The Change of Protein Intradomain Mobility on Ligand Binding: Is It a Commonly Observed Phenomenon?

AbstractAnalysis of changes in the dynamics of protein domains on ligand binding is important in several aspects: for the understanding of the hierarchical nature of protein folding and dynamics at equilibrium; for analysis of signal transduction mechanisms triggered by ligand binding, including allostery; for drug design; and for construction of biosensors reporting on the presence of target ligand in studied media. In this work we use the recently developed HCCP computational technique for the analysis of stabilities of dynamic domains in proteins, their intrinsic motions and of their changes on ligand binding. The work is based on comparative studies of 157 ligand binding proteins, for which several crystal structures (in ligand-free and ligand-bound forms) are available. We demonstrate that the domains of the proteins presented in the Protein DataBank are far more robust than it was thought before: in the majority of the studied proteins (152 out of 157), the ligand binding does not lead to significant change of domain stability. The exceptions from this rule are only four bacterial periplasmic transport proteins and calmodulin. Thus, as a rule, the pattern of correlated motions in dynamic domains, which determines their stability, is insensitive to ligand binding. This rule may be the general feature for a vast majority of proteins

Mediatization as a Factor in Increasing Interest in Modern Popular Science Media (“Science and Life” and “Popular Mechanics”)

The article is devoted to the study of multimedia tools that are used when placing content in online versions of popular science publications. The relevance of the issue is due to the need to use new convergent forms of promotion of printed publications in the field of mass communications. The novelty of the study is seen in the fact that for the first time the process of multimediaization in the popular science segment of the media is described. The role that multimedia tools play in the development of modern popular science media is shown. Particular attention is paid to topical problems of the functioning of popular science journals, possible ways to solve them, as well as, in this regard, the potential of multimedia. The content analysis and comparative research of electronic and printed versions of popular science magazines “Science and Life” and “Popular Mechanics” are presented. The question is raised about how multimedia expands the opportunities for brand promotion. On the basis of the results obtained, conclusions were formulated about the more effective use of multimedia in popular science media. The definitions of multimedia are given and the classification of the functions of multimedia elements in the publications of Internet media is proposed. It has been proven that modern popular science media does not use the entire range of multimedia elements in Internet versions, which negatively affects their popularization

IMPROVEMENT OF PHARMACEUTICAL SPECIALISTS DURING THE PERIOD OF REFORMING MEDICINE AND PHARMACY IN UKRAINE

The article describes the need to determine the main directions and priorities of the development of the pharmaceutical industry in Ukraine, which is conditioned by real political and socioeconomic processes. The main element here is the introduction and development of a formular system - a complex of management methods for the application of rational, organizational and cost-effective methods of supplying and using drugs to ensure, in specific conditions, high quality medical care and optimal use of available resources.Key words: pharmaceutical industry, pharmacist, pharmacist, higher education

Antimicrobial and therapeutic effect of probiotics in cases of experimental purulent wounds

Probiotics based on bacteria of the genus Bacillus with a multifactorial mechanism of action are considered as a possible alternative to antibiotics in the treatment of purulent wounds. The aim of the study was to determine the antimicrobial and therapeutic effect of the Arederma probiotic preparation containing probiotic strains of the genus Bacillus in an experimental model of a purulent wound in animals. The antimicrobial efficacy of the probiotic against test strains and clinical isolates of pathogenic and opportunistic microorganisms was studied using the method of delayed antagonism. Staphylococcus aureus ATCC 6538 and Streptococcus pyogenes K-7 were used to model a purulent wound. From the surface of the wounds, bacteria of Staphylococcus, Streptococcus, Enterobacteriaceae, Pseudomonas genera and Enterobacteriaceae family were sown on appropriate selective media for the cultivation and enumeration of different groups of microorganisms by generally accepted microbiological research methods. The formation of a purulent wound in rabbits caused by mechanical skin damage and subsequent double infection with Staphylococcus aureus ATCC 6538 and Streptococcus pyogenes K-7 strains was accompanied with a pronounced inflammatory process, necrosis, the formation of purulent exudate and general intoxication. Representatives of the genera Staphylococcus and Streptococcus, microscopic fungi and, to a lesser extent, members of the family Enterobacteriaceae and Pseudomonas were found on the surface of purulent wounds, which confirmed the development of the infectious-inflammatory process. Treatment of purulent wounds with a suspension of probiotic preparation once a day for 4 days led to their faster healing (gradual attenuation of the inflammatory process, reduction of edema and discharge, as well as their disappearance) compared with untreated purulent wounds (control). Representatives of the Staphylococcus and Streptococcus genera, as well as microscopic fungi, presented in purulent wounds treated with probiotic preparation in much smaller numbers than in the control, and bacteria of the Pseudomonas genus and the Enterobacteriaceae family were not detected at all. The effective antimicrobial effect of this probiotic preparation against opportunistic and pathogenic microorganisms was confirmed by in vitro studies. Therefore, the Arederma probiotic preparation showed an effective therapeutic and antimicrobial effect in the experimental model of a purulent wound in animals, so it can be recommended for further preclinical and clinical studies

