomass synthesis as a function of geochemical variables have rarely been assessed. We describe a thermodynamic model that integrates geochemical and metagenomic data for biofilms sampled at five sites along a thermal and chemical gradient in the outflow channel of the hot spring known as “Bison Pool” in Yellowstone National Park. The relative abundances of major phyla in individual communities sampled along the outflow channel are modeled by computing metastable equilibrium among model proteins with amino acid compositions derived from metagenomic sequences. Geochemical conditions are represented by temperature and activities of basis species, including pH and oxidation-reduction potential quantified as the activity of dissolved hydrogen. By adjusting the activity of hydrogen, the model can be tuned to closely approximate the relative abundances of the phyla observed in the community profiles generated from BLAST assignments. The findings reveal an inverse relationship between the energy demand to form the proteins at equal thermodynamic activities and the abundance of phyla in the community. The distance from metastable equilibrium of the communities, assessed using an equation derived from energetic considerations that is also consistent with the information-theoretic entropy change, decreases along the outflow channel. Specific divergences from metastable equilibrium, such as an underprediction of the relative abundances of phototrophic organisms at lower temperatures, can be explained by considering additional sources of energy and/or differences in growth efficiency. Although the metabolisms used by many members of these communities are driven by chemical disequilibria, the results support the possibility that higher-level patterns of chemotrophic microbial ecosystems are shaped by metastable equilibrium states that depend on both the composition of biomass and the environmental conditions.