The effects of washing a collagen sample prior to TEM examination

Transmission electron microscopy (TEM) is an important analysis technique to visualize (bio)macromolecules and their assemblies, including collagen fibers. Many protocols for TEM sample preparation of collagen involve one or more washing steps to remove excess salts from the dispersion that could hamper analysis when dried on a TEM grid. Such protocols are not standardized and washing times as well as washing solvents vary from procedure to procedure, with each research group typically having their own protocol. Here, we investigate the influence of washing with water, ethanol, but also methanol and 2-propanol, for both mineralized and unmineralized collagen samples via a protocol based on centrifugation. Washing with water maintains the hydrated collagen structure and the characteristic banding pattern can be clearly observed. Conversely, washing with ethanol results in dehydration of the fibrils, often leading to aggregation of the fibers and a less obvious banding pattern, already within 1 min of ethanol exposure. As we show, this process is fully reversible. Similar observations were made for methanol and propanol. Based on these results, a standardized washing protocol for collagenous samples is proposed.     

Effect of aluminium acetyl acetonate on the hydrogen and nitrogen permeation of carbon molecular sieves membranes

With a growing interest in hydrogen as energy carrier, the efficient purification of hydrogen from gaseous mixtures is very important. This paper addresses the separation of hydrogen using Carbon Molecular Sieves Membranes (CMSM), which show an attractive combination of high permeability, selectivity and stability. Supported CMSM containing various amounts of aluminium have been prepared from novolac and aluminium acetyl acetonate (Al(acac)₃) as carbon and alumina precursors. The thickness of the CMSM layers depend on the content of Al(acac)₃ in the dipping solution, which also has influence in the pore size and pore size distribution of the membranes. The permeation properties of the membranes against the Al content in the membrane follows a volcano shape, where the membrane containing 4 wt (%) of Al(acac)₃ has the best properties and was stable during 720 h for hydrogen at 150 °C and 6 bar pressure difference. All the CMSM have permeation properties well above the Robeson Upper limit.     

Inhibition of Autophagy Does Not Re-Sensitize Acute Myeloid Leukemia Cells Resistant to Cytarabine

Elevated activation of the autophagy pathway is currently thought to be one of the survival mechanisms allowing therapy-resistant cancer cells to escape elimination, including for cytarabine (AraC)-resistant acute myeloid leukemia (AML) patients. Consequently, the use of autophagy inhibitors such as chloroquine (CQ) is being explored for the re-sensitization of AraC-resistant cells. In our study, no difference in the activity of the autophagy pathway was detected when comparing AraC-Res AML cell lines to parental AraC-sensitive AML cell lines. Furthermore, treatment with autophagy inhibitors CQ, 3-Methyladenine (3-MA), and bafilomycin A1 (BafA1) did not re-sensitize AraC-Res AML cell lines to AraC treatment. However, in parental AraC-sensitive AML cells, treatment with AraC did activate autophagy and, correspondingly, combination of AraC with autophagy inhibitors strongly reduced cell viability. Notably, the combination of these drugs also yielded the highest level of cell death in a panel of patient-derived AML samples even though not being additive. Furthermore, there was no difference in the cytotoxic effect of autophagy inhibition during AraC treatment in matched de novo and relapse samples with differential sensitivity to AraC. Thus, inhibition of autophagy may improve AraC efficacy in AML patients, but does not seem warranted for the treatment of AML patients that have relapsed with AraC-resistant disease.     

Electrowetting-Assisted Generation of Ultrastable High Charge Densities in Composite Silicon Oxide–Fluoropolymer Electret Samples for Electric Nanogenerators

Electric nanogenerators that directly convert the energy of moving drops into electrical signals require hydrophobic substrates with a high density of static electric charge that is stable in “harsh environments” created by continued exposure to potentially saline water. The recently proposed charge-trapping electric generators (CTEGs) that rely on stacked inorganic oxide–fluoropolymer (FP) composite electrets charged by homogeneous electrowetting-assisted charge injection (h-EWCI) seem to solve both problems, yet the reasons for this success have remained elusive. Here, systematic measurements at variable oxide and FP thickness, charging voltage, and charging time and thermal annealing up to 230 °C are reported, leading to a consistent model of the charging process. It is found to be controlled by an energy barrier at the water-FP interface, followed by trapping at the FP-oxide interface. Protection by the FP layer prevents charge densities up to −1.7 mC m⁻² from degrading and the dielectric strength of SiO₂ enables charge decay times up to 48 h at 230 °C, suggesting lifetimes against thermally activated discharging of thousands of years at room temperature. Combining high dielectric strength oxides and weaker FP top coatings with electrically controlled charging provides a new paradigm for developing ultrastable electrets for applications in energy harvesting and beyond.     

Estimates of daily oxygen consumption,carbon dioxide and methane emissions,and heat production for beef and dairy cattle using spot gas sampling

