Assessment of glial scar, tissue sparing, behavioral recovery and axonal regeneration following acute transplantation of genetically modified human umbilical cord blood cells in a rat model of spinal cord contusion

©2016 Mukhamedshina et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Objective and Methods This study investigated the potential for protective effects of human umbilical cord blood mononuclear cells (UCB-MCs) genetically modified with the VEGF and GNDF genes on contusion spinal cord injury (SCI) in rats. An adenoviral vector was constructed for targeted delivery of VEGF and GDNF to UCB-MCs. Using a rat contusion SCI model we examined the efficacy of the construct on tissue sparing, glial scar severity, the extent of axonal regeneration, recovery of motor function, and analyzed the expression of the recombinant genes VEGF and GNDF in vitro and in vivo. Results Transplantation of UCB-MCs transduced with adenoviral vectors expressing VEGF and GDNF at the site of SCI induced tissue sparing, behavioral recovery and axonal regeneration comparing to the other constructs tested. The adenovirus encoding VEGF and GDNF for transduction of UCB-MCs was shown to be an effective and stable vehicle for these cells in vivo following the transplantation into the contused spinal cord. Conclusion Our results show that a gene delivery using UCB-MCs-expressing VEGF and GNDF genes improved both structural and functional parameters after SCI. Further histological and behavioral studies, especially at later time points, in animals with SCI after transplantation of genetically modified UCB-MCs (overexpressing VEGF and GDNF genes) will provide additional insight into therapeutic potential of such cells

CALIBRATION LABORATORY RISK MANAGEMENT AS AN INNOVATIVE APPROACH TO THE MANAGEMENT OF WEATHERFORD

This article describes the goals, objectives, definition and analysis of typical risks as an innovative approach to management in the verification of measuring instruments in a metrological laboratory

Подход к верификации семейства мультиагентных систем разрешения конфликтов

In the paper, we describe a verification method for families of distributed systems generated by context-sensitive network grammar of a special kind. This grammar includes special non-terminal symbols, so called quasi-terminals, which uniquely correspond to grammar terminals. These quasi-terminals specify processes which are merging of base system processes, in contrast to simple nonterminals which specify networks of parallel compositions of the processes. The method is based on model checking technique and abstraction. An abstract representative model for a family of systems depends on their specification grammar and system properties to be verified. This model simulates the behaviour of the systems in such a way that the properties which hold for the representative model are satisfied for all these systems. The properties of the representative model can be verified by model checking method. The properties of a generated system are specified by universal branching time logic ∀CTL with finite deterministic automata as atomic formulas. We show the use of this method for verification of some properties of a multiagent system for conflict resolution, in particular, for context-dependent disambiguation in ontology population. We also suggest that this approach should be used for verification of computations on sub-grids which are sub-graphs of computation grids. In particular, we consider the computation of parity of the active processes number in a sub-grid.В данной работе мы описываем метод верификации для семейств распределенных систем, которые порождаются контекстно-зависимой сетевой грамматикой специального вида. Эта грамматика содержит специальные нетерминальные символы ䷘ квази-терминалы. Квази-терминалы однозначно соответствуют терминалам грамматики и могут задавать процессы,которые определяются слиянием базовых процессов системы, в то время как нетерминалы задают сети параллельных композиций этих процессов. Данный метод верификации основан на техниках верификации моделей и абстракции. Абстрактная репрезентативная модель семейства систем зависит от задающей их грамматики и верифицируемых свойств системы. Эта модель симулирует поведение заданных систем таким образом, что свойства, которые выполняются в репрезентативной модели, также выполняются и во всех заданных системах. Проверку свойств репрезентативной модели можно осуществлять с помощью метода проверки моделей. Свойства порождаемых систем специфицируются с помощью универсальной логики ветвящегося времени ∀CTL с конечными детерминированными автоматами в качестве атомарных формул. Мы показываем, что предложенный метод верификации можно применять для проверки некоторых свойств мультиагентных систем разрешения конфликтов, в частности, систем разрешения неоднозначностей при пополнении онтологий. Также показано, что этот подход можно использовать для верификации вычислений на подрешетках, являющихся подграфами решеток вычислений, например, для вычисления четности числа работающих процессов

