Immunochemical methods for biomonitoring of chlorophenols as potential biomarkers of exposure

The thesis presents the development of immunochemical methods for detection of trichlorophenols (TCP) in environmental and biological samples. An indirect enzyme linked immunosorbent assay (ELISA) for 2,4,5-TCP has been developed after a rational design of the immunizing hapten chemical structure, and the screening of 12 competitor haptens. The effect of the conjugation degree of the competitors and their homology with the target analyte, the physicochemical parameters (pH, ionic strength), the concentration of detergent, the time of incubation and the specificity were studied. Two immunoassays (2,4,5- and 2,4,6-TCP ELISAs) were evaluated for the analysis of water, milk, serum and urine. Drinking water was analyzed directly after buffering the sample. The strong matrix effects in milk samples requires the sample clean up. Human serum can be analyzed after protein precipitation with absolute ethanol. The strong matrix effect of urine and its variability for samples from different individuals suggested the introduction of a purification step prior to ELISA. The C18- solid phase extraction (SPE) is an effective clean up method to remove an important part of the nonspecific interferences present in urine. The C18-SPE-ELISA method allows accurate quantification of TCPs in urine of occupationally exposed persons. SPE based on immunosorbents (immunoaffinity extraction, IAE) have been developed in single and 96-column formats. IAE is an effective clean up method to remove all nonspecific urinary interferences. The IAE step was optimized regarding sample volume, loading level, type of urine hydrolysis washing and elution conditions. The selectivity of the immunosorbents can be modulated by the washing conditions. The immunosorbents have sufficient capacity to effectively extract 2,4,6-TCP from urine samples of occupationally exposed persons and the general population. The HTS-IAE-ELISA method allows the processing of 100 samples/day with very good precision and accuracy. The method was validated with GC-MS and applied to the biomonitoring of three groups of population from Catalonia.A quenching fluorescence immunoassay based on the laser-induced fluorescence detection in microdroplets (LIF-microdroplet-QFIA) for 2,4,6-TCP has been developed as a novel biodetection system. This approach offers significant improvement in method detectability compared to the microplate immunoassays and is the first application urine samples that can be directly analyzed after sample dilution

Zero Liquid Discharge (ZLD) Industrial Wastewater Treatment Systems as Sustainable Development Basic Ecological Components

The problems in processing the acid ferrous-containing rinsing water and spent pickling solutions are considered. The first-ever in metallurgy zero liquid discharge (ZLD) industrial wastewater treatment systems of a transformer cold rolling plant at the Verkh-Isetsky steelworks in Sverdlovsk (now - LLC VIZ Steel is a part of Novolipetsk Steel Group of Companies) is described. The experience of the VIZ Steel, particularly in relation to industrial wastewater treatment and sludge processing, is useful for designing ZLD industrial wastewater treatment systems of metallurgical, machine-building and metalworking enterprises. The recommendations for organizing the ZLD industrial wastewater treatment systems in the pickling plants with the repeated use of the treated rinsing water and processing the spent pickling solutions and the sludges are presented. For practical calculations of clarifiers the empirical formulas are offered. Process flow diagram of acid ferrous-containing rinsing water treatment is suggested. Acid ferrous-containing rinsing water treatment includes the following processes: rinsing water neutralization to pH 9-10,5 by 5 % calcium hydroxide and partial ferrous oxidation by air; addition of anionic flocculant and active gypsum priming to the neutralized rinsing water; settling of rinsing water conditioned by reagents in clarifiers; treatment of clarified water on deep bed sand filters; reclaimed water treatment. © Published under licence by IOP Publishing Ltd

ACTUALITY OF THE PROBLEM OF LABOR PROTECTION AT WORK AT THE DESALINATION INSTALLATIONS

The main hazardous and harmful production factors associated with the operation of desalination plants are considered. Approaches to the management of labor protection and the provision of safe production at desalination complexes are proposed

THE PROBLEM OF LACK OF FRESH WATER IN THE CRIMEAN REGION: PERSPECTIVE OF DEVELOPMENT OF DESALINATION INSTALLATIONS

The perspective of development of desalination plants in the Crimean region is considered as one of the ways to solve problems with a shortage offresh water. A technological scheme of a station for the preparation of drinking wa¬ter from the waters of the Black Sea is proposed

OPTIMIZATION OF THE SELECTION OF THE METHOD OF SEA WATER DESALINATION FOR THE REGION

Criteria for the choice of a way of desalting of sea water in the Crimean region - «study and reliability», «geographical», «industrial application», «economic», «an electric power expense», «ecological safety» and others are offered. Based on these criteria we selected the most optimal method of desalination of sea water for the Crimean region

Miniaturized Protein Microarray with Internal Calibration as Point-of-Care Device for Diagnosis of Neonatal Sepsis

