Development of a comprehensive on-line multidimensional high performance column liquid chromatography system for protein and peptide mapping with integrated size selective sample fractionation

Aufbau einer kontinuierlichen, mehrdimensionalen Hochleistungs-flüssigchromatographie-Anlage für die Trennung von Proteinen und Peptiden mit integrierter größenselektiver ProbenfraktionierungEs wurde eine mehrdimensionale HPLC-Trennmethode für Proteine und Peptide mit einem Molekulargewicht von <15 kDa entwickelt.Im ersten Schritt werden die Zielanalyte von höhermolekularen sowie nicht ionischen Bestandteilen mit Hilfe von 'Restricted Access Materialien' (RAM) mit Ionenaustauscher-Funktionalität getrennt. Anschließend werden die Proteine auf einer analytischen Ionenaustauscher-Säule sowie auf Reversed-Phase-Säulen getrennt. Zur Vermeidung von Probenverlusten wurde ein kontinuierlich arbeitendes, voll automatisiertes System auf Basis unterschiedlicher Trenngeschwindigkeiten und vier parallelen RP-Säulen aufgebaut.Es werden jeweils zwei RP-Säulen gleichzeitig, jedoch mit zeitlich versetztem Beginn eluiert, um durch flache Gradienten ausreichende Trennleistungen zu erhalten. Während die dritte Säule regeneriert wird, erfolgt das Beladen der vierte Säule durch Anreicherung der Proteine und Peptide am Säulenkopf. Während der Gesamtanalysenzeit von 96 Minuten werden in Intervallen von 4 Minuten Fraktionen aus der 1. Dimension auf die RP-Säulen überführt und innerhalb von 8 Minuten getrennt, wobei 24 RP-Chromatogramme resultieren.Als Testsubstanzen wurden u.a. Standardproteine, Proteine und Peptide aus humanem Hämofiltrat sowie aus Lungenfibroblast-Zellkulturüberständen eingesetzt. Weiterhin wurden Fraktionen gesammelt und mittels MALDI-TOF Massenspektrometrie untersucht. Bei einer Injektion wurden in den 24 RP-Chromatogrammen mehr als 1000 Peaks aufgelöst. Der theoretische Wert der Peakkapazität liegt bei ungefähr 3000.

Steel and macro-synthetic self-compacting fibre reinforced concrete, experimental study on the long-term deformations

Experimental study on the long-term deformations of the fibre reinforced concrete. Steel and macro-synthetic fibers were used to evaluate the shrinkage, creep, mid-span deflection, cracking and rupture analysis of three different types of samples. At the end the main topics of ACI guidelines were analyzed in order to perform an overview of design.

Pushover analysis of an existing reinforced concrete bridge:Jamboree Road Overcrossing in Irvine, California