Innovative Approach to Company Management in the Part of Risk Management of a Calibration Laboratory

В статье описаны цели, задачи, определение и анализ типовых рисков как инновационный подход к управлению при поверке средств измерений в метрологической лаборатории.This article describes the goals, objectives, definition and analysis of typical risks as an innovative approach to management in the verification of measuring instruments in a metrological laboratory

poST

Publisher Copyright: AuthorThis paper presents the core concepts for the poST language - a process-oriented extension of the IEC 61131-3 Structured Text (ST) language which intends to provide a conceptual consistency of the PLC source code with technological description of the plant operating procedure. The poST can be seamlessly used as a textual programming language for complex PLC software in the context of IEC 61131-3 (3rd Edition). The language combines the advantages of FSM-based programming with the conventional syntax of the ST language which would facilitate its adoption. The poST language assumes that a poST-program is a set of weakly connected concurrent processes, structurally and functionally corresponding to the technological description of the plant. Each process is specified by a sequential set of states. The states are specified by a set of the ST constructs, extended by TIMEOUT operation, SET STATE operation, and START / STOP / check state operations to communicate with other processes. The paper describes the basic syntax of the poST language, demonstrates the usage of the poST language by developing control software for an elevator, and compares the development in poST with pure Structured Text.Peer reviewe

Detection of Antibodies Recognizing Puumala Virus Nucleocapsid and Glycoprotein Peptides in NE Serum

© 2016, Springer Science+Business Media New York.Nephropatia epidemica (NE), a mild form of hemorrhagic fever with renal syndrome (HFRS), is an endemic zoonosis in the Republic of Tatarstan, Russia. Humans become infected by inhaling an aerosol contaminated with Puumala virus, a member of genus Hantavirus. NE diagnosis is based on detection of anti-hantavirus antibodies using ELISA. Antibodies to hantavirus nucleocapsid (N) protein are detected early in the course of infection, suggesting that this viral protein is the most immunogenic. Several epitopes were previously identified on N protein as well as glycoproteins of Puumala viruses endemic in Europe. However, there is limited knowledge about Puumala virus N protein antigenic epitopes in NE patients in the Republic of Tatarstan. The aim of the present study is to identify N protein and glycoprotein epitopes which induce a humoral immune response in NE cases. Analysis of NE serum using an array of overlapping N protein and glycoprotein peptides identified the most immunogenic epitopes, which can then be used for developing Puumala virus-specific vaccine

Верификация алгоритмов мультиагентного анализа данных с помощью системы проверки моделей SPIN

The paper presents an approach to formal verification of multi-agent data analysis algorithms for ontology population. The system agents correspond to information items of the input data and the rule of ontology population and data processing. They determine values of information objects obtained at the preliminary phase of the analysis. The agents working in parallel check the syntactic and semantic consistency of tuples of information items. Since the agents operate in parallel, it is necessary to verify some important properties of the system related to it, such as the property that the controller agent correctly determines the system termination. In our approach, the model checking tool SPIN is used. The protocols of agents are written in Promela language (the input language of the tool) and the properties of the multi-agent data analysis system are expressed in the liner time logic LTL. We carried out several experiments to check this model in various modes of the tool and various numbers of agents.В статье представлен подход к формальной верификации алгоритмов мультиагентного анализа данных для пополнения онтологий. Агенты системы на основе входных данных устанавливают значения элементов объектов, полученных на предварительной стадии анализа. Агенты параллельно осуществляют проверку семантической и синтаксической согласованности, используя правила пополнения онтологий и обработки данных. Поскольку агенты действуют параллельно, необходимо верифицировать некоторые важные свойства системы, связанные с этим, например, свойство корректности определения завершения работы системы. В нашем подходе используется инструмент проверки моделей SPIN. Протоколы агентов записаны на языке Promela, а свойства мультиагентной системы анализа данных выражены в логике LTL. Мы провели ряд экспериментов по проверке данной модели

Results of Studies of Genetic Heterogeneity in Hantavirus RNA-Isolates, Found in the Territory of the Saratov Region

