Tumor cell nuclei soften during transendothelial migration

During cancer metastasis, tumor cells undergo significant deformation in order to traverse through endothelial cell junctions in the walls of blood vessels. As cells pass through narrow gaps, smaller than the nuclear diameter, the spatial configuration of chromatin must change along with the distribution of nuclear enzymes. Nuclear stiffness is an important determinant of the ability of cells to undergo transendothelial migration, yet no studies have been conducted to assess whether tumor cell cytoskeletal or nuclear stiffness changes during this critical process in order to facilitate passage. To address this question, we employed two non-contact methods, Brillouin confocal microscopy (BCM) and confocal reflectance quantitative phase microscopy (QPM), to track the changes in mechanical properties of live, transmigrating tumor cells in an in vitro collagen gel platform. Using these two imaging modalities to study transmigrating MDA-MB-231, A549, and A375 cells, we found that both the cells and their nuclei soften upon extravasation and that the nuclear membranes remain soft for at least 24 h. These new data suggest that tumor cells adjust their mechanical properties in order to facilitate extravasation

Two-photon interference with thermal light

The study of entangled states has greatly improved the basic understanding about two-photon interferometry. Two-photon interference is not the interference of two photons but the result of superposition among indistinguishable two-photon amplitudes. The concept of two-photon amplitude, however, has generally been restricted to the case of entangled photons. In this letter we report an experimental study that may extend this concept to the general case of independent photons. The experiment also shows interesting practical applications regarding the possibility of obtaining high resolution interference patterns with thermal sources.Comment: Added reference 1

The role of cell body density in ruminant retina mechanics assessed by atomic force and Brillouin microscopy

Cells in the central nervous system (CNS) respond to the stiffness of their environment. CNS tissue is mechanically highly heterogeneous, thus providing motile cells with region-specific mechanical signals. While CNS mechanics has been measured with a variety of techniques, reported values of tissue stiffness vary greatly, and the morphological structures underlying spatial changes in tissue stiffness remain poorly understood. We here exploited two complementary techniques, contact-based atomic force microscopy and contact-free Brillouin microscopy, to determine the mechanical properties of ruminant retinae, which are built up by different tissue layers. As in all vertebrate retinae, layers of high cell body densities ('nuclear layers') alternate with layers of low cell body densities ('plexiform layers'). Different tissue layers varied significantly in their mechanical properties, with the photoreceptor layer being the stiffest region of the retina, and the inner plexiform layer belonging to the softest regions. As both techniques yielded similar results, our measurements allowed us to calibrate the Brillouin microscopy measurements and convert the Brillouin shift into a quantitative assessment of elastic tissue stiffness with optical resolution. Similar as in the mouse spinal cord and the developing Xenopus brain, we found a strong correlation between nuclear densities and tissue stiffness. Hence, the cellular composition of retinae appears to strongly contribute to local tissue stiffness, and Brillouin microscopy shows a great potential for the application in vivo to measure the mechanical properties of transparent tissues.IPW is a recipient of an EMBO Long-Term Fellowship (ALTF 1263-2015; European Commission FP7 (Marie Curie Actions, LTFCOFUND2013, GA-2013-609409)). This work was further supported by the National Science Foundation (CMMI-1562863 to SHY and CMMI-1537027 to GS), the Human Frontier Science Program Young Investigator Grant RGY0074/2013 (GS and KF), the UK Medical Research Council Career Development Award G1100312/1 (KF), and the National Institutes of Health (R01EY025454 to SHY, K25EB015885 and R33CA204582 to GS, R21HD080585 to KF)

An RbAp48-like gene regulates adult stem cells in planarians.

