Marketing management of the territory in the aspect of the regional brand formation

The importance of managing branding of territory in terms of satisfying the demands of domestic consumers and attracting external economic agents is mediated by the realities of modern economic development in various regions of the country and the focus on the formation of accelerated development territories. These realities are characterized by unevenness. High competition between the territories in managing branding raises the importance of marketing management in territorial development and brand building. The article contains the statement that the success of the brand of the territory facilitates its sustainable development, attracting new economic agents, increasing the confidence of the population and business partners in regional business, building social and economic potential, and creating territorial advantages.peer-reviewe

Basic management principles of the formation of consumer loyalty to brands of regional companies

The problem of forming of consumer loyalty to companies that implement market activity in the regions of the Russian Federation on the scale of small and medium business is investigated. Due to the fact that such business structures do not have the opportunity to compete with large international corporations, they should identify their vector of development with a focus on the consumer, understanding that the formation of loyalty is a very important element of ensuring a stable market position. It is noted that the managers need to work towards improving the quality of not only products, but also implemented management measures in a number of areas. The customer satisfaction is an indicator of achieving a match between the subjectively expected value of the product to the potential consumer and the actual value obtained as a result of the experience of its acquisition and consumption. Thus, only in the case of positive consumer experience can the formation of loyal attitude. A list of basic practice-oriented principles that allow to attract customers, to form a loyal attitude to the company and products has been presented. It is determined that the formation of loyalty is not a one-time marketing campaign, but a complex work of a cohesive team of employees under the guidance of an effective manager. Their effective work, implemented on a permanent basis, will ensure the strengthening of the market position of the brand of the regional business structure. The presented principles will help managers working on the scale of small and medium-sized businesses to form and implement their own strategies to ensure the competitiveness of the company and the demand for products in modern turbulent times, which in turn will contribute to a positive impact on the development of the regional economy of our country

Development of the cell-ELISA test for the subtype identification of circulating influenza A(H1) and A(H3) viruses

The sensitive version of cell-ELISA was developed for the subtype-specific differentiation of current influenza A(H1N1)pdm09 and A(H3N2) viruses that are circulating in the human population. This method is based on the estimation of virus reproduction in infected MDCK cells. The detection step of this method is an interaction of the subtype-specific monoclonal antibodies (mAbs) with the viral hemagglutinin (НА) molecule. The influenza A virus strains, isolated in the 2014 epidemic season, were used to validate this method.It was shown that when using mAbs # 1/ # 2 or # 4 at a concentration of 10-15 µg/ml, the developed variant of cell-ELISA was able to detect НА protein synthesized in the infected cells of influenza A(H3N2) and A(H1N1)pdm09 viruses, respectively.The developed method can be used for the identification of modern influenza A viruses with low hemagglutination activity, which is not possible by the conventional hemagglutination inhibition test.The sensitive version of cell-ELISA was developed for the subtype-specific differentiation of current influenza A(H1N1)pdm09 and A(H3N2) viruses that are circulating in the human population. This method is based on the estimation of virus reproduction in infected MDCK cells. The detection step of this method is an interaction of the subtype-specific monoclonal antibodies (mAbs) with the viral hemagglutinin (НА) molecule. The influenza A virus strains, isolated in the 2014 epidemic season, were used to validate this method. It was shown that when using mAbs # 1/ # 2 or # 4 at a concentration of 10-15 µg/ml, the developed variant of cell-ELISA was able to detect НА protein synthesized in the infected cells of influenza A(H3N2) and A(H1N1)pdm09 viruses, respectively. The developed method can be used for the identification of modern influenza A viruses with low hemagglutination activity, which is not possible by the conventional hemagglutination inhibition test

Influence of single amino acid substitutions in the hemagglutinin on the antigenic and receptor-binding properties of influenza virus B/Florida/04/2006 of Yamagata-like evolutionary lineage