The work for the present thesis started in California, during my semester as an exchange student overseas. California is known worldwide for its seismicity and its effort in the earthquake engineering research field. For this reason, I immediately found interesting the Structural Dynamics Professor, Maria Q. Feng's proposal, to work on a pushover analysis of the existing Jamboree Road Overcrossing bridge. Concrete is a popular building material in California, and for the most part, it serves its functions well. However, concrete is inherently brittle and performs poorly during earthquakes if not reinforced properly. The San Fernando Earthquake of 1971 dramatically demonstrated this characteristic. Shortly thereafter, code writers revised the design provisions for new concrete buildings so to provide adequate ductility to resist strong ground shaking. There remain, nonetheless, millions of square feet of non-ductile concrete buildings in California. The purpose of this work is to perform a Pushover Analysis and compare the results with those of a Nonlinear Time-History Analysis of an existing bridge, located in Southern California. The analyses have been executed through the software OpenSees, the Open System for Earthquake Engineering Simulation. The bridge Jamboree Road Overcrossing is classified as a Standard Ordinary Bridge. In fact, the JRO is a typical three-span continuous cast-in-place prestressed post-tension box-girder. The total length of the bridge is 366 ft., and the height of the two bents are respectively 26,41 ft. and 28,41 ft.. Both the Pushover Analysis and the Nonlinear Time-History Analysis require the use of a model that takes into account for the nonlinearities of the system. In fact, in order to execute nonlinear analyses of highway bridges it is essential to incorporate an accurate model of the material behavior. It has been observed that, after the occurrence of destructive earthquakes, one of the most damaged elements on highway bridges is a column. To evaluate the performance of bridge columns during seismic events an adequate model of the column must be incorporated. Part of the work of the present thesis is, in fact, dedicated to the modeling of bents. Different types of nonlinear element have been studied and modeled, with emphasis on the plasticity zone length determination and location. Furthermore, different models for concrete and steel materials have been considered, and the selection of the parameters that define the constitutive laws of the different materials have been accurate. The work is structured into four chapters, to follow a brief overview of the content. The first chapter introduces the concepts related to capacity design, as the actual philosophy of seismic design. Furthermore, nonlinear analyses both static, pushover, and dynamic, time-history, are presented. The final paragraph concludes with a short description on how to determine the seismic demand at a specific site, according to the latest design criteria in California. The second chapter deals with the formulation of force-based finite elements and the issues regarding the objectivity of the response in nonlinear field. Both concentrated and distributed plasticity elements are discussed into detail. The third chapter presents the existing structure, the software used OpenSees, and the modeling assumptions and issues. The creation of the nonlinear model represents a central part in this work. Nonlinear material constitutive laws, for concrete and reinforcing steel, are discussed into detail; as well as the different scenarios employed in the columns modeling. Finally, the results of the pushover analysis are presented in chapter four. Capacity curves are examined for the different model scenarios used, and failure modes of concrete and steel are discussed. Capacity curve is converted into capacity spectrum and intersected with the design spectrum. In the last paragraph, the results of nonlinear time-history analyses are compared to those of pushover analysis.

Fiber beam-columns models with flexure-shear interaction for nonlinear analysis of reinforced concrete structures

The aim of this study was to develop a model capable to capture the different contributions which characterize the nonlinear behaviour of reinforced concrete structures. In particular, especially for non slender structures, the contribution to the nonlinear deformation due to bending may be not sufficient to determine the structural response. Two different models characterized by a fibre beam-column element are here proposed. These models can reproduce the flexure-shear interaction in the nonlinear range, with the purpose to improve the analysis in shear-critical structures. The first element discussed is based on flexibility formulation which is associated with the Modified Compression Field Theory as material constitutive law. The other model described in this thesis is based on a three-field variational formulation which is associated with a 3D generalized plastic-damage model as constitutive relationship. The first model proposed in this thesis was developed trying to combine a fibre beamcolumn element based on the flexibility formulation with the MCFT theory as constitutive relationship. The flexibility formulation, in fact, seems to be particularly effective for analysis in the nonlinear field. Just the coupling between the fibre element to model the structure and the shear panel to model the individual fibres allows to describe the nonlinear response associated to flexure and shear, and especially their interaction in the nonlinear field. The model was implemented in an original matlab® computer code, for describing the response of generic structures. The simulations carried out allowed to verify the field of working of the model. Comparisons with available experimental results related to reinforced concrete shears wall were performed in order to validate the model. These results are characterized by the peculiarity of distinguishing the different contributions due to flexure and shear separately. The presented simulations were carried out, in particular, for monotonic loading. The model was tested also through numerical comparisons with other computer programs. Finally it was applied for performing a numerical study on the influence of the nonlinear shear response for non slender reinforced concrete (RC) members. Another approach to the problem has been studied during a period of research at the University of California Berkeley. The beam formulation follows the assumptions of the Timoshenko shear beam theory for the displacement field, and uses a three-field variational formulation in the derivation of the element response. A generalized plasticity model is implemented for structural steel and a 3D plastic-damage model is used for the simulation of concrete. The transverse normal stress is used to satisfy the transverse equilibrium equations of at each control section, this criterion is also used for the condensation of degrees of freedom from the 3D constitutive material to a beam element. In this thesis is presented the beam formulation and the constitutive relationships, different analysis and comparisons are still carrying out between the two model presented.

