Laboratory of Senses

It is reported about a series of scientific and research events “Laboratory of Senses” organized in October and December 2016 at the Philological faculty of Samara State Socio-Pedagogical University. The content and purpose of “Laboratory of Senses” are characterized - teaching the analysis of literary text in literary, philological and linguistic methodology aspects with the achievements of various methodologies and scientific schools. Constructive performance of this series of activities carried out by teachers of the University with the bachelors and undergraduates is described

THE MOLECULAR GENETIC STUDY OF KRIM-SAGHYZ (Taraxacum hybernum Steven) USING SSR, RAPD AND ISSR MARKERS

Krim-saghyz (Taraxacum hybernum Steven) is an alternative to Hevea brasiliensis as a source of natural rubber. In Russia, krim-saghyz is common only in the Crimean Peninsula and is traditionally named after it. In spite of its potential for economical use, the genetic structure of the Crimean population of this plant is still unexplored. In this regard, the purpose of our work was a comparative molecular-genetic characterization of T. hybernum from various habitats of the Crimean Peninsula using SSR, RAPD and ISSR markers. According to the plan, we collected achenes, leaves and roots of krim-saghyz in 10 spots all over the Crimean Peninsula. We found the plants in the western part of the southern Crimean coast and the western part of the Crimean foothills, which are two general regions of the area of this species. Total DNA was extracted from dry leaves of krim-saghyz with cetyltrimethylammonium bromide (CTAB). For the first time 12 SSR, 3 RAPD and 3 ISSR markers were tested on krim-saghyz. To observe polymorphism of RAPD- and ISSR-fragments, we used analytical electrophoresis in 1.7 % agarose gel. To compare the length of SSR amplicons, we used gel-electrophoresis in 8 % polyacrylamide gel. We found that the Crimean population of krim-saghyz appears to be genetically homogeneous. This could be due to a small geographic range and apomictic reproduction of this species. However, the phenotypical diversity within the population of T. hybernum is well known from the literature. Consequently, the study of the DNA polymorphism of this species should be continued, in particular, with the help of high-resolution techniques

Эффективность интерферонотерапии при острых респираторных вирусных инфекциях у детей раннего возраста

The problem of choosing an effective etiotropic treatment for acute respiratory viral infections in young children is one of the urgent in pediatrics.Materials and methods. The article presents data on the mechanism of the combined antiviral and immunomodulating action of the drug recombinant interferon alpha-2b. The results of observation of 80 children aged 3 to 12 months with acute respiratory viral infections of various etiologies are presented. Results. Clinical and laboratory indicators of intoxication that occur during acute respiratory viral infections in children have been identified. It has been shown that the use of rectal suppositories of interferon alpha-2b in the treatment of young children with acute respiratory viral infections causes a rapid relief of the clinical symptoms of the disease and helps to reduce the duration of hospital treatment of patients.Проблема выбора эффективной этиотропной терапии острых респираторных вирусных инфекций у детей раннего возраста является одной из актуальных в педиатрии. Материалы и методы: приведены данные о механизме комбинированного противовирусного и иммуномодулирующего действия препарата рекомбинантного интерферона альфа-2b с высокоактивными антиоксидантами витаминами Е и С (ВИФЕРОН®). Представлены результаты наблюдения за 80 детьми в возрасте от 3 до 12 месяцев с острыми респираторными вирусными инфекциями различной этиологии. Результаты: Выявлены клинико-лабораторные показатели интоксикации, возникающие при острых респираторных вирусных инфекциях у детей. Показано, что применение ректальных свечей интерферона альфа-2b (ВИФЕРОН®) в терапии детей раннего возраста с острыми респираторными вирусными инфекциями вызывает быстрое купирование клинических симптомов болезни и способствует сокращению продолжительности стационарного лечения больных.

Pyrrolylquinoxaline-2-one derivative as a potent therapeutic factor for brain trauma rehabilitation

Traumatic brain injury (TBI) often causes massive brain cell death accompanied by the accumulation of toxic factors in interstitial and cerebrospinal fluids. The persistence of the damaged brain area is not transient and may occur within days and weeks. Chaperone Hsp70 is known for its cytoprotective and antiapoptotic activity, and thus, a therapeutic approach based on chemically induced Hsp70 expression may become a promising approach to lower post-traumatic complications. To simulate the processes of secondary damage, we used an animal model of TBI and a cell model based on the cultivation of target cells in the presence of cerebrospinal fluid (CSF) from injured rats. Here we present a novel low molecular weight substance, PQ-29, which induces the synthesis of Hsp70 and empowers the resistance of rat C6 glioma cells to the cytotoxic effect of rat cerebrospinal fluid taken from rats subjected to TBI. In an animal model of TBI, PQ-29 elevated the Hsp70 level in brain cells and significantly slowed the process of the apoptosis in acceptor cells in response to cerebrospinal fluid action. The compound was also shown to rescue the motor function of traumatized rats, thus proving its potential application in rehabilitation therapy after TBI. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.Ministry of Education and Science of the Russian Federation, Minobrnauka: 0124-2019-002Russian Foundation for Basic Research, RFBR: 20-33-70102Russian Science Foundation, RSF: 18-74-10087Funding: This research was funded by Russian Science Foundation, research project #18-74-10087 (V.F.L., E.A.D., M.A.M., E.R.M.), Russian Foundation for Basic Research, research project #20-33-70102 (I.A.U., O.N.C., V.N.C, M.?.T., I.V.G.), and by The Ministry of Education and Science of Russian Federation № 0124-2019-002 (R.V.S., N.D.A., B.A.M.)