Influenza A and B viruses use sialylated oligosaccharide chains expressed on the surface of a host cell as the cell entry receptors. The type of the bond between sialic acid (SA) and the neighboring galactose residue (Gal) is one of the main characteristics that define the type of receptor. Influenza viruses recognize SAα2-3Gal- or SAα2-6Gal-structures on the surface of the cells. Influenza A viruses of avian origin bind α2-3-sialylated glycans, while the human strains bind preferentially α2-6-sialylated ones. However, the receptor-binding specificity of influenza B viruses has not been characterized sufficiently so far. In this study, we selected the escape mutants of influenza B/Florida/04/2006 strain (Yamagata-like lineage) using monoclonal antibodies (mAb) to hemagglutinin (HA). The analysis of the amino acid sequences of mAb-induced escape mutants revealed the single amino acid substitutions 40Tyr→His, 85His→Tyr, 202Asn→Lys and 242Ser→Arg in 10F4-, 8Н11-, 8Н3- and 9А3-induced HA variants, correspondingly. It was shown that the single amino acid substitutions 202Asn→Lys and 242Ser→Arg alter the receptor-binding specificity of the influenza B virus. These findings are important for the understanding of the influence of individual amino acid residues in HA on the receptor-binding properties of influenza B Yamagata-like lineage viruses and allow us to predict the possible ways of their evolution.Influenza A and B viruses use sialylated oligosaccharide chains expressed on the surface of a host cell as the cell entry receptors. The type of the bond between sialic acid (SA) and the neighboring galactose residue (Gal) is one of the main characteristics that define the type of receptor. Influenza viruses recognize SAα2-3Gal- or SAα2-6Gal-structures on the surface of the cells. Influenza A viruses of avian origin bind α2-3-sialylated glycans, while the human strains bind preferentially α2-6-sialylated ones. However, the receptor-binding specificity of influenza B viruses has not been characterized sufficiently so far. In this study, we selected the escape mutants of influenza B/Florida/04/2006 strain (Yamagata-like lineage) using monoclonal antibodies (mAb) to hemagglutinin (HA). The analysis of the amino acid sequences of mAb-induced escape mutants revealed the single amino acid substitutions 40Tyr→His, 85His→Tyr, 202Asn→Lys and 242Ser→Arg in 10F4-, 8Н11-, 8Н3- and 9А3-induced HA variants, correspondingly. It was shown that the single amino acid substitutions 202Asn→Lys and 242Ser→Arg alter the receptor-binding specificity of the influenza B virus. These findings are important for the understanding of the influence of individual amino acid residues in HA on the receptor-binding properties of influenza B Yamagata-like lineage viruses and allow us to predict the possible ways of their evolution

Spectral Studies of Rat Bone Tissue in Modeling Osteoporosis and Effectiveness of Treatment By Hydroxyapatite

Presents the result of experiments on the study of the model of osteoporosis in rats using Raman spectroscopy and the effectiveness of its treatment with hydroxyapatite. Were revealed spectral differences between groups of samples (control group, group with the model of osteoporosis and a group with the model of osteoporosis after treatment with hydroxyapatite). In addition, optical coefficients were introduced to evaluate the effectiveness of treatment. Keywords: Raman spectroscopy, optical coefficients, osteoporosis, hydroxyapatite, collagen matri

Retrospective trial of long acting analogues detemir and degludec usage in children and adolescents to overcome glucose variability caused by dawn phenomenon and reverse dawn phenomenon

