Nitric oxide-induced activation of the AMP-activated protein kinase alpha2 subunit attenuates IKappaB kinase activity and inflammatory responses in endothelial cells

Background: In endothelial cells, activation of the AMP-activated protein kinase (AMPK) has been linked with anti-inflammatory actions but the events downstream of kinase activation are not well understood. Here, we addressed the effects of AMPK activation/deletion on the activation of NFKappaB and determined whether the AMPK could contribute to the anti-inflammatory actions of nitric oxide (NO). Methodology/Principal Findings: Overexpression of a dominant negative AMPKalpha2 mutant in tumor necrosis factor-alpha-stimulated human endothelial cells resulted in increased NFKappaB activity, E-selectin expression and monocyte adhesion. In endothelial cells from AMPKalpha2-/- mice the interleukin (IL)-1beta induced expression of E-selectin was significantly increased. DETA-NO activated the AMPK and attenuated NFKappaB activation/E-selectin expression, effects not observed in human endothelial cells in the presence of the dominant negative AMPK, or in endothelial cells from AMPKalpha2-/- mice. Mechanistically, overexpression of constitutively active AMPK decreased the phosphorylation of IKappaB and p65, indicating a link between AMPK and the IKappaB kinase (IKK). Indeed, IKK (more specifically residues Ser177 and Ser181) was found to be a direct substrate of AMPKalpha2 in vitro. The hyper-phosphorylation of the IKK, which is known to result in its inhibition, was also apparent in endothelial cells from AMPKalpha2+/+ versus AMPKalpha2-/- mice. Conclusions: These results demonstrate that the IKK is a direct substrate of AMPKalpha2 and that its phosphorylation on Ser177 and Ser181 results in the inhibition of the kinase and decreased NFKappaB activation. Moreover, as NO potently activates AMPK in endothelial cells, a portion of the anti-inflammatory effects of NO are mediated by AMPK

Biotransformation, trace analysis and effects of perfluoroalkyl and polyfluoroalkyl substances

Polyfluorierte und perfluorierte Verbindungen stellen eine Gruppe von Umweltkontaminanten dar, die aufgrund der starken C-F-Bindung besonders negative ökologische Eigenschaften besitzen. In dieser Arbeit wurde das Biotransformations-Potenzial von Fluortelomerethoxylaten (FTEO)unter aeroben Bedingungen untersucht. Nach Ermittlung der molekularen Zusammensetzung eines technischen FTEO-Gemischs mittels Elektrospray-Massenspektrometrie (ESI-MS) konnte aufgezeigt werden, dass zwei Transformationswege möglich sind, wobei einer dieser Wege über ω-oxidierte Carboxylate (FTEOCs), welche unter bestimmten Bedingungen nicht weiter abbaubar sind, verläuft. Ein anderer Abbauweg mündet in Perfluorcarboxylaten. Nach erfolgreicher Synthese zweier kurzkettiger FTEOCs konnten diese Verbindungen mithilfe einer Methode basierend auf Festphasenextraktion und HPLC-ESIMS/MS in einer Kläranlagenablaufprobe nachgewiesen und quantifiziert werden. Neben den FTEOCs konnten eine Reihe von Perfluorcarboxylaten und Perfluoralkansulfonaten in siebzehn Kläranlagenabläufen und vier Oberflächengewässern bestimmt werden. Die Untersuchung der Bioabbaubarkeit dreier potentieller Bausteine für neue, umweltfreundliche Fluortenside ergab, dass 6-(Trifluormethoxy)hexan-1-ol 100% anorganisches Fluorid freisetzte. Die homologe Verbindung 3(Trifluormethoxy)propan-1-ol hingegen setzte lediglich 15% Fluorid frei, wobei das verbleibende Fluor in der nicht weiter abbaubaren 3-(Trifluormethoxy)propansäure organisch gebunden blieb. Der dritte Baustein - 1-(2,2,3,3,4,4,4-Heptafluorbutoxy)propan-2-ol – führte erwartungsgemäß zur persistenten Verbindung Perfluorbutansäure. Die Toxizität der Perfluorbutansäure gegenüber der Algenspezies Pseudokirchneriella subcapitata und dem Kleinkrebs Chydorus sphaericus wurde anschließend ermittelt. Die toxikologischen Kenndaten ergaben keine akute Gefährdung dieser Organismen bei umweltrelevanten Konzentrationen. Weitere fluorierte Verbindungen wurden untersucht, wobei gezeigt werden konnte, dass die Toxizität mit zunehmender Kettenlänge im Allgemeinen zunimmt, die funktionelle Gruppe jedoch einen signifikanten Einfluss auf die Toxizität hat. Die massenspektrometrische Fragmentierung aller untersuchten Verbindungen wurde untersucht. Dabei ergab sich ein allgemeiner Fragmentierungsweg für Fluortelomer-basierte Verbindungen unter negativen Elektrospray-Bedingungen und stoßinduzierter Fragmentierung. Diese Erkenntnisse können in Zukunft zur Detektion neuer unbekannter Fluortelomerbasierter Verbindungen verwendet werden.Perfluoroalkyl and polyfluoroalkyl substances are environmental contaminants with adverse properties owing to the presence of the particularly strong C-F bond. In this thesis, the biotransformation potential of fluorotelomer ethoxylates (FTEOs) was assessed under aerobic conditions. After investigation of the technical FTEO mixture by electrospray-mass spectrometry (ESI-MS) techniques, it could be shown that two pathways are possible, one involving oxidation to ω-oxidized carboxylates (FTEOCs), which can be stable under certain conditions, and another pathway releasing perfluorocarboxylates, in function of the perfluoroalkyl chain length of the FTEO. After successful synthesis of two shortchain FTEOCs, these compounds were quantified in a wastewater treatment plant (WWTP) effluent by solid phase extraction and high performance liquid chromatography coupled to ESIMS/MS. Alongside the FTEOCs, a set of perfluorocarboxylates and perfluoroalkane sulfonates was determined in seventeen WWTP effluent samples and four surface water samples. The biotransformation behavior of three building blocks of potential novel environmentally friendly fluorosurfactants was investigated. It could be shown that 6-(trifluoromethoxy)hexan-1-ol released 100% inorganic fluoride, whereas 3-(trifluoromethoxy)propan-1-ol yielded only 15% fluoride. The remainder was shown to be organically bound as the non-degradable 3-trifluoromethoxy)propanoic acid. The building block 1-(2,2,3,3,4,4,4-heptafluorobutoxy)propan-2-ol expectedly released perfluorobutanoic as the persistent transformation product. The toxicity of the dead-end transformation product PFBA towards the green algae Pseudokirchneriella subcapitata and towards the Cladoceran Chydorus sphaericus was assessed with standardized tests. The results suggested no acute toxicity of these compounds at environmental levels. Other fluorinated compounds were included in the screening suggesting a general increase of toxicity with longer perfluoroalkyl chain length. However, the functional group itself showed a major effect on toxicity. The mass spectrometric fragmentation characteristics of the compounds studied in this thesis were carefully scrutinized. Comparison of different fluorotelomerbased compounds revealed a general fragmentation pattern for these compounds after negative electrospray ionization and collision-induced dissociation, which can be utilized to screen for novel fluorotelomer-based contaminants hereafter