With a view to obtain the data on intraspecific diversity of Hantaviruses circulating in the territory of the Saratov Region, carried out has been molecular-genetic testing of the field and clinical material. Using PCR assay, Hantavirus RNA has been detected in 25 samples obtained from small mammals and 15 blood samples of HFRS patients. Sequencing of 14 hantavirus RNA-isolates from rodents and 3 RNA-isolates from clinical material has made it possible to identify their taxonomic appurtenance to Puumala virus. Phylogenetic analysis of RNA-isolates has revealed that all of them are characterized by high degree of homology in between themselves (99.8 %) and substantial distinction from the known Puumala strains, frequently occurring in Russia (3.6-21 %). Therewith, verified has been circulation of Puumala virus in the territory of the Saratov Region using molecular-genetic techniques. Identified genetic differences of Saratov RNA-isolates, in particular, opens a further prospect for allocation of the sites where people can contract HFRS infection

Human Mesenchymal Stem Cells Overexpressing Interleukin 2 Can Suppress Proliferation of Neuroblastoma Cells in Co-Culture and Activate Mononuclear Cells In Vitro

High-dose recombinant interleukin 2 (IL2) therapy has been shown to be successful in renal cell carcinoma and metastatic melanoma. However, systemic administration of high doses of IL2 can be toxic, causing capillary leakage syndrome and stimulating pro-tumor immune response. One of the strategies to reduce the systemic toxicity of IL2 is the use of mesenchymal stem cells (MSCs) as a vehicle for the targeted delivery of IL2. Human adipose tissue-derived MSCs were transduced with lentivirus encoding IL2 (hADSCs-IL2) or blue fluorescent protein (BFP) (hADSCs-BFP). The proliferation, immunophenotype, cytokine profile and ultrastructure of hADSCs-IL2 and hADSCs-BFP were determined. The effect of hADSCs on activation of peripheral blood mononuclear cells (PBMCs) and proliferation and viability of SH-SY5Y neuroblastoma cells after co-culture with native hADSCs, hADSCs-BFP or hADSCs-IL2 on plastic and Matrigel was evaluated. Ultrastructure and cytokine production by hADSCs-IL2 showed modest changes in comparison with hADSCs and hADSCs-BFP. Conditioned medium from hADSC-IL2 affected tumor cell proliferation, increasing the proliferation of SH-SY5Y cells and also increasing the number of late-activated T-cells, natural killer (NK) cells, NKT-cells and activated T-killers. Conversely, hADSC-IL2 co-culture led to a decrease in SH-SY5Y proliferation on plastic and Matrigel. These data show that hADSCs-IL2 can reduce SH-SY5Y proliferation and activate PBMCs in vitro. However, IL2-mediated therapeutic effects of hADSCs could be offset by the increased expression of pro-oncogenes, as well as the natural ability of hADSCs to promote the progression of some tumors

Effect of Adenoviral Transduction of Hepatic Stellate Cells with Adv5-optHGF-RFP on their Phenotype

© 2016, Springer Science+Business Media New York.Adenovirus is a promising gene delivery vector that has a high efficiency and relative simplicity of construction. These advantages make this system attractive for diverse research applications. In this project we transduced hepatics stellatecells (HSC) with adenoviral vector containing genes of hepatocytes growth factor (HGF) and red fluorescent protein (RFP) as a reporter (Adv5-optHGF-RFP), which let us to visualize the transduced cells. Further changes of phenotype were studied by real-time PCR and immunocytochemical staining. According to our results, most of the cells were transduced and demonstrated stable expression of RFP. Effectiveness of the transduction was also confirmed by a high expression of HGF (1200 times higher than in control HSC culture). Adenoviral transduction of HSC with HGF gene did not change their morphology, but stimulated expression of HSC marker desmin, leading to HSC activation (α-smooth muscle actin) and appearance of α-fetoprotein, one of the hepatoblasts markers. During the entire experiment, there were Ki-67+ cells, meaning that proliferation of transduced HSC was not affected. Thus, adenoviral transduction of HSC with Adv5-optHGF-RFP is a good gene delivery system with a stimulating effect on HSC