Backgraund: Children with type 1 diabetes mellitus (T1DM) need more insulin late in the evening (reverse dawn phenomenon (RDP)), and adolescents need more insulin yearly in the morning (dawn phenomenon (DP)); these cause blood glucose variability. Modern long acting insulin analogues allow to achieve satisfactory glycemic control.Aims: To study the characteristics of insulin therapy in children and adolescents with T1DM using insulin analogues detemir and degludec to overcome blood glucose variability caused by DP and RDP in different age periods.Materials and methods: We analyzed medical documents of 200 patients using detemir, admitted to pediatric endocrinology department in 2013–2019, at mean age 9.0 years (5.4; 13.0), with T1DM for 1.3 years (0.5; 3.0); and medical documents of 50 patients switched to degludec in 2018–2019 at mean age 12.0 years (10.5; 14.5) with T1DM for 3.0 years (1.5; 6.0). Before degludec they were on intensive insulin therapy with glargine (22), detemir (26), or insulin pump (2); 16 patients (32%) presented with clinical characteristics of DP, and 5 (10%) — RDP.Results: 67 children of 108 (62%) aged 1–9 years had redistribution of detemir doses to daytime; 58 adolescents of 92 (63%) aged 10–17 лет — to nighttime. Patients switched to degludec demonstrated decrease in HbA1с from 8.7% (7.8; 9.9) to 8.0% (7.4; 9.0) (р<0.001); fasting blood glucose from 9.8 mmol/l (7.4; 11.7) to 7.7 mmol/l (6.4; 8.6) (р<0.001); within-day variability from 35.2% (31.6; 40.9) to 23.5% (19.7; 28.6) (р<0.001); daily insulin dose from 0.98 U/kg/day (0.82; 1.14) to 0.87 U/kg/day (0.75; 1.07) (р=0.002). Sub-groups of patients with DP and RDP demonstrated decrease in fasting blood glucose (from 11.5 mmol/l (9.8; 13.8) to 7.5 mmol/l (6.6; 9.1) (р<0.001)), and late evening blood glucose (from 11.0 mmol/l (10.2; 11.2) to 8.0 mmol/l (6.7; 9.5) (р= 0.03)) correspondently. Achieved levels of glycemic control did not differ between sub-groups of patients initially using glargine or detemir.Conclusions: Compensation of T1DM may be complicated due to DP and RDP. Switching to degludec allowed to achieve better glycemic control and lowering of blood glucose variability caused by DP and DRP

Changes in University Teacher’s Pedagogical Activity in the Context of Digitalization of Education

The purpose of this article is to consider the changes in university teacher’s pedagogical activity in the conditions of a rapid transformation of higher education and propose a theoretical and empirical substantiation of the importance of teacher’s understanding of the value-semantic attitude to pedagogical activity, which is not limited only to mastering digital competencies, but consists in mastering pedagogical knowledge that allows you to critically analyze the productivity of modern educational strategies and methods.The conducted research was based on the methodology of post-non-classical science, which implies the widespread use of qualitative research methods that reflect the convergence of explanatory and interpretative research approaches, the use of representations, opinions, attitudes, and values of the research participants as full data.The article describes the results of studying the university teachers’ attitude to changes, the value orientations of the ongoing changes, as well as the main difficulties faced by university teachers in the context of digitalization of education – the design of educational and training programs, modern educational and methodological complex, the organization of productive self-directed work and communication with students in the educational process.The main directions of overcoming the identified difficulties are given – the construction of flexible curricula within the framework of the Federal State Educational Standard of Higher Education, the development of the digital educational environment of the university and updating the content of the professional development programs for teaching staff. The conclusion is made about the increasing importance of pedagogical knowledge in teacher’s professional activity in the conditions of a modern university

УСЛОВИЯ ЭФФЕКТИВНОГО ПОДАВЛЕНИЯ ПЦР С ПОМОЩЬЮ LNA-ОЛИГОНУКЛЕОТИДОВ ДЛЯ ПРОСТОЙ И ВЫСОКОЧУВСТВИТЕЛЬНОЙ ДЕТЕКЦИИ СОМАТИЧЕСКИХ МУТАЦИЙ

