Les travaux décrits dans ce mémoire sont consacrés à la synthèse et à la caractérisation physico-chimique de nouveaux assemblages supramoléculaires à base d'un polysaccharide d'origine naturelle, le hyaluronane (HA).<br />Les assemblages résultent du mélange de deux polymères complémentaires et plus précisément de l'établissement d'interactions multivalentes spécifiques entres des molécules hôtes, les $\beta$-cyclodextrines ($\beta$-CD) et des molécules invitées, des groupements hydrophobes d'adamantane (AD), greffés sur les chaînes de HA.<br />Afin d'évaluer l'influence de l'énergie de jonction sur la stabilité des réseaux, deux systèmes de structure différente ont été préparés. Les assemblages sont stabilisés soit, par la formation de complexes d'inclusion simples $\beta$-CD/AD, soit par des paires de complexes d'inclusion.<br />Des expériences de titration calorimétrique ont permis une analyse détaillée des propriétés d'inclusion des molécules et des macromolécules hôtes et invitées. Le comportement en milieu aqueux des polymères modifiés seuls et les propriétés viscoélastiques des mélanges ont été étudiés par des mesures rhéologiques en écoulement et en régime dynamique.<br />Ces assemblages sont sensibles à la variation de paramètres externes tels que la force ionique, la concentration en polymères, la température et l'ajout de molécules hôtes compétitives. Ce sont des réseaux temporaires dont la dynamique est gouvernée principalement par le nombre et la vitesse d'échange des complexes d'inclusion efficaces et par la mobilité des chaînes de HA.

ABA type tri-block amphiphilic polyelectrolyte consisting of poly(methyl methacrylate-block-methacrylic acid-block-methyl methacrylate) (P(MMA-b-MAA-b-MMA)) was synthesized by atom transfer radical polymerization technique (ATRP) and the self-assembly behavior of the polymers in aqueous solution was studied over the course of neutralization. Combination of potentiometric and conductometric titrations along with dynamic light scattering (DLS) techniques were used to investigate the size and shape of aggregates at various degrees of neutralization. The effect of hydrophobic-hydrophilic (MMA-MAA) ratio and polymer chain length on the aggregation behavior during neutralization was studied. P(MMA-b-MAA-b-MMA) with longer MMA segment self-assembles via the close association mechanism through stronger self-entanglement of MMA chains, whereas P(MMA-b-MAA-b-MMA) with shorter MMA chain self-assembles via the open association mechanism, as confirmed by transmission electron microscopy (TEM). Conductometric titration was used to determine the counterion condensation during the course of neutralization. When the charge density of micelle approaches a critical value as neutralization progresses, counterion condensation of Na+ ions on the polymer chains occurs. The effect of counterion condensation on the aggregation behavior during neutralization was elucidated. / Singapore-MIT Alliance (SMA)

