1 ABSTRACT The diploma theses deals with marketing in practice of social services. Marketing brings positive effects in the field of client, personal and material-technical capacities to some organisations. This can bring financial as well as nonfinancial profit together with bigger independence from donors who provide means of running of social services. Theses will help to solve problems of some social services managers who don't care about marketing in the organisation sufficiently. The object of this theses is to choose marketing goals and strategies of their obtainment on the basis of marketing macromedium and site analysis of organisation and insert them into the marketing plan in the particular social service, which moves in the competitive setting of other social services. The theses output is marketing plan of social service. Key words: marketing, social service, competition, marketing plan, site analysis, management

Indiana University-Purdue University Indianapolis (IUPUI) / Lentiviral vector mediated gene therapy has made great strides in recent years with several successful clinical trials. However, adverse events encountered with some early trials have highlighted the necessity to improve upon its safety. Improvements can range from early steps in vector production to evaluation of insertion sites post-transduction. We have evaluated an FDA approved DNase for removal of residual plasmid DNA during vector production, developed novel non-integrating lentiviral vectors and employed modified insertion site analysis post-transduction to improve the safety of lentiviral vector mediated gene therapy. To prevent the exposure of gene therapy patients to HIV-1 DNA it is essential to remove residual plasmid DNA during vector production. We evaluated a recombinant human DNase which has been FDA approved for use in patients as an alternative to a bacterially derived DNase. Our results indicate this DNase is an effective alternative with a potentially safer profile for use in patients. The ability of lentiviral vectors to stably integrate their genome into a host cell’s DNA can have negative side-effects due to the risk of insertional mutagenesis. Non-integrating lentiviral vectors have been developed to alleviate this risk in applications where integration is not necessary. However, a low frequency of illegitimate integration persists when using these vectors. We have developed a novel non-integrating vector mutation and evaluated the efficacy of combining it with other mutations for reducing the frequency of illegitimate integration. We demonstrate that combining mutations that inhibit integration can further reduce the frequency of illegitimate integration. Several methodologies have been developed for evaluating the insertion sites of normal integrating lentiviral vectors. Illegitimate integration by non-integrating vectors demonstrates mechanisms which result in insertions and/or deletions at the vector-genome junction. Current methods lack the sensitivity to account for these variables in a high-throughput manner. We have adapted modifications to current methods to improve the capture of these variable insertion sites for analysis. The results of these studies improve the safety of lentiviral vector mediated gene therapy by improving the purity of the vector product, providing a safer vector for non-integrase mediated applications, and allowing more sensitive analysis of insertion sites post-transduction.

No description available.

Solid phase microextraction (SPME), needle trap device (NTD), and membrane extraction with a sorbent interface (MESI) are solvent-free sample preparation techniques that were developed to perform the rapid routine analysis of organic compounds (VOCs) in various environmental matrices by integrating sampling, extraction, preconcentration and sample introduction procedures into one step. A portable ion mobility spectrometry (IMS) analyzer has some advantages, such as small size, light weight, operability under ambient pressure, air as carrier gas, and sensitivity, all of which make IMS suitable for on-site monitoring for low concentration of analytes. The aforementioned sampling and preconcentration techniques were coupled with a portable IMS analyzer, as well as a thermal desorption unit that can accommodate SPME, NTD and MESI, which was modified and combined with IMS for on-site monitoring of volatile organic compounds (VOCs) from human breath and plant emissions. Experimental results demonstrated that low detection limits were achievable for gaseous analytes, (25 ng/L for acetone (SPME-IMS), 43 ng/mL (NTD-IMS) and 2.3 ng/mL (MESI-IMS) for α-pinene). These three analytical systems were applied for on-site rapid determination of acetone in human breath and α-pinene from plant emissions respectively. The salient features of these systems that make them suitable for on-site monitoring of volatile organic compounds in different sources are: small size, simple operation, fast and/or on-line sampling, rapid analysis.

