Dosage du polyisoprène et des résines de la biomasse de guayule (Parthenium argentatum) par spectroscopie proche infrarouge (SPIR) Méthodes d'extraction par solvant de référence Un protocole basé sur l'extraction accélérée (ASE) avec l'acétone (EA) (résine) puis l'hexane (EH) (polyisoprène, PI) a été sélectionné et optimisé (rendement maximal; adapté à l'analyse de séries) ; quantification d'abord basée sur le poids de l'extrait (gravimétrie). L'EH est maximal à 120°C après étapes avec l'acétone à 40°C (plan d'expérience). La contamination croisée a été confirmée par SEC et FTIR (5 à 29%), conduisant à une deuxième méthode basée sur résine et PI et non plus sur EA et EH, incluant le PI de faible masse molaire (Mw) de l'EA. Ces 2 méthodes de référence ont servi à calibrer la SPIR (chimiométrie/PLS, coeff. beta) afin de relier signature spectrale, PI, résine. ASE-SPIR, couplés ici pour la première fois, ont été plus performants (R² 0.96; 0.98; RPD 4.8; 4.6; EA et EH resp.) que les méthodes de la littérature, grâce aux 215 échantillons représentatifs (génotypes, saison, âge du guayule, climat). La méthode tenant compte de la contamination est moins performante (erreur exp. due aux analyses SEC et FTIR; variation de composition des résines). Ayant montré la dégradation du PI au cours de l'extraction, un autre protocole a été étudié (biomasse humide, un solvant, une seule étape, 20°C) afin d'accéder au Mw «natif », donnant 2.106 g au lieu de 6.105 avec la biomasse séchée; il faut donc être prudent face aux Mw de guayule de la littérature. Ce travail montre la nécessité de tenir compte de la complexité de la biomasse de guayule (échelle cellulaire/PI vacuolaire, résine des canaux; moléculaire/instabilité chimique) lors de l'extraction du PI pour l'analyse structurale. Ces méthodes analytiques ont contribué à produire des prototypes (gant non allergisant, pneu) à haut Mw et à l'acclimatation en Europe dans le cadre du projet EU-Pearls. / Measurement of resin and polyisoprene in Parthenium argentatum (guayule) biomass using near infrared spectroscopy (NIRS) -Associated solvent-based reference methods .A protocol based on sequential extraction with acetone (resin) and hexane (polyisoprene, PI) with accelerated solvent extraction (ASE) was optimized and selected (maximized yield; adapted to large series) instead of Soxhlet and homogenizer. Quantification was first based on extract weight (gravimetry). Hexane extract was maximized at 120°C, after acetone steps at 40°C, through an experimental design. Cross contamination was confirmed and quantified (5 to 29%; SEC and FTIR). This gave a second method based on resin and PI, instead of crude extracts, accounting for low average molar mass PI (Mw) extracted by acetone instead of hexane. Both reference methods were used for calibrating NIRS applied to powdered biomass, with chemometric tools (PLS loadings, beta coefficients) to interpret spectral bands vs PI-resin relationship. ASE, not used before as reference, is highly reliable, and calibration with gravimetry (R² 0.96; 0.98; RPD 4.8; 4.6; for acetone and hexane extract) better than published data, thanks to the 215 samples covering genotypes, harvest date, plant age, climate. The method using cross contamination was less efficient because of higher experimental error induced by additional SEC and FTIR, and change in resin composition. Having set NIRS methods, a new protocol (single solvent THF, minimized processing, 20°C, fresh biomass) was designed to avoid degradation, yielding PI extracts with Mw above 2.106g/mole, closer to in vivo structure (6.105 when using dried guayule); caution to sample preparation in literature dealing with guayule PI structure. This calls for considering the complex structure of guayule biomass (PI in cells; resin in ducts; chemical instability) when extracting PI. These methods allowed producing high PI Mw glove and tire prototypes and domesticating this new crop in Europe within the EU-Pearls project.

Master of Science / Department of Chemical Engineering / J.R. Schlup / The curing reaction for the amine epoxy resin system of phenyl glycidyl ether (PGE) with metaphenylene diamine (mPDA) was investigated using two-dimensional correlation spectroscopy in the near-infared region (2DNIR). Synchronous and asynchronous correlation maps were generated using 2Dshige© software. The characteristic NIR band assignments were made, including the identification of new peaks for the O-H combination band in the 4825-4750 cm[superscript]-1 region and the CH stretching vibration overtone at 6018 cm[superscript]-1. Finally, the data suggests the reaction proceeds as follows: the appearance of the OH groups and C-H backbone vibrations occurs before the primary amine reactions and epoxide rings disappear.

