Explore the vast range of titles available.

Conditional gene regulation in Drosophila through binary expression systems like the LexA-LexAop system provides a superb tool for investigating gene and tissue function. To increase the availability of defined LexA enhancer trap insertions, we present molecular, genetic, and tissue expression studies of 301 novel Stan-X LexA enhancer traps derived from mobilization of the index SX4 line. This includes insertions into distinct loci on the X, II, and III chromosomes that were not previously associated with enhancer traps or targeted LexA constructs, an insertion into ptc, and seventeen insertions into natural transposons. A subset of enhancer traps was expressed in CNS neurons known to produce and secrete insulin, an essential regulator of growth, development, and metabolism. Fly lines described here were generated and characterized through studies by students and teachers in an international network of genetics classes at public, independent high schools, and universities serving a diversity of students, including those underrepresented in science. Thus, a unique partnership between secondary schools and university-based programs has produced and characterized novel resources in Drosophila, establishing instructional paradigms devoted to unscripted experimental science.

Binary expression systems like the LexA-LexAop system provide a powerful experimental tool kit to study gene and tissue function in developmental biology, neurobiology, and physiology. However, the number of well-defined LexA enhancer trap insertions remains limited. In this study, we present the molecular characterization and initial tissue expression analysis of nearly 100 novel StanEx LexA enhancer traps, derived from the StanEx1 index line. This includes 76 insertions into novel, distinct gene loci not previously associated with enhancer traps or targeted LexA constructs. Additionally, our studies revealed evidence for selective transposase-dependent replacement of a previously-undetected KP element on chromosome III within the StanEx1 genetic background during hybrid dysgenesis, suggesting a molecular basis for the over-representation of LexA insertions at the NK7.1 locus in our screen. Production and characterization of novel fly lines were performed by students and teachers in experiment-based genetics classes within a geographically diverse network of public and independent high schools. Thus, unique partnerships between secondary schools and university-based programs have produced and characterized novel genetic and molecular resources in Drosophila for open-source distribution, and provide paradigms for development of science education through experience-based pedagogy.

ETV6 is an ETS family transcriptional repressor that self-associates by its PNT domain to facilitate cooperative DNA binding. Chromosomal translocations frequently generate constitutively active oncoproteins with the ETV6 PNT domain fused to the kinase domain of one of many protein tyrosine kinases. Although an attractive target for therapeutic intervention, the propensity of the ETV6 PNT domain to polymerize via the tight head-to-tail association of two relatively flat interfaces makes it challenging to identify suitable small molecule inhibitors of this protein-protein interaction. Herein we provide a comprehensive biophysical characterization of the ETV6 PNT domain interaction interfaces to aid future drug discovery efforts and help define the mechanisms by which its self-association mediates transcriptional repression. Using NMR spectroscopy, X-ray crystallography, and molecular dynamics simulations, we demonstrate that ETV6 PNT domain variants with monomerizing mutations adopt very stable helical bundle folds that do not change in conformation upon self-association. Amide hydrogen exchange and surface plasmon resonance-monitored alanine scanning mutagenesis studies identified hot spot regions within the self-association interfaces. These regions include both central hydrophobic residues and flanking salt-bridging residues. Collectively, these studies indicate that small molecules targeted to these hydrophobic or charged regions within the relatively rigid interfaces could potentially serve as orthosteric inhibitors of ETV6 PNT domain polymerization. Competing Interest Statement The authors have declared no competing interest.

With intentions to enhance team process and performance, the prevalence of technological investments has increased in organizations. Contrary to expectations, actual experiences have resulted in ambiguous performance improvements. Recognizing the need to obtain a better understanding of the team/technology interface (Grudin & Poltrack, 1997; Khoshafian & Buckiewicz, 1995), the purpose of this dissertation was to explore the impact of information technology use on team processes and performance. Given that time factors (e.g., project deadlines, schedules, coordination of efforts, etc.) dictate many aspects of team functioning, different types of team processes are expected to be critical during different phases of task execution. Moreover, it is argued that high performing teams align different functions of information technology to support the various types of team processes. A temporal model of the effects of information technology use on team process and performance was tested. Using a repeated cross sectional field sample of information systems development (ISD) teams, stakeholders, and end-users, the results of this study provided initial evidence that different team processes are important during the different phases of task execution. Mixed support was found for the relationship between information technology use and team processes warranting more research.