Доведення регіоселективності перебігу циклізації 1-етил-3-[4-6,7,8,9-тетрагідро-5Н-[1,2,4]триазоло[4,3-а]азепін-3-іл)феніл]тіосечовини з α-бромкетоном

An important step in creation of potential drugs is to confirm the structure of the compounds synthesized. This requires the use of modern physical and physico-chemical methods of research. Nowadays a promising scientific direction for searching biologically active substances is the study of 2-R-imino-1,3-thiazoline derivatives.Aim. To study regioselectivity of the cyclization reaction of 1-ethyl-3-[4-(6,7,8,9-tetrahydro-5Н-[1,2,4]triazolo[4,3-а]azepin-3-yl)phenyl]thiourea with 2-bromo-1-phenylethanone. Results. The true structure of the interaction product was determined by the methods of 1H NMR spectroscopy and X-ray analysis. Quantum chemical calculations of the electronic structure, geometry and thermodynamic parameters of the initial thiourea three tautomers were given. Activating energy of tautomer 1A belower than 1B one, the state of 1B has modest lower relative energy, consequently tautomer 1A is more credible state. Thus, the conclusion can be made that the reaction will proceed by 1-1A-3A.Experimental part. Quantum chemical calculations of the electronic structure, geometry and thermodynamic parameters of the initial thiourea three tautomers were determined by the density functional theory (DFT) methods using the GAUSSIAN W09 computer program. The effect of the solvent was considered within the framework of polarized continuum model (PCM). Conclusions. Based on the physico-chemical studies and quantum chemical calculations of the reaction cyclization direction the conclusion has been made that the cyclization reaction of 1-ethyl-3-[4-(6,7,8,9-tetrahydro-5H-[1,2,4]triazolo[4,3-а]azepin-3-yl)phenyl]thiourea 1 with 2-bromo-1-phenylethanone 2 is regioselective, and it leads to formation of more thermodynamically advantageous (stable) isomer 3A. Важным этапом создания потенциальных лекарственных средств является подтверждение структуры синтезированных соединений, что требует использования современных физических и физико-химических методов исследования. На сегодня перспективным научным направлением в плане поиска биологически активных веществ является исследование в ряду производных 2-R-имино-1,3-тиазолина.Целью работы является изучение региоселективности реакции циклизации 1-этил-3-[4-(6,7,8,9-тетрагидро-5Н-[1,2,4]триазоло[4,3-а]азепин-3-ил)фенил]тиомочевины с 2-бром-1-фенилэтаноном.Результаты и их обсуждение. Установление истинного строения продукта взаимодействия осуществлено с помощью методов 1Н ЯМР-спектроскопии и рентгеноструктурного анализа. Приведены квантово-химические расчеты электронной структуры, геометрии и термодинамических параметров трех таутомеров исходной тиомочевины. Энергия активации таутомера 1А несколько ниже, состояние 1В имеет незначительно меньшую относительную энергию, следовательно, таутомер 1А является более вероятным. Таким образом, мы можем сделать вывод, что реакция пойдет по пути 1-1А-3А.Экспериментальная часть. Квантово-химические расчеты электронной структуры, геометрии и термодинамических параметров трех таутомеров исходной тиомочевины вычислены методами теории функционала плотности (DFT) с использованием компьютерной программы GAUSSIAN 09W. Учет влияния растворителя осуществлялся в рамках модели поляризуемого континуума (PCM).Выводы. На основе проведенных физико-химических исследований и квантово-химических расчетов направления прохождения реакции конденсации 1-этил-3-[4-6,7,8,9-тетрагидро-5Н-[1,2,4]триазоло[4,3-а]азепин-3-ил)фенил]тиомочевины с 2-бромо-1-фенилэтаноном сделан вывод о ее региоселективности с образованием более термодинамически выгодного изомера 3А.Важливим етапом створення потенційних лікарських засобів є підтвердження структури синтезованих сполук, що потребує використання сучасних фізичних та фізико-хімічних методів дослідження. На сьогодні перспективним науковим напрямком при пошуку біологічно активних речовин є дослідження в ряду похідних 2-R-іміно-1,3-тіазоліну.Метою роботи є вивчення регіоселективності реакції циклізації 1-етил-3-[4-(6,7,8,9-тетрагідро-5Н-[1,2,4]триазоло[4,3-а]азепін-3-іл)феніл]тіосечовини з 2-бромо-1-фенілетаноном. Результати та їх обговорення. Встановлення істинної будови продукту взаємодії здійснено за допомогою методів 1Н ЯМР-спектроскопії та рентгеноструктурного аналізу. Наведені квантово-хімічні розрахунки електронної структури, геометрії і термодинамічних параметрів трьох таутомерів вихідної тіосечовини. Енергія активації таутомеру 1А дещо нижча, стан 1В має незначно нижчу відносну енергію, отже, таутомер 1А є більш ймовірним. Таким чином, ми можемо зробити висновок, що реакція перебігатиме шляхом 1-1А-3А.Експериментальна частина. Квантово-хімічні розрахунки електронної структури, геометрії і термодинамічних параметрів трьох таутомерів вихідної тіосечовини обчислені методами теорії функціоналу густини (DFT) з використанням комп’ютерної програми GAUSSIAN 09W. Облік впливу розчинника здійснювався в рамках моделі континууму, що поляризується (PCM). Висновки. На основі проведених фізико-хімічних досліджень і квантово-хімічних розрахунків напрямку проходження реакції конденсації 1-етил-3-[4-6,7,8,9-тетрагідро-5Н-[1,2,4]триазоло[4,3-а]азепін-3-іл)феніл]тіосечовини з 2-бромо-1-фенілетаноном зроблений висновок про її регіоселективність з утворенням більш термодинамічно вигідного ізомеру 3А.