Abstract This thesis explores the use of assistive in-situ technologies for formal caregivers in nursing homes. More specifically, focus is placed on improving context awareness and medication management. Although these topics have previously been researched for elderly care in general, few solutions targeting nursing homes have been generated. As the aging population further increases the burden on this care environment, it is important that solutions are found to help maintain care quality. The main findings in this thesis emphasise how technology can assist formal caregivers and facilitate increased patient wellbeing. The articles presented in this thesis describe our creation of a context-aware sensor system (named CARE) and a non-expert miniaturised near-infrared spectroscopy (MNIRS) solution. Both systems were designed iteratively with the help of nurses and were evaluated in a nursing home. CARE quantifies elderly residents’ behaviour, analyses the resulting data and produces valuable and actionable insights for nurses. Results from a two-month-long user study demonstrate that the system can facilitate increased awareness of patients’ needs and enhance care service. The custom MNIRS solution allows nurses to scan pharmaceuticals and obtain accurate identifications. This method significantly outperforms currently available tools in nursing homes and represents a promising solution that can reduce medication mismanagement. In the discussion section of the thesis, we revisit the research questions defined in the introduction and examine how each were answered. In addition, we discuss the augmentation of nursing home technology and various stakeholders’ perspectives. We then highlight how the work covered in this thesis was conducted in collaboration with industry and offer some conclusions, limitations and reflections. / Tiivistelmä Tämä väitöstyö tutkii paikkasidonnaisten teknologioiden käyttöä hoitajien työn helpottamiseksi vanhusten palvelukodeissa. Työn keskiössä on erityisesti kontekstitietoisuuden lisääminen ja lääkehoidon valvonta. Näitä aiheita on tutkittu laajalti aiemminkin, mutta aitoihin ympäristöihin keskittyviä ratkaisuja on vielä vain vähän. Väestön ikääntyminen aiheuttaa haasteita vanhustenhoidossa, ja siksi on tärkeää kehittää ratkaisuja hoidon laadun ylläpitoa varten. Väitöstyön löydökset painottavat teknologian roolia hoitajien avustajana sekä hoidettavien hyvinvoinnin ylläpitäjänä. Työn artikkelit kuvailevat kontekstitietoisen järjestelmän (CARE) koko kehitys- ja elinkaaren sekä lähi-infrapunaspektroskopiaan perustuvan ratkaisun lääkehoidon prosessien parantamiseksi. Molemmat järjestelmät kehitettiin iteratiivisesti hoitajien avulla ja koestettiin aidossa ympäristössä vanhainkodissa. CARE analysoi asiakkaiden liikkeitä ja toimintoja sekä visualisoi niistä korkeamman tason tietoa hoitajille. Kahden kuukauden kenttäkokeen tuloksena voidaan todeta, että järjestelmät yhdessä voivat auttaa hoitajia ymmärtämään asiakkaiden tarpeita sekä parantamaan hoidon laatua. Lähi-infrapunaspektroskopiaan perustuvalla kannettavalla ratkaisulla puolestaan hoitajat voivat varmistaa lääkehoidon oikeellisuuden. Tämä menetelmä on tarkempi ja parempi kuin tämänhetkiset käytössä olevat ratkaisut lääkkeiden oikeaksi toteamiseen. Väitöstyön keskusteluosuus palaa tutkimuskysymyksiin ja selventää, kuinka tehty työ ja saavutetut tulokset vastaavat niihin. Lisäksi keskustelu antaa yleiskuvan eri osapuolien näkemyksistä kehitetyn ratkaisun hyödyistä ja sen soveltuvuudesta käytännön työhön. Lopuksi väitöstyö luo katsauksen yhteistyön rooliin eri osa-alueiden toteuttamisessa ja esittää kriittisen näkökulman työn puutteisiin sekä yhteenvedon.

