SEARCH FOR ALTERNATIVE SOURCES OF NATURAL PLANT ANTIOXIDANTS FOR FOOD INDUSTRY

According to the data of the Food and Agriculture Organization of the United Nations, global food losses are about one third of their total output, mainly due to spoilage. Therefore, a search for safe methods of shelf life extension is an important task, especially for products for specialized nutrition. According to literature data, natural antioxidants can be alternative to existing preservatives due to its ability to inhibit oxidation of the main nutrients. Pulp, seeds and peel of quince, apple «Simirenko», feijoa, persimmon, Jerusalem artichoke, white, red and yellow onion were objects of the study. The total antioxidant capacity was determined by Oxygen Radical Absorbance Capacity (ORAC) and Ferric Reducing Antioxidant Power (FRAP) methods and expressed in µmol equiv. Trolox / g of sample and µmol equiv. Dihydroquercetin / g of sample. It was noticed that peels demonstrated higher antioxidant activity, and red onion husk possessed the highest value, which was 722.8 ± 13.9 µmol equiv. Dihydroquercetin / g of sample and 3357.5 ± 42.2 µmol equiv. Trolox / g of sample. It was shown that the use of fruit and vegetable wastes is promising for production of food-grade antioxidants. In addition, the results of the research could facilitate stimulation of rational and efficient environmental management.According to the data of the Food and Agriculture Organization of the United Nations, global food losses are about one third of their total output, mainly due to spoilage. Therefore, a search for safe methods of shelf life extension is an important task, especially for products for specialized nutrition. According to literature data, natural antioxidants can be alternative to existing preservatives due to its ability to inhibit oxidation of the main nutrients. Pulp, seeds and peel of quince, apple «Simirenko», feijoa, persimmon, Jerusalem artichoke, white, red and yellow onion were objects of the study. The total antioxidant capacity was determined by Oxygen Radical Absorbance Capacity (ORAC) and Ferric Reducing Antioxidant Power (FRAP) methods and expressed in µmol equiv. Trolox / g of sample and µmol equiv. Dihydroquercetin / g of sample. It was noticed that peels demonstrated higher antioxidant activity, and red onion husk possessed the highest value, which was 722.8 ± 13.9 µmol equiv. Dihydroquercetin / g of sample and 3357.5 ± 42.2 µmol equiv. Trolox / g of sample. It was shown that the use of fruit and vegetable wastes is promising for production of food-grade antioxidants. In addition, the results of the research could facilitate stimulation of rational and efficient environmental management

Behavioral and neuronal responses of two mouse species, Mus musculus and Mus spicilegus, to con- and heterospecific olfactory signals

The behavioral and neuronal responses of the males of two closely related species of mice (Mus musculus  wagneri, M. spicilegus) to the urine odors of estrus con- and heterospecific females were studied. In two-choice odor tests males significantly longer investigated odor of conspecific females in comparison with heterospecific ones. To investigate neuronal activation in the main and accessory olfactory bulbs (MOB and AOB), one of the methods of functional tomography – manganese-enhanced MRI ( ME-MRI) – was used. There was a significant increase in Mn2+ accumulation in the dorsal section of the posterior part of the MOB in male M. m. wagneri and M. spicilegus exposed to odor of conspecific females compared with the control group males (odor not exposed). There was a local significant increase in manganese accumulation in the dorsal region of the anterior part of the MOB in the case of the exposure of odor of a heterospecific female. The exposure of heptanone-2 to mice resulted not only in an increase in Mn2+ accumulation in certain zones, but also in a significant decrease in the accumulation of Mn2+ in the rest of the olfactory bulbs. A significant increase in the accumulation of MRI contrast in AOB was observed in males only in the case of female urine-conspecific odor exposure. The results support the previously stated assumption of a significant difference in chemical communication systems in two species of mice. A comparison of these results and results obtained previously demonstrated the absence of any differences in behavioral and neuronal responses to con- and heterospecific odors of the house mouse subspecies allopatric (M. m. wagneri) and sympatric (M. m. musculus) to M. spicilegus. This fact does not allow us to assume the effect of the mechanism of “reinforcement” in the process of evolution in the formation of precopulatory reproductive isolation between the sympatric species M. spicilegus and M. musculus