Conditional gene regulation in Drosophila through binary expression systems like the LexA-LexAop system provides a superb tool for investigating gene and tissue function. To increase the availability of defined LexA enhancer trap insertions, we present molecular, genetic and tissue expression studies of 301 novel Stan-X LexA enhancer traps derived from mobilization of the index SX4 line. This includes insertions into distinct loci on the X, II and III chromosomes that were not previously associated with enhancer traps or targeted LexA constructs, an insertion into ptc, and eleven insertions into natural transposons. A subset of enhancer traps was expressed in CNS neurons known to produce and secrete insulin, an essential regulator of growth, development and metabolism. Fly lines described here were generated and characterized through studies by students and teachers in an international network of genetics classes at public, independent high schools, and universities serving a diversity of students, including those underrepresented in science. Thus, a unique partnership between secondary schools and university-based programs has produced and characterized novel resources in Drosophila, establishing instructional paradigms devoted to unscripted experimental science.Competing Interest StatementThe authors have declared no competing interest.Footnotes* Author's list was updated. The abstract was supplemented to reflect changes in results. The results have been expanded and include two new figures, Figure 3, showcasing the expression pattern of an enhancer trap in ptc, and Figure 6, displaying novel data of enhancer trapping natural transposable elements. The supplement is expanded by Suppl. Table 2, covering split read fragments used in Figure 6, and Suppl. Figure 3 analyzing snucRNAseq data for IPCs and CC cells. The methods are updated to cover all changes.* https://www.stan-x.org/* https://stanx.squarespace.com* https://www.ncbi.nlm.nih.gov/sra/PRJNA912892

Binary expression systems like the LexA-LexAop system provide a powerful experimental tool kit to study gene and tissue function in developmental biology, neurobiology and physiology. However, the number of well-defined LexA enhancer trap insertions remains limited. In this study, we present the molecular characterization and initial tissue expression analysis of nearly 100 novel StanEx LexA enhancer traps, derived from the StanEx1 index line. This includes 76 insertions into novel, distinct gene loci not previously associated with enhancer traps or targeted LexA constructs. Additionally, our studies revealed evidence for selective transposase-dependent replacement of a previously-undetected KP element on chromosome III within the StanEx1 genetic background during hybrid dysgenesis, suggesting a molecular basis for the over-representation of LexA insertions at the NK7.1 locus in our screen. Production and characterization of novel fly lines were performed by students and teachers in experiment-based genetics classes within a geographically diverse network of public and independent high schools. Thus, unique partnerships between secondary schools and university-based programs have produced and characterized novel genetic and molecular resources in Drosophila for open-source distribution, and provide paradigms for development of science education through experience-based pedagogy.

Human induced pluripotent stem cell derived cardiac fibroblasts (hiPSC-CFs) play a critical role in modeling human cardiovascular diseases in vitro. However, current culture substrates used for hiPSC-CF differentiation and expansion, such as Matrigel and tissue culture plastic (TCPs), are tissue mismatched and may provide pathogenic cues. Here, we report that hiPSC-CFs differentiated on Matrigel and expanded on tissue culture plastic (M-TCP-iCFs) exhibit transcriptomic hallmarks of activated fibroblasts limiting their translational potential. To alleviate pathogenic activation of hiPSC-CFs, we utilized decellularized extracellular matrix derived from porcine heart extracellular matrix (HEM) to provide a biomimetic substrate for improving hiPSC-CF phenotypes. We show that hiPSC-CFs differentiated and expanded on HEM (HEM-iCFs) exhibited reduced expression of hallmark activated fibroblast markers versus M-TCP-iCFs while retaining their cardiac fibroblast phenotype. HEM-iCFs also maintained a reduction in expression of hallmark genes associated with pathogenic fibroblasts when seeded onto TCPs. Further, HEM-iCFs more homogenously integrated into an hiPSC-derived cardiac organoid model, resulting in improved cardiomyocyte sarcomere development. In conclusion, HEM provides an improved substrate for the differentiation and propagation of hiPSC-CFs for disease modeling. [Display omitted] •Common tissue culture substrates for the derivation of hiPSC-cardiac fibroblasts induce activation of pathogenic phenotypes.•Biomimetic porcine heart extracellular matrix (HEM) reduced and prevented pathogenic activation of hiPSC-CFs.•HEM did not impair the cardiac fibroblast phenotype of hiPSC-CFs.•HEM-derived hiPSC-CFs showed enhanced integration and improved cardiomyocyte development in an hiPSC cardiac organoid model.