Pore structure and column efficiency of silica monoliths in high performance liquid chromatography (HPLC)

Five different methods were critically examined to characterize the pore structure of the silica monoliths. The mesopore characterization was performed using: a) the classical BJH method of nitrogen sorption data, which showed overestimated values in the mesopore distribution and was improved by using the NLDFT method, b) the ISEC method implementing the PPM and PNM models, which were especially developed for monolithic silicas, that contrary to the particulate supports, demonstrate the two inflection points in the ISEC curve, enabling the calculation of pore connectivity, a measure for the mass transfer kinetics in the mesopore network, c) the mercury porosimetry using a new recommended mercury contact angle values. rnThe results of the characterization of mesopores of monolithic silica columns by the three methods indicated that all methods were useful with respect to the pore size distribution by volume, but only the ISEC method with implemented PPM and PNM models gave the average pore size and distribution based on the number average and the pore connectivity values.rnThe characterization of the flow-through pore was performed by two different methods: a) the mercury porosimetry, which was used not only for average flow-through pore value estimation, but also the assessment of entrapment. It was found that the mass transfer from the flow-through pores to mesopores was not hindered in case of small sized flow-through pores with a narrow distribution, b) the liquid penetration where the average flow-through pore values were obtained via existing equations and improved by the additional methods developed according to Hagen-Poiseuille rules. The result was that not the flow-through pore size influences the column bock pressure, but the surface area to volume ratio of silica skeleton is most decisive. Thus the monolith with lowest ratio values will be the most permeable. rnThe flow-through pore characterization results obtained by mercury porosimetry and liquid permeability were compared with the ones from imaging and image analysis. All named methods enable a reliable characterization of the flow-through pore diameters for the monolithic silica columns, but special care should be taken about the chosen theoretical model.rnThe measured pore characterization parameters were then linked with the mass transfer properties of monolithic silica columns. As indicated by the ISEC results, no restrictions in mass transfer resistance were noticed in mesopores due to their high connectivity. The mercury porosimetry results also gave evidence that no restrictions occur for mass transfer from flow-through pores to mesopores in the small scaled silica monoliths with narrow distribution. rnThe prediction of the optimum regimes of the pore structural parameters for the given target parameters in HPLC separations was performed. It was found that a low mass transfer resistance in the mesopore volume is achieved when the nominal diameter of the number average size distribution of the mesopores is appr. an order of magnitude larger that the molecular radius of the analyte. The effective diffusion coefficient of an analyte molecule in the mesopore volume is strongly dependent on the value of the nominal pore diameter of the number averaged pore size distribution. The mesopore size has to be adapted to the molecular size of the analyte, in particular for peptides and proteins. rnThe study on flow-through pores of silica monoliths demonstrated that the surface to volume of the skeletons ratio and external porosity are decisive for the column efficiency. The latter is independent from the flow-through pore diameter. The flow-through pore characteristics by direct and indirect approaches were assessed and theoretical column efficiency curves were derived. The study showed that next to the surface to volume ratio, the total porosity and its distribution of the flow-through pores and mesopores have a substantial effect on the column plate number, especially as the extent of adsorption increases. The column efficiency is increasing with decreasing flow through pore diameter, decreasing with external porosity, and increasing with total porosity. Though this tendency has a limit due to heterogeneity of the studied monolithic samples. We found that the maximum efficiency of the studied monolithic research columns could be reached at a skeleton diameter of ~ 0.5 µm. Furthermore when the intention is to maximize the column efficiency, more homogeneous monoliths should be prepared.rn