Methodology of adipose tissue type detection in mammals

Nowadays, an interest in studying the composition, properties and functions of adipose tissue (AT) is growing among researchers, which is conditioned by its important role in the normal functioning of the body. Due to different types of adipose tissue (AT) in mammals (white, beige, brown and pink) and different physiological tasks performed by each type of AT, rapid, correct and effective detection of an AT type is highly topical. Methods used today are labor consuming and in the case of NMR and CT expensive, which limits possibilities of scientists. In this connection, the aim of this research was to develop a methodological approach allowing rapid and effective detection of an adipose tissue type. A methodology was formed based on the concept, formalized requirements for the method, step-wise structure of investigations and interpretation of results. The concept is based on differences in the structure of the adipose cell (adipocyte) of different AT types. The method is based on extraction of heme containing proteins. To this end, solvents and parameters of extraction that facilitate their better extraction have been chosen. An AT type has been determined by the total content of iron contained in the cytochrome fragment. Our own modification was selected. This modification includes preliminary mincing of a sample with the ice-cold TES buffer (pH 8.5) in a ratio of 1:5 (g: mL), homogenization at 9,000 rpm for 2 min with the following centrifugation at 10,000 g and 4 °C for 15 min. Effectiveness of the proposed method was confirmed by the histological and electrophoretic analyses. Therefore, the new methodology of identification and differentiation of adipocytes was proposed for rapid and effective detection of an adipose tissue type.Nowadays, an interest in studying the composition, properties and functions of adipose tissue (AT) is growing among researchers, which is conditioned by its important role in the normal functioning of the body. Due to different types of adipose tissue (AT) in mammals (white, beige, brown and pink) and different physiological tasks performed by each type of AT, rapid, correct and effective detection of an AT type is highly topical. Methods used today are labor consuming and in the case of NMR and CT expensive, which limits possibilities of scientists. In this connection, the aim of this research was to develop a methodological approach allowing rapid and effective detection of an adipose tissue type. A methodology was formed based on the concept, formalized requirements for the method, step-wise structure of investigations and interpretation of results. The concept is based on differences in the structure of the adipose cell (adipocyte) of different AT types. The method is based on extraction of heme containing proteins. To this end, solvents and parameters of extraction that facilitate their better extraction have been chosen. An AT type has been determined by the total content of iron contained in the cytochrome fragment. Our own modification was selected. This modification includes preliminary mincing of a sample with the ice-cold TES buffer (pH 8.5) in a ratio of 1:5 (g: mL), homogenization at 9,000 rpm for 2 min with the following centrifugation at 10,000 g and 4 °C for 15 min. Effectiveness of the proposed method was confirmed by the histological and electrophoretic analyses. Therefore, the new methodology of identification and differentiation of adipocytes was proposed for rapid and effective detection of an adipose tissue type

Variability of Fragments of Nuclear Brca1 Gene, Exon 11, and Mitochondrial Cox1 Gene in House Mice Mus musculus

