ANIMAL WELFARE: EUROPEAN STANDARDS AS A BASIS FOR REFORMING NATIONAL LEGISLATION IN UKRAINE

ABSTRACT: Nowadays, Ukraine faces numerous challenges in animal welfare, including ensuring animal welfare, keeping and veterinary care for stray animals, protecting them from cruelty, and preserving endangered species. These areas are important and create the need to improve national legislation and its application. The problem is also becoming more urgent within the framework of Ukraine’s European integration vector, accompanied by the harmonization of national legislation with EU law and the ratification and implementation of European standards in the relevant area. The subject matter of the study necessitated the use of dialectical, systemic, structural-functional and informational approaches. The article aims to study the legal regulation of animal rights protection in Ukraine and analyze the animal treatment standards in foreign countries and international organizations. Keywords: animals; animal protection; international standards; animal cruelty; animal rights.   Bem-estar animal: Padrões europeus como base para reformar a legislação nacional na Ucrânia   RESUMO: Atualmente, a Ucrânia enfrenta inúmeros desafios na área de bem-estar animal, incluindo as garantias e manutenção dos cuidados veterinários de animais perdidos, a proteção da crueldade e preservação de espécies ameaçadas de extinção. Estas áreas são importantes e criam a necessidade de melhorar a legislação nacional e a sua aplicação. O problema também se torna mais urgente no quadro do vetor de integração europeia da Ucrânia, acompanhado pela harmonização da legislação nacional com a legislação da União Européia (EU), e consequentemente, pela ratificação e implementação das normas europeias dessa área relevante, no território nacional. O objeto do estudo exigiu o uso de abordagens dialéticas, sistêmicas, estrutural-funcionais e informacionais. O artigo visa estudar a regulamentação legal da proteção dos direitos dos animais na Ucrânia e analisar os padrões de tratamento animal em países estrangeiros e organizações internacionais da UE. Palavras-chave: animais; proteção animal; padrões internacionais; crueldade animal; direito dos animais.ABSTRACT: Nowadays, Ukraine faces numerous challenges in animal welfare, including ensuring animal welfare, keeping and veterinary care for stray animals, protecting them from cruelty, and preserving endangered species. These areas are important and create the need to improve national legislation and its application. The problem is also becoming more urgent within the framework of Ukraine’s European integration vector, accompanied by the harmonization of national legislation with EU law and the ratification and implementation of European standards in the relevant area. The subject matter of the study necessitated the use of dialectical, systemic, structural-functional and informational approaches. The article aims to study the legal regulation of animal rights protection in Ukraine and analyze the animal treatment standards in foreign countries and international organizations. Keywords: animals; animal protection; international standards; animal cruelty; animal rights.   Bem-estar animal: Padrões europeus como base para reformar a legislação nacional na Ucrânia   RESUMO: Atualmente, a Ucrânia enfrenta inúmeros desafios na área de bem-estar animal, incluindo as garantias e manutenção dos cuidados veterinários de animais perdidos, a proteção da crueldade e preservação de espécies ameaçadas de extinção. Estas áreas são importantes e criam a necessidade de melhorar a legislação nacional e a sua aplicação. O problema também se torna mais urgente no quadro do vetor de integração europeia da Ucrânia, acompanhado pela harmonização da legislação nacional com a legislação da União Européia (EU), e consequentemente, pela ratificação e implementação das normas europeias dessa área relevante, no território nacional. O objeto do estudo exigiu o uso de abordagens dialéticas, sistêmicas, estrutural-funcionais e informacionais. O artigo visa estudar a regulamentação legal da proteção dos direitos dos animais na Ucrânia e analisar os padrões de tratamento animal em países estrangeiros e organizações internacionais da UE. Palavras-chave: animais; proteção animal; padrões internacionais; crueldade animal; direito dos animais

Impact of stromal cell components of tumor microenvironment on epithelial-mesenchymal transition in breast cancer cells