Solid phase microextraction (SPME), needle trap device (NTD), and membrane extraction with a sorbent interface (MESI) are solvent-free sample preparation techniques that were developed to perform the rapid routine analysis of organic compounds (VOCs) in various environmental matrices by integrating sampling, extraction, preconcentration and sample introduction procedures into one step. A portable ion mobility spectrometry (IMS) analyzer has some advantages, such as small size, light weight, operability under ambient pressure, air as carrier gas, and sensitivity, all of which make IMS suitable for on-site monitoring for low concentration of analytes. The aforementioned sampling and preconcentration techniques were coupled with a portable IMS analyzer, as well as a thermal desorption unit that can accommodate SPME, NTD and MESI, which was modified and combined with IMS for on-site monitoring of volatile organic compounds (VOCs) from human breath and plant emissions. Experimental results demonstrated that low detection limits were achievable for gaseous analytes, (25 ng/L for acetone (SPME-IMS), 43 ng/mL (NTD-IMS) and 2.3 ng/mL (MESI-IMS) for α-pinene). These three analytical systems were applied for on-site rapid determination of acetone in human breath and α-pinene from plant emissions respectively. The salient features of these systems that make them suitable for on-site monitoring of volatile organic compounds in different sources are: small size, simple operation, fast and/or on-line sampling, rapid analysis.

La turbulence optique et son impact sur les images obtenues à partir d'instruments de mesure est un phénomène connu dans le domaine de l'astronomie. Des modèles issus de la théorie de Kolmogorov, développée pour une turbulence dynamique, ainsi que des méthodes de correction, telles que l'optique adaptative, existent pour l'analyse et la compensation des effets de cette turbulence optique. L'analyse de cette dernière dans les milieux confinés est cependant plus limitée. Les sources susceptibles de générer une turbulence optique dans ces espaces sont pourtant multiples et peuvent avoir un impact non négligeable sur les mesures des instruments installés à proximité. Ce mémoire constitue une première approche de la caractérisation de la turbulence optique dans un espace confiné. Après l'introduction des phénomènes étudiés et des outils mathématiques employés, ce mémoire présente les résultats issus d'analyses de coupoles de télescopes et de caractérisations de salles blanches employées par l'entreprise Thales Alenia Space pour l'intégration et le test d'instruments optiques. Ces résultats sont obtenus avec l'instrument INTENSE, développé durant la thèse pour la caractérisation de la turbulence optique locale via la mesure des fluctuations des angles d'arrivée de multiples faisceaux lasers. En prévision de futurs améliorations de l'analyse de la turbulence en espace confiné, un chapitre du mémoire est dédié aux travaux réalisés sur une méthode d'extraction du profil de l'énergie de la turbulence et à son application sur l'instrument INTENSE. Les conclusions et les perspectives des travaux réalisés pendant la thèse sont présentées à la fin du mémoire. / Optical turbulence and its impact on measured images is a well-known phenomenon in astronomy. Models based on the Kolmogorov theory, elaborated for a dynamical turbulence description, and methods, such as Adaptive Optics, were both developed so as to understand and correct the degradations caused by this turbulence. Analysis of the same phenomenon in indoor situation was however less investigated. The local air volume is nonetheless prone to optical perturbations sources which could have non negligible impacts on the measurements of instruments installed at proximity. This document introduces a first approach of indoor optical turbulence characterization. After the introduction of the studied phenomenon and the mathematical tools employed, this thesis present optical turbulence characterizations inside Thales Alenia Space clean rooms used for optical instrument integration and testing. Analyses inside telescope domes are also shown in this document. All the results were obtained using the INTENSE instrument which was developed during the thesis for optical turbulence characterizations using angle of arrival fluctuations of laser beams. In anticipation for future ameliorations of optical turbulence analysis methods, a chapter of this thesis is dedicated to the work made on a turbulence energy profile extraction and its application on the INTENSE instrument. Conclusions and perspectives of the work made during this thesis are presented at the end of the document.