As intervenções com neurofeedback têm tido grande difusão na área como uma alternativa possível para tratamento do Transtorno de Déficit de Atenção/Hiperatividade (TDAH). Este estudo tem o objetivo de avaliar a viabilidade da utilização de um sistema de neurofeedback de baixo custo chamado Near Infrared Spectroscopy/Hemoencephalography Neurofeedback (NIR/HEG-NF) para tratar crianças com TDAH. Método: trata-se de um estudo clínico piloto aberto, que incluiu 5 crianças (3 meninos e 2 meninas - idade 10,4 ± 0,89 anos) com TDAH-C (tipo combinado). Os indivíduos completaram um programa de 24 sessões de NIR/HEG-NF, duas ou três vezes por semana. Os seguintes desfechos foram avaliados pré e pós-tratamento: 1) sintomas de desatenção e hiperatividade; 2) desempenho neuropsicológico; 3) imagens de SPECT cerebral; 4) qualidade de vida; e 5) efeitos adversos. Resultados: houve maior resistência do que o esperado para a participação dos indivíduos devido a vários fatores, tais como logísticos para a participação nas sessões e requerimento de não uso de medicação durante o protocolo. Detectamos uma diferença significativa na comparação pré e pós-intervenção nos escores atribuídos pelos pais na dimensão de hiperatividade e impulsividade do Swanson, Nolan and Pelham Questionnaire (SNAP-IV) (p = .042; tamanho de efeito [ES] =.38) e uma tendência de melhora nos escores atribuídos pelos pais e professores na dimensão desatenção (p = .066; ES = .47 / p = .068; ES = .60) naquelas crianças que finalizaram o tratamento. Detectamos uma tendência de aumento perfusional em diferentes partes do córtex cerebral em três indivíduos. As comparações pré e pós-intervenção nas outras medidas não indicaram diferenças estatisticamente significativas. Não foram relatados efeitos adversos significativos. Discussão: Os achados iniciais sugerem que o NIR/HEG-NF como tratamento para sintomas de TDAH pode ser promissor e deve ser melhor investigado. O protocolo lança luz sobre novas abordagens para avaliar a eficácia da intervenção. Pode-se, por exemplo, implementar o programa de intervenção no ambiente escolar e com uso concomitante de medicamentos para superar as resistências na alocação de pacientes e para uma avaliação com maior validade externa. / Background and objectives: Neurofeedback interventions have been quick and extensively introduced for clinicians to treat Attention-Deficit/Hyperactivity Disorder (ADHD). This study aims to assess the feasibility of using a low cost neurofeedback system called Near Infrared Spectroscopy/ Hemoencephalography Neurofeedback (NIR/HEG-NF) for treating children with ADHD. Method: This open pilot clinical study included 5 children (3 boys and 2 girls - age 10.4 ± 0.89 years) with ADHD-C (combined type). The subjects completed a program of 24 sessions of NIR/HEG-NF, two or three times a week. The following outcomes were assessed pre and post-treatment: 1) clinical symptoms of inattention and hyperactivity; 2) neuropsychological performance; 3) brain SPECT images; 4) quality of life and; 5) adverse effects. Results: Higher resistance than expected was experienced for patient allocation due to logistic reasons to attend the sessions in the hospital and the requirement of not using medication during the protocol. We detected a significant decrease in hyperactivity/impulsivity comparing pre and post-intervention scores in the parent’s Swanson, Nolan and Pelham Questionnaire - SNAP-IV (p = .042; Effect Size [ES] r = .38) and a trend of improvement in scores attributed by parents and teachers in inattention (p = .066; r = .47 / p = .068; r = .68) . We detected a trend for increased perfusion in different parts of the entire cortex in 3 subjects. Comparisons between pre and post-intervention scores in other measures did not suggest meaningful differences. No significant adverse effects were reported. Discussion: Our initial findings suggest that NIR/HEG-NF technique might be promising and should be further investigated. The protocol shed light on new approaches to assess the effectiveness of the intervention such as the need to both implement the intervention program in a school environment and to pursue the investigation of the treatment effects with concomitant use of medications to surpass resistances to enroll patients and to have an assessment with more external validity.

La mise en place de la démarche PAT (Process Analytical Technology), initiée par la FDA (Food and Drug Administration) s’est développée au cours de ces dernières années, au sein de l’industrie pharmaceutique. Grâce à des contrôles en continu au coeur des procédés de fabrication, elle permet une meilleure compréhension et une maîtrise de la formulation et du procédé, afin d’assurer la qualité finale des médicaments.A travers ce travail, nous avons mis en place un suivi en temps réel, par spectroscopie proche infrarouge, d’une opération d’enrobage suite à l’intégration d’une sonde à l’intérieur d’une turbine d’enrobage. La quantité d’enrobage, déterminée par une simple et rapide pesée mais néanmoins soumise à la variabilité de la masse des comprimés nus, ainsi que l’épaisseur du film, obtenue avec précision par imagerie térahertz ont servi de valeurs de référence pour calibrer l’information spectrale. Dans les deux cas, ces deux attributs qualité critiques ont été prédits avec de faibles erreurs de prédiction, qui se sont révélées être similaires. Par ailleurs, la prédiction en temps réel des propriétés de dissolution de comprimés prêts à être libérés, à partir de spectres acquis in-line, a permis de déterminer l’arrêt optimal de l’opération d’enrobage.Suite à un enrobage réalisé à partir d’une dispersion