Mammary Epithelial Metastatic Phenotype Forced Through the Extracellular Matrix

Biochemical and mechanical cues of the extracellular matrix have been shown to play important roles in cell-matrix and cell-cell interactions. We have experimentally tested the combined influence of these cues on cell motility, cell-cell interaction and assembly and traction force profile in an in vitro breast cancer model. The behavior of non-tumorigenic mammary epithelial cells was observed on surfaces with varying ligand concentration and polyacrylamide gel rigidity. Our data shows that cell velocity is biphasic in both matrix rigidity and adhesiveness, which are inversely related. Traction force microscopy revealed that maximum migration velocity is reached at intermediate force of single cells. Cell-cell adhesion becomes strongly favored on softer gels with elasticity ≤1250 Pascals. This result implies an existence of a compliance threshold that promotes cell-cell over cell-matrix adhesion. On softer gels of 400 Pa, stiffness similar to pre-malignant breast tissue in vivo, cells undergo multi-cellular assembly, division and re-arrangement into 3D spherical aggregates on 2D surface. The aggregates resemble the spherical acini found in vivo and are also formed with EpH4-J3B1 mouse mammary epithelial cells at same compliance. We establish mechanical cross talk between cell-cell and cell-matrix adhesions. Our findings of mechanotransduction show cell pairs exhibit \u27tug of war\u27 competitive dynamics between cell-cell and cell-matrix traction forces. Deletion of E-cadherin binding site to β-catenin results in loss of cell-cell tension magnitude and loss of the cross talk. We are first to show force dynamics of cell division and cytokinesis in adherent mammalian cells. In normal division intercellular force goes through a maximum. Inhibition of myosin II mediated contractility with blebbistatin completely inhibits cell cytokinesis on gel surfaces. However inhibition of Rho-associated kinase ROCK does not inhibit cytokinesis, only reduces the magnitude of the forces. Therefore myosin II mediated contractility is necessary for cytokinesis on physiologically relevant substrates

Frustration of crystallisation by a liquid–crystal phase

Frustration of crystallisation by locally favoured structures is critically important in linking the phenomena of supercooling, glass formation, and liquid-liquid transitions. Here we show that the putative liquid-liquid transition in n-butanol is in fact caused by geometric frustration associated with an isotropic to rippled lamellar liquid-crystal transition. Liquid-crystal phases are generally regarded as being “in between” the liquid and the crystalline state. In contrast, the liquid-crystal phase in supercooled n-butanol is found to inhibit transformation to the crystal. The observed frustrated phase is a template for similar ordering in other liquids and likely to play an important role in supercooling and liquid-liquid transitions in many other molecular liquids

Pressure and temperature dependence of local structure and dynamics in an ionic liquid

A detailed understanding of the local dynamics in ionic liquids remains an important aspect in the design of new ionic liquids as advanced functional fluids. Here, we use small-angle X-ray scattering and quasi-elastic neutron spectroscopy to investigate the local structure and dynamics in a model ionic liquid as a function of temperature and pressure, with a particular focus on state points (P,T) where the macroscopic dynamics, i.e., conductivity, is the same. Our results suggest that the initial step of ion transport is a confined diffusion process, on the nanosecond timescale, where the motion is restricted by a cage of nearest neighbors. This process is invariant considering timescale, geometry, and the participation ratio, at state points of constant conductivity, i.e., state points of isoconductivity. The connection to the nearest-neighbor structure is underlined by the invariance of the peak in the structure factor corresponding to nearest-neighbor correlations. At shorter timescales, picoseconds, two localized relaxation processes of the cation can be observed, which are not directly linked to ion transport. However, these processes also show invariance at isoconductivity. This points to that the overall energy landscape in ionic liquids responds in the same way to density changes and is mainly governed by the nearest-neighbor interactions

Structure evolution in poly(ethylene terephthalate) (PET) - Multi-walled carbon nanotube (MWCNT) composite films during <i>in-situ</i> uniaxial deformation