Der seit vielen Jahren etablierte Summenparameter DOC (dissolved organic carbon - gelöster organischer Kohlenstoff) wird u. a. standardmäßig für die Überwachung der Wasserqualität und zur Bewertung der Effizienz von Verfahren der Wasserbehandlung eingesetzt. Für eine möglichst kurze Reaktionszeit, z. B. im Fall von Havarien oder anderen Ereignissen, sind zeitlich hoch aufgelöste und verlässliche DOC-Daten von großer Bedeutung. Die konventionellen DOC-Bestimmungsmethoden basieren in der Regel nach einer Probenahme auf Konservierung, Transport und anschließender Analyse der Probe in einem stationären Labor. Bei einer schlechten Laborinfrastruktur birgt dieser Ansatz das Risiko, durch längere Lagerung bzw. Transport der Proben falsche und verspätete Ergebnisse zu liefern. Das Ziel der vorliegenden Arbeit be-stand darin, einen tragbaren und dennoch zuverlässigen Analysator für die DOC-Analyse vor Ort zu entwickeln. Basis dieses Messgerätes bildet eine ungeteilte, gal-vanostatisch betriebene Elektrolysezelle mit bordotierter Diamantelektrode. Die anodische Generierung von •OH- und SO4•--Radikalen als Oxidationsmittel, die In- situ-Bildung des Trägergases und die Kinetik des DOC-Abbaus wurden systematisch untersucht. Zudem wurde der Einfluss wichtiger verfahrenstechnischer und hydro-chemischer Parameter betrachtet und für eine Anwendung in der Umweltanalytik optimiert. Abgeschlossen wurde die Entwicklung mit der technischen Optimierung eines tragbaren Prototyps. Dieser wurde validiert und mit etablierten Verfahren ver-glichen sowie erfolgreich im Rahmen eines aktuellen Monitoringprogramms in Nordindien eingesetzt. Die wichtigsten Merkmale des tragbaren DOC-Analysators sind die kompakte Bauweise sowie der Verzicht auf hochreine Gase, Katalysatoren oder eine Verbrennungstechnologie. Zudem weist die Bestimmungsmethode eine hohe Robustheit in Bezug auf Art und Konzentration der Analyten sowie der Matrix auf und ermöglicht eine kurze Analysezeit sowie eine zuverlässige Bestimmung des DOC vor Ort. / Dissolved organic carbon (DOC) is an indispensable analytical parameter for water quality control. This applies to both monitoring of aquatic ecosystems and evaluation of the efficiency of drinking and waste water treatment processes. The conventional DOC determination methods consist of on-site sampling and subsequent analysis in a stationary laboratory. Storage and transport of samples may lead to significant un-certainties in measurement results and delayed findings. For this reason, the availability of suitable on-site analysis techniques is particularly important in coun-tries with an insufficient laboratory infrastructure. Therefore, the objective of the present study was to develop a portable and reliable analyzer for on-site DOC analy-sis. The core element of this analyzer is an undivided electrolysis cell equipped with a boron doped diamond electrode for an in-situ generation of •OH and SO4•- radicals as oxidizing agents. The influence of process and hydrochemical parameters on the generation of in-situ oxidizing agents, in-situ carrier gas and the DOC decomposition kinetic were systematically investigated and optimized for environmental analyses. Finally, a technical optimization of the portable prototype was conducted. The result-ing prototype was validated and successfully compared with established methods. Subsequently, the developed prototype was used in an ongoing monitoring program in northern India. The key features of the portable DOC analyzer are the independ-ence from high-purity gases, catalysts or combustion technology. In addition, this determination method shows a high robustness in terms of type and concentration of analytes and matrices. The short analysis time as well as a reliable determination of the DOC on site prove the practicality of the new technical approach.