Vattenhaltsbestämning är av stor vikt i många sammanhang. T.ex. kan vattenhalten påverka utbytet av en kemisk syntes, eller ha negativ inverkan på hållbarheten av läkemedel och livsmedel. Standardmetoden för vattenhaltsbestämning är Karl Fischer-titrering, baserad på antingen volymetri eller coulometri. I den här avhandlingen presenteras nya infallsvinklar för bestämning av mycket låga halter vatten i komplexa provmatriser, som t.ex. tekniska oljor och substanser som interfererar med alkoholbaserade Karl Fischer-reagens. Vattnet avskiljs ofta från oljematrisen före titrering genom förångning. I samband med framtagningen av nya referensmaterial för vatten i olja ifrågasattes förångningsteknikernas effektivitet av National Institute of Standards and Technology (NIST). NIST menade att en fraktion av vattnet bands hårt i oljefasen och att det inte kunde frigöras och detekteras annat än med en modifierad volymetrisk metod där reagenset innehöll minst 65% kloroform. I den här avhandlingen presenteras en alternativ metod som uppfyller det ställda kravet för en fullständig upplösning av oljefasen. Med denna metod visas att det inte finns någon anledning att ifrågasätta förångningsteknikernas effektivitet och att den modifierade metoden som NIST använder ger systematiskt för höga resultat. Fördelar som enklare handhavande, kortare konditioneringstider och att endast ett reagens behövs har gjort att diafragmafri coulometri har blivit allt mer populär. Spårhaltsbestämning med denna teknik ställer dock speciellt höga krav på reagensen eftersom strömtätheten vid katoden är låg. Med anledning av detta testades olika typer av kommersiella reagensblandningar för bestämning av små vattenmängder och kritiska parametrar identifierades. Dekanol visade sig ha en gynnsam effekt på katodreaktionen i reagens modifierade med xylen enligt standardmetodbeskrivningen för bestämning av vatten i oljor. För provtyper som inte går att analysera med alkoholbaserade reagenser presenteras en ny typ baserad på N-metylformamid. Med ett sådant reagens bestämdes vattenhalten i ett reaktivt salt som används i litiumjonbatterier. Liknande alkoholfria reagens undersöktes mer utförligt i en djupare studie som även inkluderade formamid och dimetylformamid. För- och nackdelar med dessa alternativa lösningsmedel diskuteras och möjliga reaktionsförlopp föreslås. Det visade sig att läget på jämvikten mellan svaveldioxid och vätesulfit är en avgörande faktor för att förklara den stora skillnaden i reaktionshastighet i dessa lösningsmedel. / Moisture determination is of great importance in the production and use of many substances. For example, the moisture content can affect the efficiency of a chemical reaction or determine the shelf life of pharmaceuticals or foods. The standard method for moisture determination is Karl Fischer (KF) titration, based on either volumetry or coulometry. This thesis concerns new approaches to trace determination in complex sample matrices and is focused on oils and substances that interfere with alcoholic KF reagents. Moisture is frequently separated from oil matrices before titration by means of evaporation techniques. In connection with the preparation of new reference materials for moisture in oil, the National Institute of Standards and Technology (NIST) questioned the efficiency of such evaporation techniques. NIST claimed that some of the moisture was sequestered in the oil phase and that it could only be released and detected by using a modified volumetric KF method with a reagent containing at least 65% chloroform. In this thesis, an alternative KF method that meets the proposed requirement for a complete dissolution of the oil sample is presented. With this method it is shown that there is no reason to question the efficiency of the evaporation techniques and that the criticized volumetric method used by NIST is biased high. Ever since its introduction diaphragm-free coulometry has gained popularity due to its ease of use, with a single reagent and short conditioning times. Trace determination with this technique sets great demands on the reagent due to the resulting low current densities at the generator cathode. The performance of several commercial reagents is evaluated under such unfavorable conditions and critical titration parameters are identified. It is also shown that decanol has a favorable effect on the cathode process when using reagents modified with xylene according to standard methods for moisture determination in oils. For samples that are incompatible with the alcohol component in ordinary KF reagent a new reagent based on N-methylformamide is presented. It is shown that is works well for determinations of moisture in a conductive salt used in lithium-ion batteries. The concept of alcohol-free KF reagents is taken a step further in a systematic investigation, also including formamide and dimethylformamide. Advantages and disadvantages with these solvents are discussed and possible reaction paths are surveyed. It is shown that the position of the sulfur dioxide/hydrogen sulfite equilibrium is the main explanation for the large differences in the KF reaction rates in these solvents.