Combined small- and wide-angle X-ray scattering (SAXS/WAXS), mechanical and thermal techniques have been used to follow the morphology evolution in a series of poly(ethylene terephthalate) (PET) multiwall carbon nanotube (MWCNT) composite films during quasi solid-state uniaxial deformation at low strain rates. Uniaxially deformed PET-MWCNT films displayed improved mechanical properties compared with unfilled PET films. SAXS/WAXS data revealed a well oriented lamellar structure for unfilled PET films. In contrast, the PET-MWCNT composites revealed a nanohybrid shish-kebab (NHSK) morphology, with reduced orientation and crystallinity. Mechanistically, this morphology development is attributed to the MWCNTs acting as shish for the epitaxial growth of PET crystallites. Furthermore, nucleation and crystal growth occurs in the PET matrix, but MWCNTs ultimately inhibit crystallite development and hinder a final lamellar structure developing. The results show unequivocally the role MWCNTs play as nanofillers, in the morphology development, thermal and mechanical properties in composite polymer films

Programming gels over a wide pH range using multicomponent systems

Multicomponent hydrogels offer a tremendous opportunity to prepare useful and exciting materials that cannot be accessed using a single component. Here, we describe an unusual multi‐component low molecular weight gelling system that exhibits pH‐responsive behavior involving cooperative hydrogen bonding between the components, allowing it to maintain a gel phase across a wide pH range. Unlike traditional acid‐triggered gels, our system undergoes a change in the underlying molecular packing and maintains the β‐sheet structure both at acidic and basic pH. We further establish that autonomous programming between these two gel states is possible by an enzymatic reaction which allows us to prepare gels with improved mechanical properties

The structural response of the cornea to changes in stromal hydration

The primary aim of this study was to quantify the relationship between corneal structure and hydration in humans and pigs. X-ray scattering data were collected from human and porcine corneas equilibrated with polyethylene glycol (PEG) to varying levels of hydration, to obtain measurements of collagen fibril diameter, interfibrillar spacing and intermolecular spacing. Both species showed a strong positive linear correlation between hydration and interfibrillar spacing2 and a non-linear, bi-phasic relationship between hydration and fibril diameter, whereby fibril diameter increased up to approximately physiological hydration, H = 3.0, with little change thereafter. Above H = 3.0, porcine corneas exhibited a larger fibril diameter than human corneas (p < 0.001). Intermolecular spacing also varied with hydration in a bi-phasic manner but reached a maximum value at a lower hydration (H = 1.5) than fibril diameter. Human corneas displayed a higher intermolecular spacing than porcine corneas at all hydrations (p < 0.0001). Human and porcine corneas required a similar PEG concentration to reach physiological hydration, suggesting that the total fixed charge that gives rise to the swelling pressure is the same. The difference in their structural responses to hydration can be explained by variations in molecular crosslinking and intra/interfibrillar water partitioning

Evaluation of cataract formation in fish exposed to environmental radiation at Chernobyl and Fukushima

Recent studies apparently finding deleterious effects of radiation exposure on cataract formation in birds and voles living near Chernobyl represent a major challenge to current radiation protection regulations. This study conducted an integrated assessment of radiation exposure on cataractogenesis using the most advanced technologies available to assess the cataract status of lenses extracted from fish caught at both Chernobyl in Ukraine and Fukushima in Japan. It was hypothesised that these novel data would reveal positive correlations between radiation dose and early indicators of cataract formation.The structure, function and optical properties of lenses were analysed from atomic to millimetre length scales. We measured the short-range order of the lens crystallin proteins using Small Angle X-Ray Scattering (SAXS) at both the SPring-8 and DIAMOND synchrotrons, the profile of the graded refractive index generated by these proteins, the epithelial cell density and organisation and finally the focal length of each lens.The results showed no evidence of a difference between the focal length, the epithelial cell densities, the refractive indices, the interference functions and the short-range order of crystallin proteins (X-ray diffraction patterns) in lens from fish exposed to different radiation doses. It could be argued that animals in the natural environment which developed cataract would be more likely, for example, to suffer predation leading to survivor bias. But the cross-length scale study presented here, by evaluating small scale molecular and cellular changes in the lens (pre-cataract formation) significantly mitigates against this issue