PCR clamping/wild-type blocking PCR with non-extendable locked nucleic acid (LNA) oligonucleotides is used for sensitive detection of somatic mutations in tumors. Various  versions of the technique use different DNA polymerases and LNA oligonucleotides with and  without additional phosphorothioate modifications. Here we studied requirements for successful  PCR clamping with LNA oligonucleotides and Taq DNA polymerase for analysis of mutations in  KRAS and BRAF genes by means of real-time PCR and Sanger sequencing. We found that  addition of phosphorothioate linkages at the 5’-end of LNA oligonucleotide to protect from 5’- exonuclease activity of Taq DNA polymerase did not improve clamping. For most target  sequences, efficient clamping was observed at melting temperature of LNA oligonucleotide  20‑25°C above annealing/extension temperature of the PCR with a 2-step protocol. Under such  conditions, simple and sensitive detection of mutations in KRAS and BRAF genes was feasible using real-time PCR with TaqMan probes or Sanger sequencing.Специфическое блокирование амплификации аллеля дикого типа в ПЦР с помощью олигонуклеотидов, модифицированных по остатку рибозы (закрытые нуклеиновые кислоты, locked nucleic acids, LNA), используется для высокочувствительной детекции соматических мутаций в опухолях. Описаны различные версии метода анализа мутаций с использованием LNA-олигонуклеотидов как с дополнительной модификацией фосфотиоатными группами, так и без таких групп, при этом использовались  различные ДНК полимеразы. В работе проведен анализ оптимальных условий для успешного специфического блокирования ПЦР с помощью LNA-олигонуклеотидов при анализе мутаций в генах KRAS и  BRAF. Мы обнаружили, что фосфотиоатная защита на 5’-конце олигонуклеотидов не влияет на эффективность блокирования аллеля дикого типа. Выявлено, что для большинства последовательностей  эффективное блокирование наблюдается при проведении шага отжига и элонгации ПЦР при температуре на  20–25°С ниже температуры плавления LNA-олигонуклеотида. При таких условиях реакции возможна простая и высокочувствительная детекция мутаций в генах KRAS и BRAF с использованием как секвенирования по Сэнгеру, так и ПЦР в реальном времени с Taqman зондами

ВИРУСНАЯ НАГРУЗКА ПРИ ХРОНИЧЕСКОМ ГЕПАТИТЕ В: КОРРЕЛЯЦИИ С ЛАБОРАТОРНО-МОРФОЛОГИЧЕСКИМИ ПОКАЗАТЕЛЯМИ

In our study, 155 patients with chronic hepatitis B viral load was analyzed and its relationship with some of the results of laboratory and instrumental examinations. Found that half of the patients had a viral load of 4-5 log copies/ ml and above. In HBe-negative patients, the viral load is lower than in patients with HBeAg. The level of viremia had a direct correlation with the activity of ALT and AST, and eedback to the level of blood platelets. In patients with a viral load of more than 5 log copies / ml, significantly higher ALT, AST and GGT, as well as the level of red blood cells, whereas platelet counts were significantly lower than in patients with viremia <5 log copies/ml . Platelet counts inversely correlated with age. In 11% of patients with chronic hepatitis B over 40 years, had platelet counts below normal. The level of viral load and ALT and AST is straight, and the level of blood platelets inverse correlation with the severity of liver fibrosis. In patients without evidence of liver fibrosis (F0) viral load was significantly lower than in patients with signs of fibrosis of the liver at any stage of its formation.В нашем исследовании у 155 больных хроническим гепатитом В анализировалась вирусная нагрузка и ее взаимосвязь с некоторыми результатами лабораторных и инструментальных обследований. Установлено, что половина больных имели вирусную нагрузку 4–5 log копий/мл и выше. У HBe-негативных пациентов вирусная нагрузка ниже, чем у больных с HBeAg. Уровень виремии имел прямую корреляционную связь с активностью АлАТ и АсАТ и обратную связь с уровнем тромбоцитов крови. У пациентов с вирусной нагрузкой 5 и более log копий/мл статистически значимо выше активность АлАТ, АсАТ и ГГТП, а также уровень эритроцитов в крови, в то время как уровень тромбоцитов статистически значимо ниже, чем у пациентов с виремией < 5 log копий/мл. Уровень тромбоцитов имеет обратную корреляционную связь с возрастом пациентов. У 11% больных ХГВ старше 40 лет отмечается уровень тромбоцитов ниже нормы. Уровень вирусной нагрузки, а также активность АлАТ и АсАТ имеет прямую, а уровень тромбоцитов крови – обратную корреляционную связь со степенью выраженности фиброза печени. У больных без признаков фиброза печени (F0) вирусная нагрузка статистически значимо ниже, чем у пациентов с признаками фиброза печени на любой из стадий его формирования

