925 resultados para In-house Administration


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La trattazione inizia con l’analisi del concetto di “servizio” per focalizzarsi quindi sulla categoria dei “servizi pubblici”, nell’evoluzione dell’interpretazione dottrinale, fra concezione soggettiva e oggettiva, e sulle tipologie di servizio, con specifico riguardo a quelli aventi rilevanza nazionale. L’esposizione si suddivide dunque in due sezioni, rispettando la ripartizione del titolo. La prima parte è dedicata all’esame dei “ruoli”, ovvero delle parti coinvolte, a diverso titolo, in un rapporto di pubblico servizio: il “ruolo contrattuale”, rispetto al quale è stata esaminata la posizione dell’erogatore del servizio, da un lato, e dell’utente, dall’altro lato; il “ruolo di controllo”, esercitato dalle autorità amministrative indipendenti (Authorities); infine il “ruolo giurisdizionale”, dedicato alla complessa questione del riparto di giurisdizione in materia di servizi pubblici, fra giudice ordinario e giudice amministrativo, alla luce della nota sentenza della Corte Costituzionale, n. 204/2004. La seconda parte della tesi riguarda, invece, l’analisi dei “modelli contrattuali”, ovvero delle tipologie di contratto che vengono in rilievo in materia di servizi pubblici, astrattamente ascrivibili a tre distinti “gradi”. Il primo grado è rappresentato dal “contratto di servizio”, stipulato fra l’autorità pubblica che affida in gestione il servizio ed il soggetto, pubblico o privato, che se ne fa carico; l’affidamento può avere forma “diretta”, cioè prescindere da una procedura di gara (ipotesi della cd. “gestione diretta in economia” o del cd. “affidamento in house providing”) o “indiretta” (nel qual caso trova applicazione la normativa sugli appalti pubblici e le concessioni). Al secondo grado vi è il “codice di rete”, figura contrattuale che presuppone un la presenza di un servizio “a rete”. Il servizio a rete implica la presenza di una struttura o infrastruttura, concreta o astratta, ed una relazione reciproca fra gli elementi della stessa che convergono in vista della realizzazione di uno scopo e coinvolge tradizionalmente i servizi rispetto ai quali sussiste una situazione di monopolio naturale (trasporti, energia elettrica, gas, telecomunicazioni), caratterizzati dall’inopportunità pratica ed economica di creare una nuova rete. Di conseguenza, la possibilità di erogare il servizio, per gli operatori non titolari della rete, è rimessa alla stipulazione di un contratto (cd. “codice di rete) con il titolare della stessa, avente per oggetto la regolazione dello sfruttamento della rete, dietro pagamento di un corrispettivo economico. Il terzo grado è rappresentato dal “contratto di utenza”, le cui parti sono erogatore del servizio ed utente e la cui analisi è stata necessariamente preceduta dall’analisi del “rapporto di utenza”: quest’ultimo ha subito infatti una radicale trasformazione, nel corso degli ultimi decenni, soprattutto in seguito ai processi di liberalizzazione e privatizzazione che hanno coinvolto il settore de quo e condotto alla ricerca di nuovi strumenti di tutela dell’interesse generale, sotteso ad ogni pubblico servizio, contribuendo a conferire nuovo e preponderante interesse per l’utente, anche attraverso l’introduzione e l’applicazione di normative specifiche (in primis, quella in tema di clausole vessatorie e quella relativa alle azioni a tutela del consumatore-utente, oggi confluite nel codice del consumo). Per l’esame di ciascun argomento si è fatto costante riferimento alla normativa ed agli orientamenti comunitari; inoltre, si è scelto di prendere in considerazione, dopo l’esame generale delle principali tematiche sopramenzionate, il caso specifico del servizio radiotelevisivo.

