Preparation of polyclonal and monoclonal anti-idiotypic antibodies against morphine-specific immunoglobulins

The preparation and study of anti-idiotypic (secondary) antibodies (Ab2) against monoclonal primary antibodies (Ab1) specific to biologically active molecules with a known structure is of great scientific and practical importance. Due to partial antigenic similarity of Ab2 and the initial antigen structures, these antibodies can be the basis of the vaccine, if the antigen usage is not possible, or is limited by law. In particular, one may create Ab2-based preparations, designed for immunization, in order to prevent and treat the drug addiction. The value of Ab2 properties increases even more if Ab1, used to obtain them, recognize different parts of the antigen molecule, which makes it possible to obtain second-generation antibodies with a wide range of specificity. In this work, the morphine-like polyclonal and monoclonal Ab2 were obtained. In each case, as the first-generation immunoglobulins for immunization, we used two murine monoclonal antibodies (mAbs) specific to different morphine derivatives: 3K11 antibodies to 3-0-carboxymethyl (CMM) and 2-p-carboxyphenylazomethyl (FAM) derivatives, as well as 6G1 antibodies to 6-hemisuccinyl derivative (GSM). After immunization of the horse with Ab1 and development of immune response, three pools of specific polyclonal antibodies were isolated from the animal blood serum: horse anti-species antibodies to the total mouse immunoglobulins (HAM); horse anti-idiotypic antibodies against 3K11 antibodies (HAM-K11), and against 6G1 antibodies (HAM-G1). In parallel, immunization of mice with 3K11 and 6G1 antibodies and fusion of obtained lymphocytes with Sp2/0 mouse myeloma cells by the Milstein-Köhler method resulted in three producers of anti-idiotypic antibodies: a clone producing mouse monoclonal Ab2 specific for mAb-6G1 (AIG1), as well as clones producing anti-mAb-3K11 antibodies (AI-K11A and AI-K11B). The physico-chemical and antigenic properties of all the Ab2 obtained were characterized. It was shown that the horse anti-idiotypic immunoglobulins not only belong to different classes, but are also polyvalent, while all monoclonal Ab2 obtained are represented by IgM immunoglobulins, being also strictly specific to the corresponding first-generation antibodies. Subsequently, the morphine-like properties of the first domestic polyclonal and monoclonal Ab2 obtained in the work will be investigated in a cellular model. Likewise, we shall study their ability to induce Ab3 as well as morphine-specific Ab1

Mitochondrial physiology

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of data repositories of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery

New prognostistic markers of nodular forms of goiter combined with autoimmune thyroiditis

Sheremet М. І., Sydorchuk L. P., Shidlovskyi V. О., Bedenyuk A. D., Pashkovska N. V., Leonova M. O., Chorna O. O., Stankova N. I., Rybak O. V. New prognostistic markers of nodular forms of goiter combined with autoimmune thyroiditis. Journal of Education, Health and Sport. 2017;7(3):475-482. eISSN 2391-8306. DOI http://dx.doi.org/10.5281/zenodo.399322 http://ojs.ukw.edu.pl/index.php/johs/article/view/4368 The journal has had 7 points in Ministry of Science and Higher Education parametric evaluation. Part B item 1223 (26.01.2017). 1223 Journal of Education, Health and Sport eISSN 2391-8306 7 © The Author (s) 2017; This article is published with open access at Licensee Open Journal Systems of Kazimierz Wielki University in Bydgoszcz, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 21.03.2017. Revised 22.03.2017. Accepted: 23.03.2017. NEW PROGNOSTIC MARKERS OF NODULAR FORMS OF GOITER COMBINED WITH AUTOIMMUNE THYROIDITIS М. І. Sheremet, L. P. Sydorchuk, V. О. Shidlovskyi*, A. D. Bedenyuk*, N. V. Pashkovska, M. O. Leonova, O. O. Chorna, N. I. Stankova, O. V. Rybak Department of surgery, Bukovinian State Medical University * Department of surgery, I.Y. Horbachevskyi Ternopil State Medical University Teatralna Sq. 2, Chernivtsi, 58002, Ukraine Phone / fax (+38 0372) 55-37-54, e-mail: [email protected] Abstract Background: Based on the results of a histological study of removed TG tissue NGAIT was found in 10,4% of the patients [1, 2]. At the same time, we observe processes of both thyroid epithelium metaplasia and lymphoid tissue hyperplasia that, undoubtedly, can be considered as an optional precancerous condition [3-8]. The option when cytomorphological and immunocytochemical examinations are carried out sequentially, using the same smear of a puncture material is optimal for PCE [9]. Methods: We have carried out an immunohistochemical study by means of monoclonal antibodies against Fas, FasL, Bcl-2, P53 and Ki67 antigens using the thyroid gland puncture material. Results: The results showed the degree of proliferative activity in the thyroid tissue in NGAIT. We found a high proliferative activity of lymphoid tissue, moderate proliferative activity of thyroid epithelial cells in the area of lymphoid infiltration and a low one – beyond the latter. Conclusions: The pronounced expression of Fas and FasL on the thyroid epithelial cells in the area of lymphoid infiltration indirectly indicates that NGAIT causes the processes of thyroid epithelial cells apoptosis due to the immunity. Increasing the number of immunoreactive cells expressing Ki67 in the area of lymphoid infiltration and destruction of thyroid epithelial cells, are indicative of follicular epithelial regeneration as a compensatory-adaptive response of the organ. A pronounced Bsl-2 expression in lymphocytes prevents the cells from entering apoptosis and prolongs the cell survival time. There was a high expression of p53 protein in the nuclei of thyroid epithelial cells and follicular lamina, which can be explained by mutations in the gene p53, which allows the cells to find tolerance to apoptotic action of the immune system effectors. Abbreviations: NGAIT - nodular goiter combined with autoimmune thyroiditis, TG – thyroid gland, PCE – preoperative cytological examination. Key words: nodular goiter combined with autoimmune thyroiditis, fine needle biopsy, apoptosis, proliferation, thyroid gland