Optimized scheduling in real-time environments with column generation

Im Bereich sicherheitsrelevanter eingebetteter Systeme stellt sich der Designprozess von Anwendungen als sehr komplex dar. Entsprechend einer gegebenen Hardwarearchitektur lassen sich Steuergeräte aufrüsten, um alle bestehenden Prozesse und Signale pünktlich auszuführen. Die zeitlichen Anforderungen sind strikt und müssen in jeder periodischen Wiederkehr der Prozesse erfüllt sein, da die Sicherstellung der parallelen Ausführung von größter Bedeutung ist. Existierende Ansätze können schnell Designalternativen berechnen, aber sie gewährleisten nicht, dass die Kosten für die nötigen Hardwareänderungen minimal sind. Wir stellen einen Ansatz vor, der kostenminimale Lösungen für das Problem berechnet, die alle zeitlichen Bedingungen erfüllen. Unser Algorithmus verwendet Lineare Programmierung mit Spaltengenerierung, eingebettet in eine Baumstruktur, um untere und obere Schranken während des Optimierungsprozesses bereitzustellen. Die komplexen Randbedingungen zur Gewährleistung der periodischen Ausführung verlagern sich durch eine Zerlegung des Hauptproblems in unabhängige Unterprobleme, die als ganzzahlige lineare Programme formuliert sind. Sowohl die Analysen zur Prozessausführung als auch die Methoden zur Signalübertragung werden untersucht und linearisierte Darstellungen angegeben. Des Weiteren präsentieren wir eine neue Formulierung für die Ausführung mit fixierten Prioritäten, die zusätzlich Prozessantwortzeiten im schlimmsten anzunehmenden Fall berechnet, welche für Szenarien nötig sind, in denen zeitliche Bedingungen an Teilmengen von Prozessen und Signalen gegeben sind. Wir weisen die Anwendbarkeit unserer Methoden durch die Analyse von Instanzen nach, welche Prozessstrukturen aus realen Anwendungen enthalten. Unsere Ergebnisse zeigen, dass untere Schranken schnell berechnet werden können, um die Optimalität von heuristischen Lösungen zu beweisen. Wenn wir optimale Lösungen mit Antwortzeiten liefern, stellt sich unsere neue Formulierung in der Laufzeitanalyse vorteilhaft gegenüber anderen Ansätzen dar. Die besten Resultate werden mit einem hybriden Ansatz erzielt, der heuristische Startlösungen, eine Vorverarbeitung und eine heuristische mit einer kurzen nachfolgenden exakten Berechnungsphase verbindet.

The effect of the ductility on the collapse loads: The failure of a steel roof

Studio dell'effetto della duttilità sul carico di collasso: il crollo di una copertura reticolare spaziale in acciaio causato dalla compresenza di un'ingente mancanza di resistenza e duttilità nei collegamenti delle aste e possibili stati coattivi. Il carico di collasso nel caso di strutture a bassa duttilità dipende dalla presenza di stati coattivi dovuti a imperfezioni di realizzazione o cedimenti vincolari della struttura in esercizio.

Using the Natural Period of a Structure as an Indicator of the Significance of Second-Order Effects

This study investigates the feasibility of predicting the momentamplification in beam-column elements of steel moment-resisting frames using the structure's natural period. Unlike previous methods, which perform moment-amplification on a story-by-story basis, this study develops and tests two models that aim to predict a global amplification factor indicative of the largest relevant instance of local moment amplification in the structure. To thisend, a variety of two-dimensional frames is investigated using first and secondorder finite element analysis. The observed moment amplification is then compared with the predicted amplification based on the structure's natural period, which is calculated by first-order finite element analysis. As a benchmark, design moment amplification factors are calculated for each story using the story stiffness approach, and serve to demonstrate the relativeconservatism and accuracy of the proposed models with respect to current practice in design. The study finds that the observed moment amplification factors may vastly exceed expectations when internal member stresses are initially very small. Where the internal stresses are small relative to the member capacities, thesecases are inconsequential for design. To qualify the significance of the observed amplification factors, two parameters are used: the second-order moment normalized to the plastic moment capacity, and the combined flexural and axial stress interaction equations developed by AISC