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Computer simulations play an ever growing role for the development of automotive products. Assembly simulation, as well as many other processes, are used systematically even before the first physical prototype of a vehicle is built in order to check whether particular components can be assembled easily or whether another part is in the way. Usually, this kind of simulation is limited to rigid bodies. However, a vehicle contains a multitude of flexible parts of various types: cables, hoses, carpets, seat surfaces, insulations, weatherstrips... Since most of the problems using these simulations concern one-dimensional components and since an intuitive tool for cable routing is still needed, we have chosen to concentrate on this category, which includes cables, hoses and wiring harnesses. In this thesis, we present a system for simulating one dimensional flexible parts such as cables or hoses. The modeling of bending and torsion follows the Cosserat model. For this purpose we use a generalized spring-mass system and describe its configuration by a carefully chosen set of coordinates. Gravity and contact forces as well as the forces responsible for length conservation are expressed in Cartesian coordinates. But bending and torsion effects can be dealt with more effectively by using quaternions to represent the orientation of the segments joining two neighboring mass points. This augmented system allows an easy formulation of all interactions with the best appropriate coordinate type and yields a strongly banded Hessian matrix. An energy minimizing process accounts for a solution exempt from the oscillations that are typical of spring-mass systems. The use of integral forces, similar to an integral controller, allows to enforce exactly the constraints. The whole system is numerically stable and can be solved at interactive frame rates. It is integrated in the DaimlerChrysler in-house Virtual Reality Software veo for use in applications such as cable routing and assembly simulation and has been well received by users. Parts of this work have been published at the ACM Solid and Physical Modeling Conference 2006 and have been selected for the special issue of the Computer-Aided-Design Journal to the conference.

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Oggetto della ricerca è l’esame del ruolo attuale della partecipazione pubblica locale in società per l’erogazione di servizi pubblici locali di rilevanza economica, nel nuovo contesto normativo caratterizzato dalla residualità dell’autoproduzione, dalla liberalizzazione, dall’organizzazione del servizio in ambiti territoriali, e dalle esigenze di contenimento della spesa locale, acuite dalla crisi economica-finanziaria. Si sono distinte quattro tipologie di società: nel caso di servizi gestiti in regime di esclusiva, le società a capitale pubblico-privato con socio privato operativo scelto mediante gara, le società in house affidatarie dirette, e le società affidatarie in quanto selezionate in procedure ad evidenza pubblica; nel caso di servizi sottratti al regime di esclusiva, le società che eroghino il servizio liberalizzato. L’indagine si è focalizzata sulle condizioni di costituzione e di mantenimento di tali società, e si è approfondito quel particolare aspetto del loro regime giuridico costituito dai limiti operativi. L’analisi è stata condotta esaminando le nuove disposizioni, le posizioni giurisprudenziali e le letture della dottrina relative alle società disciplinate come forme di gestione di servizi pubblici locali di rilevanza economica, relative alla funzionalizzazione dell’attività degli enti territoriali e delle società da questi partecipate, e relative alla tutela della concorrenza.

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The lateral characteristics of tires in terms of lateral forces as a function of sideslip angle is a focal point in the prediction of ground loads and ground handling aircraft behavior. However, tests to validate such coefficients are not mandatory to obtain Aircraft Type Certification and so they are not available for ATR tires. Anyway, some analytical values are implemented in ATR calculation codes (Flight Qualities in-house numerical code and Loads in-house numerical code). Hence, the goal of my work is to further investigate and validate lateral tires characteristics by means of: exploitation and re-parameterization of existing test on NLG tires, implementation of easy-handle model based on DFDR parameters to compute sideslip angles, application of this model to compute lateral loads on existing flight tests and incident cases, analysis of results. The last part of this work is dedicated to the preliminary study of a methodology to perform a test to retrieve lateral tire loads during ground turning with minimum requirements in terms of aircraft test instrumentation. This represents the basis for future works.

