Microsphere‐Mediated Protein Delivery into Cells

Delivering the goods : By coupling proteins to varyingly sized polymeric microspheres, it is possible to deliver them to cells in an easy and effective way. For this study a fluorescent protein (EGFP) and a functional enzyme (β‐galactosidase) were coupled to these particles. Evaluation of the cellular uptake after “beadfection” shows that the functionality and activity of these proteins were not adversely affected through coupling to the carrier system; this shows that their functional structure is retained.

     

Effect of skim milk treated with high hydrostatic pressure on permeate flux and fouling during ultrafiltration

Ultrafiltration (UF) is largely used in the dairy industry to generate milk and whey protein concentrate for standardization of milk or production of dairy ingredients. Recently, it was demonstrated that high hydrostatic pressure (HHP) extended the shelf life of milk and improved rennet coagulation and cheese yield. Pressurization also modified casein micelle size distribution and promoted aggregation of whey proteins. These changes are likely to affect UF performance. Consequently, this study determined the effect of skim milk pressurization (300 and 600 MPa, 5 min) on UF performance in terms of permeate flux decline and fouling. The effect of HHP on milk proteins was first studied and UF was performed in total recycle mode at different transmembrane pressures to determine optimal UF operational parameters and to evaluate the effect of pressurization on critical and limiting fluxes. Ultrafiltration was also performed in concentration mode at a transmembrane pressure of 345 kPa for 130 or 140 min to evaluate the decline of permeate flux and to determine fouling resistances. It was observed that average casein micelle size decreased by 32 and 38%, whereas β-lactoglobulin denaturation reached 30 and 70% at 300 and 600 MPa, respectively. These results were directly related to UF performance because initial permeate fluxes in total recycle mode decreased by 25% at 300 and 600 MPa compared with nonpressurized milk, critical flux, and limiting flux, which were lower during UF of milk treated with HHP. During UF in concentration mode, initial permeate fluxes were 30% lower at 300 and 600 MPa compared with the control, but the total flux decline was higher for nonpressurized milk (62%) compared with pressure-treated milk (30%). Fouling resistances were similar, whatever the treatment, except at 600 MPa where irreversible fouling was higher. Characterization of the fouling layer showed that caseins and β-lactoglobulin were mainly involved in membrane fouling after UF of pressure-treated milk. Our results demonstrate that HHP treatment of skim milk drastically decreased UF performance.     

Quantitative Evaluation of the Cellular Uptake of Nanodiamonds by Monocytes and Macrophages

Fluorescent nanodiamonds (FNDs) with negative nitrogen-vacancy (NV⁻) defect centers are great probes for biosensing applications, with potential to act as biomarkers for cell differentiation. To explore this concept, uptake of FNDs (≈120 nm) by THP-1 monocytes and monocyte-derived M0-macrophages is studied. The time course analysis of FND uptake by monocytes confirms differing FND-cell interactions and a positive time-dependence. No effect on cell viability, proliferation, and differentiation potential into macrophages is observed, while cells saturated with FNDs, unload the FNDs completely by 25 cell divisions and subsequently take up a second dose effectively. FND uptake variations by THP-1 cells at early exposure-times indicate differing phagocytic capability. The cell fraction that exhibits relatively enhanced FND uptake is associated to a macrophage phenotype which derives from spontaneous monocyte differentiation. In accordance, chemical-differentiation of the THP-1 cells into M0-macrophages triggers increased and homogeneous FND uptake, depleting the fraction of cells that were non-responsive to FNDs. These observations imply that FND uptake allows for distinction between the two cell subtypes based on phagocytic capacity. Overall, FNDs demonstrate effective cell labeling of monocytes and macrophages, and are promising candidates for sensing biological processes that involve cell differentiation.     

Prediction of apparent properties with uncertain material parameters using high‐order fictitious domain methods and PGD model reduction

This contribution presents a numerical strategy to evaluate the effective properties of image‐based microstructures in the case of random material properties. The method relies on three points: (1) a high‐order fictitious domain method; (2) an accurate spectral stochastic model; and (3) an efficient model‐reduction method based on the proper generalized decomposition in order to decrease the computational cost introduced by the stochastic model. A feedback procedure is proposed for an automatic estimation of the random effective properties with a given confidence. Numerical verifications highlight the convergence properties of the method for both deterministic and stochastic models. The method is finally applied to a real 3D bone microstructure where the empirical probability density function of the effective behaviour could be obtained. Copyright © 2016 John Wiley & Sons, Ltd.     