Este trabalho refere-se ao estabelecimento de populações humanas em tempos pretéritos e aos processos de interação que podem se desenvolver entre elas, visualisados pela arqueologia. Buscamos desvendar como ocorreu a ocupação humana (anterior a colonização) no Vale do Rio Soturno, região central do estado do Rio Grande do Sul, Brasil. Os sítios arqueológicos estudados espalhados por esta micro-região foram: Várzea dos Bugres; Cerro dos Bugres; Cerro do Tope; Moacir Rossato, com remanescentes culturais atribuídos aos Guarani. Tentamos entender quais as escolhas que estes grupos fizeram em relação aos recursos ambientais locais, suas estratégias de exploração, as relações entre si e as escolhas na elaboração de seus utensílios remanescentes de uso cotidiano. Percebemos que os grupos atuantes neste espaço optaram por adotar algumas características técnicas específicas, mas no geral seguem o padrão clássico de ocupação Guarani. Realizamos ainda um trabalho com a comunidade que vive no local atualmente, utilizando a metodologia da educação patrimonial, a fim de que esse patrimônio cultural estudado se tornasse conhecido e aceito junto àqueles que serão seus guardiões. Apesar de não contarmos com um número de sítios relevante para um estudo de caráter regional, acreditamos ter contribuído para a arqueologia Guarani apresentando especificidades culturais de grupos que ocuparam um local que até então não fora estudado. / This search refers to the establishment of human populations in past times and the interaction process that can develop between them, shown by archaeology. We seek to discover how human occupation occurred (before colonization) in Sortuno river valley, in central Rio Grande do Sul, Brazil. The archaeological sites studied in this micro-region were: Várzea dos Bugres; Cerro dos Bugres; Cerro do Tope; Moacir Rossato, with the remaining cultures attributed mostly to the Guarani. We try to understand what choices these groups have in relation to local environmental resources, their exploration strategies, relations between themselves and the choices in the development of their remaining fixtures of everyday use. We realize that groups working in this area have chosen to adopt some specific technical features, but in general follow the classic pattern Guarani occupation. We also worked with the community that lives on site using the heritage education methodology, so that the cultural heritage studied could become known and accepted between those who will be their guardians. Although we could not count with a certain number of sites that study regional character, we believe to have contributed it to archaeological Guarani, showing specific cultural groups that occupied a place that until then had not been studied.

In den letzten Jahren gerieten phenolische Substanzen, wie z.B. Chlor-, Nitrophenol oder Alkylphenolethoxylate aufgrund ihrer Toxizität sowie ihres kanzerogenen und endokrinen Potentials in das Interesse der Öffentlichkeit. Diese Substanzen gelangen in großen Mengen, z.B. aus industriellen Prozessen (Papier-, Kunststoff-, oder Lederindustrie) oder als Abbauprodukte von Pflanzenschutzmitteln in die Umwelt.<br /> <br /> Ziel dieser Arbeit war es, einfache biochemische Bestimmungsmethoden für verschiedene phenolische Umweltschadstoffe auf Basis biochemischer Erkennungselemente zu entwickeln. Diese sollten als Screeningmethoden in der Vor-Ort-Analytik einsetzbar sein. Die Anwendung sollte kostengünstig und einfach durchzuführen sein, so dass die Messung kein hochwissenschaftliches Personal erfordert. Daher stand im Hintergrund der Arbeit die Integration der Analysenmethode in ein kompaktes Handgerät.<br /> <br /> Zu diesem Zweck wurde ein Biosensor entwickelt der zur direkten Messung und in Kombination mit einem Immunoassay einsetzbar ist:<br /> <br /> 1.) Elektrochemischer Biosensor<br /> Ein elektrochemischer Biosensor stellt die Verbindung zwischen einer Elektrode und der biologischen Komponente dar. Als Messprinzip wurde die Amperometrie gewählt. Hierbei wird die Präsenz des nachzuweisenden Stoffes durch die angelegte Spannung am Sensor visualisiert, da beim Vorhandensein ein Stromfluss gemessen wird.