Background: Cell and tissue homeostasis results from the dynamic balance of cell – cell and cell – extracellular component crosstalk that regulates proliferation, differentiation, and apoptosis of cells as well as secretion and activation of soluble factors and/or deposition of extracellular matrix (ECM) components. Aim: The aim of the work was to study the crosstalk between tumor cells and stromal cell components using noncontact co-cultivation in vitro system. Materials and Methods: Human and rat breast cancer (BC) cell lines, normal human fibroblasts (NHF) and endothelial cells, and aspirates of bone marrow (BM) of BC patients with different clinical course of the disease (groups “Remission” (BM-R) and “Progression” (BM-P)) were used in noncontact co-cultivation system in vitro. The cell growth, expression of epithelial-mesenchymal transition (EMT) and tumor stem cell markers (E-cadherin, vimentin, CD44), Ki-67, p21 and Slug were investigated using immunocytochemical analysis. Results: Analysis of expression of E- and N-cadherin, vimentin and Slug in BC cells has shown that T-47D and MRS-T5 cells possess mesenchymal phenotype, while MCF-7 and MRS cells possess mostly epithelial phenotype with a part of cells with mesenchymal patterns. Upon noncontact co-cultivation of fibroblasts with Т-47D or MRS-Т5 cells, BC cells acquired higher proliferative activity compared to the control cells (р < 0.05) or MCF-7 and MRS cells co-cultivated with fibroblasts. Upon noncontact co-cultivation of Т-47D cells with normal fibroblasts and BM cells from BC patients from group “Progression” there were observed increased quantity of CD44+ Т-47D cells (by 26%), decreased quantity of Е-cadherin+ Т-47D cells, and appearance of vimentin-positive cells. In co-cultivation variant Т-47D + NHF + BM-R (“Remission“) the quantity of CD44+ Т-47D cells significantly decreased (р < 0.005) and E-cadherin expression remained unaltered compared to control cells. At the same time, in NHF cell population (co-cultivation variant Т-47D + NHF + BM-P) there was detected significant increase of quantity of р21+-cells (р < 0.005), cytoplasmic localization of p21, and nuclear localization of Slug. Expression of vimentin did not alter in any variant of co-cultivation. Conclusion: The new integration cell system for investigation of the mechanisms of interaction between tumor cells and the tumor microenvironment in vitro was developed. The significant changes in proliferative activity of TC dependently on its ­ЕМT-status were detected after their interaction with fibroblasts and endothelial cells in noncontact co-cultivation system. BM cells of BC patients had different modifying influence on TC dependent on clinical BC course. The activation of ЕМT program was revealed in TC upon noncontact co-cultivation with BM cells of BC patients with progression of the disease. Key Words: breast cancer, epithelial-mesenchymal transition, microenvironment, bone marrow, co-cultivation

Effect of antitumor drugs in low concentrations on the biological, immunophenotypic and cytogenetic characteristics of human colon cancer cells in vitro

Objective: To estimate the impact of the low-dose anticancer drugs (ACD) with the different mechanisms of action and human interferon (IFN) alpha 2b on the biological properties, immunophenotypic and cytogenetic characteristics of colon cancer cells in vitro. Materials and Methods: The study was performed on human colon cancer cell lines COLO 205, HT-29 and 3C-P treated with ACD and IFN in subtoxic concentrations. Expression of CD44, N-cadherin, vimentin, β-catenin, ERCC1 and Slug was assessed by immunocytochemical method. Using cytogenetic analysis, the numbers of mitoses, cells with micronuclei, apoptotic cells and cells with nuclear protrusions were studied. Results: The prolonged exposure (up to 30 days) of colon cancer cells to low-dose ACD (0.2–0.5 µg/ml cisplatin and 0.1–0.2 µg/ml irinotecan) in combination with IFN (500–1000 IU/ml) led to 37-fold decreased colony-forming activity of these cell and 10-fold reduction of the number of cells expressing mesenchymal protein markers (N-cadherin, vimentin). Also, in COLO 205 cells treated with ACD and IFN the number of SLUG- and CD44-positive cells decreased by 92 and by 85%, respectively. Long-term cultivation of HT-29 cells in the presence of cisplatin and IFN resulted in 5-fold suppression of ERCC1 expression. The cytogenetic analysis has shown that the ACD, IFN and their combinations in subtoxic concentrations caused significant genotoxic effect, suppression of cell proliferation and accumulation of cells with micronuclei. The sensitivity of colon cancer cells to ACD in standard cytotoxic concentrations did not change after prolonged low-dose exposure. Conclusion: The data showed that the prolonged action of the low doses of ACD on human colon cancer cells resulted in the suppression of cell proliferation, colony-forming activity in soft agar, expression of epithelialmesenchymal transition-associated markers and significant cytogenetic changes

Inhibition of malignant potential and expression of proteins associated with epithelial-mesenchymal transition in Lewis lung carcinoma cells transduced with murine ifn-β gene in recombinant baculovirus

Aim: To analyze biological characteristics. malignant potential and expression of proteins associated with epithelial mesenchynal transition in murine lung carclnoma cells transduced with intertcron-beta (ifn-β) gene in baculovirus vector. Materials and Methods: The study was perforated on Lewis lung carcinoma (LL) cells transduced with ifn-β gene in recombinant baculovirus vector. Biological characterlstics of the LL cells were studied with the use of standard cell culture method, cytogenetic and immunocytochemical assays. Resutes: Recombinant baculmirus-mediated transduction of LL cells with ifn-β gene resulted in siguficant decrease of cell growth rate and density both in complete and serum-foee medium. Also. LL cells transduction with ifn-β gene significantly inhfltited cell migration in vitro. Transduction of LL cells by baculovirus vector with or withoot ifn-β gene caused significant genotoxic effect in these cells. Furthermore. ifn-β gene transfer to lung carcinoma cells resulted in significant increase of nuclear expression of p19ARE (p < 0.01). p21WAFI (p < 0.001). cytoplasmic expression of E-cadherin (p < 0.005) and inhibition of transcription factors of epithelial-mesenchymal transition (EMT) Tmist (p < 0.005) and Slug (p < 0.00 I) expression. Conclusions; Transduction with ifn-β gene of LL cells in recombinant baculovirus resulted in aquirement of less malignant phenotype in vitro and suppressed expression ofproteins associated with EMT