MosA is a protein found in <i>Sinorhizobium meliloti</i> L5-30 and has been suggested to be responsible for the biosynthesis of the rhizopine 3-O-methyl-scyllo-inosamine (3-MSI) from scyllo-inosamine (SI). However, we have shown MosA is a dihydrodipicolinate synthase (DHDPS) catalyzing the condensation of pyruvate with aspartate-β-semialdehyde (ASA). Since the DHDPS reaction occurs through a Schiff base aldol-type mechanism it was proposed that MosA could be an O-methyltransferase utilizing 2-oxo-butyrate (2-OB) as a novel methyl donor. This interesting yet unlikely possibility would explain MosA's role in the biosynthesis of 3-MSI without ignoring its similarity to DHDPS. Alternatively, MosA may have two catalytic domains one of which possesses a novel binding motif for S-Adenosyl methionine (SAM) to account for methyltransfer activity. In vitro demonstration of MosAs methyltransferase activity is required to resolve this apparent contradiction.<p>This dissertation describes the chemical synthesis of the rhizopines, investigation into whether MosA has a direct role in rhizopine biosynthesis and the thermodynamic characterization of compounds interacting with MosA as observed by isothermal titration calorimetry. <p>Initial investigation into MosAs methyltransferase activity began with 2-OBs interaction with the enzyme. Inhibition experiments determined 2-OB is a competitive inhibitor with respect to pyruvate of the DHDPS reaction of MosA. Furthermore, protein mass spectrometry of MosA in the presence of 2-OB and sodium borohydride indicated that a Schiff base enzyme intermediate was indeed being formed providing evidence that the proposed mechanism may exist. However, neither of the rhizopines had any effect on the DHDPS activity and HPLC assays determined that no 3-MSI was being produced by MosA in the presence of SI and 2-OB. Furthermore, HPLC assays failed to detect methyl transfer activity by MosA utilizing the SAM as a methyl donor. <p>Isothermal titration calorimetry provided thermodynamic characterization of the pyruvate and 2-OB Schiff base intermediates formed with MosA. In addition, ITC provided insight into the nature and thermodynamics of (S)-lysines inhibition of MosA. ITC failed to detect any interactions between the rhizopines or SAM with MosA. These results indicate that MosA is only a DHDPS and does not catalyze the formation of 3-MSI from SI as hypothesized in the literature.

The aim of this work is to evaluate the fuzzy system for different types of patients for levodopa infusion in Parkinson Disease based on simulation experiments using the pharmacokinetic-pharmacodynamic model. Fuzzy system is to control patient’s condition by adjusting the value of flow rate, and it must be effective on three types of patients, there are three different types of patients, including sensitive, typical and tolerant patient; the sensitive patients are very sensitive to drug dosage, but the tolerant patients are resistant to drug dose, so it is important for controller to deal with dose increment and decrement to adapt different types of patients, such as sensitive and tolerant patients. Using the fuzzy system, three different types of patients can get useful control for simulating medication treatment, and controller will get good effect for patients, when the initial flow rate of infusion is in the small range of the approximate optimal value for the current patient’ type.

MosA is a protein found in <i>Sinorhizobium meliloti</i> L5-30 and has been suggested to be responsible for the biosynthesis of the rhizopine 3-O-methyl-scyllo-inosamine (3-MSI) from scyllo-inosamine (SI). However, we have shown MosA is a dihydrodipicolinate synthase (DHDPS) catalyzing the condensation of pyruvate with aspartate-β-semialdehyde (ASA). Since the DHDPS reaction occurs through a Schiff base aldol-type mechanism it was proposed that MosA could be an O-methyltransferase utilizing 2-oxo-butyrate (2-OB) as a novel methyl donor. This interesting yet unlikely possibility would explain MosA's role in the biosynthesis of 3-MSI without ignoring its similarity to DHDPS. Alternatively, MosA may have two catalytic domains one of which possesses a novel binding motif for S-Adenosyl methionine (SAM) to account for methyltransfer activity. In vitro demonstration of MosAs methyltransferase activity is required to resolve this apparent contradiction.<p>This dissertation describes the chemical synthesis of the rhizopines, investigation into whether MosA has a direct role in rhizopine biosynthesis and the thermodynamic characterization of compounds interacting with MosA as observed by isothermal titration calorimetry. <p>Initial investigation into MosAs methyltransferase activity began with 2-OBs interaction with the enzyme. Inhibition experiments determined 2-OB is a competitive inhibitor with respect to pyruvate of the DHDPS reaction of MosA. Furthermore, protein mass spectrometry of MosA in the presence of 2-OB and sodium borohydride indicated that a Schiff base enzyme intermediate was indeed being formed providing evidence that the proposed mechanism may exist. However, neither of the rhizopines had any effect on the DHDPS activity and HPLC assays determined that no 3-MSI was being produced by MosA in the presence of SI and 2-OB. Furthermore, HPLC assays failed to detect methyl transfer activity by MosA utilizing the SAM as a methyl donor. <p>Isothermal titration calorimetry provided thermodynamic characterization of the pyruvate and 2-OB Schiff base intermediates formed with MosA. In addition, ITC provided insight into the nature and thermodynamics of (S)-lysines inhibition of MosA. ITC failed to detect any interactions between the rhizopines or SAM with MosA. These results indicate that MosA is only a DHDPS and does not catalyze the formation of 3-MSI from SI as hypothesized in the literature.