<br /> <br /> Um die Signalintensität zu erhöhen können Enzyme als Katalysatoren genutzt werden, die in der Lage sind die Rückreaktion der Elektrodenreaktion zu realisieren. In diesem Fall wurde Glucose-Dehydrogenase (GDH) verwendet, die oxidierte phenolische Verbindungen reduzieren kann. Zusammen mit der Oxidation an der Sensoroberfläche bildet sich ein Verstärkungszyklus aus, der das ursprüngliche Signal vielfach erhöht.<br /> <br /> Wir waren in der Lage, GDH durch Einbetten in ein Polymerennetzwerk auf der Oberfläche einer gedruckten Platin-Dickschicht-Elektrode zu immobilisieren. Als Resultat erhielten wir einen sehr empfindlichen und äußerst stabilen Biosensor. Seine schnelle Ansprechzeit ermöglicht den Einsatz in automatisierten Fließsystemen zur Messung großer Probenzahlen. Der Einsatz in einem manuell betriebenen Handgerät konnte ebenfalls realisiert werden und brachte nur geringe Beeinträchtigungen in bezug auf die Empfindlichkeit der Messung. Die erfolgreiche Implementierung des Biosensors in das Handgerät wurde in Rahmen eines internationalen Workshops in Barcelona, anhand der Überprüfung der Reinigungsleistung von Klärwerken, gezeigt.<br /> <br /> 2.) Kombination mit Immunoassays<br /> Der Einsatzbereich der GDH-Biosensoren lässt sich durch die Kombination mit anderen Techniken erweitern, wobei der Sensor zur Visualisierung der Nachweisreaktion dient. In diesem Fall kann der Sensor zur Bestimmung der Enzymaktivität von ß?Galactosidase (ßGal) verwendet werden. Der Nachweis geringster Enzymmengen wurde realisiert. Die ßGal wird zur Markierung eines Analytanalogen in Immunoassays verwendet, um die Bindung von Antikörper und Analytmolekül sichtbar zu machen. Im Immunoassay bildet sich ein Gleichgewicht zwischen Antikörper, unmarkiertem Analyt und markiertem Analytanalog (Tracer) aus. Über die Bestimmung der Enzymaktivität kann man die Analytkonzentration in der Probe errechnen.<br /> <br /> Wir haben unseren GDH-Biosensor erfolgreich mit zwei Techniken kombiniert. Zum Einen mit einem Assay zur Bestimmung von Nitrophenol, der in einem automatisiertem Fließsystem realisiert wurde. Hier wird die Mischung aus Antikörpern, Analyt und Tracer über eine Säule gegeben und gespült. Die gebundenen Bestandteile werden durch den GDH-Biosensor quantifiziert.<br /> <br /> Zum Anderen wurde ein Kapillarimmunoassay entwickelt, der in das Handgerät integriert werden kann. Dabei wird der Antikörper direkt an der Kapillare fixiert. Die Probe wird mit Tracer vermischt und in die Kapillare gegeben. Dort bildet sich das Gleichgewicht aus und weitere Probenbestandteile werden im Spülschritt eliminiert. Die Analytkonzentration wird durch die Bestimmung des gebunden Tracers (Aktivität der ßGal) mit Hilfe des GDH-Biosensors realisiert. / The development of fast and reliable biochemical tools for on-site screening in environmental analysis was the main target of the present work. Due to various hazardous effects such as endocrine disruption and toxicity phenolic compounds are key analytes in environmental analysis and thus were chosen as model analytes.<br /> <br /> Three different methods were developed:<br /> <br /> For the enzymatic detection of phenols in environmental samples an enzyme-based biosensor was developed. In contrast to reported work using tyrosinase or peroxidases, we developed a biosensor based on glucose dehydrogenase as biorecognition element. This biosensor was devoted for an application in a laboratory flow system as well as in a portable device for on-site measurements.<br /> <br /> This enzymatic detection is applicable only for a limited number of phenols due to substrate specificity of the enzyme. For other relevant compounds based on a phenolic structure (i.e. nitrophenol, alkylphenols and alkylphenol ethoxylates) immunological methods had to be developed. The electrochemical GDH-biosensor was used as the label detector in these immunoassays.<br /> <br /> Two heterogeneous immunoassays were developed where ßGal was used as the label. An electrochemical method for the determination of the marker enzyme activity was processed. The separation step was realized with protein A/G columns (laboratory flow system) or by direct immobilization of the antibodies in small disposable capillaries (on-site analysis). <br /> <br /> All methods were targeted on the contemporary analysis of small numbers of samples.

