Expression of Tumor Assosiated and Epithelial-mesenchymal Transition Markers in 2d and 3d Cell Cultures of Mcf-7

The target effects on the expression of epithelial-mesenchymal transition regulation molecules are promising for cancer therapy, including breast cancer. 3D cell culture is a model for studying epithelial-mesenchymal transition in vitro and may become a test system for anticancer therapy.Aim of research. The aim of this research was to evaluate and compare the expression of tumor associated and epithelial-mesenchymal transition markers in tumor cells of breast adenocarcinoma (MCF-7 cell line) in 2D and 3D cell culture.Methods. For realization of the aim MCF-7 cell line (breast adenocarcinoma) was chosen as an experimental model in vitro. The monolayer cell culture was cultured in standard conditions (37 0C, 5 % CO2, humidity 95 %). The initial density of inoculated cells was 2 x 104 cells/cm2. The cells were incubated for two days before their use in the experiment. For the initial generation of spheroids the monolayer cell culture was removed off the substrate after the four days of incubation, using 0,25 % Trypsin-EDTA, and placed in nutrient medium with 5 % carboxymethyl cellulose (Bio-Rad, USA) at concentration of 5 x 105 cells/ml. Then the plates were incubated on an orbital shaker (Orbital shaker, PSU-10i, Biosan, Latvia) at 50 rpm for 3–5 hours. Half of culture medium was replenished every 3 days. A spheroid culture was maintained for 14 days. Detection of markers (ER, p53, EpCAM, vim, AE1/AE3, panCK, EGFR) in 2D and 3D cell culture was performed using immunohistochemistry method with primary monoclonal antibodies. Histological samples of cells were photographed to compare the morphological characteristics and the expression of proteins in monolayer and spheroid cultureResults. The results demonstrated that the percentage of tumor marker positive cells (ER+, EGFR+, EpCAM+, panCK+, AE1/AE3+) in monolayer culture is 1.25–2 times than more in spheroid culture. In contrast, tumor spheroids consist of fewer cells with the expression of epithelial markers such as EpCAM and AE1/AE3, but they contain a large number of cells that expressed mesenchymal marker vimentin by 5 % and p53 by 10 %. This may indicate that the cells acquire a mesenchymal phenotype. However, tumor cells of monolayer cell culture were not expressed vimentin.Conclusions. Our results demonstrated the differences of expression of tumor associated and epithelial-mesenchymal transition markers in 2D and 3D breast cancer cell cultures. Thus, the percentage of epithelial markers (Cytokeratines and epithelial cell adhesion molecule) in tumor spheroids is less than in cells of monolayer however spheroids cells begin expressing a mesenchymal marker – vimentin. In 3D cell culture only the outer cell layers expressed tumor associated proteins unlike 2D cell culture in which all of cells showed equally expression. Reduced of manifestation of tumor associated markers in 3D cell culture may indicate an increase of stem properties. These data showed that 3D cell culture more than 2D cell culture characterized processes of epithelial-mesenchymal transition