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Im Rahmen dieser Arbeit wurde die Methylierung von Quecksilber in Intestinaltrakt des Kompostwurms Eisenia foetida untersucht. Des Weiteren wurden aerobe und anaerobe Mikroorganismen aus dem Darmtrakt von Eisenia fotida isoliert, identifiziert und auf ihr Potential zur Methylierung von Quecksilber getestet. Die Bestimmung von Methylquecksilber erfolgte mittels GC-ICPMS (Gaschromatographie mit induktiv gekoppelter Plasma-Massenspektrometrie) und GC-AFS (Gaschromatographie- Atomfluoreszenzspektrometrie). Für die GC-ICPMS erfolgte die Quantifizierung des Methylquecksilbers mittels der Isotopenverdünnungsmethode. Die Extraktion des Methylquecksilbers aus dem Wurmgewebe erfolgte durch einen alkalischen Aufschluss mit TMAH (Tetramethylammoniumhydroxid) und anschließender Derivatisierung des Methylquecksilbers durch Natriumtetrapropylborat. Für die Extraktion des gebildeten Methylquecksilbers aus Bakterienkulturen wurde eine Extraktion mit einer methanolischen Kaliumhydroxidlösung verwendet. Wie bei dem Wurmgewebe wurde das Methylqueckilsber ebenfalls mit Natriumtetrapropylborat derivatisiert.rnrnFür die Untersuchung einer in vivo Methylquecksilberbildung in bodenlebenden Invertebraten wurde der Kompostwurm Eisenia foetida als Modellorganismus verwendet. Die Tiere wurden aus einer Kultur in einen Boden überführt, der mit anorganischem Quecksilber versetzt wurde. Nach zehn Tagen Inkubationszeit wurden die Würmer entnommen und das Methylquecksilber extrahiert. Um eine mögliche Methylierung von Quecksilber durch Bodenorganismen auszuschließen wurde sowohl steriles als auch unsteriles Bodenmaterial verwendet. In den Wurmproben aus dem unsterilen Bodenmaterial konnte eine Konzentration an Methylquecksilber von 17,4 ng/g Trockengewicht (Boden ohne Zugabe von Quecksilber) und 62,4 ng/g Trockengewicht (Boden mit Quecksilberzugabe). Bei den Wurmproben aus sterilem Bodenmaterial lag die Konzentration an Methylquecksilber bei 17,2 ng/g Trockengewicht (Boden ohne Zugabe von Quecksilber) und 51,9 ng/g Trockengewicht (Boden mit Quecksilberzugabe).rnrnBei den Bakterienkulturen konnte in Reinkulturen keine Methylierung von Quecksilber nachgewiesen werden. In einer fakultativ anaeroben Mischkultur konnte eine Methylierung von Quecksilber beobachtet werden. Für die Identifizierung der Mikroorganismen wurde die 16s rDNA mittels PCR amplifiziert und anschließend über eine DGGE aufgetrennt. Die Banden wurden ausgeschnitten und sequenziert. Dabei konnten drei Enterobacteriaceen identifiziert werden.rn

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Flow features inside centrifugal compressor stages are very complicated to simulate with numerical tools due to the highly complex geometry and varying gas conditions all across the machine. For this reason, a big effort is currently being made to increase the fidelity of the numerical models during the design and validation phases. Computational Fluid Dynamics (CFD) plays an increasing role in the assessment of the performance prediction of centrifugal compressor stages. Historically, CFD was considered reliable for performance prediction on a qualitatively level, whereas tests were necessary to predict compressors performance on a quantitatively basis. In fact "standard" CFD with only the flow-path and blades included into the computational domain is known to be weak in capturing efficiency level and operating range accurately due to the under-estimation of losses and the lack of secondary flows modeling. This research project aims to fill the gap in accuracy between "standard" CFD and tests data by including a high fidelity reproduction of the gas domain and the use of advanced numerical models and tools introduced in the author's OEM in-house CFD code. In other words, this thesis describes a methodology by which virtual tests can be conducted on single stages and multistage centrifugal compressors in a similar fashion to a typical rig test that guarantee end users to operate machines with a confidence level not achievable before. Furthermore, the new "high fidelity" approach allowed understanding flow phenomena not fully captured before, increasing aerodynamicists capability and confidence in designing high efficiency and high reliable centrifugal compressor stages.