Calibration methods for quantitative on-site sampling using solid phase microextraction (SPME) were developed based on diffusion mass transfer theory. This was investigated using adsorptive polydimethylsiloxane/divinylbenzene (PDMS/DVB) and Carboxen/polydimethylsiloxane (CAR/PDMS) SPME fiber coatings with volatile aromatic hydrocarbons (BTEX: benzene, toluene, ethylbenzene, and o-xylene) as test analytes. Parameters that affected the extraction process (sampling time, analyte concentration, water velocity, and temperature) were investigated. Very short sampling times (10-300 s) and sorbents with a strong affinity and large capacity were used to ensure a 'zero sink' effect calibrate process. It was found that mass uptake of analyte changed linearly with concentration. Increase of water velocity increased mass uptake, though the increase is not linear. Temperature did not affect mass uptake significantly under typical field sampling conditions. To further describe rapid SPME analysis of aqueous samples, a new model translated from heat transfer to a circular cylinder in cross flow was used. An empirical correlation to this model was used to predict the mass transfer coefficient. Findings indicated that the predicted mass uptake compared well with experimental mass uptake. The new model also predicted rapid air sampling accurately. To further integrate the sampling and analysis processes, especially for on-site or <i>in-vivo</i> investigations where the composition of the sample matrix is very complicated and/or agitation of the sample matrix is variable or unknown, a new approach for calibration was developed. This involved the loading internal standards onto the extraction fiber prior to the extraction step. During sampling, the standard partially desorbs into the sample matrix and the rate at which this process occurs, was for calibration. The kinetics of the absorption/desorption was investigated, and the isotropy of the two processes was demonstrated, thus validating this approach for calibration. A modified SPME device was used as a passive sampler to determine the time-weighted average (TWA) concentration of volatile organic compounds (VOCs) in air. The sampler collects the VOCs by the mechanism of molecular diffusion and sorption on to a coated fiber as collection medium. This process was shown to be described by Fick's first law of diffusion, whereby the amount of analyte accumulated over time enable measurement of the TWA concentration to which the sampler was exposed. TWA passive sampling with a SPME device was shown to be almost independent of face velocity, and to be more tolerant of high and low analyte concentrations and long and short sampling times, because of the ease with which the diffusional path length could be changed. Environmental conditions (temperature, pressure, relative humidity, and ozone) had little or no effect on sampling rate. When the SPME device was tested in the field and the results compared with those from National Institute of Occupational Health and Safety (NIOSH) method 1501 good agreement was obtained. To facilitate the use of SPME for field sampling, a new field sampler was designed and tested. The sampler was versatile and user-friendly. The SPME fiber can be positioned precisely inside the needle for TWA sampling, or exposed completely outside the needle for rapid sampling. The needle is protected within a shield at all times hereby eliminating the risk of operator injury and fiber damage. A replaceable Teflon cap is used to seal the needle to preserve sample integrity. Factors that affect the preservation of sample integrity (sorbent efficiency, temperature, and sealing materials) were studied. The use of a highly efficient sorbent is recommended as the first choice for the preservation of sample integrity. Teflon was a good material for sealing the fiber needle, had little memory effect, and could be used repeatedly. To address adsorption of high boiling point compounds on fiber needles, several kinds of deactivated needles were evaluated. RSC-2 blue fiber needles were the more effective. A preliminary field sampling investigation demonstrated the validity of the new SPME device for field applications.