aqueuse de polymère une étape supplémentaire de traitement thermique ou curing est généralement nécessaire afin de stabiliser le film d’enrobage. Un travail de caractérisation mené à partir de techniques innovantes a permis d’apporter un nouvel éclairage sur la compréhension des phénomènes impliqués dans la formation du film au cours du curing. La caractérisation approfondie de la structure d’enrobage de comprimés soumis à un curing en turbine (conditions dynamiques) a mis en évidence la diminution de la porosité, couplée à l’évaporation de l’eau et à une meilleure organisation des chaînes de polymère au cours du curing. L’étude de comprimés soumis à un curing de référence en étuve durant 24 h (conditions statiques) a confirmé l’obtention d’un film stable après 4 h de curing dynamique. De nouveaux phénomènes, indépendants du curing, liés à la cristallisation et à la migration de l’alcool cétylique, couplée à la migration du lauryl sulfate de sodium, au sein de la couche d’enrobage ont été détectés au cours de la conservation des comprimés enrobés. / Implementation of PAT (Process Analytical Technology) approach has recently been promoted by the FDA (Food and Drug Administration) within the pharmaceutical industry. A desired goal of the PAT framework is to enhance understanding and control of the manufacturing process through timely measurements, during processing, to ensure final product quality. Real-time monitoring of a coating operation was performed from in-line Near Infrared (NIR) measurements inside a pan coater. Mass of coating materials, determined by simple and fast weighing but depending on core tablet weight uniformity, and film coating thickness, obtained from accurate and non-destructive Terahertz Pulsed Imaging (TPI) measurements, were used as reference values to calibrate NIR spectral information. In both cases, these two critical quality attributes were predicted with low predictive errors, which were found to be similar. In addition, real-time predictions of drug release from cured tablets were carried out by in-line NIR measurements. The coating operation was successfully stopped when desired dissolution criteria were achieved. A post-coating thermal treatment, known as curing, is generally required to stabilize film coating from aqueous polymer dispersion. Innovative techniques were jointly used to elucidate the underlying mechanisms of film formation along the curing process. This study provided a new insight into the tablet coating structure, highlighting a reduced internal coating porosity, a decrease in water content and showing a better structural rearrangement of polymer chains, with dynamic curing. All investigated techniques confirmed that a stabilized state was reached after a 4 h dynamic curing in comparison with a reference curing carried out in an oven for 24 h. Interestingly even prior to curing, new findings were pointed out, during coated tablets storage, related to the crystallisation and the upward migration of cetyl alcohol, coupled to the downward migration of sodium lauryl sulfate within the coating layer.

Este estudo tem como objetivo analisar a qualidade do lenho das árvores de eucalipto geneticamente modificadas e seus respectivos controles (clones convencionais). Foram selecionadas 70 árvores de eucalipto, ao 12º mês de 14 tratamentos (10 eventos modificados e 4 clones convencionais) representando 5 árvores/tratamento. As amostras do lenho foram coletadas do tronco das árvores com sonda de Pressler para análise das suas características anatômicas, densidade aparente do lenho (microdensitometria dos raios X) e propriedades químicas (espectroscopia no infravermelho próximo). As propriedades anatômicas e a densidade aparente do lenho indicaram poucas diferenças significativas entre as árvores dos tratamentos. Nas análises da espectroscopia no infravermelho próximo (NIR) foram encontrados modelos satisfatórios para a densidade aparente do lenho, a fração parede e para o teor de celulose. O lenho das árvores dos eventos modificados e dos clones convencionais mostrou pequenas diferenças nas características anatômicas e físicas, sendo caracterizado como lenho juvenil; assim as árvores devem ser analisadas em idades mais avançadas, com lenho de transição e adulto. Os resultados constituem-se parte da análise das árvores de eucalipto geneticamente modificadas, subsidiando os estudos genéticos visando obter altos níveis de produtividade. / This study aims to analyze the quality of the wood of genetically modified eucalyptus trees and their respective controls (conventional clones). 70 eucalyptus trees were selected at 12 months of 14 treatments (10 modified events and 4 conventional clones) representing 5 trees / treatment. The wood samples were collected of the trunk of the eucalyptus trees with Pressler increment borer for the analysis of their anatomical features, density of wood (microdensitometry of X-rays) and chemical properties (near infrared spectroscopy). The anatomical properties and wood density of eucalypts trees showed few significant differences between treatments. In the analysis of near infrared spectroscopy (NIR) have been found satisfactory models for the apparent density of the wood, the wall portion and the cellulose content. The wood from the trees of the modified events and conventional clones showed small differences in the anatomical and physical characteristics, being characterized as juvenile wood; so the trees must be analyzed at older ages, with wood transition and adult. The results are a part of the analysis of genetically modified eucalyptus trees, supporting genetic studies to obtain high levels of productivity.

