Recombinant short TNF-BD protein from smallpox virus is pharmacologically active in an experimental septic shock model

Tumor necrosis factor (TNF) is one among the key cytokines that mediate the immune system to protect humans against viral infections. Throughout evolution, anthropogenic Variola virus (VARV) has developed effective mechanisms to overcome human defense reactions. The viral genome encodes soluble proteins imitating the structure of cellular cytokine receptors. These proteins compete with cellular receptors for cytokine binding, thus blocking the antiviral immune response. In particular, the G2R gene of VARV encodes the TNF decoy receptor, VARV-CrmB protein. This protein consists of N-ended TNF-biding (TNF-BD) and C-ended chemokine binding (Ch-BD) domains. Recombinant VARV-CrmB protein has been produced in insect cells using molecular cloning methods and its TNF neutralizing activity has been shown in vitro and in vivo. To decrease the immunogenicity of this protein, a recombinant plasmid coding for shortened TNF-BD protein of VARV in Escherichia coli cells has been constructed. Using the method of immobilized metal affinity chromatography, recombinant TNF-BD protein corresponding to the TNF-biding domain of VARV-CrmB protein was purified from E. coli cells. The therapeutic potential of TNF-BD was studied using an experimental model of LPS-induced septic shock. After septic shock induction, several doses of recombinant TNF-BD were injected and the mortality of experimental animals was observed during 7 days. All mice not injected with TNF-BD had been dead by day 3 of the experiment, but 30, 40 and 60 % of the experimental animals, who received different TNF-BD doses, survived in a dose-dependent manner. Data obtained demonstrate that recombinant TNF-BD protein is pharmacologically active in the experimental model of LPS-induced septic shock

The results of studying of the dust factor in copper ore concentration

The results of investigation of dispersion, chemical composition and concentrations of dust in the working area at various stages of copper ore concentration are presented. It’s shown that the overwhelming number of dust particles have sizes 2,1-5,0 mm, which determines the sustained presence of the dust in the working area, as well as the duration of its stay in the deepest parts of the respiratory system. Crystalline silicon dioxide and copper predominate in the dust. Concentration of crystalline silicon dioxide in the occupation air of crushing compartment exceeds MAC in 1,2 times and reagent one – 1,3 times. The presence of crystalline silicon dioxide as well as arsenic in the dust determines its carcinogenic danger.Приведены результаты изучения дисперсности, химического состава и концентраций пыли в воздухе рабочей зоны на различных этапах обогащения медьсодержащего сырья. Показано, что преобладающее число пылинок имеют размеры 2,1–5,0 мкм, что определяет устойчивый характер присутствия данной пыли в воздухе рабочей зоны и длительное ее нахождение в глубоких отделах органов дыхания. В составе пыли преобладают кремний диоксид кристаллический и медь. Концентрации кремния диоксида кристаллического в воздухе рабочей зоны дробильного отделения превышают ПДК в 1,2 раза, а реагентного – в 1,3 раза. Присутствие в пыли кремния диоксида кристаллического и мышьяка определяет ее канцерогенную опасность

ЛУЧЕВАЯ ДИАГНОСТИКА БЦЖОСТИТА ГРУДИНЫ У РЕБЕНКА

BCG osteitis is an uncommon complication of the antitubercular vaccination. We review some information about the epidemiology, clinical presentation and diagnostic methods  of BCG or tuberculous osteitis of sternum. We report a case of BCG osteitis, that was detected with the multispiral computer tomography. We discurse the diagnostic sings of this post-vaccination complication.  The diagnosis was verified by laboratory studies,  which allows to confirm the etiology of the process.БЦЖ-остит — редко встречающееся  осложнение после вакцинации против туберкулеза. В работе приведена краткая  информация  об эпидемиологии, клинической картине и методах диагностики остеомиелита  грудины туберкулезной или БЦЖ-этиологии. Представлено  собственное клиническое наблюдение пациента с оститом грудины БЦЖ-этиологии. Для  его идентификации была  применена  спиральная  компьютерная  томография. Описана  лучевая  семиотика  данного  поствакцинального  осложнения.  Окончательный  диагноз  установлен на основании лабораторных методов исследования, которые позволили подтвердить этиологию процесса