EXPRESSION OF TUMOR ASSOSIATED AND EPITHELIAL-MESENCHYMAL TRANSITION MARKERS IN 2D AND 3D CELL CULTURES OF MCF-7

The target effects on the expression of epithelial-mesenchymal transition regulation molecules are promising for cancer therapy, including breast cancer. 3D cell culture is a model for studying epithelial-mesenchymal transition in vitro and may become a test system for anticancer therapy. Aim of research. The aim of this research was to evaluate and compare the expression of tumor associated and epithelial-mesenchymal transition markers in tumor cells of breast adenocarcinoma (MCF-7 cell line) in 2D and 3D cell culture. Methods. For realization of the aim MCF-7 cell line (breast adenocarcinoma) was chosen as an experimental model in vitro. The monolayer cell culture was cultured in standard conditions (37 0C, 5 % CO2, humidity 95 %). The initial density of inoculated cells was 2 x 104 cells/cm2. The cells were incubated for two days before their use in the experiment. For the initial generation of spheroids the monolayer cell culture was removed off the substrate after the four days of incubation, using 0,25 % Trypsin-EDTA, and placed in nutrient medium with 5 % carboxymethyl cellulose (Bio-Rad, USA) at concentration of 5 x 105 cells/ml. Then the plates were incubated on an orbital shaker (Orbital shaker, PSU-10i, Biosan, Latvia) at 50 rpm for 3–5 hours. Half of culture medium was replenished every 3 days. A spheroid culture was maintained for 14 days. Detection of markers (ER, p53, EpCAM, vim, AE1/AE3, panCK, EGFR) in 2D and 3D cell culture was performed using immunohistochemistry method with primary monoclonal antibodies. Histological samples of cells were photographed to compare the morphological characteristics and the expression of proteins in monolayer and spheroid culture Results. The results demonstrated that the percentage of tumor marker positive cells (ER+, EGFR+, EpCAM+, panCK+, AE1/AE3+) in monolayer culture is 1.25–2 times than more in spheroid culture. In contrast, tumor spheroids consist of fewer cells with the expression of epithelial markers such as EpCAM and AE1/AE3, but they contain a large number of cells that expressed mesenchymal marker vimentin by 5 % and p53 by 10 %. This may indicate that the cells acquire a mesenchymal phenotype. However, tumor cells of monolayer cell culture were not expressed vimentin. Conclusions. Our results demonstrated the differences of expression of tumor associated and epithelial-mesenchymal transition markers in 2D and 3D breast cancer cell cultures. Thus, the percentage of epithelial markers (Cytokeratines and epithelial cell adhesion molecule) in tumor spheroids is less than in cells of monolayer however spheroids cells begin expressing a mesenchymal marker – vimentin. In 3D cell culture only the outer cell layers expressed tumor associated proteins unlike 2D cell culture in which all of cells showed equally expression. Reduced of manifestation of tumor associated markers in 3D cell culture may indicate an increase of stem properties. These data showed that 3D cell culture more than 2D cell culture characterized processes of epithelial-mesenchymal transition

EXPRESSION OF TUMOR ASSOSIATED AND EPITHELIAL-MESENCHYMAL TRANSITION MARKERS IN 2D AND 3D CELL CULTURES OF MCF-7

The target effects on the expression of epithelial-mesenchymal transition regulation molecules are promising for cancer therapy, including breast cancer. 3D cell culture is a model for studying epithelial-mesenchymal transition in vitro and may become a test system for anticancer therapy. Aim of research. The aim of this research was to evaluate and compare the expression of tumor associated and epithelial-mesenchymal transition markers in tumor cells of breast adenocarcinoma (MCF-7 cell line) in 2D and 3D cell culture. Methods. For realization of the aim MCF-7 cell line (breast adenocarcinoma) was chosen as an experimental model in vitro. The monolayer cell culture was cultured in standard conditions (37 0C, 5 % CO2, humidity 95 %). The initial density of inoculated cells was 2 x 104 cells/cm2. The cells were incubated for two days before their use in the experiment. For the initial generation of spheroids the monolayer cell culture was removed off the substrate after the four days of incubation, using 0,25 % Trypsin-EDTA, and placed in nutrient medium with 5 % carboxymethyl cellulose (Bio-Rad, USA) at concentration of 5 x 105 cells/ml. Then the plates were incubated on an orbital shaker (Orbital shaker, PSU-10i, Biosan, Latvia) at 50 rpm for 3–5 hours. Half of culture medium was replenished every 3 days. A spheroid culture was maintained for 14 days. Detection of markers (ER, p53, EpCAM, vim, AE1/AE3, panCK, EGFR) in 2D and 3D cell culture was performed using immunohistochemistry method with primary monoclonal antibodies. Histological samples of cells were photographed to compare the morphological characteristics and the expression of proteins in monolayer and spheroid culture Results. The results demonstrated that the percentage of tumor marker positive cells (ER+, EGFR+, EpCAM+, panCK+, AE1/AE3+) in monolayer culture is 1.25–2 times than more in spheroid culture. In contrast, tumor spheroids consist of fewer cells with the expression of epithelial markers such as EpCAM and AE1/AE3, but they contain a large number of cells that expressed mesenchymal marker vimentin by 5 % and p53 by 10 %. This may indicate that the cells acquire a mesenchymal phenotype. However, tumor cells of monolayer cell culture were not expressed vimentin. Conclusions. Our results demonstrated the differences of expression of tumor associated and epithelial-mesenchymal transition markers in 2D and 3D breast cancer cell cultures. Thus, the percentage of epithelial markers (Cytokeratines and epithelial cell adhesion molecule) in tumor spheroids is less than in cells of monolayer however spheroids cells begin expressing a mesenchymal marker – vimentin. In 3D cell culture only the outer cell layers expressed tumor associated proteins unlike 2D cell culture in which all of cells showed equally expression. Reduced of manifestation of tumor associated markers in 3D cell culture may indicate an increase of stem properties. These data showed that 3D cell culture more than 2D cell culture characterized processes of epithelial-mesenchymal transition

