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Abstract The oncogene RAS, extensively studied for decades, presents persistent gaps in understanding, hindering the development of effective therapeutic strategies due to a lack of precise details on how RAS initiates MAPK signaling with RAF effector proteins at the plasma membrane. Recent advances in X-ray crystallography, cryo-EM, and super-resolution fluorescence microscopy offer structural and spatial insights, yet the molecular mechanisms involving protein-protein and protein-lipid interactions in RAS-mediated signaling require further characterization. This study utilizes single-molecule experimental techniques, nuclear magnetic resonance spectroscopy, and the computational Machine-Learned Modeling Infrastructure (MuMMI) to examine KRAS4b and RAF1 on a biologically relevant lipid bilayer. MuMMI captures long-timescale events while preserving detailed atomic descriptions, providing testable models for experimental validation. Both in vitro and computational studies reveal that RBDCRD binding alters KRAS lateral diffusion on the lipid bilayer, increasing cluster size and decreasing diffusion. RAS and membrane binding cause hydrophobic residues in the CRD region to penetrate the bilayer, stabilizing complexes through β-strand elongation. These cooperative interactions among lipids, KRAS4b, and RAF1 are proposed as essential for forming nanoclusters, potentially a critical step in MAP kinase signal activation.

Background: Stratify new lung cancer patients based on the risk of in-hospital mortality rate after diagnosis. Methods: 522,941 lung cancer cases with available data on the Surveillance, Epidemiology, and End Results (SEER) were analyzed for the predicted probability based on six fundamental variables including age, gender, tumor size, T, N, and AJCC stages. The patients were randomly assigned to the training (n = 115,145) and validation datasets (n = 13,017). The remaining cohort with missing values (n = 394,779) was then combined with the primary lung tumour datasets (n = 1018) from The Cancer Genome Atlas, Lung Adenocarcinoma and Lung Squamous Cell Carcinoma projects (TCGA-LUAD & TCGA-LUSC) for external validation and sensitivity analysis. Results: Receiver Operating Characteristic (ROC) analyses showed high discriminatory power in the training and internal validation cohorts (Area under the curve [AUC] of 0.78 (95%CI = 0.78–0.79) and 0.78 (95%CI = 0.77–0.79), respectively), whereas that of the model on external validation data was 0.759 (95%CI = 0.757–0.761). We developed a static nomogram, a web app, and a risk table based on a logistic regression model using algorithm-selected variables. Conclusions: Our model can stratify lung cancer patients into high- and low-risk of in-hospital mortality to assist clinical further planning.

(1) Objective: This study evaluates the effects of simulation education at our institute on cardiac arrest resuscitation regarding knowledge, attitude, and practice (KAP) over a five-year period (2016–2020). (2) Subjects: Staff responded to the annual survey questionnaires followed by monthly training in Basic Life Support/Advanced Cardiovascular Life Support (BLS/ACLS) and Immediate Cardiac Life Support (ICLS) of the Japanese Association for Acute Medicine (JAAM) in 2016, 2017, and 2018. Additionally, in-house training was implemented in 2019 without post-assessment followed by training suspension in 2020 due to the COVID-19 pandemic. The last delivery of the survey questionnaires was in late 2020 for KAP retention measurement. (3) Measurements and Results: The self-efficacy level of BLS/ACLS/ICLS KAP of the survey respondents was analyzed using a five-point Likert scale. The mean self-efficacy level of BLS/ACLS/ICLS KAP increased over time, and that of the trained people was three-fold that of the untrained people. Trainees that had no cardiopulmonary resuscitation (CPR) experience gained the BLS/ACLS/ICLS key-point self-efficacy level, which we call the “Grip 14” in this study, as high as their untrained counterparts who had three-time CPR experience. Training suspension lessened the BLS/ACLS/ICLS KAP in both groups. (4) Conclusions: Continuous training enhances not only the BLS/ACLS/ICLS KAP of trainees but also of their untrained colleagues. The training likely had the same efficacy as the CPR experience.

RAS proteins are mutated in approximately 20% of all cancers and are generally associated with poor clinical outcomes. RAS proteins are localized to the plasma membrane and function as molecular switches, turned on by partners that receive extracellular mitogenic signals. In the on-state, they activate intracellular signal transduction cascades. Membrane-bound RAS molecules segregate into multimers, known as nanoclusters. These nanoclusters, held together through weak protein–protein and protein–lipid associations, are highly dynamic and respond to cellular input signals and fluctuations in the local lipid environment. Disruption of RAS nanoclusters results in downregulation of RAS-mediated mitogenic signaling. In this review, we discuss the propensity of RAS proteins to display clustering behavior and the interfaces that are associated with these assemblies. Strategies to therapeutically disrupt nanocluster formation or the stabilization of signaling incompetent RAS complexes are discussed.

