Gel express: a novel frugal method quantifies gene relative expression in conventional RT-PCR

Background Real-time PCR system is a valuable scientific mainstream needed for quantifying specific gene expression. Nevertheless, compared with conventional PCR, the real-time PCR system is extremely expensive and not affordable for limited or mid-budget research laboratories. Here, a novel, doable and low-cost recipe (referred to as gel express) is developed to quantify gene expression using conventional RT-PCR assay. The novelty of the gel express method is based on replacing crossing point (CP) values with integrated density (IntDen) values of PCR amplicon bands in real-time PCR regular mathematical formulas. Results In this work, gene expression profiles of two different rice stress-marker genes (OsCYP94C2a and OsLOX8) were quantified in response to mechanical wounding at different time points (0, 30, 60, and 150 min). In the gel express method, the free software ImageJ was employed to measure integrated density (IntDen) values of PCR amplicon bands in agarose gel images. IntDen values were then used instead of crossing point (CP) values according to the following modified formula: [EIntDen(ref)/EIntDen(target)]sample ÷ [EIntDen(ref)/EIntDen(target)]control. Gene relative expression profiles (dynamic expression pattern) quantified by gel express method in both genes were highly comparable with real-time RT-PCR. R2 values were 0.9976 and 0.9975 in OsCYP94C2a and OsLOX, respectively. PCR amplification efficiency (E) for all studied genes could be calculated depending on IntDen values through experimentally designed calibration curves. PCR amplification efficiencies with all studied genes obtained by gel express were all in the accepted range. For better-visualized PCR amplicons thus detectable biological effects between treatments, the number of PCR cycles applied in gel express method (IntCyc) was experimentally estimated to be 29 cycles. Conclusions Gel express is a novel, cost-effective and feasible recipe for quantifying gene relative expression in conventional RT-PCR. The expression pattern quantified by gel express is highly comparable and fits the expression data revealed by the used real-time PCR system.

Methodological Approach to Designing Fermented Dairy Products with Optimal Biological Value

The development of food products implies ensuring the optimal composition and ratio of the basic components, as well as their technological compatibility. A priori, the quality of raw materials, the optimal formula of the product and the efficiency of the technological process determine the quality of products, including biological value. The use of whole-cell sensors such as infusoria Tetrahymena pyriformis is most productive for screening biological studies. At present, for a comprehensive assessment there are no data on the use of simplest technology of fermented dairy products and the design of their biological value. The purpose of this research is to develop a methodology for creating whole-milk products of optimal biological value using the express method of biotesting. The research object was yogurt with the ratio of the mass fraction of fat and protein in the range of 0.36 ÷ 1.5, sucrose in the range of 5 ÷ 10%. An express method for determining the relative biological value of fermented dairy products using test organisms and an original methodology for creating whole-milk products of optimal biological value have been developed. A software has been developed to calculate formula of the product optimized for the following indicators: the relative biological value of the product, the cost of raw material and basic materials. The methodology is a tool to assist industry organizations in improving production technologies and quality management systems.

A new express method for determining the number of cycles of freezing and thawing poultry meat

Temperature fluctuations cause significant harm to the quality of poultry meat and its food safety, both at various stages of storage and transportation, as well as sales. Repeated repeated defrosting and freezing lead to a violation of the integrity of cells or protein denaturation, which is accompanied by a change in the ratio of the forms of moisture binding to the product. In this regard, there is an urgent task of establishing the facts of thawing and freezing of poultry meat, determining the number of cycles of repeated freezing. This article is devoted to the development of a new express method for determining the number of cycles of freezing – thawing of a broiler at the stages of the life cycle of poultry products. As the number of «freeze-defrost» cycles increases, the peak area of the differential scanning calorimetry curve (DSC) decreases from 206.4 to 192.6 kJ/mol with electric stunning and from 168.6 to 151.5 kJ/mol with gas stunning, the melting peak temperature also decreases from 5.7 to 5.2 oC with electric stunning and from 5.0 to 4.0 oC with gas stunning. The DSC temperature program and recommendations for its use for monitoring broiler processing and storage processes accompanied by phase transformations of water have been developed.

EXPRESS METHOD FOR CALCULATING THE SPECIFIC HEAT OF EXPLOSIVE TRANSFORMATION OF CHNO CONDENSED EXPLOSIVES

Разработан экспресс-метод расчета теплоты взрыва СаHbNcOdконденсированных взрывчатых веществ с различным кислородным балансом от резко отрицательного до положительного. Предложенный метод использует минимальный набор входных данных, состоящих из элементного состава, плотности энтальпий образований исходного взрывчатого вещества и его продуктов детонации. Расчет теплоты взрыва основывается на корреляционной связи между минимальной и максимальной теплотами взрыва с плотностью высокоэнергетического соединения. В статье подробно приведены реакции разложения взрывчатых веществ для случаев с минимальной и максимальными теплотами взрыва. Проведены расчеты теплоты взрыва по новому способу и методу Пепекина по представленной в статье базы взрывчатых веществ, а также приведены результаты сравнения, которые показали большую точность (в 2,3 раза) предложенного метода. An express method has been developed for calculating the explosion heat of cahbncod condensed explosives with different oxygen balance from sharply negative to positive. The proposed method uses a minimal set of input data consisting of the elemental composition, enthalpy density of the formations of the initial explosive and its detonation products. The calculation of the heat of explosion is based on the correlation between the minimum and maximum heat of explosion with the density of a high-energy compound. The article describes in detail the decomposition reactions of explosives for cases with minimum and maximum explosion heats. Calculations of the heat of explosion according to the new method and the pepekin method are carried out according to the explosives database presented in the article, and comparison results are also presented, which showed a better accuracy (2.3 times) of the proposed method.