Application of 5S in Context of Lean Management

Práce se zabývá praktickou případovou studií navržení aplikace a zavedení nástroje 5S ve vybraném podniku. V teoretické části jsou podrobně popsány principy a metody lean přístupu, jednotlivé pilíře a principy nástroje 5S a cíle aplikace nástroje 5S. Teoretická část je zakončena analýzou rozdílnosti přístupů vybraných literárních zdrojů k nástroji 5S. Praktická část se zabývá analýzou stavu pracoviště před zavedením nástroje 5S, samotným navržením a implementací nástroje 5S na analyzovaném pracovišti s následným vyhodnocením výsledků této implementace a jejích přínosů. Součástí této práce je vytvořený univerzální manuál 5S sloužící k implementaci nástroje 5S na ostatních pracovištích vybraného podniku.The thesis deals with a practical case study of designing the implementation and implementation of the 5S tool in the selected company. The theoretical part describes in detail the principles and methods of the lean approach, the individual pillars and principles of the 5S and the objectives of the implementation of the 5S. The theoretical part ends with an analysis of the differences in the attitudes of selected literary sources to the 5S tool. The practical part deals with the analysis of the state of the workplace before the implementation of the 5S, with the design and implementation of the 5S at the analyzed workplace with the subsequent evaluation of the results of this implementation and its benefits. Part of this thesis is a universal 5S manual proposed for implementation of the 5S tool in other workplaces of the selected company.639 - Katedra managementu kvalityvýborn

Reverse Engineering of mechanical part

katedra: KVS; přílohy: 1 CD ROM; rozsah: 46 s.This Bachelor´s thesis deals with the 3D digitizing of a real mechanical component using a non-contact scanner. At the beginning the notions of digitization and reverse engineering are explained. Then following distribution and description of machine for 3D digitizing. Another part deals with the digitization of real 3D engineering component using 3D scanner REVscan. The output from this part are digital data in the form of the so-called cloud of points. The last part deals with optimization of measured data using polygon network in the program Geomagic Studio.Tato bakalářská práce se zabývá 3D digitalizací reálné strojní součásti pomocí bezdotykového skeneru. Na začátku jsou vysvětleny pojmy digitalizace a reverzní inženýrství. Následuje rozdělení a popis zařízení sloužících k 3D digitalizaci. Další část se zabývá 3D digitalizací reálné strojní součásti s využitím 3D skeneru REVscan. Výstupem z této části jsou digitální data ve formě tzv. mraku bodů. Poslední část se zabývá optimalizací naměřených dat pomocí polygonové sítě v programu Geomagic Studio

Bound States of Heavy Flavor Hyperons

Several realistic phenomenological nucleon-nucleon interaction models are employed to investigate the possibility of bound deuteron-like states of such heavy flavor hyperons and nucleons, for which the interaction between the light flavor quark components is expected to be the most significant interaction. The results indicate that deuteron-like bound states are likely to form between nucleons and the $\Xi_c^{'}$ and $\Xi_{cc}$ charm hyperons as well as between $\Xi$ hyperons and double-charm hyperons. Bound states between two $\Sigma_c$ hyperons are also likely. In the case of beauty hyperons the corresponding states are likely to be deeply bound.Comment: 25 pages, 3 figures. Accepted Nucl. Phys.

The Concept of the Implementation of Present Evidence-based Knowledge and Technology into the Preparation of Sport Professionals

AbstractThe study assesses the feasibility of the concept of professional preparation of sport teachers and instructors that attempts to connect education and research activities with the emphasis on presenting evidence-based knowledge and new technologies. The seven-year-long international study involved 670-850 university students annually. Students participated directly in the research activities that were in compliance with the curricula of sport education study programs. The presented educational model describes possibilities and major limits how to provide dozens evidence-based knowledge of prospective sport professionals in the areas of physical activity monitoring, self-assessment of physical fitness and evaluation of sport preferences

Führer zum Material über Ostafrika im Evangelisch-Lutherischen Missionsblatt 1906-1910

This volume is a sequel to Volumes 4 and 17. All three volumes list articles and photographs in the Leipzig Mission's main journal relating to what are now northern Tanzania and southern Kenya, where the Mission worked. This volume covers the years 1906-1910