Spatial Gradients of E-Cadherin and Fibronectin in TGF-ß1-Treated Epithelial Colonies Are Independent of Fibronectin Fibril Assembly.

Epithelial to Mesenchymal Transition (EMT) is a dynamic, morphogenetic process characterized by a phenotypic shift in epithelial cells towards a motile and often invasive mesenchymal phenotype. We have previously demonstrated that EMT is associated with an increase in assembly of the extracellular matrix protein fibronectin (FN) into insoluble, viscoelastic fibrils. We have also demonstrated that Transforming Growth Factor-ß1 (TGF-ß1) localizes to FN fibrils, and disruption of FN assembly or disruption of TGF-ß1 localization to FN fibrils attenuates EMT. Previous studies have shown that TGF-ß1 induces spatial gradients of EMT in mammary epithelial cells cultured on FN islands, with cells at free edges of the island preferentially undergoing EMT. In the current work, we sought to investigate: (a) whether FN fibril assembly is also spatially patterned in response to TGF-ß1, and (b) what effects FN fibril inhibition has on spatial gradients of E-Cadherin and FN fibrillogenesis. We demonstrate that mammary epithelial cells cultured on square micropatterns have fewer E-Cadherin-containing adherens junctions and assemble more FN fibrils at the periphery of the micropattern in response to increasing TGF-ß1 concentration, indicating that TGF-ß1 induces a spatial gradient of both E-Cadherin and FN fibrils. Inhibition of FN fibril assembly globally diminished E-Cadherin-containing adherens junctions and FN fibrillogenesis, but did not eliminate the spatial gradient of either. This suggests that global inhibition of FN reduces the degree of both FN fibrillogenesis and E-Cadherin-containing adherens junctions, but does not eliminate the spatial gradient of either, suggesting that spatial gradients of EMT and FN fibrillogenesis are influenced by additional factors.

Diabetic hyperglycemia promotes primary tumor progression through glycation-induced tumor extracellular matrix stiffening.

Diabetes mellitus is a complex metabolic disorder that is associated with an increased risk of breast cancer. Despite this correlation, the interplay between tumor progression and diabetes, particularly with regard to stiffening of the extracellular matrix, is still mechanistically unclear. Here, we established a murine model where hyperglycemia was induced before breast tumor development. Using the murine model, in vitro systems, and patient samples, we show that hyperglycemia increases tumor growth, extracellular matrix stiffness, glycation, and epithelial-mesenchymal transition of tumor cells. Upon inhibition of glycation or mechanotransduction in diabetic mice, these same metrics are reduced to levels comparable with nondiabetic tumors. Together, our study describes a novel biomechanical mechanism by which diabetic hyperglycemia promotes breast tumor progression via glycating the extracellular matrix. In addition, our work provides evidence that glycation inhibition is a potential adjuvant therapy for diabetic cancer patients due to the key role of matrix stiffening in both diseases.

Teaching mindfulness-based stress management techniques to medical learners through simulation.

Acutely traumatic clinical events can exacerbate stress and burnout amongst healthcare providers. The Simulated Training for Resilience in Various Environments (STRIVE) course may provide a useful framework for medical educators to teach stress management skills to promote resilience amongst physician trainees. The course introduces the Big Four+ techniques (goal setting, visualization, self-talk, progressive muscular relaxation, attention control and tactical breathing) created by the Canadian Armed Forces using clinical scenarios. This framework can be easily adapted across other training contexts to equip future clinicians with a foundational skill set to optimize their response and recovery following critically stressful incidents.

Un événement clinique très traumatisant peut exacerber le stress et l'épuisement professionnel vécus par les soignants. Le cours « Simulated Training for Resilience in Various Environments ¼ ou STRIVE (formation par simulation pour développer la résilience dans divers environnements) offre un cadre utile aux enseignants en médecine pour initier les apprenants aux stratégies de gestion du stress afin de renforcer leur résilience. Le cours présente, à l'aide de scénarios cliniques, les principales techniques (les « Big Four+ ¼) créées par les Forces armées canadiennes, à savoir la fixation d'objectifs, la visualisation, le dialogue intérieur, la relaxation musculaire progressive, le contrôle de l'attention et la respiration tactique. Ce cadre peut être facilement adapté à d'autres contextes de formation afin de doter les futurs cliniciens d'un ensemble de compétences fondamentales pour optimiser leur capacité de réaction et de rétablissement face à un incident particulièrement stressant.

A hybrid model of intercellular tension and cell-matrix mechanical interactions in a multicellular geometry.

Epithelial cells form continuous sheets of cells that exist in tensional homeostasis. Homeostasis is maintained through cell-to-cell junctions that distribute tension and balance forces between cells and their underlying matrix. Disruption of tensional homeostasis can lead to epithelial-mesenchymal transition (EMT), a transdifferentiation process in which epithelial cells adopt a mesenchymal phenotype, losing cell-cell adhesion and enhancing cellular motility. This process is critical during embryogenesis and wound healing, but is also dysregulated in many disease states. To further understand the role of intercellular tension in spatial patterning of epithelial cell monolayers, we developed a multicellular computational model of cell-cell and cell-substrate forces. This work builds on a hybrid cellular Potts model (CPM)-finite element model to evaluate cell-matrix mechanical feedback of an adherent multicellular cluster. Cellular movement is governed by thermodynamic constraints from cell volume, cell-cell and cell-matrix contacts, and durotaxis, which arises from cell-generated traction forces on a finite element substrate. Junction forces at cell-cell contacts balance these traction forces, thereby producing a mechanically stable epithelial monolayer. Simulations were compared to in vitro experiments using fluorescence-based junction force sensors in clusters of cells undergoing EMT. Results indicate that the multicellular CPM model can reproduce many aspects of EMT, including epithelial monolayer formation dynamics, changes in cell geometry, and spatial patterning of cell-cell forces in an epithelial tissue.