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Lo scopo di questa tesi è presentare un progetto per la realizzazione di una piattaforma DBAAS per il database Jackrabbit. Per piattaforma DBAAS si intende un sistema in grado di offrire ai clienti un database su richiesta (DataBase As A Service), e di amministrare automaticamente i database per conto dei clienti. Il progetto viene realizzato in un primo momento tramite un prototipo della piattaforma, in cui vengono effettuati test intensivi per controllare ogni funzionalità della piattaforma. Il prototipo viene sviluppato in un singolo host, nel quale è possibile simulare tutte le operazioni base della piattaforma DBAAS, come se fosse il sistema finale. In un secondo momento, il progetto sarà realizzato nella sua versione finale, con tutto l'hardware necessario a garantire un servizio ottimale. Questa tesi contiene la base per realizzare il sistema finale, che può essere realizzato in modi diversi a seconda delle scelte che farà il progettista. Ad esempio, è possibile comprare tutto l'hardware necessario e implementare la piattaforma DBAAS in-house, oppure si può scegliere di affidare tutta la gestione hardware ad un servizio di hosting.

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Der atmosphärische Kreislauf reaktiver Stickstoffverbindungen beschäftigt sowohl die Naturwissenschaftler als auch die Politik. Dies ist insbesondere darauf zurückzuführen, dass reaktive Stickoxide die Bildung von bodennahem Ozon kontrollieren. Reaktive Stickstoffverbindungen spielen darüber hinaus als gasförmige Vorläufer von Feinstaubpartikeln eine wichtige Rolle und der Transport von reaktivem Stickstoff über lange Distanzen verändert den biogeochemischen Kohlenstoffkreislauf des Planeten, indem er entlegene Ökosysteme mit Stickstoff düngt. Die Messungen von stabilen Stickstoffisotopenverhältnissen (15N/14N) bietet ein Hilfsmittel, welches es erlaubt, die Quellen von reaktiven Stickstoffverbindungen zu identifizieren und die am Stickstoffkeislauf beteiligten Reaktionen mithilfe ihrer reaktionsspezifischen Isotopenfraktionierung genauer zu untersuchen. rnIn dieser Doktorarbeit demonstriere ich, dass es möglich ist, mit Hilfe von Nano-Sekundärionenmassenspektrometrie (NanoSIMS) verschiedene stickstoffhaltige Verbindungen, die üblicherweise in atmosphärischen Feinstaubpartikeln vorkommen, mit einer räumlichen Auflösung von weniger als einem Mikrometer zu analysieren und zu identifizieren. Die Unterscheidung verschiedener stickstoffhaltiger Verbindungen erfolgt anhand der relativen Signalintensitäten der positiven und negativen Sekundärionensignale, die beobachtet werden, wenn die Feinstaubproben mit einem Cs+ oder O- Primärionenstrahl beschossen werden. Die Feinstaubproben können direkt auf dem Probenahmesubstrat in das Massenspektrometer eingeführt werden, ohne chemisch oder physikalisch aufbereited zu werden. Die Methode wurde Mithilfe von Nitrat, Nitrit, Ammoniumsulfat, Harnstoff, Aminosären, biologischen Feinstaubproben (Pilzsporen) und Imidazol getestet. Ich habe gezeigt, dass NO2 Sekundärionen nur beim Beschuss von Nitrat und Nitrit (Salzen) mit positiven Primärionen entstehen, während NH4+ Sekundärionen nur beim Beschuss von Aminosäuren, Harnstoff und Ammoniumsalzen mit positiven Primärionen freigesetzt werden, nicht aber beim Beschuss biologischer Proben wie z.B. Pilzsporen. CN- Sekundärionen werden beim Beschuss aller stickstoffhaltigen Verbindungen mit positiven Primärionen beobachtet, da fast alle Proben oberflächennah mit Kohlenstoffspuren kontaminiert sind. Die relative Signalintensität der CN- Sekundärionen ist bei kohlenstoffhaltigen organischen Stickstoffverbindungen am höchsten.rnDarüber hinaus habe ich gezeigt, dass an reinen Nitratsalzproben (NaNO3 und KNO3), welche auf Goldfolien aufgebracht wurden speziesspezifische stabile Stickstoffisotopenverhältnisse mithilfe des 15N16O2- / 14N16O2- - Sekundärionenverhältnisses genau und richtig gemessen werden können. Die Messgenauigkeit auf Feldern mit einer Rastergröße von 5×5 µm2 wurde anhand von Langzeitmessungen an einem hausinternen NaNO3 Standard als ± 0.6 ‰ bestimmt. Die Differenz der matrixspezifischen instrumentellen Massenfraktionierung zwischen NaNO3 und KNO3 betrug 7.1 ± 0.9 ‰. 23Na12C2- Sekundärionen können eine ernst zu nehmende Interferenz darstellen wenn 15N16O2- Sekundärionen zur Messung des nitratspezifischen schweren Stickstoffs eingesetzt werden sollen und Natrium und Kohlenstoff im selben Feinstaubpartikel als interne Mischung vorliegt oder die natriumhaltige Probe auf einem kohlenstoffhaltigen Substrat abgelegt wurde. Selbst wenn, wie im Fall von KNO3, keine derartige Interferenz vorliegt, führt eine interne Mischung mit Kohlenstoff im selben Feinstaubpartikel zu einer matrixspezifischen instrumentellen Massenfraktionierung die mit der folgenden Gleichung beschrieben werden kann: 15Nbias = (101 ± 4) ∙ f − (101 ± 3) ‰, mit f = 14N16O2- / (14N16O2- + 12C14N-). rnWird das 12C15N- / 12C14N- Sekundärionenverhältnis zur Messung der stabilen Stickstoffisotopenzusammensetzung verwendet, beeinflusst die Probematrix die Messungsergebnisse nicht, auch wenn Stickstoff und Kohlenstoff in den Feinstaubpartikeln in variablen N/C–Verhältnissen vorliegen. Auch Interferenzen spielen keine Rolle. Um sicherzustellen, dass die Messung weiterhin spezifisch auf Nitratspezies eingeschränkt bleibt, kann eine 14N16O2- Maske bei der Datenauswertung verwendet werden. Werden die Proben auf einem kohlenstoffhaltigen, stickstofffreien Probennahmesubstrat gesammelt, erhöht dies die Signalintensität für reine Nitrat-Feinstaubpartikel.