Characterization of Coupled Ground State and Excited State Equilibria by Fluorescence Spectral Deconvolution

Fluorescence probes with multiparametric response based on the relative variation in the intensities of several emission bands are of great general utility. An accurate interpretation of the system requires the determination of the number, positions and intensities of the spectral components. We have developed a new algorithm for spectral deconvolution that is applicable to fluorescence probes exhibiting a two-state ground-state equilibrium and a two-state excited-state reaction. Three distinct fluorescence emission bands are resolved, with a distribution of intensities that is excitation-wavelength-dependent. The deconvolution of the spectrum into individual components is based on their representation as asymmetric Siano-Metzler log-normal functions. The application of the algorithm to the solvation response of a 3-hydroxychromone (3HC) derivative that exhibits an H-bonding-dependent excited-state intramolecular proton transfer (ESIPT) reaction allowed the separation of the spectral signatures characteristic of polarity and hydrogen bonding. This example demonstrates the ability of the method to characterize two potentially uncorrelated parameters characterizing dye environment and interactions

The cardio- and endothelial protective effects of ethyl methyl hydroxyl pyridine malate in modeling L-name induced nitric oxide deficiency

Currently, endothelial dysfunction is considered as a predictor of a number of pathologies, including arterial hypertension, ischemic heart disease, chronic heart failure, and is also a pathogenetic component of organ damage in diabetes, hypo estrogenic and other conditions. The purpose of the study is an experimental study of the cardio and vasoprotective effects of etoxidol under conditions of endothelial dysfunctio

Blood platelets in chronic graft-versus-host disease: association with Th1/Th2 ratio