To clarify genetic differences between subspecies of the house mouse Mus musculus, their distribution, and hybridization, we first conducted a comparative analysis of variability of nucleotide sequences of fragments of the nuclear gene Brca1, exon 11 (2331 bp), and mitochondrial gene Cox1 (1260 bp) in 40 house mice from West and East Europe, Transcaucasia, Siberia, and Central and South Asia. Brca1 genotypes were divided into five main groups, which differed in a number of fixed substitutions. Genotypes of each group are characteristic for the certain geographical region and the following subspecies: M. m. musculus, M. m. domesticus, M. m. castaneus, and M. m. wagneri together with M. m. gansuensis; a fifth group corresponds to an unidentified subspecies or a distinct genetic form of M. musculus from India (Sikkim State). Besides the homozygous specimens, we revealed mice, which were heterozygous for all diagnostic loci simultaneously; these specimens were determined as hybrid. Hybrid mice were mainly found in the zones of contact of subspecies, but in some cases, quite far from one of the parent subspecies (possibly, due to transportation). In two hybrid mice (from Bakhtiari Province of Iran and Transbaikalia of Russia), unique Brca1 haplotypes were detected. It cannot be ruled out that, at least partly, they may be characteristic of the M. m. bactrianus and M. m. gansuensis subspecies, respectively. Thus, the results of the study showed that the nuclear Brca1 gene is a promising molecular genetic marker for the analysis of variability, differentiation, and hybridization of house mice as well for subspecific identification of M. musculus specimens. Despite more rapid evolution of the Cox1 gene, it is not well suited for discrimination of M.m. musculus, M. m. wagneri, M. m. gansuensis specimens and Transcaucasian representatives of M.m. domesticus due to introgression and long-term maintenance of foreign mitochondrial DNA in populations. However, Cox1 gene analysis (along with the diagnostics of animals by nuclear DNA) may be useful for estimation of population differences in M. m. castaneus and M.m. domesticus subspecies

ИЗУЧЕНИЕ ВОЗМОЖНОСТИ ПРИМЕНЕНИЯ МЕТОДОВ IN VITRO ДЛЯ ОЦЕНКИ БИОКОРРИГИРУЮЩИХ СВОЙСТВ БИОЛОГИЧЕСКИ АКТИВНЫХ ВЕЩЕСТВ ЖИВОТНОГО ПРОИСХОЖДЕНИЯ

Slaughter products are good source of bioactive substances with different biological effects. Nowadays, approximately 220 functional peptides consisting of 2–10 amino acid residues have been identified. In this study, the biological activity of medium-molecular-weight substances (5–30 kDa, Mw) and low-molecular-weight substances (less than 5 kDa, Mw) extracted from aorta of Sus scrofa were investigated in vitro. Medium-molecular-weight and low-molecular-weight fractions possessed a tissue specific effect on organotypic tissue cultures of chicken embryos and laboratory rats. Biological activity depended on molecular weight. Medium-molecular-weight fractions possessed higher effect on aortas explants of chicken embryo: area index of cellular monolayer was 56.6±5.2 % (area index of the monolayer was 37.47±3.27 % for low-molecularweight fraction) in comparison with control explants. The low-molecular-weight fractions stimulated aortic tissues explants of aging rats — area index exceeded area index formedium-molecularweight fractions by more than 30%.Известно, что продукты убоя являются источником большого количества биологически активных веществ, обладающих различными биологическими эффектами. На настоящий момент выявлено порядка 220 функциональных пептидов, большинство из которых состоят из 2–10 аминокислотных остатков. В данном исследовании изучена биологическая активность среднемолекулярных (Мм 5–30 кДа) и  низкомолекулярных (Мм менее 5 кДа) веществ, содержащихся в  аорте Sus scrofa, с  применением методов in vitro. Установлено, что среднемолекулярные и  низкомолекулярные фракции, обладают тканеспецифическим действием на органотипические культуры тканей куриных эмбрионов и лабораторных крыс, при этом наблюдается выраженная зависимость биологической активности от молекулярной массы исследуемых образцов. Показано, что, на эксплантаты аорты куриного эмбриона более выраженное действие оказывают среднемолекулярные фракции — индекс площади образующегося монослоя клеток составил 56,6±5,2  % (индекс площади монослоя при добавлении низкомолекулярных фракций составил 37,47±3,27 %), по отношению к контрольным эксплантатам. Низкомолекулярные фракции оказывали стимулирующее влияние на эксплантаты тканей аорты стареющих крыс — индекс площади превышал значения среднемолекулярных фракций более чем на 30%

Применение метода тонкослойной хроматографии для анализа антиоксидантной активности

