Methodology for the identification of bioactive and marker peptides in the organs of cattle and pigs

The development of general conception methodology for the meat-based functional food compositions is especially relevant today due to the growing consumers’ interest and attention to their health. This category of these food-products is intended for personal  ized nutrition of various age groups in the population, taking into account fortification of the food with nutraceuticals and with functional and metabolically active ingredients obtained from animal and vegetable source. Therefore, it was necessary to develop a certain tool for reliable identification of free peptides from the offals (by-products like hearts and aorta from Sus scrofa and Bos taurus) and from the ready-to-consume meat food (canned food) based on the free peptides, which food is potentially targeted to help with some issues in the human body. The authors proposed the methodology for identification of peptides weighing less than 5 kDa. This methodology has a row of significant advantages, such as a short time of analysis (90 minutes) and the possibility to prepare a large number of samples simultaneously (n=16). Analysis of bioactive peptides (BAPs) was performed by liquid chro  matography combined with time-of-flight mass spectrometry (Agilent 6545XT AdvanceBio LC/Q-TOF). The marker peptides were detected by a triple quadrupole mass spectrometer (Agilent 6410 Triple Quadrupole LC/MS). All peptide sequences were defined with the help of mass spectrometric data processing databases like PepBank, BioPep, AHTPDB. In this work from 39 to 269 peculiar soluble peptides were found, with an extraction level of 0.17–0.23%. The main fraction consisted of short peptides less than 1000 Da (71.0–98.0%). In experimental samples of pork hearts and arteries 7 peculiar marker peptides were identified. FFESFGDL  SNADAVMGNPK peptide obtained from the β-hemoglobin protein is of a special interest, as this peptide showed the maximum intensity of a signal. Presumably, this peptide can serve as an indicator of the blood presence in the finished food product. So it can serve as an assessment tool of bleeding degree of meat raw. For pork aortas a specific peptide TVLGNFAAFVQK was isolated from serum albumin, which turned out to be stable during heat treatment. This is also important for assessment of meat food that are subjected to high thermal exposure.The development of general conception methodology for the meat-based functional food compositions is especially relevant today due to the growing consumers’ interest and attention to their health. This category of these food-products is intended for personal  ized nutrition of various age groups in the population, taking into account fortification of the food with nutraceuticals and with functional and metabolically active ingredients obtained from animal and vegetable source. Therefore, it was necessary to develop a certain tool for reliable identification of free peptides from the offals (by-products like hearts and aorta from Sus scrofa and Bos taurus) and from the ready-to-consume meat food (canned food) based on the free peptides, which food is potentially targeted to help with some issues in the human body. The authors proposed the methodology for identification of peptides weighing less than 5 kDa. This methodology has a row of significant advantages, such as a short time of analysis (90 minutes) and the possibility to prepare a large number of samples simultaneously (n=16). Analysis of bioactive peptides (BAPs) was performed by liquid chro  matography combined with time-of-flight mass spectrometry (Agilent 6545XT AdvanceBio LC/Q-TOF). The marker peptides were detected by a triple quadrupole mass spectrometer (Agilent 6410 Triple Quadrupole LC/MS). All peptide sequences were defined with the help of mass spectrometric data processing databases like PepBank, BioPep, AHTPDB. In this work from 39 to 269 peculiar soluble peptides were found, with an extraction level of 0.17–0.23%. The main fraction consisted of short peptides less than 1000 Da (71.0–98.0%). In experimental samples of pork hearts and arteries 7 peculiar marker peptides were identified. FFESFGDL  SNADAVMGNPK peptide obtained from the β-hemoglobin protein is of a special interest, as this peptide showed the maximum intensity of a signal. Presumably, this peptide can serve as an indicator of the blood presence in the finished food product. So it can serve as an assessment tool of bleeding degree of meat raw. For pork aortas a specific peptide TVLGNFAAFVQK was isolated from serum albumin, which turned out to be stable during heat treatment. This is also important for assessment of meat food that are subjected to high thermal exposure

ИДЕНТИФИКАЦИЯ И КОЛИЧЕСТВЕННОЕ ОПРЕДЕЛЕНИЕ МЫШЕЧНОЙ ТКАНИ НА ОСНОВЕ КОНТРОЛЯ ПРОТОТИПИЧЕСКИХ ПЕПТИДОВ С ИСПОЛЬЗОВАНИЕМ МЕТОДА МОНИТОРИНГА ЗАДАННЫХ РЕАКЦИЙ