As found in clinical and laboratory studies, platelets not only play a key role in the processes of coagulation and thrombosis, but are also able to actively participate in other pathophysiological processes, including the development of immune reactions. It has been shown that changes in the immune system leading to systemic lupus erythematosus (SLE) are often accompanied by changes in the number of platelets and their activity in the peripheral blood of SLE patients, which correlate with the severity of the clinical manifestations of the disease. Earlier we have studied the standard experimental model of SLE in detail, based on the induction of chronic graft-versus-host disease (cGVHD) in the semi-allogeneic system DBA/2 → (C57Bl/6 x DBA/2)F1. However, the participation of platelets in this immunopathological process has not been studied. There are no data in the literature on the behavior of platelets in cGVHD or on their relationship with the state of Th1/Th2 balance. It can been expected that the platelet count changes according to the development of cGVHD in the used experimental model by analogy with the development of SLE in humans.In the experiments, we used female mice of the DBA/2 strain and (C57Bl/6 × DBA/2)F1 hybrids. Chronic GVHD in a semi-allogeneic system was induced by injecting DBA/2 mouse splenocytes into B6D2F1 hybrid mice: 60-70 × 106 cells intravenously twice with an interval of 6 days. The studied parameters were evaluated three months after the start of the experiment and the formation of lupus-like glomerulonephritis in animals with Th2-dependent cGVHD variant.A decrease in the number of erythrocytes and hemoglobin, a decrease in hematocrit and a parallel increase in the number of reticulocytes in the blood of mice with cGVHD are in good agreement with our earlier conclusion that these animals have autoimmune hemolytic anemia. It was found that, platelets increase significantly with the development of cGVHD unlike other blood cells. Secondary thrombocytosis is observed in the case of the Th2-dependent variant of сGVHD in this model of SLE, while in the group with the Th1-dependent variant of сGVHD, the average number of platelets in the blood does not differ from the control group

Extracellular DNA in blood: an index of <i>in vivo</i> inflammatory response

Increased concentration of cell-free DNA (cfDNA) in the circulating blood of humans and animals is a sign of inflammatory conditions and a distinctive characteristic of various pathophysiological processes in the body. The aim of the present study was to investigate the possible role of tumor necrosis factor (TNFα) in changes of cfDNA contents in peripheral blood as a response to experimentally induced systemic inflammation.We used 40 female hybrid mice (C57Bl/6xDBA/2) F1 at the age of 6-8 weeks. The concentration of cfDNA and its individual fractions was determined using a PicoGreen fluorescent dye. The dynamics of inflammatory process was evaluated after 4, 8, 11 and 24 hours following LPS injection. A significant increase in the blood plasma cfDNA levels was shown under the action of E. coli lipopolysaccharide (LPS), along with simultaneous decreased levels of cfDNA, associated with cell surface. The ratio of cell surface-bound cfDNA to the total cfDNA contents was reduced in dose-dependent manner as early as 4 hours after LPS injection to the animals, thus allowing us to consider this ratio a characteristic sign of netosis of neutrophilic granulocytes during the development of acute inflammation. The described effects are significantly suppressed with co-injection of recombinant TNFα neutralizing protein along with LPS, whereas increased intake of neutrophils in the tissues is determined by some other factors which are not directly related to the production of this cytokine.Based on the obtained data, we proposed a following hypothesis: induction of netosis by inflammatory stimuli causes an increase in the concentration of cfDNA in blood plasma not only due to de novo emerging extracellular DNA by neutrophil netosis, but also by the release of distinct cfDNA fraction that was previously firmly bound to cell membranes in multiple body tissues under the action of proteases released during netosis

Full Multiple Scattering Analysis of XANES at the Cd L 3- and O K- Edges in CdO Films Combined with a Soft-X-Ray Emission Investigation

X-ray absorption near edge structure (XANES) at the cadmium L3 and oxygen K edges for CdO thin films grown by pulsed laser deposition method, is interpreted within the real-space multiple scattering formalism, FEFF code. The features in the experimental spectra are well reproduced by calculations for a cluster of about six and ten coordination shells around the absorber for L3 edge of Cd and K edge of O, respectively. The calculated projected electronic density of states is found to be in good agreement with unoccupied electronic states in experimental data and allows to conclude that the orbital character of the lowest energy of the conductive band is Cd 5s-O 2p[sigma]*. The charge transfer has been quantified and not purely ionic bonding has been found. Combined XANES and resonant inelastic x-ray scattering measurements allow us to determine the direct and indirect band gap of investigated CdO films to be ~2.4 eV and ~0.9 eV, respectively