Human dendritic cells process and present Listeria antigens for in vitro priming of autologous CD4+ T lymphocytes

The role of human dendritic cells (DC) in the immune response toward intracellularly growing Listeria was analyzed under in vitro conditions using several morphological and functional methods. DC incubated with Listeria innocua and L. monocytogenes, respectively, readily phagocytosed the bacteria. Listeria did not impair viability and immunogenic potential of human DC. Listerial antigens were found to be processed within the lysosomal compartment of DC and colocalized with major histocompatibility complex (MHC) class II molecules, as shown by fluorescence and transmission electron microscopy. DC challenged with apathogenic L. innocua were highly effective in priming autologous naïve T cells (mainly CD4+) in vitro. The T cells strongly proliferated in the presence of DC incubated with L. innocua, which could be significantly inhibited by anti-MHC II mAb. L. innocua-primed T cells were also successfully stimulated by DC harboring the pathogenic L. monocytogenes, either the wild-type strain EGD or the p60 reduced mutant strain RIII. From our results, we conclude that human DC infected with nonpathogenic intracellular bacteria are able to efficiently prime naïve T cells, which are then suitable for recognition of antigens derived from related virulent bacterial species. This in vitro human model provides an interesting tool for basic research in infectious immunology and possibly for a new immunotherap

Cholesterol in Negatively Charged Lipid Bilayers Modulates the Effect of the Antimicrobial Protein Granulysin

The release of granulysin, a 9-kDa cationic protein, from lysosomal granules of cytotoxic T lymphocytes and natural killer cells plays an important role in host defense against microbial pathogens. Granulysin is endocytosed by the infected target cell via lipid rafts and kills subsequently intracellular bacteria. The mechanism by which granulysin binds to eukaryotic and prokaryotic cells but lyses only the latter is not well understood. We have studied the effect of granulysin on large unilamellar vesicles (LUVs) and supported bilayers with prokaryotic and eukaryotic lipid mixtures or model membranes with various lipid compositions and charges. Binding of granulysin to bilayers with negative charges, as typically found in bacteria and lipid rafts of eukaryotic cells, was shown by immunoblotting. Fluorescence release assays using LUV revealed an increase in permeability of prokaryotic, negatively charged and lipid raft-like bilayers devoid of cholesterol. Changes in permeability of these bilayers could be correlated to defects of various sizes penetrating supported bilayers as shown by atomic force microscopy. Based on these results, we conclude that granulysin causes defects in negatively charged cholesterol-free membranes, a membrane composition typically found in bacteria. In contrast, granulysin is able to bind to lipid rafts in eukaryotic cell membranes, where it is taken up by the endocytotic pathway, leaving the cell intac

Modelling of Fuzzy Expert System for an Assessment of Security Information Management System UIS (University Information System)

Several methodologies based on the international standard ISO/IEC 27001 have been developed for modelling information security management systems within higher education. This paper transformed the ISO/IEC 27001 standard into a questionnaire, which was sent digitally to about 100 universities in Bosnia and Herzegovina, and to the EU, Norway and the USA. The questions are arranged by levels, and the levels have their numerical weights, derived from individual questions in the levels themselves. Otherwise, the questions are asked with Yes or No and thus are reduced to binary variables. The rules necessary for the functioning of the system have been calculated. The fuzzy logic method represents a new approach to the problems of managing complex systems, which is very difficult to describe with a certain mathematical model, as well as in systems with a large number of inputs and outputs where there are unclear interactions. Risk assessment is a major part of the ISMS process. Traditional risk calculation models are based on the application of probability and classical set theory. Here, we have converted the risk assessment into a system rating of 5 to 10 numerically or from five to ten descriptively. We perform fuzzy optimization by finding the values of the input parameters of a complex simulated system, which results in the desired output. We use the fuzzy logic controller to execute fuzzy inference rules from the fuzzy rule database in determining congestion parameters, obtaining warning information and appropriate action. Simulating the situation of an advanced system that evaluates the protection quality of such a system with fuzzy logic, we use MATLAB. The paper combines the original Visual Basic programming language and MATLAB\u27s Fuzzy Toolbox, to solve the complex problem of assessing compliance with the ISO/IEC 27001 standard, as one of the main standards for information systems security modelling. University information systems were used, but it is also applicable to all other information systems. The evaluation has been done for several universities and it has been proven that the system evaluates correctly, but these universities must not be publicly named. There was no such approach in the use of fuzzy logic and on such systems, and that is the originality of this work

Automation and Artificial Intelligence in Software Engineering: Experiences, Challenges, and Opportunities

Automation and Artificial Intelligence have a transformative influence on many sectors, and software engineers are the actors who engineer this transformation. On the other hand, there is little knowledge of how automation and Artificial Intelligence impact software engineering practice. To answer this question, we conducted semi-structured interviews with experienced software practitioners across frontend and backend development, DevOps, R&D, integration, and leadership positions. Our findings reveal 1) automation to appear as micro-automation in the sense of automation of tiny and specific tasks, 2) automation as a side product of work, and bottom-up driven in software engineering, and 3) automation as a possible cause for cognitive overhead due to automatically generated notifications. Furthermore, we notice that our interview participants do not expect automation and artificial intelligence tools to substantially change software engineering\u27s essence in the foreseeable future

Efikasnost fungicida Vivando (A.S. Metrafenon) u suzbijanju pepelnice vinove loze u Crnoj Gori