One of the main problems of modern production of meat products is the quality of raw meat, which depends on various factors including genetic components, conditions of transportation, production and processing. The most important components of meat are proteins, the total content, structure and functional state of which in this complex biological system with many interacting components is constantly changing. In order to study the interspecific and intraspecific features of meat proteins, their transformations in the process of maturation and processing requires modern analytical technologies based on the systematic approach to the analysis. Proteomics, as a methodology for studying proteins in a certain system and at a certain time, opens wide opportunities in this direction, allowing to identify and develop accurate analytical methods for searching biomarkers and identifying unfair practices. Given the high added value and multicomponent composition, finished meat products are among the most susceptible to adulteration. Current paper presents a technique of high-performance liquid chromatography with mass spectrometric detector (HPLC-MS) adapted for the detection and quantification of two different types of meat (beef and pork) in a complex biological matrix such as structureless minced meat. After the protein isolation and trypsin cleavage, species-specific peptide markers were selected for each animal species for the quantification. The technique was tested on model samples of a mixture of muscle tissue of the two species of animals. A good sensitivity was established with the possibility of quantitative determination of muscle tissue of each animal species using special calibration graphs. The developed technique can find wide application at the supervising organizations aimed at counteracting the discrepancies in the hidden replacement of types of meat by cheaper or low-grade raw materials.Keywords: Biomarker, prototypical peptides, HPLC-MS, species identification(Russian) DOI: http://dx.doi.org/10.15826/analitika.2019.23.4.012D.V. Khvostov1, N.L. Vostrikova1, A.V. Zherdev2, E.A. Zvereva2, A.A. Kurzova1  1V.M. Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences, 109316 Moscow, Russian Federation 2Federal State Institution «Federal Research Centre «Fundamentals of Biotechnology» of the Russian Academy of Sciences», Leninsky prospect, 33, build. 2, 119071 Moscow, Russian FederationОдной из основных проблем современного производства мясных продуктов является качество мясного сырья, которое зависит от разных факторов, включая генетические составляющие, условия транспортирования, производства и переработки. К наиболее значимым компонентам мяса относятся белки, общее содержание, структура и функциональное состояние которых в составе этой сложной биологической системы, с большим количеством взаимодействующих составляющих, постоянно изменяется. Для изучения межвидовых и внутривидовых особенностей белков мяса, трансформации их в процессе созревания и технологической обработки требуются современные аналитические технологии, основанные на системном подходе к анализу. Широкие возможности в этом направлении открывает протеомика, как методологии изучения белков в определенной системе и в определенное время, позволяющая идентифицировать и разрабатывать точные аналитические методы поиска биомаркеров и выявления недобросовестных практик. Учитывая высокую добавленную стоимость и многокомпонентность состава, готовые мясные продукты относятся к числу наиболее подверженных фальсификации. В работе представлена методика высокоэффективной жидкостной хроматографии с масс-спектрометрическим детектором (ВЭЖХ-МС), адаптированная для обнаружения и количественного определения двух различных видов мяса (говядина и свинина) в сложной биологической матрице, такой как бесструктурные фарши. После выделения белков и расщепления их трипсином были выбраны видоспецифичные пептидные маркеры для каждого вида животного с целью количественного определения. Методика была апробирована на модельных образцах смеси мышечной ткани двух видов животных. Установлена хорошая чувствительность с возможностью количественного определения мышечной ткани каждого вида животного, при использовании специальных градуировочных графиков. Разработанная методика может найти широкое применение у контролирующих организаций, направленных на противодействие несоответствиям по скрытой замене видов мяса более дешевым или низкосортными сырьем.Ключевые слова: Биомаркер, прототипические пептиды, ВЭЖХ-МС, видовая идентификацияDOI: http://dx.doi.org/10.15826/analitika.2019.23.4.01

Use of perovskite in solar energy

The article examine relatively new material for solar energy – perovskite. It dealt with physical properties. Data are given about developments in this field Russian and foreign scientists. It is analyzed coefficients of efficiency used in industry materials.В статье рассмотрен относительно новый материал для солнечной энергетики – перовскит. Дано описание его физических свойств, приведены разработки в этой отрасли как российских, так и зарубежных ученых. В работе проведен сравнительный анализ материалов для изготовления солнечных элементов по коэффициентам полезного действия

The morphofunctional and biochemical characteristics of opisthorchiasis-associated cholangiocarcinoma in a Syrian hamster model