A simulation study was conducted to examine accuracy of estimating daily O₂ consumption, CO₂ and CH₄ emissions, and heat production (HP) using a spot sampling technique and to determine optimal spot sampling frequency (FQ). Data were obtained from 3 experiments where daily O₂ consumption, emissions of CO₂ and CH₄, and HP were measured using indirect calorimetry (respiration chamber or headbox system). Experiment 1 used 8 beef heifers (ad libitum feeding; gaseous exchanges measured every 30 min over 3 d in respiration chambers); Experiment 2 used 56 lactating Holstein-Friesian cows (restricted feeding; gaseous exchanges measured every 12 min over 3 d in respiration chambers); Experiment 3 used 12 lactating Jersey cows (ad libitum feeding; gaseous exchanges measured every hour for 1 d using headbox style chambers). Within experiment, averages of all measurements (FQALL) and averages of measurements selected at time points with 12, 8, 6, or 4 spot sampling FQ (i.e., sampling every 2, 3, 4, and 6 h in a 24-h cycle, respectively; FQ12, FQ8, FQ6, and FQ4, respectively) were compared. Within study a mixed model was used to compare gaseous exchanges and HP among FQALL, FQ12, FQ8, FQ6, and FQ4, and an interaction of dietary treatment by FQ was examined. A regression model was used to evaluate accuracy of spot sampling within study [i.e., FQALL (observed) vs. FQ12, FQ8, FQ6, or FQ4 (estimated)]. No interaction of diet by FQ was observed for any variables except for CH₄ production in experiment 1. No FQ effect was observed for gaseous exchanges and HP except in experiment 2 where CO₂ production was less (5,411 vs. 5,563 L/d) for FQ4 compared with FQALL, FQ12, and FQ8. A regression analysis between FQALL and each FQ within study showed that slopes and intercepts became farther from 1 and 0, respectively, for almost all variables as FQ decreased. Most variables for FQ12 and FQ8 had root mean square prediction error (RMSPE) less than 10% of the mean and concordance correlation coefficient (CCC) greater than 0.80, and RMSPE increased and CCC decreased as FQ decreased. When a regression analysis was conducted with combined data from the 3 experiments (mixed model with study as a random effect), results agreed with those from the analysis for the individual studies. Prediction errors increased and CCC decreased as FQ decreased. Generally, all the estimates from FQ12, FQ8, FQ6, and FQ4 had RMSPE less than 10% of the means and CCC greater than 0.90 except for FQ6 and FQ4 for O₂ consumption and CH₄ production. In conclusion, the spot sampling simulation with 3 indirect calorimetry experiments indicated that FQ of at least 8 samples (every 3 h in a 24-h cycle) was required to estimate daily O₂ consumption, CO₂ and CH₄ production, and HP and to detect changes in those in response to dietary treatments. This sampling FQ may be considered when using techniques that measure spot gas exchanges such as the GreenFeed and face mask systems.     

Symmetric and asymmetric membranes based on La0.5Sr0.5Fe0.7Ga0.3O3-δ perovskite with high oxygen semipermeation flux performances and identification of the rate-determining step of oxygen transport

This work focuses on identifying the rate-determining step of oxygen transport through La_0.5Sr_0.5Fe_0.7Ga_0.3O_3-δ membranes with symmetric and asymmetric architectures. The best oxygen semipermeation fluxes are 3.4 10⁻³ mol. m^-2.s^-1 and 6.3 10⁻³ mol. m^-2.s^-1 at 900 °C for the symmetric membrane and asymmetric membrane with a modified surface. The asymmetric membrane with a modified surface leads to an increase of approximately 7 times the oxygen flux compared to that obtained with the La_0.5Sr_0.5Fe_0.7Ga_0.3O_3-δ dense membrane without surface modification. This work also shows that the oxygen flux is mainly governed by gaseous oxygen diffusion through the porous support of asymmetric La_0.5Sr_0.5Fe_0.7Ga_0.3O_3-δ membranes.     

Material encapsulation in poly(methyl methacrylate) shell: A review

Material encapsulation is a relatively new technique for coating a micro/nanosize particle or droplet with polymeric or inorganic shell. Encapsulation technology has many applications in various fields including drug delivery, cosmetic, agriculture, thermal energy storage, textile, and self-healing polymers. Poly(methyl methacrylate) (PMMA) is widely used as shell material in encapsulation due to its high chemical stability, biocompatibility, nontoxicity, and good mechanical properties. The main approach for micro/nanoencapsulation of materials using PMMA as shell comprises emulsion-based techniques such as emulsion polymerization and solvent evaporation from oil-in-water emulsion. In the present review, we first focus on the encapsulation techniques of liquid materials with PMMA shell by analyzing the effective processing parameters influencing the preparation of PMMA micro/nanocapsules. We then describe the morphology of PMMA capsules in emulsion systems according to thermodynamic relations. The techniques to investigation of mechanical properties of capsule shell and the release mechanisms of core material from PMMA capsules were also investigated. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 48039.     

Structural analysis and optimization of a tethered swept wing for airborne wind energy generation

In this paper, we present an aero‐structural model of a tethered swept wing for airborne wind energy generation. The carbon composite wing has neither fuselage nor actuated aerodynamic control surfaces and is controlled entirely from the ground using three separate tethers. The computational model is efficient enough to be used for weight optimisation at the initial design stage. The main load‐bearing wing component is a nontypical “D”‐shaped wing‐box, which is represented as a slender carbon composite shell and further idealised as a stack of two‐dimensional cross section models arranged along an anisotropic one‐dimensional beam model. This reduced 2+1D finite element model is then combined with a nonlinear vortex step method that determines the aerodynamic load. A bridle model is utilised to calculate the individual forces as a function of the aerodynamic load in the bridle lines that connect the main tether to the wing. The entire computational model is used to explore the influence of the bride on the D‐box structure. Considering a reference D‐box design along with a reference aerodynamic load case, the structural response is analysed for typical bridle configurations. Subsequently, an optimisation of the internal geometry and laminate fibre orientations is carried out using the structural computation models, for a fixed aerodynamic and bridle configuration. Aiming at a minimal weight of the wing structure, we find that for the typical load case of the system, an overall weight savings of approximately 20% can be achieved compared with the initial reference design.