Collagen-hyaluronic acid scaffolds for adipose tissue engineering.

Three-dimensional (3-D) in vitro models of the mammary gland require a scaffold matrix that supports the development of adipose stroma within a robust freely permeable matrix. 3-D porous collagen-hyaluronic acid (HA: 7.5% and 15%) scaffolds were produced by controlled freeze-drying technique and crosslinking with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride. All scaffolds displayed uniform, interconnected pore structure (total porosity approximately 85%). Physical and chemical analysis showed no signs of collagen denaturation during the formation process. The values of thermal characteristics indicated that crosslinking occurred and that its efficiency was enhanced by the presence of HA. Although the crosslinking reduced the swelling of the strut material in water, the collagen-HA matrix as a whole tended to swell more and show higher dissolution resistance than pure collagen samples. The compressive modulus and elastic collapse stress were higher for collagen-HA composites. All the scaffolds were shown to support the proliferation and differentiation 3T3-L1 preadipocytes while collagen-HA samples maintained a significantly increased proportion of cycling cells (Ki-67+). Furthermore, collagen-HA composites displayed significantly raised Adipsin gene expression with adipogenic culture supplementation for 8 days vs. control conditions. These results indicate that collagen-HA scaffolds may offer robust, freely permeable 3-D matrices that enhance mammary stromal tissue development in vitro.This was supported by the Biotechnology and Biological Sciences Research Council

The Australian Census Longitudinal Dataset: using record linkage to create a longitudinal sample from a series of cross-sections

© 2017 Australian Statistical Publishing Association Inc. Published by John Wiley & Sons Australia Pty Ltd. The Australian Bureau of Statistics is creating a longitudinal sample, called the Australian Census Longitudinal Dataset (ACLD), by linking person records across its five-yearly Census of Population and Housing. This paper proposes a Multi-Panel framework for selecting and weighting records in the ACLD. This framework can be applied more generally to selecting longitudinal samples from a series of cross-sectional administrative files. The proposed framework avoids some significant limitations of the popular ‘Top-Up’ sampling approach to maintaining the cross-sectional and longitudinal representativeness of a sample over time

A data extraction system for underwater particle holography

Pulsed laser holography is an extremely powerful technique for the study of particle fields as it allows instantaneous, noninvasive high-resolution recording of substantial volumes. By replaying the real image one can obtain the size, shape, position and - if multiple exposures are made - velocity of every object in the recorded field. Manual analysis of large volumes containing thousands of particles is, however, an enormous and time-consuming task, with operator fatigue an unpredictable source of errors. Clearly the value of holographic measurements also depends crucially on the quality of the reconstructed image: not only will poor resolution degrade size and shape measurements, but aberrations such as coma and astigmatism can change the perceived centroid of a particle, affecting position and velocity measurements. For large-scale applications of particle field holography, specifically the in situ recording of marine plankton with 'HoloCam,' we have developed an automated data extraction system that can be readily switched between the in-line and off-axis geometries and provides optimised reconstruction from holograms recorded underwater. As a videocamera is automatically stepped through the 200 by 200 by 1000mm sample volume, image processing and object tracking routines locate and extract particle images for further classification by a separate software module

Ambient humidity control for maximising replay intensity and resolution in aberration-compensated off-axis holograms of underwater objects

In hologrammetry it is usually more desirable to reconstruct the real image than the virtual image, since the latter must be viewed at a distance through the window of the holographic plate itself. In applications where the recorded scene was in water but with replay into air it is necessary to correct for the refractive index difference. This can be done by reconstructing the image with shorter wavelength illumination combined with a change in beam angle to satisfy the grating equation, but these changes mean that the Bragg condition may no longer be satisfied during replay, reducing the diffraction efficiency and making the reconstructed images difficult to see. Changing the replay beam angle to better satisfy the Bragg condition makes the images brighter, but also renders them unrecognizable by introducing severe optical aberrations. A possible solution is to alter the Bragg properties of the hologram. In particular, the emulsion thickness can be conveniently controlled by altering the humidity of the atmosphere surrounding the hologram without causing any long-term changes or damage to the holographic plate. The validity of using humidity change to tune the Bragg properties of emulsions during replay has been demonstrated by measuring the brightness and perceived resolution of a reconstructed real image from a hologram over a wide range of humidities. The results have been compared with a simple model based on the Flory-Huggins theory of polymer swelling