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In the last years, the European countries have paid increasing attention to renewable sources and greenhouse emissions. The Council of the European Union and the European Parliament have established ambitious targets for the next years. In this scenario, biomass plays a prominent role since its life cycle produces a zero net carbon dioxide emission. Additionally, biomass can ensure plant operation continuity thanks to its availability and storage ability. Several conventional systems running on biomass are available at the moment. Most of them are performant either in the large-scale or in the small power range. The absence of an efficient system on the small-middle scale inspired this thesis project. The object is an innovative plant based on a wet indirectly fired gas turbine (WIFGT) integrated with an organic Rankine cycle (ORC) unit for combined heat and power production. The WIFGT is a performant system in the small-middle power range; the ORC cycle is capable of giving value to low-temperature heat sources. Their integration is investigated in this thesis with the aim of carrying out a preliminary design of the components. The targeted plant output is around 200 kW in order not to need a wide cultivation area and to avoid biomass shipping. Existing in-house simulation tools are used: They are adapted to this purpose. Firstly the WIFGT + ORC model is built; Zero-dimensional models of heat exchangers, compressor, turbines, furnace, dryer and pump are used. Different fluids are selected but toluene and benzene turn out to be the most suitable. In the indirectly fired gas turbine a pressure ratio around 4 leads to the highest efficiency. From the thermodynamic analysis the system shows an electric efficiency of 38%, outdoing other conventional plants in the same power range. The combined plant is designed to recover thermal energy: Water is used as coolant in the condenser. It is heated from 60°C up to 90°C, ensuring the possibility of space heating. Mono-dimensional models are used to design the heat exchange equipment. Different types of heat exchangers are chosen depending on the working temperature. A finned-plate heat exchanger is selected for the WIFGT heat transfer equipment due to the high temperature, oxidizing and corrosive environment. A once-through boiler with finned tubes is chosen to vaporize the organic fluid in the ORC. A plate heat exchanger is chosen for the condenser and recuperator. A quasi-monodimensional model for single-stage axial turbine is implemented to design both the WIFGT and the ORC turbine. The system simulation after the components design shows an electric efficiency around 34% with a decrease by 10% compared to the zero-dimensional analysis. The work exhibits the system potentiality compared to the existing plants from both technical and economic point of view.