Use of new similarities Bl and Blturb. to optimize the calculation and selection of heat exchange equipment for working with nanofluid coolants

The problem of correct, exact calculation and selection of the optimal heat exchange equipment at use in it of nanoliquid heat carriers was investigated in the work. Classical numerical equations, which are widely used in the calculation and selection of heat exchangers with nanofluids, especially at temperatures above 50 °C, give an error of (15–20) % or more. This leads to the fact that the selected heat exchange equipment may not work efficiently with excessive consumption of thermal energy. A new approach to heat transfer processes is considered, taking into account the theory of J. Businesque, which gives an idea of turbulent viscosity and thermal conductivity, as well as comparing the resistance of the coolant flow to the nanoparticle with surface forces and considering turbulent fluid as Newtonian. It is shown that the consideration of the behavior of a nanoparticle in a turbulent liquid coolant without taking into account surface forces is inaccurate and erroneous. The physical content of the previously obtained new numbers of similarity Bl and Blturb is considered and the possibility of their effective application in the new numerical equation obtained by us for the calculation of heat exchangers using nanofluid coolants is shown. The existing express method of estimating the efficiency of nanorluids use in heat exchangers on the basis of classical numerical equations is analyzed and a new express method on the basis of a new numerical equation and new numbers of similarity Bl and Blturb is proposed. The proposed express calculation method shows that a mixture of H2O + EG (60:40) improves the heat transfer properties of water by + 12.86 %, and a mixture of (H2O + EG (60:40) + 1.5 % TiO2) and (milk) + 0.5 % pumpkin seed oil) – by +16.75 %, which corresponds to the experiments and our calculations, and the known express method based on classical numerical equations shows a deterioration of – 4.5 % and, accordingly, by – 1.2 %. An example of calculating the optimal shell-and-tube heat exchanger according to the new algorithm when heating milk with hot water with the addition of mixtures (H2O + EG (60:40) + 1.5 % TiO2) and accordingly (milk + 0.5 % pumpkin seed oil) fully confirms the correctness of the new express –method.

MONITORING OF THE FERMENTATION PROCESS IN VARIOUS TYPES OF WINE

A classical approach to monitoring the fer-mentation of grape must based on the use of the areometric method is considered. Based on the processing of the data of measuring the refractive index and density of grape must, as well as data on the physicochemical composi-tion of grape must during fermentation, meth-odological approaches have been developed to create methods for monitoring the composi-tion of the fermenting must using refractomet-ric and refractodensimetric methods of analy-sis. The results of the work can become the basis for the development of a non-destructive express method for determining the volume fraction of ethyl alcohol and the mass concentration of sugars in fermenting must based on the standard equipment of the laboratory of wine-making enterprises, and for the improvement of technical specifications for the manufacture of a portable device for determining the concentration of alcohol and sugar content, based on the simultaneous measuring the refractive index and density of liquid media.

The short-term tsunami forecast

A brief overview of the methods of a tsunami early warning in the Kuril Islands, which turned out to be ineffective during recent events, is presented. A hydrophysical method for short-term tsunami forecasting based on information about a tsunami in the ocean, used in the United States, and an express method, also using information about a tsunami in the ocean, are briefly described. The results of the retrospective forecast of the tsunami that occured on March 11, 2011, by the express method are presented.

Metal chips preparation for utilization using advanced reagents

The article touches upon the problems of utilization of metal chips containing residues of cutting fluids. The research results are shown for nickel alloy chips washing to remove cutting fluids with the use of various reagents available on the current market. In the course of the research, an original colorimetric express method for assessing the contamination of metal chips was developed, which allows quickly and efficiently establishing the quantitative drop in the cutting fluids content in the chips after the washing. Conclusions are made as to the most effective reagent from the point of view of economics and quality of cutting fluid removal.

Express method of assessment of compliance of the object of supervision with fire safety requirements

Introduction. The paper considers the new requirements for the activities of the state fire supervision bodies. The ways of solving the problems arising during the implementation of these innovations are given. One of such solutions is the developed by the authors algorithm of the express method for assessing the compliance of the object of supervision with fire safety requirements. The article analyzes the work of inspectors of the state fire supervision during inspections of low-rise retail facilities of small area, taking into account the time spent on them, and the work of the inspector using a new express method of conducting inspections. Problem Statement. The purpose of the study is to increase the efficiency of managerial decision-making in ensuring fire safety based on the criterion of "fire risk", taking into account the minimum fire safety requirements determined by the calculation. Conclusions. The developed algorithm for supporting managerial decision-making on the compliance of the protection of commercial facilities with fire safety requirements will allow inspectors of the state fire supervision during inspections to assess the existence of a threat to human life and health and decide on further actions depending on the presence or absence of this threat.