Development of three-dimensional collagen scaffolds with controlled architecture for cell migration studies using breast cancer cell lines

Cancer is characterized by cell heterogeneity and the development of 3D $\textit{in vitro }$ assays that can distinguish more invasive or migratory phenotypes could enhance diagnosis or drug discovery. 3D collagen scaffolds have been used to develop analogues of complex tissues $\textit{in vitro }$ and are suited to routine biochemical and immunological assays. We sought to increase 3D model tractability and modulate the migration rate of seeded cells using an ice-templating technique to create either directional/anisotropic or non-directional/isotropic porous architectures within cross-linked collagen scaffolds. Anisotropic scaffolds supported the enhanced migration of an invasive breast cancer cell line MDA-MB-231 with an altered spatial distribution of proliferative cells in contrast to invasive MDA-MB-468 and non-invasive MCF-7 cells lines. In addition, MDA-MB-468 showed increased migration upon epithelial-to-mesenchymal transition (EMT) in anisotropic scaffolds. The provision of controlled architecture in this system may act both to increase assay robustness and as a tuneable parameter to capture detection of a migrated population within a set time, with consequences for primary tumour migration analysis. The separation of invasive clones from a cancer biomass with in vitro platforms could enhance drug development and diagnosis testing by contributing assay metrics including migration rate, as well as modelling cell-cell and cell-matrix interaction in a system compatible with routine histopathological testing.The authors gratefully acknowledge the financial support of the ERC Advanced Grant 320598 3D-E and the Newton Trust. A.H. held a Daphne Jackson Fellowship funded by the University of Cambridge for part of the work. R.D.H. is funded through a NC3Rs studentship

Optimising replay intensity and resolution in aberration-compensated

In hologrammetry it is desirable to reconstruct the real image rather than the virtual image as the latter must be viewed at a distance through the window of the holographic plate itself. When a scene is located in water but the image is replayed in air, it is necessary to correct for the refractive index difference by reconstructing the image with shorter wavelength illumination and changing the beam angle to satisfy the grating equation. However this means that the Bragg condition may no longer be satisfied during replay, reducing the diffraction efficiency and decreasing the signal-to-noise ratio of the reconstructed images. Changing the replay beam angle to satisfy better the Bragg condition makes the images brighter but also renders them unusable by increasing the optical aberrations. Our solution is to alter the Bragg properties of the hologram by altering the humidity of the surrounding atmosphere. This approach has been experimentally demonstrated for Agfa 8E56HD emulsions by measuring the brightness and resolution of a reconstructed real image from an off-axis hologram over a humidity range from 6 to 93 percent. The emulsion swelling and its effect on the Bragg properties of the hologram were modelled using the Flory-Huggins theory of polymer swelling

The development and sea trials of a subsea holographic camera for large volume in-situ recording of marine organisms

We describe the development, construction and sea testing of an underwater holographic camera (HoloCam) for in situ recording of marine organisms and particles in large volumes of sea water. HoloCam comprises a laser, power supply, holographic recording optics and plate holders, a water-tight housing and a support frame. Added to this are control electronics such that the entire camera is remotely operable and controllable from ship or dock-side. Uniquely the camera can simultaneously record both in-line and off-axis holograms using a pulsed frequency doubled Nd-YAG laser. In-line holography is capable of producing images of organisms with a resolution of better than 10 Pm (at concentrations up to a few thousand per cubic centimetre at the smallest sizes). Off-axis holograms of aquatic systems of up to 50,000 cm3 volume, have been recorded. Following initial laboratory testing, the holo-camera was evaluated in an observation tank and ultimately was tested in Loch Etive, Scotland. In-line and off-axis holograms were recorded to a depth of 100 m. We will present results on the test dives and evaluation of the camera performance

Positive association between serum silicon levels and bone mineral density in female rats following oral silicon supplementation with monomethylsilanetriol.