EXPRESSION OF mRNA FOR CYTOKINES COMPARED TO THEIR CONCENTRATIONS IN CULTURE SUPERNATES OF U937 CELLS EXPOSED TO POLYCLONAL ACTIVATORS

Investigation of the cytokine expression dynamics as well as the cytokine-producing potential of immune-competent cells allows extensive studies of their functional characteristics. mRNAs encoding a number of cytokine genes are relatively stable, thus their level may be used as a marker for assessing the levels of activation and proliferation of immunocompetent cells as well as for evaluating the cytokine-producing potential of immunocompetent cells.In our work, we assessed correlations between the levels of mRNA expression specific for IL-10, TNF-α, GM-CSF cytokines determined in a culture of differentiated macrophage U937 cells, and protein concentrations of the same cytokines as measured in supernates of U937 cell cultures, without and after exposure to polyclonal activators. The IL-10, TNF-α, GM-CSF mRNA expression was determined by real-time quantitative polymerase chain reaction. Protein concentrations of IL-10, TNF-α, GM-CSF cytokines in the culture supernatant of U937 cells were measured by an enzyme immunoassay. The use of an initially homogeneous cell culture in the study is convenient due to the identical conditions in all experimental variants.The most pronounced effect of polyclonal activators is exerted upon production of GM-CSF mRNA, as well as protein concentration of this cytokine in the cell culture supernatants, thus actually coinciding with RT-qPCR results.  The TNF-α mRNA level decreased under the influence of polyclonal activators, whereas concentration of this cytokine was decreased in the cell supernate. The TNF-α protein in a culture medium did not reflect temporal changes in the cellular TNF-α mRNA expression, probably, due to potential decrease of cellular mRNA occurring by the feedback inhibitory mechanism. While the cytokines can accumulate and remain in the supernatant, the mRNA-related events leading to cytokine formation may be completed earlier. Therefore, the signalling pathways and cytokine release kinetics should be studied after establishing the time dependence at short time intervals, which may be individual for each cytokine.Thus, the results of a study using polyclonal activators suggest that polyclonal activators applied as mitogens, have a significant effect upon the concentration of secreted IL-10, TNFα and GM-CSF. In this case, polyclonal activators affect the levels of mRNA encoded by cytokine genes, thus indicating transcriptional mechanisms of its action. But, in view of the fact that the data are ambiguous, in order to achieve greater correspondence between the changes in the studied proteins and specific mRNAs, a detailed description of the time dependence is required for the changes in mRNA contents

Cytokinesproduction by blood immune cells in patients of different age groups with invasive ductal carcinoma of no special type and lymphatic metastases

The aim of this study was to evaluate production of cytokines by the blood immune cells in patients of different age groups with invasive non-specified type mammary ductal carcinoma, with or without metastases. Production of cytokines by periphery blood immunocompetent cells, either spontaneous and stimulated with polyclonal activators, was assessed in 82 patients with invasive mammary ductal carcinoma. The concentrations of IL-6, IL-8, IL-10, IL-17, IL-18, IL-1p, IL-1ra, TNFa, IFNy, G-CSF, GM-CSF, VEGF и MCP-1 were determined by solid-phase enzyme-linked immunosorbent assay. The patients were divided into 2 age groups: the first group consisted of 48 patients from 45 to 60 years old; the second group consisted of 34 older patients (61 to 75 years old). Metastases in local lymph nodes were documented in twenty patients from the first group and nine patients from the second group. The younger patients (45-60 years old) with metastases in local lymph nodes showed higher polyclonal activation index of IL-4 and IL-1ra production, when compared to the patients without lymphatic metastases. As for the older patients (61 to 75 years old), their polyclonal activation index of IL-6, IL-8, IL-1ra, G-CFS, GM-CSF production was significantly lower in cases of local lymph nodes metastases. The latter was due to higher level of spontaneous production, which suppressed the influence of polyclonal activators. The index of polyclonal activation upon production of cytokines in patients with lymphatic metastases was shown to be significantly higher for the age group of 45 to 60 years than in the age group of 61 to 75 years old. This fact suggested a highly stimulating effect of polyclonal activators in patients younger than 60 years. It was found that correlation between the index of polyclonal activation of cytokine production, and the indexes of ER, PR, HER2/NEU and Ki-67 receptor expression, which are used for the determination of molecular genetic subtype of the tumor, differ significantly for the distinct age groups