During infections with the malaria parasites Plasmodium vivax, patients exhibit rhythmic fevers every 48 h. These fever cycles correspond with the time the parasites take to traverse the intraerythrocytic cycle (IEC). In other Plasmodium species that infect either humans or mice, the IEC is likely guided by a parasite-intrinsic clock [Rijo-Ferreiraet al., Science 368, 746–753 (2020); Smith et al., Science 368, 754–759 (2020)], suggesting that intrinsic clock mechanisms may be a fundamental feature of malaria parasites. Moreover, because Plasmodium cycle times are multiples of 24 h, the IECs may be coordinated with the host circadian clock(s). Such coordination could explain the synchronization of the parasite population in the host and enable alignment of IEC and circadian cycle phases. We utilized an ex vivo culture of whole blood from patients infected with P. vivax to examine the dynamics of the host circadian transcriptome and the parasite IEC transcriptome. Transcriptome dynamics revealed that the phases of the host circadian cycle and the parasite IEC are correlated across multiple patients, showing that the cycles are phase coupled. In mouse model systems, host–parasite cycle coupling appears to provide a selective advantage for the parasite. Thus, understanding how host and parasite cycles are coupled in humans could enable antimalarial therapies that disrupt this coupling.

Biological factors such as genetics contribute to nutrition-related outcomes, but nutritional epidemiological studies often lack consideration of genetics despite evidence of their functional impacts on health and cognition. genetic variation has been hypothesized to influence health and cognitive outcomes due to evidence of metabolic perturbations in -phenylalanine to -tyrosine hydroxylation, including plausible downstream effects on catecholamine neurotransmitters among not only individuals with phenylketonuria (PKU) [homozygotes for mutations] but also PKU carriers [heterozygotes]. Related to these metabolic perturbations, diminished executive functioning has been observed in individuals with PKU, even when treated, but research is lacking exploring this outcome in PKU carriers. The present study aims to detail methods for stratifying populations based on genetic variation, for use in epidemiological precision nutrition research. It further provides an exploratory exemplar of such research through population stratification by genetic variation (i.e., PKU carriers vs. non-carriers), while providing the first descriptive data on executive functioning skills using the validated Executive Skills Questionnaire-Revised (ESQ-R) tool with -genetically stratified groups (PKU carriers and non-carriers). Participants were ≥18 years of age and heterozygotes (PKU carriers) or non-carriers. Levels of executive functioning were self-reported anonymously online and included the validated Executive Skills Questionnaire-Revised (ESQ-R) tool. Data were analyzed using t-tests, chi-square tests, ANOVAs, and ANCOVAs. Respondents (n = 99, n = 79 carriers and n = 20 non-carriers) consisted of males (22.2%) and females (77.8%), primarily of European ancestry. There were no significant differences between groups (carriers vs. non-carriers) for total scores (mean ± SD ESQ-R score carriers = 17.41 ± 14.01; non-carriers = 14.95 ± 10.00), but carriers scored significantly worse than non-carriers for the ESQ-R item \"I have trouble making a plan\" in the adjusted model. This study provides a methodological exemplar for exploring genetically stratified subpopulations in epidemiological precision nutrition research.

The Center for Epidemiologic Studies Depression Scale – Revised (CESD-R) is a popular self-report screening measure for depression. A 20-item questionnaire with scores ranging from 0 to 4 for each item, the CESD-R can produce total scores ranging from 0 to 80. However, the typical scoring protocol for the CESD-R restricts the range of possible scores to between 0 and 60 to retain the same range and clinical cutoff scores as the original CES-D. Despite the widespread adoption of this scoring approach, the psychometric impact has never been systematically examined. In an undergraduate and community adult sample (n = 869), item response theory analyses indicated that scoring the CESD-R with all 5 response options (CESD-R5opt) provided nearly twice as much information about a person’s latent depression for individuals with high levels of depression than did scoring the CESD-R with 4 response options per item (CESD-R4opt). The CESD-R5opt retained the strong reliability and factor structure of the CESD-R4opt and was more sensitive to individual differences for participants at high levels of depression compared to the CESD-R4opt. Results provide preliminary evidence that researchers and clinicians should score the CESD-R using the full 0-to-80 scale and a clinical cutoff score of 29.