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recombinant activated factor VII (rFVIIa) is used off-label for massive bleeding. There is no convincing evidence of the benefits of this practice and the minimal effective dose is unknown. The aim of the study was to evaluate our in-house guideline recommending a low dose of 60 μg/kg for off-label use of rFVIIa.

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OBJECTIVE: Dynamic ventilation (3)He-MRI is a new method to assess pulmonary gas inflow. As differing airway diameters throughout the ventilatory cycle can influence gas inflow this study intends to investigate the influence of volume and timing of a He gas bolus with respect to the beginning of the tidal volume on inspiratory gas distribution. MATERIALS AND METHODS: An ultrafast 2-dimensional spoiled gradient echo sequence (temporal resolution 100 milliseconds) was used for dynamic ventilation (3)He-MRI of 11 anesthetized and mechanically ventilated pigs. The applied (3)He gas bolus was varied in volume between 100 and 200 mL. A 150-mL bolus was varied in its application time after the beginning of the tidal volume between 0 and 1200 milliseconds. Signal kinetics were evaluated using an in-house developed software after definition of parameters for the quantitative description of (3)He gas inflow. RESULTS: The signal rise time (time interval between signal in the parenchyma reaches 10% and 90% of its maximum) was prolonged with increasing bolus volume. The parameter was shortened with increasing delay of (3)He application after the beginning of the tidal volume. Timing variation as well as volume variation showed no clear interrelation to the signal delay time 10 (time interval between signal in the trachea reaches 50% of its maximum and signal in the parenchyma reaches 10% of its maximum). CONCLUSIONS: Dynamic ventilation (3)He-MRI is able to detect differences in bolus geometry performed by volume variation. Pulmonary gas inflow as investigated by dynamic ventilation (3)He-MRI tends to be accelerated by an increasing application delay of a (3)He gas bolus after the beginning of the tidal volume.

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The reserves of gasoline and diesel fuels are ever decreasing, which plays an important role in the technological development of automobiles. Numerous countries, especially the United States, wish to slowly decrease their fuel dependence on other countries by producing in house renewable fuels like biodiesels or ethanol. Therefore, the new automobile engines have to successfully run on a variety of fuels without significant changes to their designs. The current study focuses on assessing the potential of ethanol fuels to improve the performance of 'flex-fuel SI engines,' which literally means 'engines that are flexible in their fuel requirement.' Another important area within spark ignition (SI) engine research is the implementation of new technologies like Variable Valve Timing (VVT) or Variable Compression Ratio (VCR) to improve engine performance. These technologies add more complexity to the original system by adding extra degrees of freedom. Therefore, the potential of these technologies has to be evaluated before they are installed in any SI engine. The current study focuses on evaluating the advantages and drawbacks of these technologies, primarily from an engine brake efficiency perspective. The results show a significant improvement in engine efficiency with the use of VVT and VCR together. Spark ignition engines always operate at a lower compression ratio as compared to compression ignition (CI) engines primarily due to knock constraints. Therefore, even if the use of a higher compression ratio would result in a significant improvement in SI engine efficiency, the engine may still operate at a lower compression ratio due to knock limitations. Ethanol fuels extend the knock limit making the use of higher compression ratios possible. Hence, the current study focuses on using VVT, VCR, and ethanol-gasoline blends to improve overall engine performance. The results show that these technologies promise definite engine performance improvements provided both their positive and negative potentials have been evaluated prior to installation.