UNLABELLED: Observational (epidemiological) studies suggest the positive association between dietary silicon intake and bone mineral density may be mediated by circulating estradiol level. Here, we report the results of a silicon supplementation study in rats that strongly support these observations and suggest an interaction between silicon and estradiol. INTRODUCTION: Epidemiological studies report strong positive associations between dietary silicon (Si) intake and bone mineral density (BMD) in premenopausal women and indicate that the association may be mediated by estradiol. We have tested this possibility in a mixed-gender rodent intervention study. METHODS: Tissue samples were obtained from three groups of 20-week-old Sprague Dawley rats (five males and five females per group) that had been supplemented ad libitum for 90 days in their drinking water with (i) <0.1 mg Si/L (vehicle control), (ii) 115 mg Si/L (moderate dose) or (iii) 575 mg Si/L (high dose). All rats received conventional laboratory feed, whilst supplemental Si was in the form of monomethylsilanetriol, increasing dietary Si intakes by 18 and 99 %, for the moderate- and high-dose groups, respectively. RESULTS: Fasting serum and tissue Si concentrations were increased with Si supplementation (p < 0.05), regardless of gender. However, only for female rats was there (i) a trend for a dose-responsive increase in serum osteocalcin concentration with Si intervention and (ii) strong significant associations between serum Si concentrations and measures of bone quality (p < 0.01). Correlations were weaker or insignificant for tibia Si levels and absent for other serum or tibia elemental concentrations and bone quality measures. CONCLUSIONS: Our findings support the epidemiological observations that dietary Si positively impacts BMD in younger females, and this may be due to a Si-estradiol interaction. Moreover, these data suggest that the Si effect is mediated systemically, rather than through its incorporation into bone

HoloCam: A subsea holographic camera for recording marine organisms and particles

The HoloCam system is a major component of a multi-national multi-discipline project known as HoloMar (funded by the European Commission under the MAST III initiative). The project is concerned with the development of pulsed laser holography to analyse and monitor the populations of living organisms and inanimate particles within the world's oceans. We describe here the development, construction and evaluation of a prototype underwater camera, the purpose of which is to record marine organisms and particles, in-situ. Recording using holography provides several advantages over conventional sampling methods in that it allows non-intrusive, non-destructive, high-resolution imaging of large volumes (up to 10^5 cm^3) in three dimensions. The camera incorporates both in-line and off-axis holographic techniques, which allows particles from a few micrometres to tens of centimetres to be captured. In tandem with development of the HoloCam, a dedicated holographic replay system and an automated data extraction and image processing facility are being developed. These will allow, optimisation of the images recorded by the camera, identification of species and particle concentration plotting

GRB 180620A: Evidence for Late-time Energy Injection

The early optical emission of gamma-ray bursts (GRBs) gives an opportunity to understand the central engine and first stages of these events. About 30% of GRBs present flares whose origin is still a subject of discussion. We present optical photometry of GRB 180620A with the COATLI telescope and RATIR instrument. COATLI started to observe from the end of prompt emission at T + 39.3 s and RATIR from T + 121.4 s. We supplement the optical data with the X-ray light curve from Swift/XRT. We observe an optical flare from T + 110 s to T + 550 s, with a temporal index decay α O,decay = 1.32 ± 0.01, and Δt/t = 1.63, which we interpret as the signature of a reverse shock component. After the initial normal decay the light curves show a long plateau from T + 500 s to T + 7800 s in both X-rays and the optical before decaying again after an achromatic jet break at T + 7800 s. Fluctuations are seen during the plateau phase in the optical. Adding to the complexity of GRB afterglows, the plateau phase (typically associated with the coasting phase of the jet) is seen in this object after the "normal" decay phase (associated with the deceleration phase of the jet), and the jet break phase occurs directly after the plateau. We suggest that this sequence of events can be explained by a rapid deceleration of the jet with t d ≲ 40 s due to the high density of the environment (≈100 cm-3) followed by reactivation of the central engine, which causes the flare and powers the plateau phase