Chan Jacklynn = 摻近紅外染料溶凝膠薄膜之光學及激光性質研究 / 陳在琳. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 137-152). / Text in English; abstracts in English and Chinese. / Chan Jacklynn = Shan jin hong wai ran liao rong ning jiao bo mo zhi guang xue ji ji guang xing zhi yan jiu / Chen Zailin. / Abstract --- p.i / Acknowledgements --- p.v / Publications --- p.vii / Table of Contents --- p.viii / List of Figures --- p.xi / List of Tables --- p.xiv / Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Sol-gel Glass Waveguides --- p.3 / Chapter 1.2 --- Ellipsometry --- p.7 / Chapter 1.3 --- Aim of this Research Work and Organization of Thesis --- p.12 / Chapter 2 --- Basics on Ellipsometry --- p.16 / Chapter 2.1 --- Principle of Ellipsometry --- p.17 / Chapter 2.1.1 --- Definition of Ellipsometric Parameters --- p.17 / Chapter 2.1.2 --- Configurations of an Ellipsometer --- p.22 / Chapter 2.2 --- Mathematical Representation of Ellipsometry --- p.26 / Chapter 2.2.1 --- Bulk Material Structure --- p.26 / Chapter 2.2.2 --- Single Layer Structure --- p.28 / Chapter 2.3 --- Parameterization of Optical Functions --- p.31 / Chapter 2.3.1 --- Optical Functions --- p.34 / Chapter 2.3.2 --- Regression Analysis --- p.38 / Chapter 3 --- Modification of Rotating-Polarizer-Analyzer Ellipsometer --- p.40 / Chapter 3.1 --- Principle of RPA System --- p.41 / Chapter 3.2 --- Optical Setup of RPA System --- p.45 / Chapter 3.3 --- Components of the System and Modification for Infrared Measurement --- p.47 / Chapter 3.3.1 --- Light Source --- p.47 / Chapter 3.3.2 --- Monochromator --- p.52 / Chapter 3.3.3 --- Polarizers and Analyzer --- p.53 / Chapter 3.3.4 --- Detector and Data Reduction --- p.54 / Chapter 3.4 --- Optical Alignment --- p.59 / Chapter 3.5 --- Alignment of the Azimuthal Angles of the Polarizers --- p.61 / Chapter 3.6 --- Calibration Procedure --- p.63 / Chapter 3.6.1 --- Calibration by Gold Film on Silicon --- p.63 / Chapter 3.6.2 --- Calibration by Silicon Dioxide Film on Silicon --- p.67 / Chapter 4 --- Near Infrared Dyes in Sol-gel Waveguides --- p.72 / Chapter 4.1 --- Sol-gel Process --- p.73 / Chapter 4.2 --- Near Infrared Dyes in Sol-gel Waveguides --- p.77 / Chapter 4.2.1 --- Development of Near Infrared Dyes --- p.77 / Chapter 4.2.2 --- Studies on Near Infrared Dye Solid State Laser --- p.79 / Chapter 5 --- Optical Properties of Dye-doped Sol-gel Waveguides --- p.83 / Chapter 5.1 --- Experimental Procedure --- p.84 / Chapter 5.1.1 --- Preparation of Samples --- p.84 / Chapter 5.1.1.1 --- Materials --- p.84 / Chapter 5.1.1.2 --- Procedure --- p.86 / Chapter 5.1.2 --- Discussion on Synthesis of Infrared Dye-doped Sol-gel Waveguides --- p.88 / Chapter 5.1.2.1 --- Choice of Solvents --- p.89 / Chapter 5.1.2.2 --- Thermal Treatment and Lifetime --- p.93 / Chapter 5.1.2.3 --- Necessity of Addition of GLYMO --- p.94 / Chapter 5.1.3 --- Sample Characterization --- p.96 / Chapter 5.2 --- Surface Morphology --- p.97 / Chapter 5.3 --- Optical Properties of Dye-doped Zirconia Organically Modified Silicate Waveguides --- p.100 / Chapter 5.3.1 --- Modeling of Ellipsometric Values --- p.101 / Chapter 5.3.2 --- Interpretation of the Modeling Results --- p.107 / Chapter 6 --- Amplified Spontaneous Emission Based on Sol-gel Waveguides --- p.109 / Chapter 6.1 --- Experimental Setup --- p.110 / Chapter 6.2 --- Features of ASE and Fluorescence --- p.112 / Chapter 6.3 --- Prolonged Thermal Treatment Effect on Light Emission --- p.119 / Chapter 6.3.1 --- Difference in Preparation of Samples --- p.121 / Chapter 6.3.2 --- Light Emission of Samples under Extended Thermal Treatment --- p.123 / Chapter 6.3.3 --- Directionality of the Emission --- p.125 / Chapter 6.3.4 --- Some Features of the Emission --- p.127 / Chapter 7 --- Conclusion and Recommendation --- p.133 / Chapter 7.1 --- Conclusion --- p.133 / Chapter 7.2 --- Recommendation --- p.135 / Bibliography --- p.137

The goal of this dissertation is to demonstrate the utility of portable, small-scale diffuse optical spectroscopic (DOS) systems for the diagnosis and treatment monitoring of various diseases. These systems employ near-infrared light (wavelength range of 650nm to 950nm) to probe human tissue and are sensitive to changes in scattering and absorption properties of tissues. The absorption is mainly influenced by the components of blood, namely oxy- and deoxy-hemoglobin (HbO2 and Hb) and parameters that can be derived from them (e.g. total hemoglobin concentration [THb] and oxygen saturation, StO2). Therefore, I focused on diseases in which these parameters change, which includes vascular diseases such as Peripheral Atrial Disease (PAD) and Infantile Hemangiomas (IH) as well as musculoskeletal autoimmune diseases such as Rheumatoid Arthritis (RA). In each of these specific diseases, current monitoring techniques are limited by their sensitivity to disease progression or simply do not exist as a quantitative metric. As part of this project, I first designed and built a wireless handheld DOS device (WHDD) that can perform DOS measurements at various tissue depths. This device was used in a 15-patient pilot study for infantile hemangiomas (IH) to differentiate diseased skin from normal skin and monitor the vascular changes during intervention. In another study, I compare the ultra-small form- factor WHDD’s ability to monitor synovitis and disease progression during a patient’s treatment of RA against the capabilities of a proven frequency domain optical tomographic (FDOT) system that has shown to differentiate patients with and without RA. Learning from clinical utility of the WHDD from these two studies, I adapted the WHDD technology to develop a compact multi- channel DOS measurement system to monitor perfusion changes in the lower extremities before and after surgical intervention for patients with peripheral artery disease (PAD). Using this multi- channel system, which we called the vascular optical spectroscopic measurement (VOSM) system, our group conducted a 20-subject pilot study to quantify its ability to monitor blood perfusion before and after revascularization of stenotic arteries in the lower extremities. This proof-of- concept study demonstrated how DOS may help vascular surgeons perform revascularization procedures in the operating room and assists in post-operative treatment monitoring of vascular diseases.

