Assessment of Hypoxia Inducible Factor Levels in Cancer Cell Lines upon Hypoxic Induction Using a Novel Reporter Construct

Hypoxia Inducible Factor (HIF) signaling pathway is important for tumor cells with limited oxygen supplies, as it is shown to be involved in the process of proliferation and angiogenesis. Given its pivotal role in cancer biology, robust assays for tracking changes in HIF expression are necessary for understanding its regulation in cancer as well as developing therapies that target HIF signaling. Here we report a novel HIF reporter construct containing tandem repeats of minimum HIF binding sites upstream of eYFP coding sequence. We show that the reporter construct has an excellent signal to background ratio and the reporter activity is HIF dependent and directly correlates with HIF protein levels. By utilizing this new construct, we assayed HIF activity levels in different cancer cell lines cultured in various degrees of hypoxia. This analysis reveals a surprising cancer cell line specific variation of HIF activity in the same level of hypoxia. We further show that in two cervical cancer cell lines, ME180 and HeLa, the different HIF activity levels observed correlate with the levels of hsp90, a cofactor that protects HIF against VHL-independent degradation. This novel HIF reporter construct serves as a tool to rapidly define HIF activity levels and therefore the therapeutic capacity of potential HIF repressors in individual cancers

Non-local first-order modelling of crowd dynamics: a multidimensional framework with applications

In this work a physical modelling framework is presented, describing the intelligent, non-local, and anisotropic behaviour of pedestrians. Its phenomenological basics and constitutive elements are detailed, and a qualitative analysis is provided. Within this common framework, two first-order mathematical models, along with related numerical solution techniques, are derived. The models are oriented to specific real world applications: a one-dimensional model of crowd-structure interaction in footbridges and a two-dimensional model of pedestrian flow in an underground station with several obstacles and exits. The noticeable heterogeneity of the applications demonstrates the significance of the physical framework and its versatility in addressing different engineering problems. The results of the simulations point out the key role played by the physiological and psychological features of human perception on the overall crowd dynamics.Comment: 26 pages, 17 figure

The glycolytic enzyme phosphofructokinase-1 assembles into filaments.

Despite abundant knowledge of the regulation and biochemistry of glycolytic enzymes, we have limited understanding on how they are spatially organized in the cell. Emerging evidence indicates that nonglycolytic metabolic enzymes regulating diverse pathways can assemble into polymers. We now show tetramer- and substrate-dependent filament assembly by phosphofructokinase-1 (PFK1), which is considered the "gatekeeper" of glycolysis because it catalyzes the step committing glucose to breakdown. Recombinant liver PFK1 (PFKL) isoform, but not platelet PFK1 (PFKP) or muscle PFK1 (PFKM) isoforms, assembles into filaments. Negative-stain electron micrographs reveal that filaments are apolar and made of stacked tetramers oriented with exposed catalytic sites positioned along the edge of the polymer. Electron micrographs and biochemical data with a PFKL/PFKP chimera indicate that the PFKL regulatory domain mediates filament assembly. Quantified live-cell imaging shows dynamic properties of localized PFKL puncta that are enriched at the plasma membrane. These findings reveal a new behavior of a key glycolytic enzyme with insights on spatial organization and isoform-specific glucose metabolism in cells

Listening to a conversation with aggressive content expands the interpersonal space

The distance individuals maintain between themselves and others can be defined as ‘interpersonal space’. This distance can be modulated both by situational factors and individual characteristics. Here we investigated the influence that the interpretation of other people interaction, in which one is not directly involved, may have on a person’s interpersonal space. In the current study we measured, for the first time, whether the size of interpersonal space changes after listening to other people conversations with neutral or aggressive content. The results showed that the interpersonal space expands after listening to a conversation with aggressive content relative to a conversation with a neutral content. This finding suggests that participants tend to distance themselves from an aggressive confrontation even if they are not involved in it. These results are in line with the view of the interpersonal space as a safety zone surrounding one’s body

Enhancing Perceived Safety in Human–Robot Collaborative Construction Using Immersive Virtual Environments

Advances in robotics now permit humans to work collaboratively with robots. However, humans often feel unsafe working alongside robots. Our knowledge of how to help humans overcome this issue is limited by two challenges. One, it is difficult, expensive and time-consuming to prototype robots and set up various work situations needed to conduct studies in this area. Two, we lack strong theoretical models to predict and explain perceived safety and its influence on human–robot work collaboration (HRWC). To address these issues, we introduce the Robot Acceptance Safety Model (RASM) and employ immersive virtual environments (IVEs) to examine perceived safety of working on tasks alongside a robot. Results from a between-subjects experiment done in an IVE show that separation of work areas between robots and humans increases perceived safety by promoting team identification and trust in the robot. In addition, the more participants felt it was safe to work with the robot, the more willing they were to work alongside the robot in the future.University of Michigan Mcubed Grant: Virtual Prototyping of Human-Robot Collaboration in Unstructured Construction EnvironmentsPeer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/145620/1/You et al. forthcoming in AutCon.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/145620/4/You et al. 2018.pdfDescription of You et al. 2018.pdf : Published Versio

Engineering influenza HA nanoparticles to better understand their immune responses and improve vaccine design

Thesis (Ph.D.)--University of Washington, 2023Protein nanoparticle-based vaccines are a promising platform, shown to be efficacious in preventing disease against a variety of pathogens. Building on their success, increasing our understanding of how nanoparticle vaccines elicit immune responses will aid both in our general understanding of immunology and also in designing improved vaccines. This work centers on influenza hemagglutinin (HA) nanoparticle vaccine design. It starts with the design of HA ‘trihead’ nanoparticle vaccines, that incorporate several layers of immune refocusing to increase breadth and potency in responses elicited against the HA head. Trihead design was then adapted onto several strains of HA, with the ultimate goal of creating a new platform for seasonal influenza vaccines. Lastly, the interaction of antibodies and HA ectodomain nanoparticles was explored as a means to elucidate mechanisms of their immunogenicity

Factors Associated With Climacteric Reaction.

PhDPsychologyUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/178494/2/7907061.pd