The Adaptive Marine Policy (AMP) toolbox: Supporting policy-makers developing adaptive policies in the Mediterranean and Black Sea

Adaptive management is essential to the practical application of the Ecosystem-Based Approach (EBA). Despite there are frequent assertions that adaptive management is being used, evidence on its success is still limited. Indeed, it is difficult to bring the different elements of adaptive management together in a robust way and to choose the appropriate tools to do it. Therefore, it is necessary to provide a practical framework for adaptive policy action, consistent with the EBA. Accordingly, to operationalize the design and implementation of adaptive policies on the basis of the EBA, the Adaptive Marine Policy toolbox has been developed. The objective of the toolbox is to provide policy-makers a practical framework to design and implement adaptive policies. To show the functionality of the toolbox, the guidelines and resources provided within the toolbox have been applied to the marine litter issue in the Mediterranean and Black Sea as an example. The example application has shown that the toolbox is a useful and operational framework to build a science-policy interface according to the EBA. Despite some resources could be missing from the toolbox, they provide a practical and useful starting point to support the application of the different steps and key activities

Background study of the AMoRE-pilot experiment

We report a study on the background of the Advanced Molybdenum-Based Rare process Experiment (AMoRE), a search for neutrinoless double beta decay (\znbb) of $^{100}$Mo. The pilot stage of the experiment was conducted using $\sim$1.9 kg of \CAMOO~ crystals at the Yangyang Underground Laboratory, South Korea, from 2015 to 2018. We compared the measured $\beta/\gamma$ energy spectra in three experimental configurations with the results of Monte Carlo simulations and identified the background sources in each configuration. We replaced several detector components and enhanced the neutron shielding to lower the background level between configurations. A limit on the half-life of $0\nu\beta\beta$ decay of $^{100}$Mo was found at $T_{1/2}^{0\nu} \ge 3.0\times 10^{23}$ years at 90\% confidence level, based on the measured background and its modeling. Further reduction of the background rate in the AMoRE-I and AMoRE-II are discussed

Radioassay of the materials for AMoRE-II experiment

The AMoRE-II experiment will search for the 0νββ decay of 100Mo nuclei using molybdate crystal scintillators, operating at milli-Kelvin (mK) temperatures, with a total of 80 kg of 100Mo. The background goal for the experiment is 10–4 counts/keV/kg/year in the region of interest around the 0νββ decay Q-value of 3,034 keV. To achieve this level, the rate of background signals arising from emissions produced by decays of radioactive impurities in the detector and shielding materials must be strictly controlled. To do this, concentrations of such impurities are measured and are controlled through materials selection and purification. In this paper, we describe the design and the construction materials used to build the AMoRE-II detector and shielding system, including active and passive shielding, the cryostat, and the detector holders and instrumentation, and we report on measurements of radioactive impurities within candidate and selected materials

RECOMBINANT PROTEIN OF VARIOLA VIRUS ABOLISHES THE EFFECTS OF TUMOR NECROSIS FACTOR UPON MARROW HEMATOPOIESIS IN BALB/С MICE

We studied the effect of a recombinant murine TNF (rmTNF) and TNF-binding protein of Variola virus (VARV-CrmB) upon colony-forming ability of bone marrow cells (BMC) from Balb/c mice. BMC were grown in semisolid methylcellulose medium supplemented with murine growth factors in the absence or presence of mrTNF along at concentrations ranging from 2 to 40 ng/ml, VARV-CrmB (2 to 24 ng/ml), as well as in presence of both mrTNF (2 ng/ml) and VARV-CrmB (2 to 12 ng/ml). Hematopoietic colonies (BFU-E, CFU-E, and CFU-GM) were scored on day 14. VARV-CrmB protein didn't influence clonogenicity of the bone marrow progenitors. rmTNF inhibited growth of erythroid cells (BFU-E+CFU-E) at all concentrations tested, and stimulated growth of granulocyte-macrophage progenitors (CFU-GM) at concentrations of 2 ng/ml and 10 ng/ml. Combined effect of rmTNF and VARV-CrmB resulted into abolition of rmTNF-induced reduction of BFU-E+CFU-E formation and rmTNF-induced increase of CFU-GM number up to basic levels. Thеse results clearly demonstrate the anti-TNF activity of recombinant viral VARV-CrmB protein