Metro Kaohsiung with high percentage (6-10 %) of poor air quality (PSI>100) has been announced officially by Taiwan Environmental Protection Administration (TEPA) as the worst air quality region among seven Air Quality Zones (AQZ) in Taiwan. Ozone is one of two major air pollutants that are responsible for the poor air quality. In this study, the vertical concentration profiles of ozone and its precursors (NOX and VOCs) at eight sites were measured by tethered balloons with air pumps and tedlar sampling bags. This method was used to investigate the vertical profile and the tempospatial distribution of ozone and its precursors in offshore/inland regions. This study further investigated ozone formation mechanism and air mass trajectory via simultaneous air quality sampling around the coastal region of metro Kaohsiung. This study sampled the vertical concentration profiles of ozone and its precursors at both inland and offshore sites during eight intensive sampling periods on August 16-17 and November 2-3, 2006, January 24-25, March 6-7 and May 2-3, 2007, October 30-31, 2008, and March 11-12 and July 15-16, 2009. Eight sampling periods were divided into the sea-land breeze period, the northeast monsoon period, and the mixing wind field period. During the sea-land breeze period, the wind direction changed 90˚ and more between daytime and nighttime, and the wind speeds of the sea breezes varied significantly than those of the land breezes. During the northeast monsoon period, prevailing wind blew from the north (300~60˚) with the average wind speeds of 1~4 m/s. During the mixing wind field period, the wind direction varied significantly from 270˚ to 90˚ with the average wind speeds of 1~3 m/s. Results obtained from the vertical profiles showed that O3 concentration appeared stratification phenomenon at 40 out of 64 sampling sites, in which its precursors (NOX or VOCs) demonstrated stratification phenomenon at 30 sampling sites, accounting for 75 % of total O3 stratification. It suggested that ozone and its precursors had strong correlation with each other. The linear slope of the titration effect showed that the intensity of titration effect at night during the northeast monsoon period was larger and had higher correlation (R> 0.7), and followed by the mixing wind field period and the sea-land breeze period. This phenomenon correlated closely with meteorological conditions, the concentrations of O3 precursors, and solar radiation intensity. Therefore, O3 concentration at night during the northeast monsoon period was lower than those of the sea-land breeze period. Results obtained from VOCs measurement indicated that the major species of VOCs was acetone which accounted for 16.25~64.05 % of total TVOCs-C2 in the offshore region. High concentration of TVOCs-C2 was affected by the usage of organic solvents. While, the major species of VOCs in the inland region was toluene which accounted for 6.41~43.77 % of total TVOCs-C2. Furthermore, results obtained from backward trajectory showed that air pollutants emited from land sources could transport to the offshore region, resulting in high concentration of oversea NOX and VOCs. Major species of VOCs for high O3 formation potential were aromatics and vinyls at the height of 0~500 m around the coastal region of metro Kaohsiung. The control of O3 precursors concentration showed that the ratio of [TVOCs-C2]/[NOX] in the offshore region was higher, indicating that O3 formation was NOX-limited. Therefore, NOX must be controlled for reducing O3 formation. However, the ratio of [TVOCs-C2]/[NOX] in the inland region was lower, some cases even below 4, showing that O3 formation was VOCs-limited. Thus, VOCs must be controlled for reducing O3 formation.