Hybrid atomistic�Ccontinuum simulations of fluid flows involving interfaces

In this work, we use an hybrid atomistic–continuum (HAC) simulation method to study transient and steady isothermal flows of Lennard-Jones fluids near interfaces. Our hybrid method is based on a domain decomposition algorithm. The flow domain is composed of two overlapping regions: an atomistic region described by molecular dynamics, and a continuum region described by a finite volume discretization of the incompressible Navier–Stokes equations. To show the interest of such an hybrid method to compute flows near fluid/solid interface, we first applied our hybrid scheme to the classical Couette flow, where the moving wall is modelled at the atomistic scale. In addition, we also studied an oscillatory shear flow. Then, to compute flows near fluid/fluid interface, we applied our method to a two-phase Couette flow (liquid/gas), where the interface is modelled at the molecular scale. We show that hybrid results can sometimes differ from those provided by analytical solutions deduced from continuum mechanics equations combined with usual boundary/interface relations. For the Couette and oscillatory shear flows, a good agreement is found between hybrid simulations and macroscopic analytical solutions, however, we noticed that the fluid in contact with the wall can be more entailed than what expected. For the liquid/gas Couette flow, the hybrid simulation exhibits an unexpected jump of the velocity in the interfacial region, corresponding to a partial slip between the two fluid phases. Those interesting results highlight the interest of using an HAC method to deal with systems for which surfaces/interfaces effects are important.     

Metal redistribution by surface casting of four earthworm species in sandy and loamy clay soils

Bioturbation of metal contaminated soils contributes considerably to redistribution and surfacing of contaminated soil from deeper layers. To experimentally measure the contribution of Allolobophora chlorotica, Aporrectodea caliginosa, Lumbricus rubellus and L. terrestris to soil surface casting, a time-course experiment was performed under laboratory conditions. Earthworms were incubated in perspex columns filled with sandy soil (2% organic matter, 2.9% clay) or loamy clay soil (15% organic matter, 20% clay), and surface casts were collected after up to 80 days. On the sandy soil, A. caliginosa and L. rubellus brought approximately 7.1-16 g dry wt. casts/g fresh wt. earthworm to the surface, which is significantly more than A. chlorotica and L. terrestris (2.5-5.0 g dry wt./g fresh wt.). A. caliginosa was the only species that produced significantly more surface casts in the sandy soil than in the loamy clay soil. In the loamy clay soil, no differences in biomass-corrected casting rates were found among the species. Surface casting rates tended to decrease after 20 days. Considering the densities of the different species in a Dutch floodplain area Afferdensche and Deestsche Waarden, surface cast production is estimated to amount to 2.0 kg dry soil/m² after 80 days, which could be extrapolated to 2.7-9.1 kg/m² per year. These amounts correspond to a surface deposition of a layer of approximately 1.9-6.5 mm/year, which is of the same order or even slightly higher than the sedimentation rate and much higher than the amount of soil brought to the soil surface by bioturbating small mammals.     

Contribution of Yeast Studies to the Understanding of BCL-2 Family Intracellular Trafficking

BCL-2 family members are major regulators of apoptotic cell death in mammals. They form an intricate regulatory network that ultimately regulates the release of apoptogenic factors from mitochondria to the cytosol. The ectopic expression of mammalian BCL-2 family members in the yeast Saccharomyces cerevisiae, which lacks BCL-2 homologs, has been long established as a useful addition to the available models to study their function and regulation. In yeast, individual proteins can be studied independently from the whole interaction network, thus providing insight into the molecular mechanisms underlying their function in a living context. Furthermore, one can take advantage of the powerful tools available in yeast to probe intracellular trafficking processes such as mitochondrial sorting and interactions/exchanges between mitochondria and other compartments, such as the endoplasmic reticulum that are largely conserved between yeast and mammals. Yeast molecular genetics thus allows the investigation of the role of these processes on the dynamic equilibrium of BCL-2 family members between mitochondria and extramitochondrial compartments. Here we propose a model of dynamic regulation of BCL-2 family member localization, based on available evidence from ectopic expression in yeast.     

Dissolution and geopolymerization of synthetic fly ash glass

Journal of Materials Science - Two fly ash glasses were synthesized and dissolved in KOH at a water/solid ratio of?≤?0.3. We measured reaction kinetics, gel formation, and...