Liu, Haijing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 112-116). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iii / 摘要 --- p.v / Table of contents --- p.vi / List of figures --- p.xii / List of tables --- p.xv / List of abbreviations --- p.xvi / Chapter / Chapter 1 --- Introduction / Chapter 1.1 --- Fermentation --- p.1 / Chapter 1.1.1 --- Biopharmaceutical production --- p.1 / Chapter 1.1.2 --- History of fermentation --- p.1 / Chapter 1.1.3 --- Fermentation Technology --- p.2 / Chapter 1.1.3.1 --- Fermentation process --- p.2 / Chapter 1.1.3.2 --- Fermenter --- p.4 / Chapter "1,1.3.3" --- Mode of fermentation process --- p.5 / Chapter 1.1.3.4 --- Bacterial growth --- p.6 / Chapter 1.1.4 --- High cell density fermentation for plasmid DNA production --- p.7 / Chapter 1.1.4.1 --- Porcine growth hormone releasing factor (pGRF) --- p.8 / Chapter 1.1.4.2 --- H5N1 avian influenza viruses DNA vaccine --- p.9 / Chapter 1.1.4.3 --- Fed-batch high-cell-density fermentation --- p.9 / Chapter 1.1.5 --- Fermentation process monitoring --- p.11 / Chapter 1.2 --- Near-infrared spectroscopy (NIRS) --- p.12 / Chapter 1.2.1 --- Basic near-infrared spectroscopy --- p.13 / Chapter 1.2.1.1 --- Rationale of near-infrared spectroscopy --- p.13 / Chapter 1.2.1.2 --- NIR spectra acquisition --- p.16 / Chapter 1.2.1.3 --- Interpretation of NIR spectra --- p.18 / Chapter 1.2.2 --- Multivariate calibration --- p.20 / Chapter 1.2.2.1 --- Why multivariate calibration --- p.22 / Chapter 1.2.2.2 --- The problem of collinearity --- p.25 / Chapter 1.2.2.3 --- Spectral range selection --- p.26 / Chapter 1.2.2.4 --- Signal optimization --- p.26 / Chapter 1.2.2.5 --- Spectral pretreatment --- p.27 / Chapter 1.2.2.6 --- Parameter selection --- p.28 / Chapter 1.2.3 --- Applications of NIRS in high cell density E. coli fermentation --- p.31 / Chapter 1.2.4 --- Adaptive calibration strategy --- p.33 / Chapter 1.3 --- Aims of study --- p.34 / Chapter 2 --- Materials and methods / Chapter 2.1 --- High cell density fermentation of plasmid DNA --- p.35 / Chapter 2.1.1 --- Fermentation system --- p.35 / Chapter 2.1.2 --- pGRF fermentation --- p.38 / Chapter 2.1.2.1 --- Prepare seed flask --- p.38 / Chapter 2.1.2.2 --- Reagents for bacterial culture by fermenter --- p.38 / Chapter 2.1.2.2.1 --- LB based complex fermentation medium --- p.38 / Chapter 2.1.2.2.2 --- Batch fermentation medium --- p.39 / Chapter 2.1.2.2.3 --- Feeding medium-1 --- p.41 / Chapter 2.1.2.2.4 --- Base feed --- p.41 / Chapter 2.1.2.3 --- Fermentation methods --- p.42 / Chapter 2.1.2.3.1 --- Fermenter set-up --- p.42 / Chapter 2.1.2.3.2 --- Inoculate seed culture into the fermenter --- p.43 / Chapter 2.1.2.3.3 --- Transfer the feeding medium --- p.44 / Chapter 2.1.2.3.4 --- Heat induction --- p.44 / Chapter 2.1.2.3.5 --- Fermentation harvest --- p.44 / Chapter 2.1.2.4 --- Scale-up fermentation and large scale production 45 / Chapter 2.1.3 --- H5N1 fermentation --- p.45 / Chapter 2.1.4 --- Reference analytical testing --- p.45 / Chapter 2.1.4.1 --- Optical density --- p.45 / Chapter 2.1.4.2 --- Cell dry weight --- p.45 / Chapter 2.1.4.3 --- Specific plasmid DNA yield --- p.46 / Chapter 2.1.4.3.1 --- Plasmid DNA isolation --- p.46 / Chapter 2.1.4.3.2 --- Plasmid DNA measurement --- p.46 / Chapter 2.1.4.3.3 --- Plasmid DNA identification --- p.46 / Chapter 2.1.4.4 --- Analysis of glycerol and acetate --- p.46 / Chapter 2.1.4.4.1 --- Standard samples --- p.46 / Chapter 2.1.4.4.2 --- Enzymatic test kits --- p.47 / Chapter 2.1.4.4.3 --- Automatic biochemistry analyzer --- p.47 / Chapter 2.1.5 --- Summary of all fermentation batches --- p.48 / Chapter 2.2 --- Full factorial design and semi-synthetic samples --- p.49 / Chapter 2.2.1 --- Adaptive calibration samples --- p.49 / Chapter 2.2.1.1 --- Matrix 1 calibration samples --- p.49 / Chapter 2.2.1.2 --- Matrix 2 calibration samples --- p.50 / Chapter 2.2.1.3 --- Matrix 3 calibration samples --- p.50 / Chapter 