All cells generate an electrical potential across their plasma membrane driven by a concentration gradient of charged ions. A typical resting membrane potential ranges from −40 to −70 mV, with a net negative charge on the cytosolic side of the membrane. Maintenance of the resting membrane potential depends on the presence of two-pore-domain potassium “leak” channels, which allow for outward diffusion of potassium ions along their concentration gradient. Disruption of the ion gradient causes the membrane potential to become more positive or more negative relative to the resting state, referred to as “depolarization” or “hyperpolarization,” respectively. Changes in membrane potential have proven to be pivotal, not only in normal cell cycle progression but also in malignant transformation and tissue regeneration. Using polystyrene nanoparticles as a model system, we use flow cytometry and fluorescence microscopy to measure changes in membrane potential in response to nanoparticle binding to the plasma membrane. We find that nanoparticles with amine-modified surfaces lead to significant depolarization of both CHO and HeLa cells. In comparison, carboxylate-modified nanoparticles do not cause depolarization. Mechanistic studies suggest that this nanoparticle-induced depolarization is the result of a physical blockage of the ion channels. These experiments show that nanoparticles can alter the biological system of interest in subtle, yet important, ways.

Understanding intracellular redox chemistry requires new tools for the site-specific visualization of intracellular oxidation. We have developed a spatially-resolved intracellular sensor of hydrogen peroxide, HyPer-Tau, for time-resolved imaging in live cells. This sensor consists of a hydrogen peroxide-sensing protein tethered to microtubules. We demonstrate the use of the HyPer-Tau sensor for three applications; dose-dependent response of human cells to exogenous hydrogen peroxide, a model immune response of mouse macrophages to stimulation by bacterial toxin, and a spatially-resolved response to localized delivery of hydrogen peroxide. These results demonstrate that HyPer-Tau can be used as an effective tool for tracking changes in spatially localized intracellular hydrogen peroxide and for future applications in redox signaling.

Background The purpose of this study is to describe human epidermal growth factor 2 (HER2) overexpression in head and neck squamous cell carcinoma (HNSCC) and re‐evaluate its potential as a target for HER2‐directed immunotherapies. Methods A retrospective cohort of patients with HNSCC receiving curative treatment was identified, and HER2 expression evaluated in archival tissue by immunohistochemistry and correlated with clinicopathological characteristics. HER2 expression data were also determined for HNSCC patients in The Cancer Genome Atlas. Results Nineteen percent of HNSCC and 39% of oropharyngeal HNSCC (OPSCC) were HER2 positive. HER2 expression positively correlated with nodal metastasis (p = 0.035). Patients with HER2‐positive tumors had decreased overall survival (p = 0.012), including within the human papilloma virus‐positive OPSCC subgroup (p = 0.007). Conclusions A substantial fraction of HNSCC overexpresses HER2 protein, suggesting it may be a suitable target for antigen‐directed immunotherapy. HER2 expression and its correlation with survival vary across HNSCC subsites, making it unsuitable as a prognostic marker.