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Squeeze film damping effects naturally occur if structures are subjected to loading situations such that a very thin film of fluid is trapped within structural joints, interfaces, etc. An accurate estimate of squeeze film effects is important to predict the performance of dynamic structures. Starting from linear Reynolds equation which governs the fluid behavior coupled with structure domain which is modeled by Kirchhoff plate equation, the effects of nondimensional parameters on the damped natural frequencies are presented using boundary characteristic orthogonal functions. For this purpose, the nondimensional coupled partial differential equations are obtained using Rayleigh-Ritz method and the weak formulation, are solved using polynomial and sinusoidal boundary characteristic orthogonal functions for structure and fluid domain respectively. In order to implement present approach to the complex geometries, a two dimensional isoparametric coupled finite element is developed based on Reissner-Mindlin plate theory and linearized Reynolds equation. The coupling between fluid and structure is handled by considering the pressure forces and structural surface velocities on the boundaries. The effects of the driving parameters on the frequency response functions are investigated. As the next logical step, an analytical method for solution of squeeze film damping based upon Green’s function to the nonlinear Reynolds equation considering elastic plate is studied. This allows calculating modal damping and stiffness force rapidly for various boundary conditions. The nonlinear Reynolds equation is divided into multiple linear non-homogeneous Helmholtz equations, which then can be solvable using the presented approach. Approximate mode shapes of a rectangular elastic plate are used, enabling calculation of damping ratio and frequency shift as well as complex resistant pressure. Moreover, the theoretical results are correlated and compared with experimental results both in the literature and in-house experimental procedures including comparison against viscoelastic dampers.

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PCR tests for the rapid and valid detection of methicillin-resistant Staphylococcus aureus (MRSA) are now available. We evaluated the costs associated with contact screening for MRSA carriage in a tertiary-care hospital with low MRSA endemicity. Between 1 October 2005 and 28 February 2006, 232 patients were screened during 258 screening episodes (644 samples) for MRSA carriage by GenoType MRSA Direct (Hain Lifescience GmbH, Nehren, Germany). Conventional culture confirmed all PCR results. According to in-house algorithms, 34 of 258 screening episodes (14.7%) would have qualified for preemptive contact isolation, but such isolation was not done upon negative PCR results. MRSA carriage was detected in 4 (1.5%) of 258 screening episodes (i.e., in four patients), of which none qualified for preemptive contact isolation. The use of PCR for all 258 screening episodes added costs (in Swiss francs [CHF]) of CHF 104,328.00 and saved CHF 38,528.00 (for preemptive isolation). The restriction of PCR screening to the 34 episodes that qualified for preemptive contact isolation and screening all others by culture would have lowered costs for PCR to only CHF 11,988.00, a savings of CHF 38,528.00. Therefore, PCR tests are valuable for the rapid detection of MRSA carriers, but high costs require the careful evaluation of their use. In patient populations with low MRSA endemicity, the broad use of PCR probably is not cost-effective.

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DNA ligases are important enzymes which catalyze the joining of nicks between adjacent bases of double-stranded DNA. NAD1-dependent DNA ligases (LigA) are essential in bacteria and are absent in humans. They have therefore been identified as novel, validated and attractive drug targets. Using virtual screening against an in-house database of compounds and our recently determined crystal structure of the NAD1 binding domain of the Mycobacterium tuberculosis LigA, we have identified N1, Nn-bis-(5-deoxy-a-D-xylofuranosylated) diamines as a novel class of inhibitors for this enzyme. Assays involving M.tuberculosis LigA, T4 ligase and human DNA ligase I show that these compounds specifically inhibit LigA from M.tuberculosis. In vitro kinetic and inhibition assays demonstrate that the compounds compete with NAD1 for binding and inhibit enzyme activity with IC50 values in the mM range. Docking studies rationalize the observed specificities and show that among several glycofuranosylated diamines, bis xylofuranosylated diamines with aminoalkyl and 1, 3-phenylene carbamoyl spacers mimic the binding modes of NAD1 with the enzyme. Assays involving LigA-deficient bacterial strains show that in vivo inhibition of ligase by the compounds causes the observed antibacterial activities. They also demonstrate that the compounds exhibit in vivo specificity for LigA over ATPdependent ligase. This class of inhibitors holds out the promise of rational development of new anti-tubercular agents.