2.2.2 --- Summary of all samples --- p.51 / Chapter 2.3 --- NIR sample presentation and spectral acquisition --- p.52 / Chapter 2.3.1 --- NIR spectrophotometers --- p.52 / Chapter 2.3.2 --- Cuvettes for transmittance spectral acquisition --- p.53 / Chapter 2.3.3 --- Bottles for reflectance spectral acquisition --- p.54 / Chapter 2.3.4 --- Spectral acquisition --- p.55 / Chapter 2.3.4.1 --- Transmittance --- p.55 / Chapter 2.3.4.2 --- Reflectance --- p.55 / Chapter 2.4 --- Multivariate calibration and validation --- p.56 / Chapter 2.4.1 --- Spectral preprocessing --- p.56 / Chapter 2.4.2 --- Multivariate calibration --- p.57 / Chapter 2.4.3 --- Model validation --- p.57 / Chapter 3 --- Results and discussion / Chapter 3.1 --- Sample presentation and NIR spectrum --- p.59 / Chapter 3.1.1 --- Transmission measurement --- p.60 / Chapter 3.1.2 --- Reflectance measurement --- p.63 / Chapter 3.1.3 --- Spectral responses and pre-treatment method --- p.64 / Chapter 3.1.4 --- Design of experiments for calibration sample preparation --- p.67 / Chapter 3.1.5 --- Summary --- p.68 / Chapter 3.2 --- Adaptive calibration --- p.69 / Chapter 3.2.1 --- Selection of Multivariate calibration model --- p.70 / Chapter 3.2.1.1 --- Matrix 1 calibration models --- p.70 / Chapter 3.2.1.2 --- Matrix 2 calibration models --- p.74 / Chapter 3.2.1.3 --- Matrix 3 calibration models --- p.76 / Chapter 3.2.1.4 --- Summary --- p.78 / Chapter 3.2.2 --- Model validation --- p.79 / Chapter 3.2.2.1 --- Performance of Reference analytical methods --- p.79 / Chapter 3.2.2.1.1 --- Enzymatic test kits --- p.79 / Chapter 3.2.2.1.2 --- Automatic biochemistry anaylyzer (Bioprofile) --- p.81 / Chapter 3.2.2.1.3 --- Summary --- p.83 / Chapter 3.2.2.2 --- Model validation using external test samples --- p.83 / Chapter 3.2.2.2.1 --- Matrix 1 models --- p.83 / Chapter 3.2.2.2.2 --- Matrix 2 models --- p.87 / Chapter 3.2.2.2.3 --- Matrix 3 models --- p.89 / Chapter 3.2.2.2.4 --- Overall NIR measurement errors --- p.90 / Chapter 3.2.2.2.5 --- Summary --- p.91 / Chapter 3.3 --- Use of calibrated NIRS in at-line monitoring and control of fermentation --- p.93 / Chapter 3.3.1 --- Analysis of small-scale fermentation --- p.94 / Chapter 3.3.1.1 --- pGRF plasmid DNA production --- p.94 / Chapter 3.3.1.2 --- H5N1 plasmid DNA production --- p.95 / Chapter 3.3.1.3 --- Summary --- p.97 / Chapter 3.3.2 --- Analysis of large scale fermentation --- p.97 / Chapter 3.3.2.1 --- 30 L clinical production of H5N1 plasmid DNA --- p.97 / Chapter 3.3.2.2 --- 80 L scale-up production of H5N1 plasmid DNA --- p.99 / Chapter 3.3.2.3 --- Summary --- p.100 / Chapter 3.3.3 --- Effective control of fermentation production using at-line NIR analysis --- p.101 / Chapter 3.3.3.1 --- At-line monitoring of Batch 11 --- p.101 / Chapter 3.3.3.2 --- At-line monitoring of Batch 12 X --- p.102 / Chapter 3.3.3.3 --- Summary --- p.104 / Chapter 3.4 --- General discussion and conclusion --- p.105 / Chapter 3.5 --- Future prospects --- p.108 / References --- p.112 / Appendix 1 --- p.117 / Appendix 2 --- p.118 / Appendix 3 --- p.123 / Appendix 4 --- p.131 / Appendix 5 --- p.134

A classificação e caracterização do petróleo cru em tempo real é um dos mais importantes desafios atuais da indústria petroquímica e agencias ambientais. Um método rápido e econômico antes da destilação do petróleo iria a fornecer informações químicas de grande importância para o ajuste em tempo real dos parâmetros críticos de uma refinaria, permitindo a otimização do processo, resultando em benefícios econômicos e ambientais. Uma variedade de técnicas espectroscópicas está sendo utilizadas nas ultima décadas para as analises, classificação e caracterização de petróleo cru nos campos de perfuração, para as análises dos produtos petrolíferos e para detecção de derrames. As vantagens do uso desta técnica incluem uma rápida resposta, requerimento mínimo de preparação da amostra e equipamentos relativamente econômicos. As