Retinoblastoma-associated proteins 46 and 48 (RbAp46 and RbAp48) are factors that are components of different chromatin-modelling complexes, such as polycomb repressive complex 2, the activity of which is related to epigenetic gene regulation in stem cells. To date, no direct findings are available on the in vivo role of RbAp48 in stem-cell biology. We recently identified DjRbAp48 — a planarian ( Dugesia japonica ) homologue of human RBAP48 — expression of which is restricted to the neoblasts, the adult stem cells of planarians. In vivo silencing of DjRbAp48 induces lethality and inability to regenerate, even though neoblasts proliferate and accumulate after wounding. Despite a partial reduction in neoblast number, we were always able to detect a significant number of these cells in DjRbAp48 RNAi animals. Parallel to the decrease in neoblasts, a reduction in the number of differentiated cells and the presence of apoptotic-like neoblasts were detectable in RNAi animals. These findings suggest that DjRbAp48 is not involved in neoblast maintenance, but rather in the regulation of differentiation of stem-cell progeny. We discuss our data, taking into account the possibility that DjRbAp48 might control the expression of genes necessary for cell differentiation by influencing chromatin architecture

Nature of light correlations in ghost imaging

We investigate the nature of correlations in Gaussian light sources used for ghost imaging. We adopt methods from quantum information theory to distinguish genuinely quantum from classical correlations. Combining a microscopic analysis of speckle-speckle correlations with an effective coarse-grained description of the beams, we show that quantum correlations exist even in `classical'-like thermal light sources, and appear relevant for the implementation of ghost imaging in the regime of low illumination. We further demonstrate that the total correlations in the thermal source beams effectively determine the quality of the imaging, as quantified by the signal-to-noise ratio.Comment: 12 pages, 5 figures. To appear in Scientific Reports (NPG

Recent progress and current opinions in Brillouin microscopy for life science applications

This is the final version. Available on open access from Springer via the DOI in this recordMany important biological functions and processes are reflected in cell and tissue mechanical properties such as elasticity and viscosity. However, current techniques used for measuring these properties have major limitations, such as that they can often not measure inside intact cells and/or require physical contact-which cells can react to and change. Brillouin light scattering offers the ability to measure mechanical properties in a non-contact and label-free manner inside of objects with high spatial resolution using light, and hence has emerged as an attractive method during the past decade. This new approach, coined "Brillouin microscopy," which integrates highly interdisciplinary concepts from physics, engineering, and mechanobiology, has led to a vibrant new community that has organized itself via a European funded (COST Action) network. Here we share our current assessment and opinion of the field, as emerged from a recent dedicated workshop. In particular, we discuss the prospects towards improved and more bio-compatible instrumentation, novel strategies to infer more accurate and quantitative mechanical measurements, as well as our current view on the biomechanical interpretation of the Brillouin spectra.COST Action BioBrilloui

On the Application of a Monolithic Array for Detecting Intensity-Correlated Photons Emitted by Different Source Types

It is not widely appreciated that many subtleties are involved in the accurate measurement of intensity-correlated photons; even for the original experiments of Hanbury Brown and Twiss (HBT). Using a monolithic 4x4 array of single-photon avalanche diodes (SPADs), together with an off-chip algorithm for processing streaming data, we investigate the difficulties of measuring second-order photon correlations g2 in a wide variety of light fields that exhibit dramatically different correlation statistics: a multimode He-Ne laser, an incoherent intensity-modulated lamp-light source and a thermal light source. Our off-chip algorithm treats multiple photon-arrivals at pixel-array pairs, in any observation interval, with photon fluxes limited by detector saturation, in such a way that a correctly normalized g2 function is guaranteed. The impact of detector background correlations between SPAD pixels and afterpulsing effects on second-order coherence measurements is discussed. These results demonstrate that our monolithic SPAD array enables access to effects that are otherwise impossible to measure with stand-alone detectors.Comment: 17 pages, 6 figure

Bisphenol A and Bisphenol S Induce Endocrine and Chromosomal Alterations in Brown Trout