The validity of experimental models of pathologies is one of the key challenges in translational medicine. Cholangiocarcinoma, or bile duct cancer, ranks second among oncological diseases of the liver. There is a strong association between bile duct cancer and parasitic infestation of the liver caused by trematodes in the family Opisthorchiidae. We have recently demonstrated that cholangiocarcinoma can develop in Syrian hamsters (Mesocricetus auratus) infected by Opisthorchis felineus and administered with dimethylnitrosamine. However, there is still no description of how this experimental model can possibly be used in translational research. The aim of this work was to study the morphological, functional and biochemical characteristics during cholangiocarcinoma development in Syrian hamsters infected by O. felineus and administered with dimethylnitrosamine. The experi­ment lasted 30 weeks with combined exposure to dimethylnitrosamine in drinking water at a dose of 12.5 ppm and a single injection of 50 metacercariae O. felineus. It was shown that the development of cholangiocarcinoma (18 weeks) increased the total number of basophils, eosinophils and monocytes, the relative number of granulocytes, the amount of total and direct bilirubin, and cholesterol and ALT levels, but reduced the relative number of lymphocytes. Based on pathological, morphometric and biochemical analyses, our model has characteristics similar to those in patients with opisthorchiasisassociated cholangiocarcinoma. Thus, this model can be used to test anticancer drugs, to study the mechanisms of cholangiocarcinogenesis and to search for molecular markers for early diagnosis of bile duct cancer

METHODS OF ASSESSING THE DEGREE OF DESTRUCTION OF RUBBER PRODUCTS USING COMPUTER VISION ALGORITHMS

For technical inspection of rubber products are essential methods of improving video scopes analyzing the degree of destruction and aging of rubber in an aggressive environment. The main factor determining the degree of destruction of the rubber product, the degree of coverage is cracked, which can be described as the amount of the total area, perimeter cracks, geometric shapes and other parameters. In the process of creating a methodology for assessing the degree of destruction of rubber products arises the problem of the development of machine vision algorithm for estimating the degree of coverage of the sample fractures and fracture characterization. For the development of image processing algorithm performed experimental studies on the artificial aging of several samples of products that are made from different rubbers. In the course of the experiments it was obtained several samples of shots vulcanizates in real time. To achieve the goals initially made light stabilization of array images using Gaussian filter. Thereafter, for each image binarization operation is applied. To highlight the contours of the surface damage of the sample is used Canny algorithm. The detected contours are converted into an array of pixels. However, a crack may be allocated to several contours. Therefore, an algorithm was developed by combining contours criterion of minimum distance between them. At the end of the calculation is made of the morphological features of each contour (area, perimeter, length, width, angle of inclination, the At the end of the calculation is made of the morphological features of each contour (area, perimeter, length, width, angle of inclination, the Minkowski dimension). Show schedule obtained by the method parameters destruction of samples of rubber products. The developed method allows you to automate assessment of the degree of aging of rubber products in telemetry systems, to study the dynamics of the aging process of polymers to obtain initial information for mathematical modeling of aging processes of polymers, to predict changes in the state of rubber products over time

Managing of the baking bread process based on mathematical model of change crust color dynamics

In this article a support system of making decision about control of baking process of bread on the basis of estimating the quality ( the colors of the product surface and the mathematical model of the dynamics of change in this indicator) is offered

IDENTIFICATION OF THE MASS DISTRIBUTIONS OF THE MILK FAT PHASE USING UNIVERSAL PEARSON DISTRIBUTIONS

In this paper we consider the problem of approximating the experimental values of the coefficient of attenuation of ultrasonic oscillations and the mass distribution of the fat globules in the milk and milk products. The analysis of experimental data in terms of the choice of the method of approximation was done. A approximating dependence is based on the solution of Pearson differential equations. The advantages of the proposed method for the type of approximation of the experimental data obtained. An algorithm for constructing a mathematical model describing the relaxation spectrum and mass distribution of the fat globules in the milk and milk products was implemented. As a result, a family of Pearson approximation curves of the experimental data shows the ability to qualitatively correctly describe the change in the distribution of the fat phase in the process of homogenization. It estimates the error of approximating dependence, which amounted to 18 %. It is shown that during of the process of homogenization of dairy products changes shape of the curve describing the distribution of the fat globules, in view of the fact that there is a local extremum, caused by the presence of the non-homogenized fat globules. The accuracy of the selected mathematical model is significantly reduced. At the same time, it loses its physical meaning and its parameters. To address the identified deviations in the proposed mass distribution of fat globules as a function with two modes. It is proved that the complexity of the model is not only doubles the number of its parameters, but also complicates the interpretation of measurement results in a control system, and makes it difficult to analyze the obtained parameters of approximation by decision-maker. As a result of approximation of experimental data suggested to use statistical moments of the distribution for problem decision