LYELL`S SYNDROME. PRACTICAL CASE

In the article the interdisciplinary problem of drug allergy being analysed, namely its systemic symptoms. There have been shown the variants of systemic allergic reactions and the characteristics of the toxic epidermal necrolysis or Lyell`s syndrome as one of the most serious and rare form of illness. There has been carried out the analysis of the clinical case of the Lyell`s syndrome with diagnostic difficulties in the beginning phase of the disease and favorable outcome. There has been proved the dynamics of clinical signs and their reversed development

BIOLOGICAL EFFECTS OF TNF-BINDING VARIOLAVIRUS RECOMBINANT PROTEIN

Abstract. This review presents a summary of our data concerning in vivo and in vitro effects of recombinant TNF-binding protein from variola virus (VARVCrmB) upon TNF-induced functional changes of human and murine cells. VARV-CrmB protein blocks TNF-induced production of IL-1β and IL-6 by human mononuclear cells, and their in vitro oxidation-related metabolic (OM) activity. VARV-CrmB protein restores TNF-induced reduction of BFU-E+CFU-E colony-forming activity and normalizes TNF-induced effects upon CFU-GM formation in a colony-forming model of human and murine bone morrow cells (BMC). VARV-CrmB protein displays a pronounced in vivo alleviation of LPS-induced endotoxic shock symptoms in SPF BALB/C mice thus significantly increasing survival of experimental animals. Moreover, VARV-CrmBprotein decreases intensity of collagen-induced arthritis at early terms. Application of VARV-CrmB protein results in normalization of TNF-induced increase of migratory and OM activity of murine leukocytes, and exerts corrective effects upon colony-forming ability of murine hematopoietic precursors. Skin application of VARV-CrmB protein decreases leukocyte migration from a skin scrap in afferent phase of DNCB-induced contact reaction, as well as “ear oedema” index. Our results demonstrate TNF-blocking properties of VARVCrmB protein. In summary, our data allow to consider a recombinant variola virus VARV-CrmB as a new potential TNF-antagonist. Its effects can be explained by its ability of neutralizing TNF-induced activation of oxidation-related metabolic, cytokine-producing and migratory functions of effector cells in therapy of pathological inflammatory processes

EFFICIENCY OF RECOMBINANT TNF-BINDING PROTEIN FROM VARIOLA VIRUS IN A MODEL OF COLLAGEN-INDUCED ARTHRITIS

Abstract. This paper presents the results of the research on the effectiveness of recombinant TNF-binding protein of variola virus (VARV-CrmB) in a model of collagen-induced arthritis (CIA) in mice (CBAxC57Bl6) F1. The introduction of VARV-CrmB and polyclonal antibody to recombinant mouse TNF (poly-AbMuTNF) led to an improvement of clinical manifestations of CIA by reducing the swelling and increasing the mobility of mice limbs. The introduction of VARV-CrmB and poly-AbMuTNF reduced the number of neutrophilic granulocytes and granulocytic precursors. The introduction of VARV-CrmB and poly-AbMuTNF into mice decreased collagenolysis in the blood serum and the content of glycosaminoglycans at the early stages of experimentation. Treatment with VARV-CrmB and poly-AbMuTNF of mice with CIA significantly decreased the chemiluminescence response of blood leukocytes. VARV-CrmB exerted more pronounced inhibitory effect on the production of reactive oxygen metabolites by blood leukocytes of mice with CIA than poly-AbMuTNF. Improvement of clinical condition of the mice with CIA has a more prolonged effect following introduction of the VARV-CrmB than after injection of poly-AbMuTNF. The results suggest the recombinant viral protein VARVCrmB to be a new potential TNF-antagonist