A Toxicology Suite Adapted for Comparing Parallel Toxicity Responses of Model Human Lung Cells to Diesel Exhaust Particles and Their Extracts

Epidemiological studies have shown that exposure to airborne particulate matter (PM) can be an important risk factor for some common respiratory diseases. While many studies have shown that PM exposures are associated with inflammatory reactions, the role of specific cellular responses in the manifestation of primary hypersensitivities and the progression of respiratory diseases remains unclear. In order to better understand mechanisms by which PM can exert adverse health effects, more robust approaches to support in vitro studies are warranted. In response to this need, a group of accepted toxicology assays was adapted to create an analytical suite for screening and evaluating the effects of important, ubiquitous atmospheric pollutants on two model human lung cell lines (epithelial and immature macrophage). To demonstrate the utility of this suite, responses to intact diesel exhaust particles (DEP) and mass-based equivalent doses of their organic extracts were examined. Results suggest that extracts have the potential to induce greater biological responses than those associated with their colloidal counterpart. Additionally, macrophage cells appear to be more susceptible to the cytotoxic effects of both intact DEP and their organic extract, than epithelial cells tested in parallel. As designed, the suite provided a more robust basis for characterizing toxicity mechanisms than the analysis of any individual assay. Findings suggest that cellular responses to PM are cell line dependent, and show that the collection and preparation of PM and/or their extracts have the potential to impact cellular responses relevant to screening fundamental elements of respiratory toxicity.

Copyright 2015 American Association for Aerosol Research     

Influence of branching regularity on the behavior of hyperbranched polymers in solution

A carbosilane pseudo‐dendrimer with irregular structure and degree of branching DB = 1 was synthesized and fractionated. The molar masses of the obtained fractions ranged from 5000 to 490 000 g mol^?1. The hydrodynamic and conformational properties were studied by the methods of light scattering, sedimentation‐diffusion analysis and viscometry in dilute hexane and toluene solutions. The Mark ? Kuhn ? Houwink dependences were obtained. The results for the pseudo‐dendrimer are compared with the data for the initial hyperbranched poly(diallylcarbosilane) with DB = 0.5 and carbosilane dendrimer. It is shown that the solution properties of the pseudo‐dendrimer are close to those for the irregular hyperbranched polymer and differ significantly from the dendrimer behavior. Hence, it is branching regularity rather than the degree of branching that determines the conformational and hydrodynamic properties of hyperbranched polymers with a high degree of branching DB ≥ 0.5. © 2014 Society of Chemical Industry     

Interleukins 6 and 15 Levels Are Higher in Subcutaneous Adipose Tissue,but Obesity Is Associated with Their Increased Content in Visceral Fat Depots

Excess adiposity is associated with chronic inflammation, which takes part in the development of obesity-related complications. The aim of this study was to establish whether subcutaneous (SAT) or visceral (VAT) adipose tissue plays a major role in synthesis of pro-inflammatory cytokines. Concentrations of interleukins (IL): 1β, 6, 8 and 15 were measured at the protein level by an ELISA-based method and on the mRNA level by real-time PCR in VAT and SAT samples obtained from 49 obese (BMI > 40 kg/m²) and 16 normal-weight (BMI 20–24.9 kg/m²) controls. IL-6 and IL-15 protein concentrations were higher in SAT than in VAT for both obese (p = 0.003 and p < 0.0001, respectively) and control individuals (p = 0.004 and p = 0.001, respectively), while for IL-1β this was observed only in obese subjects (p = 0.047). What characterized obese individuals was the higher expression of IL-6 and IL-15 at the protein level in VAT compared to normal-weight controls (p = 0.047 and p = 0.016, respectively). Additionally, obese individuals with metabolic syndrome had higher IL-1β levels in VAT than did obese individuals without this syndrome (p = 0.003). In conclusion, concentrations of some pro-inflammatory cytokines were higher in SAT than in VAT, but it was the increased pro-inflammatory activity of VAT that was associated with obesity and metabolic syndrome.     

Asymptotic Properties of a Probabilistic‐Statistical Model of Particle Classification in Hydrocyclones

The peculiarities of practical implementation of a probabilistic‐statistical model for a hydrodynamic stage of particle classification process of liquid‐solid polydisperse systems in cylinder‐conic hydrocyclones of small size have been investigated. Within reasonable assumptions, stationary solutions of the Fokker‐Planck‐Kolmogorov kinetic equation were obtained for the considered separation process. In order to describe changes in characteristics of suspension separation in hydrocyclones it was proposed to use stationary distributions, which parameters depend not only on hydraulic and dynamic features of flows inside an apparatus, but also are determined by relative magnitudes of the impact of particle classification and centrifugal forces in comparison with the intensity of random perturbations.     