The oncogene RAS, extensively studied for decades, presents persistent gaps in understanding, hindering the development of effective therapeutic strategies due to a lack of precise details on how RAS initiates MAPK signaling with RAF effector proteins at the plasma membrane. Recent advances in X-ray crystallography, cryo-EM, and super-resolution fluorescence microscopy offer structural and spatial insights, yet the molecular mechanisms involving protein-protein and protein-lipid interactions in RAS-mediated signaling require further characterization. This study utilizes single-molecule experimental techniques, nuclear magnetic resonance spectroscopy, and the computational Machine-Learned Modeling Infrastructure (MuMMI) to examine KRAS4b and RAF1 on a biologically relevant lipid bilayer. MuMMI captures long-timescale events while preserving detailed atomic descriptions, providing testable models for experimental validation. Both in vitro and computational studies reveal that RBDCRD binding alters KRAS lateral diffusion on the lipid bilayer, increasing cluster size and decreasing diffusion. RAS and membrane binding cause hydrophobic residues in the CRD region to penetrate the bilayer, stabilizing complexes through β-strand elongation. These cooperative interactions among lipids, KRAS4b, and RAF1 are proposed as essential for forming nanoclusters, potentially a critical step in MAP kinase signal activation.RAS binding to RAF’s RBDCRD domain on lipid bilayers cooperatively induces nanoclustering, slowing complex diffusion—an essential mechanism in initial signal activation. [ABSTRACT FROM AUTHOR]

The 2019 African swine fever (ASF) outbreaks in Vietnam imposed considerable impacts on the pig sector in Vietnam, resulting in the death or culling of nearly six million pigs, or more than 20% of the country's pig population. In order to assess the magnitude of the outbreak at sector level (both on farm and at value chain level), on livelihoods, and on the broader national economy, a comprehensive impact assessment was conducted using a mixed methods approach that integrated a value chain assessment with the use of quantitative modeling tools at sector and national levels. The results showed that the outbreak caused severe direct and indirect economic losses among farmers, particularly medium- and large-farmers whose livelihoods are largely derived from pig production. The outbreaks also affected other value chain actors due to a halving in the volume of pigs traded. At sector level, the outbreaks posed adverse impacts on the domestic supply and demand for pork, especially in the traditional sector. Meanwhile, the modern sector with higher levels of biosecurity and high technology growth was less likely to be affected and even benefited from the outbreak, which was evidenced by increased supply and income throughout the simulation period in this sector. At national level, different model simulation scenarios showed a sharp reduction in total gross domestic product (GDP) and a substantial loss of jobs. Improvements in the system of ASF compensation scheme are needed, both in terms of its administration, but also in its targeting, with greater emphasis needed on developing improved risk-sharing and funding mechanisms across national and local levels.

Over the last four years, a series of outreach programs have been offered through the Colorado School of Mines to middle school teachers from eleven school districts in the State of Colorado in the United States. Each of these programs is designed to illustrate through hands-on activities the application of mathematics to science and engineering. Each also has an academic year follow-up such that a faculty member, an expert teacher, or a graduate student assists the teachers in the classroom. An expected outcome of this effort is the improvement of instruction in mathematics and science in the participating middle schools; an unexpected outcome has been the impact of these projects on the culture of the participating schools, both middle schools and university. Based on our assessment efforts, this article describes the qualitative and quantitative outcomes of this sequence of projects on middle school students, teachers, graduate students, professors, and college curriculum. [ABSTRACT FROM AUTHOR]

SummaryTurnip crinkle virus encodes two proteins, p8 and p9, that are both required for cell-to-cell movement. The p8 movement protein has been demonstrated to bind RNA in a cooperative manner, although, similar to many other plant virus movement proteins, it contains no canonical RNA binding domain(s). However, three positively charged regions of p8 may potentially form ionic interactions with the RNA backbone. To identify functional regions of p8, a series of alanine and deletion scanning mutations were produced. The effects of these mutations were analysed using both in vitro RNA binding assays and in vivo infections of susceptible (Di-3) and resistant (Di-17) Arabidopsis thaliana plants. Several mutants that have reduced RNA binding ability were also demonstrated to be movement deficient and replication competent. Based on these results, there appear to be two regions, located between amino acids 18 and 31, and 50 and 72, that are required for RNA binding. Furthermore, additional regions (amino acids 12–15, and 34–37) appear to play a role in vivo unrelated to in vitro RNA binding activity. [ABSTRACT FROM AUTHOR]

RAS proteins are mutated in approximately 20% of all cancers and are generally associated with poor clinical outcomes. RAS proteins are localized to the plasma membrane and function as molecular switches, turned on by partners that receive extracellular mitogenic signals. In the on-state, they activate intracellular signal transduction cascades. Membrane-bound RAS molecules segregate into multimers, known as nanoclusters. These nanoclusters, held together through weak protein–protein and protein–lipid associations, are highly dynamic and respond to cellular input signals and fluctuations in the local lipid environment. Disruption of RAS nanoclusters results in downregulation of RAS-mediated mitogenic signaling. In this review, we discuss the propensity of RAS proteins to display clustering behavior and the interfaces that are associated with these assemblies. Strategies to therapeutically disrupt nanocluster formation or the stabilization of signaling incompetent RAS complexes are discussed. [ABSTRACT FROM AUTHOR]