Особливості комплексного маркетингову підходу в менеджменті туристичної індустрії

Introduction. The tourist industry is a priority in the strategic development of a region. The main problems of research in the field of marketing tourism services are the essence and role of marketing in the development of the tourism industry, including the process of marketing management, types of strategies and marketing plans. The purpose of marketing tourism services is to attract as many customers as possible to the business entities, while providing them with the highest quality of service and guaranteeing their full satisfaction as part of the basic and expanded product. Aim and tasks. The purpose of this article is to study specifics and the complex nature of marketing in the tourism industry. Results. In the modern period of functioning of the economy, the service industry is developing very dynamically. In connection with this, the specificity of marketing used in travel services is also changing. The article analyzes the models of state regulation of the tourism industry. World practice separates the four models of state regulation of tourism industry. Eligible for the development of the tourist industry of Ukraine is the third "European" and fourth "Mixed" model, based on solid cooperation between the state and private business, i.e. the presence of central executive authority, recognition tourism is a priority area for the country. It is proved that the main positive point in introducing these models is that the State acts as the coordinator for the development of the tourism industry. The essence and features of the concept of "tourist demand" and "tourist offer" are also analyzed. Established in the tourist marketing system of continuous coordination of services in the marketing process with those that are in demand in the market, and which are planned to be introduced to the market with greater profits than competitors do. Conclusions. After analyzing the current concepts of marketing in tourism, we came to the conclusion that it is necessary to consider them through the concept of marketing interaction, that is, when using social and communication techniques to enhance marketing activities in tourism. Having reviewed the model of state regulation of tourism development, it was noted that it is European model of state participation in the development of the tourism industry is acceptable for Ukraine. It would be wise to develop a model tourism management in Ukraine and its regions that will meet the peculiarities of country and will make it possible to work effectively at this stage of development of tourism in Ukraine. With regard to public relations tools as a general form of promotion of travel services, they are quite popular among tourists. However, the state needs to pay special attention to the development of recreational areas, and other areas of tourism in Ukraine.Проблема. Туристична індустрія є пріоритетною у питанні стратегічного розвитку регіону. Основними проблемами досліджень у сфері послуг маркетингового туризму є сутність і роль маркетингу в розвитку туристичної галузі, в тому числі, процес управління маркетингом, види стратегій та маркетингові плани. Метою маркетингових послуг туризму є залучення суб'єктами господарювання як можна більше клієнтів, в той час надаючи їм найвищу якість обслуговування та гарантуючи повне задоволення їх потреб, як частини базового та розширеного продукту. Мета і завдання. Метою статті є дослідження специфіки та комплексного характеру маркетингу в туристичній галузі. Результати. У сучасний період функціонування економіки індустрія послуг розвивається дуже динамічно. У зв’язку з цим змінюється також специфіка маркетингу, що використовується в туристичних сервісах. Проведено аналіз моделей державного регулювання розвитку туристичної галузі. Світова практика відокремлює чотири моделі державного регулювання розвитку туристичної галузі. Прийнятними для розвитку туристичної галузі України є моделі, які ґрунтуються на щільній взаємодії держави і приватного бізнесу, тобто наявності централізованого органу виконавчої влади, визнання туризму пріоритетною сферою для країни із координацією з боку держави у розвитку індустрії туризму. Проаналізовано сутність та особливості поняття «туристичного попиту» та «туристичної пропозиції». Визначено в туристичному маркетингу систему безперервного узгодження послуг в процесі маркетингової діяльності з тими, які користуються попитом на ринку, і які планується виводити на ринок з більшим прибутком, ніж це роблять конкуренти. Висновки. Проаналізувавши сучасні поняття маркетингу в туризмі ми дійшли до висновку, що необхідно розглядати їх через концепцію маркетингу взаємодії, тобто під час використання соціальних та комунікаційних прийомів для підвищення маркетингової діяльності в туризмі. Розглянувши існуючі моделі державного регулювання розвитку туристичної діяльності, відзначено, що саме Європейська модель участі держави у розвитку індустрії туризму є прийнятною для України. Доцільно розробити модель управління туризмом в Україні та її регіонах, яка буде відповідати особливостям держави та дасть змогу ефективно працювати на даному етапі розвитку галузі туризму в Україні. Що стосується інструментів взаємодії із громадськістю як загальної форми просування туристичних послуг, вони є досить популярними серед мешканців туристичних напрямків. Проте державі потрібно звернути особливу увагу на розвиток рекреаційних зон, та інших напрямків туризму на Україні

