Preparation of single cell suspensions enriched in mouse brain vascular cells for single-cell RNA sequencing.

Cerebral blood vessels supply oxygen and nutrients, remove metabolic waste, and play a critical role in maintaining brain homeostasis. Cerebrovasculature is composed of heterogeneous populations of brain vascular cells (BVCs). A major challenge in effective cerebrovascular transcriptional profiling is high-quality BVC procurement, permitting high sequencing depth. Here, we establish cell isolation procedures for glio-vascular cell-enriched single-cell RNA sequencing enabling unbiased characterization of BVC transcriptional heterogeneity. Our approach can be used to address vascular-specific contribution to brain diseases. For complete details on the use and execution of this protocol, please refer to Yamazaki et al. (2021).

Development of Novel Bead Milling Technology with Less Metal Contamination by pH Optimization of the Suspension Medium.

To develop novel contamination-less bead milling technology without impairing grinding efficiency, we investigated the effect of the formulation properties on the grinding efficiency and the metal contamination generated during the grinding process. Among the various formulations tested, the combination of polyvinylpyrrolidone and sodium dodecyl sulfate was found to be suitable for efficiently pulverizing phenytoin. However, this stabilization system included a relatively strong acid, which raised the concern of possible corrosion of the zirconia beads. An evaluation of the process clearly demonstrated that acidic pH promoted bead dissolution, suggesting that this could be suppressed by controlling the pH of the suspension. Among the various pH values tested, the metal contamination generated during the grinding process could be significantly reduced in the optimized pH range without significant differences in the particle size of the phenytoin suspension after pulverization. In addition, the contamination reduction by pH optimization in the presence of physical contact among the beads was approximately 10-times larger than that without bead contact, suggesting that pH optimization could suppress not only bead dissolution but also the wear caused by bead collisions during the grinding process. These findings show that pH optimization is a simple but effective approach to reducing metal contamination during the grinding process.

Persistence of Severe Acute Respiratory Syndrome Coronavirus 2 in Aerosol Suspensions.

We aerosolized severe acute respiratory syndrome coronavirus 2 and determined that its dynamic aerosol efficiency surpassed those of severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome. Although we performed experiment only once across several laboratories, our findings suggest retained infectivity and virion integrity for up to 16 hours in respirable-sized aerosols.

Separation of suspensions and emulsions via ultrasonic standing waves - a review.

Ultrasonic standing waves (USW) separation is an established technology for micro scale applications due to the excellent control to manipulate particles acoustically achieved when combining high frequency ultrasound with laminar flow in microchannels, allowing the development of numerous applications. Larger scale systems (pilot to industrial) are emerging; however, scaling up such processes are technologically very challenging. This paper reviews the physical principles that govern acoustic particle/droplet separation and the mathematical modeling techniques developed to understand, predict, and design acoustic separation processes. A further focus in this review is on acoustic streaming, which represents one of the major challenges in scaling up USW separation processes. The manuscript concludes by providing a brief overview of the state of the art of the technology applied in large scale systems with potential applications in the dairy and oil industries.

Bio-inspired particle separator design based on the food retention mechanism by suspension-feeding fish.

A new particle separator is designed using a crossflow filtration mechanism inspired by suspension-feeding fish in this study. To construct the model of the bio-inspired particle separator, computational fluid dynamics techniques are used, and parameters related to separator shape, fluid flow and particle properties that might affect the performance in removing particles from the flow, are varied and tested. The goal is to induce a flow rotation which enhances the separation of particles from the flow, reduce the particle-laden flow that exits via a collection zone at the lower/posterior end of the separator, while at the same time increase the concentration of particles in that flow. Based on preliminary particle removal efficiency tests, an exiting flow through the collection zone of about 8% of the influent flow rate is selected for all the performance tests of the separator including trials with particles carried by air flow instead of water. Under this condition, the simulation results yield similar particle removal efficiencies in water and air but with different particle properties. Particle removal efficiencies (percentage of influent particles that exit through the collection zone) were determined for particles ranging in size from 1 to 1500 µm with a density between 1000 and 1150 kg m(-3) in water and 2 and 19 mm and 68 and 2150 kg m(-3) in air. As an example, removal efficiencies are 66% and 64% for 707 µm diameter particles with a density of 1040 kg m(-3) in water and for 2 mm particles with a density of 68 kg m(-3) in air, respectively. No significant performance difference is found by geometrically scaling the inlet diameter of the separator up or down in the range from 2.5 to 10 cm.

Effect of particle size on skin permeation and retention of piroxicam in aqueous suspension.

The effects of particle size on the skin permeation and retention of piroxicam (PXC) in an aqueous suspension were investigated. PXC particles of about 23 nm, 173 nm, and 2.1 microm in size were prepared by the cogrinding of PXC/polyvinylpyrrolidone (PVP) K12/sodium dodecyl sulfate physical mixture (mean particle size, 9.6 microm) using a vibrational rod and ball mills. Particles were stable after storage in 0.1 m acetate buffer (pH 4.5) for 24 h. The amount of PXC that permeated and was retained in hairless mouse skin increased with the reduction of particle size up to 23 nm. Amorphous PXC was formed when PXC was coground with PVP, though the amorphous formation did not affect the amount of PXC permeated. Reduction of particle size to less than 50 nm, and the subsequent increase in surface area of PXC nanocrystals appeared to affect skin permeation and retention behavior.

Scale-down of continuous filtration for rapid bioprocess design: Recovery and dewatering of protein precipitate suspensions.

The early specification of bioprocesses often has to be achieved with small (tens of millilitres) quantities of process material. If extensive process discovery is to be avoided at pilot or industrial scale, it is necessary that scale-down methods be created that not only examine the conditions of process stages but also allows production of realistic output streams (i.e., streams truly representative of the large scale). These output streams can then be used in the development of subsequent purification operations. The traditional approach to predicting filtration operations is via a bench-scale pressure filter using constant pressure tests to examine the effect of pressure on the filtrate flux rate and filter cake dewatering. Interpretation of the results into cake resistance at unit applied pressure (alpha) and compressibility (n) is used to predict the pressure profile required to maintain the filtrate flux rate at a constant predetermined value. This article reports on the operation of a continuous mode laboratory filter in such a way as to prepare filter cakes and filtrate similar to what may be achieved at the industrial scale. Analysis of the filtration rate profile indicated the filter cake to have changing properties (compressibility) with time. Using the insight gained from the new scale-down methodology gave predictions of the flux profile in a pilot-scale candle filter superior to those obtained from the traditional batch filter used for laboratory development.

Direct influent filtration as a pretreatment step for more sustainable wastewater treatment systems.

Within the scope of a research programme into wastewater treatment scenarios based on physical-chemical pretreatment, the application of direct influent filtration (DIF) as a first treatment step in a wastewater treatment system was investigated. The aim of the experimental research was to investigate the feasibility of DIF as a pretreatment step for advanced particle removal. With a large scale pilot-plant filter at WWTP Leiden-Noord, The Netherlands the removal characteristics for suspended and colloidal material were investigated as well as operational conditions of eight different filter configurations. From the experimental research it was concluded that filtration of raw wastewater is possible, notwithstanding the relatively short run-times due to clogging. In general, the filters produced a filtrate with a constant quality with low concentrations of solids and low turbidity. Without addition of chemicals hardly any colloids and solubles were retained, but only suspended particulates were removed. After dosage of iron or polymer, it was possible to remove more suspended matter and a high proportion of colloidal material. Finally it was concluded that DIF could be applied as a compact treatment system to produce a high quality primary effluent with a constant composition, but for practical application further research has to be done.