Underestimated importance and status of the specialist at the stage of pre-trial investigation

Злочинність є соціальною закономірністю, наслідком взаємовідносин суб’єктів соціуму, зі своїми передумовами і причинами, то і дослідженням цього феномену, його природи, суті да змісту, як правило, займаються соціальні та гуманітарні науки. Разом із тим, прояв кожного окремого злочину, тобто його зовнішня сторона в загальному розумінні є предметом саме природничих наук, які в подальшому виокремлювались в окремі відносно незалежні, а іноді і взагалі незалежні криміналістичні науки.he current Criminal Procedure Code of Ukraine is among certain participants in criminal proceedings who have or are obliged to have and possess scientific, technical and special knowledge and skills in the use of technical or other means of a specialist and expert. Unfortunately, the CPC of Ukraine does not have special norms, enshrined general and consolidated provisions that would regulate the activities of both the specialist and the expert as participants in the pre-trial investigation. We have only fragmentary references to their procedural rights and responsibilities. The only clearly defined emphasis is the emphasis on the various goals of the involved specialist and expert, as well as on the procedural significance of the results obtained during the implementation of these special knowledge by these specialists. The thesis on the definition of the obligatory participation of a specialist in the conduct of investigative actions, during which special knowledge and skills of using technical or other means are used, is discussed

High levels of proinflammatory cytokines tumor necrosis factor alpha, interleukin 1β and interleukin 6 in bone marrow and peripheral blood of breast cancer patients as predictors of relapse

Breast cancer provides a typical example of an inflammation-linked malignant disease. The inflammatory components present in the tumor microenvironment contribute to the further progression of the disease. The present study was conducted to evaluate the correlation between levels of proinflammation cytokines tumor necrosis factor α, interleukin 1β, interleukin 6, C-reactive protein and tumor recurrence in breast cancer patients. Seventy two breast cancer patients with histologically proven diagnosis and 15 healthy donors were enrolled into study. Thirty one patients with progression of the disease and 41 patients with clinical stabilization (conditional remission) were included to “progression” and “remission” groups respectively. This division of breast cancer patients was made during the 3 years study. The levels of proinflammation cytokines – tumor necrosis factor α, interleukin 1β, interleukin 6 and C-reactive protein were revealed. This cytokines in bone marrow and peripheral blood act as a specific microenvironment and was found that the elevated levels of this cytokines were strongly associated with progression of breast cancer. The data showed that most prominent predictive markers of tumor recurrence are high levels of indicated cytokines in combination with other markers (C-reactive protein; disseminated tumor cells in bone marrow). Determination of the high levels of tumor necrosis factor, interleukin 1β, interleukin 6 in bone marrow and peripheral blood of breast cancer patients are important to establish the features of bone marrow microenvironment for prediction of metastasis and correction antitumor therapy

Broad-spectrum Anti-tumor and Anti-metastatic DNA Vaccine Based on p62- encoding Vector

Abstract Autophagy plays an important role in neoplastic transformation of cells and in resistance of cancer cells to radio and chemotherapy. p62 (SQSTM1) is a key component of autophagic machinery which is also involved in signal transduction. Although recent empirical observations demonstrated that p62 is overexpressed in variety of human tumors, a mechanism of p62 overexpression is not known. Here we report that the transformation of normal human mammary epithelial cells with diverse oncogenes (RAS, PIK3CA and Her2) causes marked accumulation of p62. Based on this result, we hypothesized that p62 may be a feasible candidate to be an anti- cancer DNA vaccine. Here we performed a preclinical study of a novel DNA vaccine encoding p62. Intramuscularly administered p62-encoding plasmid induced anti-p62 antibodies and exhibited strong antitumor activity in three models of allogeneic mouse tumors – B16 melanoma, Lewis lung carcinoma (LLC), and S37 sarcoma. P62-encoding plasmid has demonstrated its potency both as a preventive and therapeutic vaccine. Importantly, p62 vaccination drastically suppressed metastasis formation: in B16 melanoma where tumor cells where injected intravenously, and in LLC and S37 sarcoma with spontaneous metastasis. Overall, we conclude that a p62-encoding vector(s) constitute(s) a novel, effective broad-spectrum antitumor and anti- metastatic vaccine feasible for further development and clinical trials