Prospects for the Tax Reform «10-10-10» in Ukraine

In the wake of the war in Ukraine and the rapid spread of the COVID-19 pandemic, many businesses have either suspended or closed their operations. The introduction of martial law requires rapid adaptation of tax legislation to new realities. To do this, the State must, in particular, distribute its burden to promote the survival of businesses in the wartime conditions. One of such plans is to carry out the tax reform «10-10-10», which provides for amendments to tax legislation, revision of preferential taxation of certain business segments and reduction of basic tax rates. Making adjustments to these tax rates will lead to serious changes in the State budget itself. The main taxes that will be changed during the reform are VAT, personal income tax and business income tax. Analysts argue that reducing them to 10% and strengthening tax control over entrepreneurs and enterprises will lead to the withdrawal of a significant part of business from the «shadow». The important point is that, in contrast to reducing the rates on basic taxes, the authors of the reform propose to strengthen control over the activities of not only registered entrepreneurs, but also the population that receives income without registering as a business entity. This will entail an increase in fines for violating tax legislation. Among the important shortcomings of the reform are the potential increase in the budget deficit resulting from a significant reduction in taxes, as well as the conditions of the European Directive 112, which Ukraine has to adhere to within the framework of European integration and according to which the VAT rate cannot be less than 15%. In turn, the advantages are strengthening tax control, de-shadowing the economy and reducing the tax burden on the wage fund, improving the standard of living of the population, supporting business and attracting foreign investment. Thus, after a detailed consideration of the reform, familiarization with the opinions of leading experts and a brief analysis, it can be argued that in such a difficult situation in which Ukraine finds itself now, it is appropriate to make fundamental changes to the tax system in order to improve the financial condition of the population, enterprises and the economy in general

Orally Administered Activated Charcoal as a Medical Countermeasure for Acute Radiation Syndrome in Rats

Activated charcoal (AC) can be taken orally as enterosorbent for treatment of pathological states related to exogenous and endogenous intoxications. Synthesized granulated AC with a highly developed active surface (SBET ~2700 m2/g) was used as a medical countermeasure (MCM) to acute radiation sickness (ARS) in rats after total body X-ray irradiation. AC demonstrates positive results in ARS treatment, as expressed in, (i) a decrease in body weight loss, (ii) a protection of bone marrow (BM) cells colony formation capacity, (iii) a reduction of BM chromosomal aberrations and small intestine and spleen tissue damage, (iv) an amelioration of white blood cell count, and (v) a mitigation of superoxide ion generation rate in the liver. AC oral prescription seems to be perspective modality of ARS treatment.This research was funded by H2020-MSCA-RISE-2016 project, grant number 734641 acronym NanoMed

On hereditary reducibility of 2-monomial matrices over commutative rings

A 2-monomial matrix over a commutative ring $R$ is by definition any matrix of the form $M(t,k,n)=\Phi\left(\begin{smallmatrix}I_k&0\\0&tI_{n-k}\end{smallmatrix}\right)$, $0<k<n$, where $t$ is a non-invertible element of $R$, $\Phi$ the compa\-nion matrix to $\lambda^n-1$ and $I_k$ the identity $k\times k$-matrix. In this paper we introduce the notion of hereditary reducibility (for these matrices) and indicate one general condition of the introduced reducibility

Methodological instructions for laboratory work No.5(1) «The rotational motion of a solid body»

