Dynamics of proteins: Light scattering study of dilute and dense colloidal suspensions of eye lens homogenates

We report a dynamic light scattering study on protein suspensions of bovine lens homogenates at conditions (pH and ionic strength) similar to the physiological ones. Light scattering data were collected at two temperatures, 20 oC and 37 oC, over a wide range of concentrations from the very dilute limit up to the dense regime approaching to the physiological lens concentration. A comparison with experimental data from intact bovine lenses was advanced revealing differences between dispersions and lenses at similar concentrations. In the dilute regime two scattering entities were detected and identified with the long-time, self-diffusion modes of alpha-crystallins and their aggregates, which naturally exist in lens nucleus. Self-diffusion coefficients are temperature insensitive, whereas the collective diffusion coefficient depends strongly on temperature revealing a reduction of the net repulsive interparticle forces with lowering temperature. While there are no rigorous theoretical approaches on particle diffusion properties for multi-component, non-ideal hard-sphere, polydispersed systems, as the suspensions studied here, a discussion of the volume fraction dependence of the long-time, self-diffusion coefficient in the context of existing theoretical approaches was undertaken. This study is purported to provide some insight into the complex light scattering pattern of intact lenses and the interactions between the constituent proteins that are responsible for lens transparency. This would lead to understand basic mechanisms of specific protein interactions that lead to lens opacification (cataract) under pathological conditions.Comment: To appear in J. Chem. Phy

Dynamic light scattering study on phase separation of a protein-water mixture: Application on cold cataract development in the ocular lens

We present a detailed dynamic light scattering study on the phase separation in the ocular lens emerging during cold cataract development. Cold cataract is a phase separation effect that proceeds via spinodal decomposition of the lens cytoplasm with cooling. Intensity auto-correlation functions of the lens protein content are analyzed with the aid of two methods providing information on the populations and dynamics of the scattering elements associated with cold cataract. It is found that the temperature dependence of many measurable parameters changes appreciably at the characteristic temperature ~16+1 oC which is associated with the onset of cold cataract. Extending the temperature range of this work to previously inaccessible regimes, i.e. well below the phase separation or coexistence curve at Tcc, we have been able to accurately determine the temperature dependence of the collective and self-diffusion coefficient of proteins near the spinodal. The analysis showed that the dynamics of proteins bears some resemblance to the dynamics of structural glasses where the apparent activation energy for particle diffusion increases below Tcc indicating a highly cooperative motion. Application of ideas developed for studying the critical dynamics of binary protein/solvent mixtures, as well as the use of a modified Arrhenius equation, enabled us to estimate the spinodal temperature Tsp of the lens nucleus. The applicability of dynamic light scattering as a non-invasive, early-diagnostic tool for ocular diseases is also demonstrated in the light of the findings of the present paper

The Thermal Structural Transition of α-Crystallin Inhibits the Heat Induced Self-Aggregation

-crystallin, the major constituent of human lens, is a member of the heat-shock proteins family and it is known to have a quaternary structural transition at . The presence of calcium ions and/or temperature changes induce supramolecular self-aggregation, a process of relevance in the cataractogenesis. Here we investigate the potential effect of the bovine -crystallin's structural transition on the self-aggregation process. Along all the temperatures investigated, aggregation proceeds by forming intermediate molecular assemblies that successively aggregate in clusters. The final morphology of the aggregates, above and below , is similar, but the aggregation kinetics are completely different. The size of the intermediate molecular assemblies, and their repulsive energy barrier show a marked increase while crossing . Our results highlight the key role of heat modified form of -crystallin in protecting from aggregation and preserving the transparency of the lens under hyperthermic conditions

Different experimental approaches in modelling cataractogenesis: An overview of selenite-induced nuclear cataract in rats

Cataract, the opacification of eye lens, is the leading cause of blindness worldwide. At present, the only remedy is surgical removal of the cataractous lens and substitution with a lens made of synthetic polymers. However, besides significant costs of operation and possible complications, an artificial lens just does not have the overall optical qualities of a normal one. Hence it remains a significant public health problem, and biochemical solutions or pharmacological interventions that will maintain the transparency of the lens are highly required. Naturally, there is a persistent demand for suitable biological models. The ocular lens would appear to be an ideal organ for maintaining culture conditions because of lacking blood vessels and nerves. The lens in vivo obtains its nutrients and eliminates waste products via diffusion with the surrounding fluids. Lens opacification observed in vivo can be mimicked in vitro by addition of the cataractogenic agent sodium selenite (Na2SeO3) to the culture medium. Moreover, since an overdose of sodium selenite induces also cataract in young rats, it became an extremely rapid and convenient model of nuclear cataract in vivo. The main focus of this review will be on selenium (Se) and its salt sodium selenite, their toxicological characteristics and safety data in relevance of modelling cataractogenesis, either under in vivo or in vitro conditions. The studies revealing the mechanisms of lens opacification induced by selenite are highlighted, the representatives from screening for potential anti-cataract agents are listed

Residues of veterinary medicinal products and coccidiostats in eggs – causes, control and results of surveillance program in Poland

The use of veterinary medicinal products in food producing animals for a variety of purposes causes that their residues may be presented in edible tissues. As a result, in concern of public health, European Union Countries establish each year monitoring plans and they control the levels of harmful substances in food of animal origin. This paper presents survey of residues of veterinary medicinal products and coccidiostats in eggs for Poland and European Union in years 2007-2010. Despite the decrease in reported non-compliant results for coccidiostats, the numbers were still higher than those for veterinary medicines. The most often determined coccidiostats were: nicarbazin, dinitrocarbanilide, salinomycin and lasalocid, and the most often reported non-compliant results for veterinary medicines were: antimicrobials, enrofloxacin and doxycycline

WWG multi-voltage mining fans manufactured by Eko-Win

Przedstawiono konstrukcje typoszeregu wielonapięciowych wentylatorów górniczych typu WWG firmy Eko-Win, z omówieniem ich charakterystyk, parametrów technicznych oraz pola zastosowań. Przedstawiono sposób wyznaczania charakterystyk wentylatorów na stanowisku badawczym.A design of series of types of WWG multi-voltage mining fans manufactured by Eko-Win is presented and their characteristics, technical parameters and fields of application are discussed. A method for determination of characteristics of fans at the test stand is given

A dual validation approach to detect anthelmintic residues in bovine liver over an extended concentration range

peer-reviewedThis paper describes a method for the detection and quantification of 38 residues of the most widely used anthelmintics (including 26 veterinary drugs belonging to the benzimidazole, macrocyclic lactone and flukicide classes) in bovine liver using two different protocols for MRL and non-MRL levels. A dual validation approach was adopted to reliably quantify anthelmintic residues over an extended concentration range (1–3000 gkg−1). Sample extraction and purification was carried out using a modified QuEChERS method. A concentration step was included when analysing in the low gkg−1 range. Rapid analysis was carried out by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS), which was capable of detecting residues to <2 gkg−1. The method has been single-laboratory validated according to the 2002/657/EC guidelines and met acceptability criteria in all but a few cases. The inclusion of 19 internal standards, including 14 isotopically labelled internal standards, improved accuracy, precision, decision limit (CC˛) and detection capability (CCˇ).This research was funded by the Irish Department of Agriculture and Food under the Food Institutional Research Measure as part of the National Development Plan (Projects 06RDTAAFRC479) and by EU Sixth Framework programme on Food Quality and Safety, ProSafeBeef (Project FOOD-CT-405 2006-36241)