Fibronectin fibrils regulate TGF-ß1-induced Epithelial-Mesenchymal Transition.

Epithelial-Mesenchymal Transition (EMT) is a dynamic process through which epithelial cells transdifferentiate from an epithelial phenotype into a mesenchymal phenotype. Previous studies have demonstrated that both mechanical signaling and soluble growth factor signaling facilitate this process. One possible point of integration for mechanical and growth factor signaling is the extracellular matrix. Here we investigate the role of the extracellular matrix (ECM) protein fibronectin (FN) in this process. We demonstrate that inhibition of FN fibrillogenesis blocks activation of the Transforming Growth Factor-Beta (TGF-ß) signaling pathway via Smad2 signaling, decreases cell migration and ultimately leads to inhibition of EMT. Results show that soluble FN, FN fibrils, or increased contractile forces are insufficient to independently induce EMT. We further demonstrate that inhibition of latent TGF-ß1 binding to FN fibrils via either a monoclonal blocking antibody against the growth factor binding domain of FN or through use of a FN deletion mutant that lacks the growth factor binding domains of FN blocks EMT progression, indicating a novel role for FN in EMT in which the assembly of FN fibrils serves to localize TGF-ß1 signaling to drive EMT.

The YsrS Paralog DygS Has the Capacity To Activate Expression of the Yersinia enterocolitica Ysa Type III Secretion System.

UNLABELLED: The Yersinia enterocolitica Ysa type III secretion system (T3SS) is associated with intracellular survival, and, like other characterized T3SSs, it is tightly controlled. Expression of the ysa genes is only detected following growth at low temperatures (26°C) and in high concentrations of sodium chloride (290 mM) in the medium. The YsrSTR phosphorelay (PR) system is required for ysa expression and likely responds to NaCl. During our investigations into the Ysr PR system, we discovered that genes YE3578 and YE3579 are remarkably similar to ysrR and ysrS, respectively, and are probably a consequence of a gene duplication event. The amino acid differences between YE3578 and ysrR are primarily clustered into two short regions. The differences between YE3579 and ysrS are nearly all located in the periplasmic sensing domain; the cytoplasmic domains are 98% identical. We investigated whether these paralogs were capable of activating ysa gene expression. We found that the sensor paralog, named DygS, is capable of compensating for loss of ysrS, but the response regulator paralog, DygR, cannot complement a ysrR gene deletion. In addition, YsrR, but not DygR, interacts with the histidine phosphorelay protein YsrT. Thus, DygS likely activates ysa gene expression in response to a signal other than NaCl and provides an example of a phosphorelay system in which two sensor kinases feed into the same regulatory pathway. IMPORTANCE: All organisms need mechanisms to promote survival in changing environments. Prokaryotic phosphorelay systems are minimally comprised of a histidine kinase (HK) that senses an extracellular stimulus and a response regulator (RR) but can contain three or more proteins. Through gene duplication, a unique hybrid HK was created. We show that, while the hybrid appears to retain all of the phosphorelay functions, it responds to a different signal than the original. Both HKs transmit the signal to the same RR, which activates a promoter that transcribes a set of genes encoding a type III secretion system (T3SS) whose function is not yet evident. The significance of this work lies in finding that two HKs regulate this T3SS, highlighting its importance.

Introduction and testing of an alternative control approach for a robotic prosthetic arm.

Commercially available robotic prosthetic arms currently use independent joint control. An alternative controller involving only control of the hand in a Cartesian frame rather than controlling each joint independently is proposed and tested. An experimental 4DOF robotic arm was used as the platform for testing the proposed control approach. As opposed to joint control, Cartesian control requires the solution to the inverse kinematics problem. The inverse kinematics solution was developed for the robotic arm using the extended Jacobian method. The two control methodologies, joint control and Cartesian control, were tested on five able-bodied human subjects. Improvement of one control methodology over the other was measured by the time it took for the subjects to complete a simple motor task. The timed trial results indicated that Cartesian control was both more intuitive and more effective than joint control. So, the results suggest that much improvement can be achieved by using the proposed Cartesian control methodology.

Ultrafine sputter-deposited Pt nanoparticles for triiodide reduction in dye-sensitized solar cells: impact of nanoparticle size, crystallinity and surface coverage on catalytic activity.

This paper presents a detailed electrochemical impedance spectroscopy and cyclic voltammetry (CV) investigation into the electrocatalytic activity of ultrafine (i.e., smaller than 2 nm) platinum (Pt) nanoparticles generated on a fluorine-doped tin oxide (FTO) surface via room temperature tilted target sputter deposition. In particular, the Pt-decorated FTO electrode surfaces were tested as counter electrode candidates for triiodide (I3(-)) reduction in dye-sensitized solar cells (DSSCs). We observed a direct correlation between size-dependent Pt nanoparticle crystallinity and the I3(-) reduction activity underlying DSSC performance. CV analysis confirmed the higher electrocatalytic activities of sputter-deposited crystalline Pt nanoparticles (1-2 nm) compared with either sub-nanometre Pt clusters or a continuous Pt thin film. While the low catalytic activity and DSSC performance of Pt clusters smaller in size than 1 nm is believed to arise from their non-crystalline nature and charge-trapping attributes, we attribute the high catalytic performance of larger Pt nanoparticles in the 1-2 nm regime to their well-defined crystallinity and fast electron transfer kinetics. For DSSC applications, the optimized Pt loading was calculated to be ~2.54 × 10(-7) g cm(-2), which corresponds to surface coverage by ~1.6 nm sized Pt nanoparticles.