A Rapid Human Lung Tissue Dissociation Protocol Maximizing Cell Yield and Minimizing Cellular Stress.

The human lung is a complex organ that comprises diverse populations of epithelial, mesenchymal, vascular, and immune cells, which gains even greater complexity during disease states. To effectively study the lung at a single-cell level, a dissociation protocol that achieves the highest yield of viable cells of interest with minimal dissociation-associated protein or transcription changes is key. Here, we detail a rapid collagenase-based dissociation protocol (Col-Short) that provides a high-yield single-cell suspension that is suitable for a variety of downstream applications.

Gold-nanoparticle-embedded hydrogel droplets with enhanced fluorescence for imaging and quantification of proteins in cells.

Conventional cellular protein detection techniques such as immunocytochemistry and flow cytometry require abundant cells, posing multiple challenges, including difficulty and cost for obtaining enough cells and the potential for clogging the instrument when using flow cytometry. Also, it is challenging to conduct cellular protein imaging and quantification simultaneously from a single experiment. We present a novel 3D platform, which integrates highly biocompatible cell-entrapped alginate hydrogel droplet array with gold-nanoparticle (AuNP)-based metal enhanced fluorescence (MEF), to achieve simultaneous imaging and quantification of proteins in intact cells in a sensitive manner.

Assessment of left atrial fibrosis progression in canines following rapid ventricular pacing using 3D late gadolinium enhanced CMR images.

Background: Atrial fibrillation (AF) is associated with extracellular matrix (ECM) remodelling and often coexists with myocardial fibrosis (MF); however, the causality of these conditions is not well established.

Objective: We aim to corroborate AF to MF causality by quantifying left atrial (LA) fibrosis in cardiac magnetic resonance (CMR) images after persistent rapid ventricular pacing and subsequent AF using a canine model and histopathological validation.

Methods: Twelve canines (9 experimental, 3 control) underwent baseline 3D LGE-CMR imaging at 3T followed by insertion of a pacing device and 5 weeks of rapid ventricular pacing to induce AF (experimental) or no pacing (control).

Influence of Aqueous Phase Composition on Double Emulsion Stability and Colour Retention of Encapsulated Anthocyanins.

Water-in-oil-in-water (W/O/W) emulsions (double emulsions) have often been used for the encapsulation of bioactive compounds such as anthocyanins. Instability of both anthocyanins and double emulsions creates a need for a tailored composition of the aqueous phase. In this work, double emulsions with a gelled internal water phase were produced and monitored over a 20-day storage period.

The effect of emulsifier type on the secondary crystallisation of monoacylglycerol and triacylglycerols in model dairy emulsions.

Dairy emulsions contain an intrinsically heterogeneous lipid phase, whose components undergo crystallisation in a manner that is critical to dairy product formulation, storage, and sensory perception. Further complexity is engendered by the diverse array of interfacially-active molecules naturally present within the serum of dairy systems, and those that are added for specific formulation purposes, all of which interact at the lipid-serum interface and modify the impact of lipid crystals on dairy emulsion stability. The work described in this article addresses this complexity, with a specific focus on the impact of temperature cycling and the effect of emulsifier type on the formation and persistence of lipid crystals at lipid-solution interfaces.

Lysozyme uptake into pharmaceutical grade fucoidan/chitosan polyelectrolyte multilayers under physiological conditions.

Polyelectrolyte multilayers composed of pharmaceutical grade fucoidan and chitosan have been assembled and studied in terms of their response to physiological solution conditions and the presence of lysozyme. The influence of phosphate buffered saline (PBS) solution on the multilayer was interrogated using attenuated total reflectance (ATR) Fourier transform infrared (FTIR) spectroscopy and atomic force microscopy (AFM). The combination of the techniques reveal that the polyelectrolyte multilayers swell when exposed to PBS after build-up and may include a small degree of mass loss as the film swells.

Odd-even effects on hydration of natural polyelectrolyte multilayers: An in situ synchrotron FTIR microspectroscopy study.

Hypothesis: Odd-even effects in polysaccharide polyelectrolyte multilayers influence their hydration content and the chemical environment of the water within them.

Experiments: Polysaccharide polyelectrolyte multilayers (PEMs) composed of pharmaceutical grade fucoidan and chitosan were studied under confinement using synchrotron FTIR microspectroscopy at increasing pressure, in order to isolate and measure infrared spectra of water within the PEM, without interference from bulk water. Complementary studies of the PEMs were carried out using lab-based in situ attenuated total reflectance Fourier transform spectroscopy (ATR FTIR) and quartz crystal microbalance with dissipation monitoring (QCM-D), as well as zeta potential measurements, to determine the quantity of adsorbed polymer, hydration content, film thickness, viscoelastic properties and surface charge during layer-by-layer deposition.

Fractionation and characterisation of hard milk fat crystals using atomic force microscopy.

The hard milk fat (HMF) fraction of milk fat was isolated via dry, thermal fractionation, followed by a solvent washing process. The resulting HMF crystals were visibly free of entrapped liquid fat, and subsequently characterised by thermal analysis, X-ray diffraction, and electron microscopy. The HMF crystals were found to be mostly β' and β' crystalline structures, with a lamellar thickness of 42.

A Novel Soft Contact Piezo-Controlled Liquid Cell for Probing Polymer Films under Confinement using Synchrotron FTIR Microspectroscopy.

Soft polymer films, such as polyelectrolyte multilayers (PEMs), are useful coatings in materials science. The properties of PEMs often rely on the degree of hydration, and therefore the study of these films in a hydrated state is critical to allow links to be drawn between their characteristics and performance in a particular application. In this work, we detail the development of a novel soft contact cell for studying hydrated PEMs (poly(sodium 4-styrenesulfonate)/poly(allylamine hydrochloride)) using FTIR microspectroscopy.

Analysis of ionic strength effects on the adsorption of simple amino acids.

Hypothesis: Amino acid adsorption by metals and/or oxide surfaces is important in many biomedical and industrial processes, however limited information exists discussing ionic strength influences on the mechanism of adsorption. A comparison of pure water solution and added 1:1 electrolyte should highlight the effects of electrolyte on amount adsorbed. ATR spectroscopy of the adsorbed phase should demonstrate the effects on the mechanism of adsorption.

Infrared spectrum analysis of the dissociated states of simple amino acids.

In this work, we present detailed analyses of the dissociation of dilute aqueous solutions of glycine and of lysine over the range 1 View Article and Find Full Text PDF

Why adolescent boys dream of becoming professional athletes.

A review of studies which investigated drawings of the ideal man and the occupational aspirations of boys (ages 11-18 years) from several countries indicated that becoming a professional athlete was a popular but unrealistic aspiration for many adolescent boys. Boys who were athletes and nonathletes from diverse ethnic groups and nationalities dreamed of becoming professional athletes. In two additional studies in the United States of America, adolescents were asked why they thought boys most often selected professional athlete as a possible future occupation.