Practical Application of the Probabilistic‐Statistical Model of the Suspension Separation in Hydrocyclones

An effective practical approach that allows not only a significant reduction in the scope of practical experiments in the course of studying suspension separation processes in hydrocyclones, but also makes it possible to assess the intensity of random components of the processes and define the interrelation between such components and hydrodynamics of flows in a hydrocyclone is presented. Within the frames of the developed probabilistic‐statistical model of suspension separation in hydrocyclones on the basis of statistical self‐similarity properties, a relationship was found between determined and random components of the processes. This allowed transitioning from three‐parameter probability density functions for suspension particles in hydrocyclones to two‐parameter functions; thus significantly improving the efficiency of practical application of the developed model.     

A Kinetic Study on the Heat-Induced Changes of Whey Proteins Concentrate at Two pH Values

Whey protein concentrate (WPC) is used as food ingredients due to their commercially important functional properties. The effects of heat treatment on the components of milk are very important for the final product character, since they undergo modifications that affect sensorial and nutritional quality of milk. The heat-induced changes on dispersions of whey proteins concentrate were monitored by measurement of thiol availability, protein solubility, and turbidity at pH 6.6 and 7.5. The fractional conversion model was used to quantitatively describe the effect of different temperature–time combination on denaturation mechanism. The results demonstrate that heat-induced changes of WPC greatly influence their solubility, expressed as degree of denaturation at pH 4.6 and were related to the heating conditions. The denaturation mechanism involved a number of consecutive conformational changes in the molecules. A curvature in Arrhenius plots was observed around 75 °C, indicating changes in the reaction mechanism. The deflection of Arrhenius plot reflects the generally accepted two-step denaturation/aggregation process of whey proteins.     

Influence of B-site cations ordering on magnetization and dielectric relaxations in Li–Ni spinel ferrites

Ho-substituted Li–Ni ferrites with composition L i_1.2Ni_0.4Ho_xFe_2-xO₄; 0≤ x ≤ 0.15 were synthesized by a self-ignited sol-gel process. An annealing temperature of 950 °C is estimated via thermal-gravimetric (TGA) analysis. X-ray diffraction (XRD) scans have confirmed the formation of the ferrite phase with a spinel structure in all samples. Substitution of Ho ions on the B-site significantly reduced the porosity from 38 -to 23% and the crystallite size from 23.4 -to 21.7 nm. Microstructural analysis revealed a denser structure with an increase in Ho content. Dielectric results showed that both the dielectric loss and dielectric constant depict a nonlinear variation with the addition of Ho. Complex impedance behavior with a single semicircle for all samples suggests the predominant effect of the grain boundary mechanism. The substitution of Ho ions in place of Fe ions significantly decreased the electrical conductivity. The anisotropic Ho³⁺ ions reinforce the L-S coupling which consequently enhanced the coercive force from 145 -to 389 Oe, and thus the anisotropy constant.     

Lack of GABARAP-Type Proteins Is Accompanied by Altered Golgi Morphology and Surfaceome Composition

GABARAP (γ-aminobutyric acid type A receptor-associated protein) and its paralogues GABARAPL1 and GABARAPL2 comprise a subfamily of autophagy-related Atg8 proteins. They are studied extensively regarding their roles during autophagy. Originally, however, especially GABARAPL2 was discovered to be involved in intra-Golgi transport and homotypic fusion of post-mitotic Golgi fragments. Recently, a broader function of mammalian Atg8s on membrane trafficking through interaction with various soluble N-ethylmaleimide-sensitive factor-attachment protein receptors (SNAREs) was suggested. By immunostaining and microscopic analysis of the Golgi network, we demonstrate the importance of the presence of individual GABARAP-type proteins on Golgi morphology. Furthermore, triple knockout (TKO) cells lacking the whole GABARAP subfamily showed impaired Golgi-dependent vesicular trafficking as assessed by imaging of fluorescently labelled ceramide. With the Golgi apparatus being central within the secretory pathway, we sought to investigate the role of the GABARAP-type proteins for cell surface protein trafficking. By analysing the surfaceome composition of TKOs, we identified a subset of cell surface proteins with altered plasma membrane localisation. Taken together, we provide novel insights into an underrated aspect of autophagy-independent functions of the GABARAP subfamily and recommend considering the potential impact of GABARAP subfamily proteins on a plethora of processes during experimental analysis of GABARAP-deficient cells not only in the autophagic context.