Plants are a rich source of effective non-toxic biologically active substances. Various physicochemical methods of analysis are used for evaluation of plant antioxidant activity. Composition of ethanol extracts of red, yellow and white onion husks, dried rosemary, basil, and chaga were studied by high performance thin layer chromatography (HPTLC) method. The antioxidant activity of the obtained fractions on a chromatographic plate was assessed by subsequent DPPH screening. The extracts red and yellow onion husk and rosemary demonstrated the highest antioxidant activity, variability of the qualitative composition and similarity of antioxidant profiles, while extract of white onion husks did not contain any antioxidant classes. Intensive spots with Rf of 0.13-0.97 were observed along the whole chromatogram track corresponding to red onion husks. It was also found that all tested extract, excepting white onion husk and chaga, contained spots with varying degrees of intensity in the Rf range of 0.96-0.98, which corresponded quercetin Rf value.Для исследования антиоксидантной активности растительного сырья, которое является богатым источником эффективных нетоксичных биологически активных веществ, используются различные физикохимические методы анализа. В данной работе был применен метод высокоэффективной тонкослойной хроматографии (ВЭТСХ) для разделения биологически активных веществ, входящих в этанольные экстракты шелухи красного, желтого и белого лука, сушенных розмарина, базилика и чаги. Антиоксидантную активность полученных фракций на хроматографической пластине оценивали последующим DPPH-скринингом. Наибольшими показателями антиоксидантной активности и вариабельностью качественного состава характеризовались экстракты шелухи красного и желтого лука, а также розмарина, причем для этих образцов была отмечена схожесть антиоксидантных профилей. В экстракте белого лука не было обнаружено каких-либо классов антиоксидантов. Для шелухи красного лука было отменено наличие значительно выраженных пятен по всему треку, значения Rf которых расположены в диапазоне 0,13-0,97. Доказано, что все экстракты, кроме шелухи белого лука и чаги, в разной степени интенсивности имеют пятна с коэффициентом замедления в диапазоне 0,96-0,98, что соответствует полученному значению Rf кверцетина

ВЛИЯНИЕ АВТОЛИЗА НА ПРОТЕОМНО-ПЕПТИДНЫЙ ПРОФИЛЬ СЕРДЕЧНОЙ МЫШЦЫ И АОРТЫ Bos taurus и Sus scrofa

The article presents the results of autolytic processes impact on the protein-peptide profile of Bos taurus and Sus scrofa cardiac muscle and aorta. The results of tissue-specific protein identification are also presented as well as the effect of autolysis. Apolipoprotein A-1 involved in the formation of high-density lipoproteins, peroxiredoxin-1 involved in the suppression of oxidative stress, galectin-1 induced apoptosis of T-lymphocytes, as well as number of heat shock proteins with molecular weight less than 30 kDa were identified in Sus scrofa aorta tissue. It was discovered that functional proteins with molecular weight less than 30 kDa are retained during the freezing process, but destroyed under the action of autolytic enzymes. This work was supported by the Russian Science Foundation (project No. 16–16–10073).В статье представлены результаты изучения влияния автолитических процессов на белково-пептидный состав сердечной мышцы и аорты Bos taurus и Sus scrofa, приведены результаты идентификации тканеспецифичных белковых молекул и влияние автолиза на их сохранность. В тканях аорты Sus scrofa были обнаружены аполипопротеин А-1, участвующий в образовании липопротеинов высокой плотности, пероксиредоксин-1, участвующий в подавлении окислительного стресса, галектин-1, индуцирующий апоптоз Т-лимфоцитов, а также ряд белков теплового шока, имеющих молекулярную массу менее 30 кДа. Было установлено, что функциональные белковые вещества с  молекулярной массой менее 30 кДа сохраняются в процессе заморозки, но разрушаются под действием ферментов сырья при автолизе. Исследование выполнено за счет гранта Российского научного фонда (проект №16–16–10073)