АНАЛИЗ МУТАЦИЙ В ГЕНАХ KRAS И BRAF ПРИ РАКЕ ТОЛСТОЙ И ПРЯМОЙ КИШКИ В РОССИЙСКОЙ ПОПУЛЯЦИИ

Mutations in KRAS and BRAF genes in 80 colorectal cancer (CRC) samples from Russian patients were tested using two methods: 1) allele-specific real-time PCR (as-rt PCR) and 2) wild-type blocking PCR with Sanger sequencing (WTBS). Material and methods. Sections of fresh frozen or formalin-fixed paraffin embedded tumor tissue from 80 patients were used in the study. Tumor tissue content was determined on H&E stained sections. Samples were first tested by as-rtPCR for common mutations of the KRAS gene (G12C, G12S, G12R, G12V, G12D, G12A, G13D) and mutations BRAFV600E. After that samples were evaluated in PCR with oligonucleotide blocking amplification of wild-type DNA for enrichment with mutant allele followed by Sanger sequencing of the PCR DNA (WTBS method). Results. In 5 (6.3 %) cases samples had low tumor tissue content (<20 %). In reconstruction experiments both methods detected 1–5 % mutant allele. Mutations of KRAS and BRAF genes were found in 37 (46 %) and 3 (3.8 %) of the clinical cases, respectively. Classification in to wild-type and mutant samples by both methods was in agreement in 79 (98.8 %) cases. A single case with rare mutation KRASG13R was detected by WTBS, but was missed by as-rtPCR since this mutation is not included in the test. Of note, KRAS mutations were detected by both tests in two cases with low tumor content. Two cases were found with multiple mutations: one with KRASG12V and G13D, and one with KRASG13D and BRAFV600E. Conclusion. The frequency of mutations in CRC was 46 % for mutations in the KRAS gene, and 3.8 % for the BRAF. We showed 98.8% agreement in KRAS mutation detection by sensitive Sanger sequencing and as-rt PCR. The data on the frequencies of mutations are in agreement with studies in other countries. This in the first study to discover CRC case with multiple mutations KRASG13D and BRAFV600E.Образцы опухоли толстой и прямой кишки (РТПК) от российских пациентов на наличие мутаций вгенах KRAS и BRAF исследовали с помощью двух высокочувствительных методов анализа: аллельспецифической ПЦР в режиме реального времени (ас-рв ПЦР) и секвенирования по Сэнгеру с блокированием аллеля дикого типа. Материал и методы. В качестве материала для исследования использовали срезы свежезамороженной и фиксированной в формалине и заключенной в парафин ткани опухоли от 80 пациентов. На гистологическом исследовании было определено содержание опухолевых клеток в каждом образце. Образцы были протестированы методом ас-рв ПЦР на мутации в гене KRAS (G12C, G12S, G12R, G12V, G12D, G12A, G13D) и мутации BRAFV600E. Затем была проведена амплификация ДНК образцов в присутствии олигонуклеотида, блокирующего амплификацию аллеля KRAS дикого типа с дальнейшем секвенированием ДНК по Сэнгеру. Результаты. По данным гистологического заключения из 80 образцов низкое содержание опухолевых клеток (<20 %) было обнаружено в 5 (6 %) случаях. В модельных экспериментах оба метода анализа позволяли детектировать 5 % аллеля с мутацией. Мутации в генах KRAS и BRAF были обнаружены в 37 (46 %) и 3 (3,8 %) случаях соответственно. Совпадение результатов анализа с помощью обоих методов произошло в 79 (98,8 %) случаях. Расхождение результатов анализа для одного образца связано с тем, что секвенирование ДНК по Сэнгеру обнаружило мутацию KRASG13R, тогда как ас-рв ПЦР не включала методику детекции этой мутации. Кроме того, мутация в гене KRAS была обнаружена в 2 образцах с низким содержанием опухолевых клеток. Обнаружены 2 случая с несколькими мутациями: 1 образец с мутациями в гене KRASG12V и G13D, а также 1 образец с мутациями KRASG13D и BRAFV600E. Заключение. Частота мутаций при РТПК для мутаций в гене KRAS составила 46 %, для BRAF – 3,8 %. Показана сходимость результатов анализа с помощью ас-рв ПЦР и секвенирования по Сэнгеру в 98,8 %. Полученные данные о частотах мутаций согласуются с исследованиями в других странах. В исследовании впервые обнаружен образец РТПК, одновременно содержащий мутации KRASG13D и BRAFV600E