Nanoparticles are promising platforms for biomedical applications ranging from diagnostic tools to therapeutic delivery agents. During the course of these applications, nanoparticles are exposed to a complex mixture of extracellular serum proteins that nonspecifically adsorb onto the surface. The resulting protein layer, or protein "corona," creates an interface between nanoparticles and the biological environment. Protecting the nanoparticle surface can reduce protein adsorption, but complete inhibition remains a challenge. As a result, the corona, rather than the nanoparticle itself, mediates the cellular response to the nanoparticle. The following dissertation describes the fundamental characterization of the cellular binding of charged nanoparticles, interactions of protein-nanoparticle complexes with cellular receptors, and the structural and thermodynamic properties of adsorbed corona proteins.

The tumor suppressor protein, p53, is mutated or dysregulated in nearly all human cancers(1). The amino terminal domains are essential for transcriptional activation in stressed cells and play a vital role in cell cycle regulation, apoptosis and senescence. The transactivation (TAD) and proline rich domains in this region are dynamic and intrinsically disordered; lacking stable secondary or tertiary structure. This region contains multiple binding sites; arguably, the most significant of these is for p53's negative regulator, the E3 ligase, MDM2. An important, but less understood interaction involving the single stranded DNA binding protein, RPA70A, is hypothesized to be involved in maintaining genome integrity(2-4). Additionally, the amino terminus contains an important single nucleotide polymorphism that has demonstrated different affinity for MDM2 and is of significant biological importance in the induction of apoptosis (5). Isothermal titration calorimetry (ITC) and nuclear magnetic resonance (NMR) spectroscopy were employed to investigate how the thermodynamics and the inherent flexibility of the amino terminus of p53 play a role in complex formation with the MDM2 or RPA70 proteins. Understanding the structure, dynamics, and function of p53 is paramount in the fight against cancer.

Scanning electrochemical microscopy (SECM) was used for the study and characterization of catalytic and electrocatalytic processes occurring at electrodes. The Surface Interrogation mode (SI-SECM) was introduced for the detection and quantification of adsorbed intermediates and products of catalyzed chemical and electrochemical reactions at noble metals (Pt, Au). In SI-SECM two micro electrodes (i.e. an SECM tip and a substrate of the desired material) are aligned concentrically at a micrometric distance where SECM feedback effects operate. A contrast mechanism based on feedback effects allows for the detection of reactive adsorbed intermediates at the substrate: the SECM tip generates a reactive homogeneous species that “micro-titrates” the substrate adsorbates to yield an electrochemical signal that contains information about the amount of intermediate and about its kinetics of reaction with the redox mediator. The technique was used for the study of the reactivity of three model small adsorbates: 1) the reactivity of adsorbed oxygen on Au and Pt with a reducing mediator was explored and suggested the detection of “incipient oxides” at these surfaces; kinetic parameters of the reactivity of Pt oxides with mediators were obtained, fit to theory and used to explain observations about the electrocatalytic behavior of Pt under anodizing conditions; 2) the reactivity of oxidizing mediators with adsorbed hydrogen on Pt was studied and showed the cation of N,N,N,N-tetramethyl-p-phenylenediamine (TMPD) to be a successful interrogation agent, the detection of hydrogen generated by the decomposition of formic acid on Pt at open circuit was investigated; 3) electrogenerated bromine was used to catalytically interrogate carbon monoxide at Pt, this reaction was previously unreported. The mentioned applications of SECM were validated through the use of digital simulations of diffusion in the complex SECM geometry through flexible commercial finite element method software. / text