Neonatal sepsis is still a leading cause of death among newborns. Therefore a protein-microarray for point-of-care testing that simultaneously quantifies the sepsis associated serum proteins IL-6, IL-8, IL-10, TNF alpha, S-100, PCT, E-Selectin, CRP and Neopterin has been developed. The chip works with only a 4 μL patient serum sample and hence minimizes excessive blood withdrawal from newborns. The 4 μL patient samples are diluted with 36 μL assay buffer and distributed to four slides for repetitive measurements. Streptavidin coated magnetic particles that act as distinct stirring detection components are added, not only to stir the sample, but also to detect antibody antigen binding events. We demonstrate that the test is complete within 2.5 h using a single step assay. S-100 conjugated to BSA is spotted in increasing concentrations to create an internal calibration. The presented low volume protein-chip fulfills the requirements of point-of-care testing for accurate and repeatable (CV < 14%) quantification of serum proteins for the diagnosis of neonatal sepsis

Метод импедансной спектроскопии для тестирования увлажненных зерен пшеницы

The authors presented the results of a study of the electrical and dielectric characteristics of wetted wheat grains by measuring their complex electrical resistance (impedance Z) in a wide frequency range (from 1 Hz to 100 MHz). The results of electrical impedance measurements of grains with surface or volumetric moisture content under different experimental conditions can provide useful information on the properties of the biological tissues of grain crops. These results can also be used to develop a new type of impedance sensor for testing grain quality and moisture content. The authors used well-dried wheat grains and grains saturated with moisture and saline as objects of research. A major problem in grain impedance measurements is the selection of a suitable electrode material to be placed on the end surfaces of the samples. The electrodes must ensure reliable contact with the grain and have a minimum transient resistance. The end surfaces of the pressed samples were reinforced with a protective dielectric ring to prevent transverse deformation. These contacts provided a transition resistance between 1-2 ohms. The authors have identified processes of accumulation of electric charges near the surface of metal electrodes at low frequencies and on internal grain structures, leading to an increase in the dielectric permittivity and dissipation factor. The behavior of the active and reactive components of the impedance at higher frequencies is determined by dielectric relaxation processes. The obtained impedance spectra were compared with the spectra of the most suitable equivalent electrical circuits. The radio components of the circuits provide information about the basic mechanisms of alternating electric current flow through the complex inhomogeneous structure of the grain. The authors found that moistening the grain with saline water enhances the process of accumulation of electric charges and affects the dispersion of the real and imaginary components of the impedance.Представлены результаты исследования электрических и диэлектрических характеристик увлажнённых зёрен пшеницы методом измерений их комплексного электрического сопротивления (импеданса Z) в широком диапазоне частот (от 1 Гц до 100 МГц). Результаты измерений электрического импеданса зерна с поверхностным или объёмным содержанием влаги в разных условиях эксперимента могут дать полезную информацию о свойствах биологических тканей зерновых культур и использоваться для разработки нового типа импедансных датчиков для тестирования качества зерна и его влажности. В качестве объекта исследований использовались хорошо высушенные зёрна пшеницы и зёрна, насыщенные влагой и солевым раствором. Серьёзной проблемой при измерениях импеданса зёрен является выбор подходящего материала электродов, накладываемых на торцевые поверхности образцов. Электроды должны обеспечивать надёжный контакт с зерном и обладать минимальным переходным сопротивлением. Для исключения поперечной деформации торцевые поверхности прессованных образцов укрепляли защитным диэлектрическим кольцом. Такие контакты обеспечивали переходное сопротивление в пределах 1–2 Ом. В области низких частот выявлены процессы накопления электрических зарядов вблизи поверхности металлических электродов и на внутренних структурах зерна, приводящие к увеличению диэлектрической проницаемости и тангенса угла потерь. В области более высоких частот поведение активной и реактивной компонент импеданса определяется процессами диэлектрической релаксации. Полученные спектры импеданса сопоставлялись со спектрами наиболее подходящих эквивалентных электрических схем, радиотехнические компоненты которых позволяют понять основные механизмы прохождения переменного электрического тока через сложную неоднородную структуру зерна. Установлено, что увлажнение зерна подсоленной водой усиливает процесс накопления электрических зарядов и влияет на дисперсию действительной и мнимой компонент импеданса

Facile Synthesis of Amine-Functionalized Eu3+-Doped La(OH)3 Nanophosphors for Bioimaging

Here, we report a straightforward synthesis process to produce colloidal Eu3+-activated nanophosphors (NPs) for use as bioimaging probes. In this procedure, poly(ethylene glycol) serves as a high-boiling point solvent allowing for nanoscale particle formation as well as a convenient medium for solvent exchange and subsequent surface modification. The La(OH)3:Eu3+ NPs produced by this process were ~3.5 nm in diameter as determined by transmission electron microscopy. The NP surface was coated with aminopropyltriethoxysilane to provide chemical functionality for attachment of biological ligands, improve chemical stability and prevent surface quenching of luminescent centers. Photoluminescence spectroscopy of the NPs displayed emission peaks at 597 and 615 nm (λex = 280 nm). The red emission, due to 5D0 → 7F1 and 5D0 → 7F2 transitions, was linear with concentration as observed by imaging with a conventional bioimaging system. To demonstrate the feasibility of these NPs to serve as optical probes in biological applications, an in vitro experiment was performed with HeLa cells. NP emission was observed in the cells by fluorescence microscopy. In addition, the NPs displayed no cytotoxicity over the course of a 48-h MTT cell viability assay. These results suggest that La(OH)3:Eu3+ NPs possess the potential to serve as a luminescent bioimaging probe