Efficacy of the fungicide based on metrafenone (Vivando) in control of the pathogenic fungus Erysiphe necator, the causal agent of grapevine powdery mildew, has been studied at the experimental field of the Biotechnical Faculty in Podgorica (Podgorica's vineyards area). The experiments were conducted in 2011 and 2012, and were set according to the EPPO methods. The assessment of efficacy was performed on leaves and clusters of grapes. During 2011, nine treatments were performed, and in 2012 five. The obtained efficiency in both years amounted 100%, although the intensity of the disease varied in different experimental years. In 2011, on control plots disease intensity was high - 98.25% on leaves and 98.03% on clusters, and in 2012 was quite lower - 65.83% on leaves and 62.05% on clusters. The active substance metrafenone exhibited high efficiency at different, very high, intensities of the disease.Na oglednom imanju Biotehničkog fakulteta u Podgorici (Podgoričko vinogorje) ispitivana je efikasnost fungicida na bazi metrafenona (Vivando) u suzbijanju fitopatogene gljive Erysiphe necator, prouzrokovača pepelnice vinove loze. Eksperimenti su izvedeni tokom 2011. i 2012. godine, a postavljeni su po metodama EPPO. Ocjena efikasnosti obavljena je na listovima i grozdovima. Tokom 2011. obavljeno je devet tretiranja, a u 2012. pet. Dobijena efikasnost u obe godine je iznosila 100%, iako se intenzitet oboljenja razlikovao u eksperimentalnim godinama. Tokom 2011. na kontrolnim parcelama iznosio je visokih 98,25% na listovima i 98,03 na grozdovima, a u 2012. bio je dosta niži - na listovima 65,83%, a na grozdovima 62,50%. Aktivna supstanca metrafenon ispoljila je visoku efikasnost pri različitim, vrlo visokim, intenzitetima oboljenja

Supressão de crescimento de fungos patogênicos de plantas usando extratos de briófitas

Chemicals are often used in attempts to control diseases caused by plant pathogenic fungi during food production. However, chemicals can have adverse effects not just on food, but they also remain active for a long time within ecosystems, and thus are not environmentally friendly. Therefore, development of bio-treatment and avoiding use of chemicals are urgently needed. With the aim of studying and developing new environmentally-friendly treatments, we tested extracts from selected bryophyte species (Porella platyphylla, Cinclidotus fontinaloides and Anomodon viticulosus) on five plant pathogenic fungi under controlled conditions. The fungi (Botryosphaeria dothidea, Phomopsis viticola, Calosphaeria sp., Colletotrichum acutatum and Monilinia laxa) were selected based on common diseases they cause on fruits and grapevine. They were isolated in cultures and treated with bryophyte extracts. Bryophyte extracts were shown to be effective in suppression of certain plant pathogenic fungi growth and to have a huge potential in development of novel biotechnological treatments and biofungicides. The best results were achieved in inhibition of B. dothidea, P. viticola and Calosphaeria sp.Fungos fitopatogênicos são controlados com produtos químicos para combater doenças causadas por eles durante a produção de alimentos. Tais produtos são ruins não apenas para a alimentação, mas também podem permanecer por muito tempo nos ecossistemas, portanto, não são ecologicamente corretos. Desta forma, os biotratamentos e a prevenção de usos químicos são urgentemente necessários no futuro próximo. Com o objetivo de estudar e desenvolver nova alternative decontrole de doenças de plantas, testamos os extratos de espécies de briófitas selecionadas (Porella platyphylla, Cinclidotus fontinaloides e Anomodon viticulosus)  em cinco fungos fitopatogênicos (Botryosphaeria dothidea, Phomopsis viticola, Calosphaeria sp., Colletotrichum acutatum e Monilinia laxa) em condição controlada. Estes fungos, selecionados com base nas doenças que causam em frutos e videiras, foram isolados em culturas puras e tratados com os respectivos extratos de briófitas. Os extratos de briófitas demonstraram ser eficazes na supressão de certos fungos fitopatogênicos e têm um enorme potencial no desenvolvimento de novos biofungicidas e tratamentos biotecnológicos. Os mais interessantes resultados foram obtidos na inibição de B. dothidea, P. viticola e Calosphaeria sp

Posture-induced changes in peripheral nerve stiffness measured by ultrasound shear-wave elastography

Introduction: Peripheral nerves slide and stretch during limb movements. Changes in nerve stiffness associated with such movements have not been examined in detail but may be important in understanding movement-evoked pain in patients with a variety of different musculoskeletal conditions. Methods: Shear-wave elastography was used to examine stiffness in the median and tibial nerves of healthy individuals during postures used clinically to stretch these nerves. Results: Shear-wave velocity increased when limbs were moved into postures that are thought to increase nerve stiffness (mean increase: median nerve = 208% in arm, 236% in forearm; tibial nerve = 136%). There was a trend toward a negative correlation between age and shear-wave velocity (r = 0.58 for tibial nerve). Conclusions: Shear-wave elastography provides a tool for examining nerve biomechanics in healthy individuals and patients. However, limb position, age, and effects of nerve tension on neural architecture should be taken into consideration

Volatility spillover effect in Western Balkans

This article examines volatility spillover among Western Balkan’s stock markets and selected developed markets. If there is an evidence of weak linkage between various markets, then there are potential benefits that could arise from international diversification. However, if we analyse the relationship between two markets that are different in terms of their economic development, and if there is a strong connection between them, market shocks from the developed markets can have an impact on the frontier/emerging markets. Market integration can be indicated with returns linkage and transmission of shocks and volatility between markets. Hence, this can have implications for investment strategies. It is found that there is statistically significant regional spillover between countries of the Western Balkan region. Also, there is global spillover between developed markets and this region as well. Furthermore, there is evidence that Western Balkan’s markets are late in response to important market events, and that can be used when formulating investment strategy