AUTOMATION OF QUALITY CONTROL OF MILK HOMOGENIZATION BY ULTRASONIC SPECTROSCOPY METHODS

The paper deals with the possibility of determining homogenization degree of milk and dairy products using ultrasonic vibrations absorption spectra. Advantages of this method application in automated manufacturing systems were examined. Theoretical background of the method, as well as the possibility of determining the distribution of the fat globules in milk, depending on their sizes were substantiated. We derived mathematical equations, showing the relationship between the homogenization degree of dairy products and the acoustic properties of the medium, such as the propagation velocity of ultrasonic vibrations in the medium tested and the absorption coefficient in the medium at a specific frequency of the ultrasonic impact and medium temperature. It was shown that the measurement of the propagation of the milk acoustic properties in frequency, evaluation of their density function of the relaxation times spectrum, and then from the relaxation times to the particles masses allow operational control of the fat globules masses distribution in fractions. Theoretical studies were confirmed by experimental research carrying out whose results demonstrate clearly the dependence of absorption degree of ultrasonic vibrations on the degree of milk homogenization. The dependence between the estimates of the relaxation spectra and the first two moments of the statistical distribution of milk fat globules, which demonstrated their relationship was studied. Possible improvements the method under consideration to increase the reliability of the results obtained were proposed

Computational experiment for the purpose of determining the probabilistic and temporal characteristics of information security systems against unauthorized access in automated information systems

The article is devoted to the method of experimental estimation of parameters of functioning of standard information protection systems from unauthorized access, certified, widely used in organizations operating automated information systems. In the course of the experiment, statistical data were evaluated in the dynamics of the functioning of information security systems against unauthorized access in automated information systems. Registration of the parameters for the execution time of protective protection functions was carried out using a special utility called ProcessMonitor from the Sysinternals suite of utilities used to filter processes and threads. The loading of the processor and main memory of the computer with the use of special software, specially designed for performing experimental research, simulates the operation of GIS in real-world work for its intended purpose. A special software for simulating the work of a system with high load is developed in "VisualStudio 2015" within the framework of "ConsoleApplication". At the same time, the processor is loaded at a level of 50-70% and 60-80% of the operative memory. The obtained values of the time of implementation of protective functions in conditions of high utilization of resources of computer facilities for their intended purpose will allow us to assess the conflict and dynamic properties of the GIS. In the future, the obtained experimental estimates can be used to develop a model of information security in automated information systems, as well as in the formation of quality requirements (resource intensity, response time to the user's request, availability, etc.). Also, the results of the computational experiment in the future can be used to develop a software package for assessing the dynamic performance of information security systems against unauthorized access in automated information system

AUTOMATED PROCESSING OF DAIRY PRODUCT MICROPHOTOS USING IMAGEJ AND STATISTICA

Summary. The article discusses the construction of algorithms for automated processing of microphotos of dairy products. Automated processing of micro photos of dairy products relevant in the study of the degree of homogenization. Microphotos of dairy products contain information about the distribution of fat globules in the mass fractions. Today, there are some of software products, offering image processing and relieving researchers from routine operations manual data processing. But it need to be adapted for performing the processing of microphotos of dairy products. In this paper we propose to use for processing the application package ImageJ for processing image files taken with digital microscope, and to calculate the statistical characteristics of the proposed use of the software package Statistica. Processing algorithm consists of successive stages of conversion to gray scale, scaling, filtering, binarization, object recognition and statistical processing of the results of recognition. The result of the implemented data processing algorithms is the distribution function of the fat globules in terms of volume or mass fraction, as well as the statistical parameters of the distribution (the mathematical expectation, variance, skewness and kurtosis coefficients). For the inspection of the algorithm and its debugging experimental studieswere carried out. Carries out the homogenization of farm milk at different pressures of homogenization. For each sample were made microphoto sand image processing carried out in accordance with the proposed algorithm. Studies have shown the effectiveness and feasibility of the proposed algorithm in the form of java script for ImageJ and then send the data to a file for the software package Statistica