Bisphenol A is a widely used compound found in large amount of consumer products. As concerns have been raised about its toxicological and public health effect, the use of alternatives to bisphenol A are now increasing. Bisphenol S is one of the analogues being used as a replacement for bisphenol A despite the fact that little is known about the effects of bisphenol S on living organisms. In this study, we investigated the potential endocrine and genotoxic effects of bisphenol A and bisphenol S in juvenile brown trout (Salmo trutta). The fish were exposed to the compounds for either 2 weeks or 8 weeks via sustained-release cholesterol implants containing doses of 2 mg/kg fish or 20 mg/kg fish of the substances. The effects on the thyroid hormone levels and the estrogenic disrupting marker vitellogenin were evaluated, along with the genotoxic markers micronucleated cells and erythrocyte nuclear abnormalities. An increase in plasma vitellogenin was observed in fish exposed to the high dose of bisphenol A for 2 weeks. At this experimental time the level of the thyroid hormone triiodothyronine (T3) in plasma was elevated after bisphenol S exposure at the high concentration, and paralleled by an increase of micronucleated cells. Moreover, bisphenol A induced an increase of micronuclei frequency in fish erythrocytes after the exposure at the lowest dose tested. Taken together the results indicate that both bisphenol A and its alternative bisphenol S cause endocrine disrupting and genotoxic effects in brown trout, although suggesting two different mechanisms of damage underlying bisphenol A and bisphenol S activity

Jaws from the deep: biological and ecological insights on the kitefin shark Dalatias licha from the Mediterranean Sea

Due to their late maturation, extreme longevity, low fecundity and slow growth rates, deep-sea Chondrichthyes are extremely vulnerable to human impacts. Moreover, assessing the impact of deep-sea fisheries is difficult, as many species (including sharks) are part of the bycatch and are often discarded at sea, and/or landed under generic commercial-species codes. The lack of this information on fishery data sets and the limited availability of species-specific life history data make challenging the management of deep-sea Chondrichthyes. The kitefin shark Dalatias licha is a cosmopolitan elasmobranch, mainly found on continental and insular shelf-breaks and slopes in warm-temperate and tropical waters. This species is a common by-catch of the deep-sea trawling, considered as “Endangered” by the IUCN Red List for all European waters, Mediterranean Sea included. Here we present the results of a study based on a total of 78 specimens of kitefin shark collected over 3 years in the Ligurian Sea (NW Mediterranean) as by-catch from deep-water fisheries. Total length ranged from 380 to 1164 mm, and individual weight ranged from 198 to 8000 g. Immature and mature individuals showed a sex ratio dominated by males. Adult males were observed throughout the year, while mature females were observed only in spring-summer. These data lead to hypothesise a spatial segregation between genders. The kitefin shark diet was dominated by bony fish (mainly Macrouridae) and other small sharks (e.g., Galeus melastomus and Etmopterus spinax), but their gut included plastic items and parasites. Data reported here underline the rarity, complex ecology and the threat for this shark species and support the urgency of promoting initiatives for their monitoring and conservation

PMS53 Association Between Teriparatide Adherence and Health Care Utilization and Costs in Real World United States Kyphoplasty/Vertebroplasty Patients

The Gulf of Follonica (Italy) is impacted by the chemical pollution from ancient mining activity and present industrial processes. This study was aimed to determine the bioavailability of dioxin-like compounds (DLCs) in coastal marine environment and to assess the genotoxic potential of waste waters entering the sea from an industrial canal. Moderately high levels of DCLs compounds ( 11 PCDDs + PCDFs 2.18\u201329.00 pg/g dry wt) were detected in Mytilus galloprovincialis transplanted near the waste waters canal and their corresponding Toxic Equivalents (TEQs) calculated. In situ exposed mussels did not show any genotoxic effect (by Comet and Micronucleus assay). Otherwise, laboratory exposure to canal waters exhibited a reduced genomic template stability (by RAPD-PCR assay) but not DNA or chromosomal damage. Our data reveal the need to focus on the levels and distribution of DLCs in edible species from the study area considering their potential transfer to humans through the consumption of sea food