У посібнику викладено методику проведення лабораторної роботи, присвяченої вивченню фізичних законів, що описують обертальний рух твердого тіла. Пропонований метод заснований на експериментальному дослідженні руху системи твердих тіл, закріпленої на маятнику Обербека. Публікація дає короткий опис теоретичних даних, які необхідні для опису явища, яке вивчається в лабораторній роботі. Наведено теоретичні розрахунки, необхідні для інтерпретації відповідних експериментальних даних і підтвердження відомих теоретичних закономірностей. Також представлена ​​покрокова інструкція з проведення експериментальних вимірювань і відповідних розрахунків фізичних величин. Призначено для студентів вищих навчальних закладів з технічним профілем.The tutorial outlines a methodology for carrying out laboratory work devoted to the study of physical laws that describe the rotational motion of a solid. The proposed method is based on an experimental study of the motion of a system of solids fixed on the Oberbeck pendulum. The publication gives a brief description of the theoretical data, which are needed to describe the phenomenon that is being studied in laboratory work. The theoretical calculations that are necessary for the interpretation of the corresponding experimental data and confirmation of the known theoretical regularities are given. Also the step-by-step instruction for conducting experimental measurements, and corresponding calculations of physical quantities is presented. For the students of higher educational institutions with technical profile.В пособии изложена методика проведения лабораторной работы, посвященной изучению физических законов, описывающих вращательное движение твердого тела. Предлагаемый метод основан на экспериментальном исследовании движения системы твердых тел, закрепленной на маятнике Обербека. Публикация дает краткое описание теоретических данных, которые необходимы для описания явления, которое изучается в лабораторной работе. Приведены теоретические расчеты, необходимые для интерпретации соответствующих экспериментальных данных и подтверждения известных теоретических закономерностей. Также представлена пошаговая инструкция по проведению экспериментальных измерений и соответствующих расчетов физических величин. Предназначено для студентов высших учебных заведений с техническим профилем

Methodological instructions for laboratory work No.7(1) «Investigation of the rotational motion of a solid body and determination of the velocity of the bullet with the help of a torsion ballistic pendulum»

В посібнику викладена методика проведення лабораторної роботи, що присвячена вивченню фізичних закономірностей, які описують обертальний рух твердого тіла. Запропонована методика базується на експериментальному дослідженні руху твердого тіла за допомогою крутильного балістичного маятника. В публікації наведено короткий виклад теоретичних даних, які необхідні для опису явища, яке вивчається в лабораторній роботі. Представлені теоретичні розрахунки необхідні для інтерпретації відповідних експериментальних даних та підтвердження відомих теоретичних закономірностей. Також представлена покрокова інструкція проведення експериментальних вимірювань, та відповідних розрахунків фізичних величин.The tutorial outlines a methodology for carrying out laboratory work devoted to the study of physical laws that describe the rotational motion of a solid. The proposed method is based on an experimental study of the motion of a solid body with the help of a torsion ballistic pendulum. The publication gives a brief description of the theoretical data, which are needed to describe the phenomenon that is being studied in laboratory work. The theoretical calculations that are necessary for the interpretation of the corresponding experimental data and confirmation of the known theoretical laws are given. Also the step-by-step instruction for conducting experimental measurements, and corresponding calculations of physical quantities is presented. For the students of higher educational institutions with technical profile

Specifics of the Surface Structure of Steel Products After Various Laser Alloying Methodics

In modern conditions, volumetric alloying of steels is becoming an increasingly less economically viable process. However, the level of performance properties of unalloyed steels cannot cover the current industry needs. One of the ways to solve this problem – usage of various surface alloying methods of steels, makes it possible to obtain the required properties of the working surface of the alloyed part with minimal consumption of expensive alloying elements. Over the course of the study, combined method of laser-plasma alloying of steel surfaces was analyzed, its technological capabilities were determined, and the structural-phase state of surface layers, formed during laser and hybrid laser-plasma processing was compared. During comparative studies of samples obtained by both methods of surface alloying, it was found that in the case of laser-plasma alloying, the observed structure and carbide phases are smaller in size, with a low density and uniform distribution of dislocations in the metal of the alloyed layer. After various analysis, it was established that during both laser and laser-plasma methods of surface alloying, the crack formation tendencies was mostly attributed to various structural and concentration changes associated with the redistribution of elements, leading to the formation of sharp grain-boundary concentration gradients. An increase in the number of cracks is observed in regimes with higher heating temperatures, increased duration of exposure to high temperatures and reduced cooling rates