Investigation of the Sorption Properties of Ore Materials for the Removal of Sulfur Dioxide from Exhaust Flue Gases of Power Plants

The prospects of using a natural material – ferromanganese nodules (FMN) from the Gulf of Finland – as the SO2 adsorbent are discussed. The starting material was studied as pellets and powder using X-ray fluorescence spectroscopy, XRD, BET, and mechanical strength analysis; dependences of physicochemical parameters of the material on heat treatment at 100-1000 ºC were found. FMN samples were tested in the process of SO2 sorption. The sorptive capacity of FMN samples for SO2 was found to increase with humidity of adsorbents; FMN samples with a humidity above 40% were shown to be promising for the removal of sulfur dioxide from gase

Blood biomarkers and Ki-67 proliferation marker in breast cancer

Metastasis is the leading cause of death in patients with breast cancer (BC). It is known that the lesion of regional lymph nodes by tumor cells is more common in tumors with higher proliferative activity. Moreover, there is literature evidence on effects of cytokines and proteins upon the migration potential of the tumor. The aim of our work was to study the correlation between the concentrations of cytokines, proteins, and expression of Ki-67 proliferation marker in breast cancer with histology of non-specific invasive carcinoma.On the basis of pathological findings, 16 patients had metastases in regional lymph nodes (group I), and 18 patients had no detectable metastases (group II). Solid-phase enzyme immunoassay was used to determine concentrations of 14 cytokines in the supernatants of immunocompetent blood cells, i.e., IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1ra, TNFα, IFNγ, G-CSF, GM-CSF, VEGF and MCP-1, and concentrations of 6 proteins were determined in blood serum: estrogen and progesterone receptors, cadherin-E (CDH1), plasminogen activator type 1 (PAI-1), mucin 1 (MUC1), heat shock protein 90αA1 (HSP90αA1). Immunohistochemical study of Ki-67 expression was performed in paraffin sections of tumors using monoclonal antibodies.The study showed that Ki-67 expression in tumor tissues and blood concentrations of IL-6, IL-8, IL-1β and TNFα were higher in group I patients. On the contrary, blood concentrations of CDH1 and PAI-1 were higher in group II patients. It was found that Ki-67 showed both inverse correlations with CDH1 and PAI1, and direct correlations with IL-8 and TNFα. CDH1 had a direct correlation with PAI1, and inverse correlations with IL-6, IL-1β and TNFα. The studied cytokines showed direct correlations with each other. The analysis of ROC curves showed good quality and optimal values of the cut-off points for Ki-67 expression, cytokine and protein concentrations, thus allowing best prediction for detectable lymphatic metastasis.On the basis of these results, a quotient was proposed, which represents a ratio of CDH1 contents to the sum of IL-1β and TNFα concentrations in blood samples, which can help identification of the patients with breast cancer at risk for lymphatic metastasis

Perspectives of using Illumina MiSeq for identification of arbuscular mycorrhizal fungi

Arbuscular mycorrhiza fungi (AMF) form one of the most common symbiosis with the majority of land plants. AMF supply the plant with various mineral elements, primarily phosphorus, and improve the water supply. The search for the most effective AMF strains for symbiosis and the creation of microbial preparations on that basis is an important task for modern biology. Owing to the difficulties of cultivation without a host plant and their high genetic polymorphism, identifying AMF is very difficult. The high number of cryptic species often also makes morphological identification unreliable. Recent years have seen growth in the number of AMF biodiversity studies performed by modern NGS-based methods, Illumina MiSeq in particular. Currently, there are still many questions that remain for the identification of AМF. The most important are whether conservative or variable sequences should be used to select a marker for barcoding and whether universal primers or those specific to AMF should be used. In our work, we have successfully used universal primers ITS3 and ITS4 for the sequencing in Illumina MiSeq of the 5.8S rDNA – ITS2 region of the 35S rRNA genes, which contain both a conservative and variable regions. The molecular genetic approach for AMF identification was quite effective and allowed us to reliably identify eight of nine isolates to the species level: five isolates of Rhizophagus irregularis, and one isolate of R. invermaius, Paraglomus laccatum, and Claroideoglomus etunicatum, respectively. For all five R. irregularis isolates high variability in the ITS region and the absence of ecotopic-related molecular characters in the ITS2 region were demonstrated. The NCBI data is still insufficient for accurate AMF identification of Acaulospora sp. isolates from the genus to the species level