técnicas espectroscópicas envolvidas na tese (NIR) mostraram o maior potencial no campo. No entanto, os estudos realizados com todas estas técnicas não foram enfocados em aplicações em linha. O NIR é tradicionalmente a técnica espectroscópica mais utilizada para a análise de petróleo cru. Mas os problemas típicos que esta técnica apresenta são sobreposição das diferentes bandas de absorção e saturação do sinal devido à alta absorbância característico do petróleo cru. A Fluorescência é um fenômeno mais complexo do que a absorção (NIR) e efeitos como quenchinge transferência de energia têm que ser consideradas. No entanto, a espectroscopia de fluorescência pode fornecer sinais bidimensionais com muito mais informação do que uma emissão unidimensional do espectro de absorção. Enquanto a espectrometria de massa, a técnica MALDI TOF faz possível a ionização de macromoléculas que são difíceis de serem ionizadas por não ser facilmente decompostas. Neste trabalho, as vantagens e limitações das técnicas disponíveis são analisadas e o potencial do analises remoto em linha do petróleo cru é avaliado. A pesar da boa analise e classificação obtida com algumas técnicas, nem sempre é possível integrá-los em um sistema em linha quando uma preparação previa da amostra é necessária ou quando o sistema é demasiado complexo. Para a extração da informação química contida nos espectros foram utilizados métodos matemáticos e estatísticos (quimiométricos) nos espectros: Mínimos Quadrados Parciais (PLS). / Online remote characterization and real-time classification of crude petroleum is one of the most important current challenges faced by the petrochemical industry and environmental agencies. A rapid and inexpensive method for the remote analysis and classification of petroleum prior to distillation of the crude would provide chemical information of great importance for real time adjustment of the critical parameters of a refinery, permitting an optimization of the process and resulting in economic and environmental benefits. A variety of spectroscopic techniques have been used over the last few decades for the analysis, characterization and classification of crude oil in drilling fields, for the analysis of petroleum products and for the detection of oil-spills. The advantages of using these techniques include rapid response, the requirement of minimal sample preparation and relatively inexpensive equipment costs. The spectroscopic techniques involved in the thesis (NIR, fluorescence and mass spectrometry) showed the greatest potential in the field. Nevertheless, the studies carried out with these techniques typically do not focus on online applications. NIR is traditionally the spectroscopic techniques most used for crude petroleum analysis. Overlap of the different absorption bands and saturation of the signal due to the high absorbance of crude petroleum are typical problems encountered with this technique. Fluorescence is a more complex phenomenon than absorption (NIR) and effects such as quenching and energy transfer have to be considered. However, fluorescence spectroscopy can provide two-dimensional signals with much more information than a one-dimensional emission or absorption spectrum. While mass spectrometry, MALDI TOF technique makes possible the ionization of macromolecules that are difficult to be ionized not be easily broken. In this work, the advantages and limitations of the available techniques are analyzed and the potential for remote on-line analysis of crude petroleum is evaluated. In spite of the good analysis and classification obtained with some of the techniques, it is not always possible to integrate them into a remote on-line analytical system when some prior sample preparation is required or when the system is too complex. For the extraction of chemical information contained in the spectra were used mathematical and statistical (chemometric) in the spectra: Partial Least Squares (PLS).