京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第19189号 / 工博第4066号 / 新制||工||1627(附属図書館) / 32181 / 京都大学大学院工学研究科合成・生物化学専攻 / (主査)教授 北川 進, 教授 濵地 格, 教授 森 泰生 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

No description available.

The interaction of carboxylic acids with copper is a phenomenon found both outdoors and, more commonly, indoors. The influence on copper of some carboxylic acids (formic, acetic, propionic, and butyric) have so far been studied at concentrations levels at least three or four orders of magnitude higher than actual indoor conditions (< 20 ppb, volume parts per billion), and with only limited emphasis on any mechanistic approach. In this licentiate study a unique analytical setup has been successfully applied for in situ characterization and quantification of corrosion products formed during initial atmospheric corrosion of copper in the presence of acetic, formic or propionic acid. The setup is based on monitoring mass changes by the quartz crystal microbalance (QCM) and simultaneously identifying the chemical species by infrared reflection-absorption spectroscopy (IRAS). Post-analysis of corrosion products was performed by coulometric reduction (mass of copper (I) oxide formed), grazing incidence x-ray diffraction (phase identification) and atomic force microscopy (surface topography). The absolute amounts of mass of individual constituents in the corrosion products, mainly copper (I) oxide or cuprite, copper (II) carboxylate and water or hydroxyl groups, have been deduced in situ during exposure in 120 ppb of carboxylic acid concentration, 95% relative humidity and 20ºC. An overall result is the consistency of analytical information obtained. For copper (I) oxide the quantified data estimated from IRAS, QCM or coulomeric reduction agrees with a relative accuracy of 12 % or better. The interaction of copper with the carboxylic acids seems to follow two spatially separated main pathways. A proton-induced dissolution of cuprous ions followed by the formation of copper (I) oxide, and a carboxylate-induced dissolution followed by the formation of copper (II) carboxylate. The first pathway is initially very fast but levels off with a more uniform growth over the surface. This pathway dominates in acetic acid. The second pathway exhibits a more constant growth rate and localized growth, and dominates in formic acid. Propionic acid exhibits low rates for both pathways. The difference between the carboxylic acids with respect to both total corrosion rate and carboxylate-induced dissolution can be attributed to differences in acid dissociation constant and deposition velocity.

Tesis por compendio / [ES] Esta tesis doctoral se enmarca en uno de los retos demandados por la sociedad relativo a la seguridad y calidad alimentaria, establecido en los actuales planes estatales y europeos de investigación científica, técnica y de innovación. En particular, la investigación realizada en esta tesis doctoral, contribuye a la validación de una técnica de análisis novedosa para la detección de pesticidas y antibióticos de interés en la miel. Esta técnica está basada en el uso combinado de la tecnología de sensores acústicos de alta resolución (High Fundamental Frequency Quartz Crystal Microbalance - HFF-QCM) funcionalizados con anticuerpos monoclonales, como elemento de reconocimiento biológico, para la detección de dos pesticidas (carbaril y DDT) y un antibiótico (sulfatiazol), en la matriz alimentaria miel. La metodología desarrollada se presenta como una técnica de análisis, alternativa y/o complementaria a las ya existentes, que resuelve algunos inconvenientes de las mismas; con la ventaja de requerir bajo consumo de muestra, ser fiable, simple, de bajo coste y de fácil manejo, por lo que no necesita de personal altamente cualificado. Además, es una técnica rápida que proporciona los resultados en pocos minutos, y está basada en instrumentación portátil, lo que facilita su implantación en la línea de trabajo. Los límites de detección alcanzados en miel por los inmunosensores HFF-QCM para carbaril, DDT y sulfatiazol fueron: 7, 24 y 0.10 µg/kg, respectivamente. Por su parte, los límites de cuantificación para los mismos analitos fueron: 17, 52 y 2 µg/kg, respectivamente. Los valores obtenidos con esta nueva tecnología son del mismo orden de magnitud que las técnicas de referencia actuales (cromatografía, SPR y ELISA), y confirman que cumple con los LMRs establecidos en el marco normativo actual. Respecto a la validación de esta nueva tecnología, el inmunosensor HFF-QCM desarrollado para la detección de sulfatiazol cumplió los requisitos de precisión y exactitud establecidos por la guía SANCO 12571/2013 para concentraciones de sulfatiazol en miel iguales o superiores a 10 ¿g/kg. Los inmunosensores HFF-QCM desarrollados para la detección de carbaril y DDT se aproximaron a los criterios de validación recomendados por la guía SANCO 12571/2013. Los resultados obtenidos con la técnica propuesta para el análisis de mieles reales han sido corroborados mediante cromatografía. En este sentido, ninguna de las muestras de miel reales analizadas mediante cromatografía y HFF-QCM reveló la presencia de pesticidas o de antibióticos en la miel, lo que confirma la fiabilidad de la técnica para la detección de estos contaminantes. Los hallazgos obtenidos en esta tesis son prometedores y establecen un punto de partida para futuros desarrollos encaminados a lograr la detección simultánea de varios analitos en un único análisis y muestra. Esto podría lograrse con un array de sensores HFF-QCM. En definitiva, la investigación realizada pone de manifiesto que los inmunosensores basados en la tecnología HFF-QCM son una alternativa seria a las técnicas actuales, para la cuantificación in situ de pesticidas y antibióticos en la industria de envasado de la miel. / [CA] Aquesta tesi doctoral s'emmarca en un dels reptes demandats per la societat relatiu a la seguretat i qualitat alimentària, establit en els actuals plans estatals i europeus d'investigació científica, tècnica i d'innovació. En particular, la investigació realitzada en aquesta tesi doctoral, contribuïx a la validació d'una tècnica d'anàlisi nova per a la detecció de pesticides i antibiòtics d'interés en la mel. Esta tècnica està basada en l'ús combinat de la tecnologia de sensors acústics d'alta resolució (High Fudamental Frequency Quartz Crystal Microbalance - HFF-QCM) functionalitzats amb anticossos monoclonals, com a element de reconeixement biològic, per a la detecció de dos pesticides (carbaril i DDT) i un antibiòtic (sulfatiazol), en la matriu alimentària mel. La metodologia desenvolupada es presenta com una tècnica d'anàlisi, alternativa y/o complementària a les ja existents, que resol alguns inconvenients de les mateixes; amb l'avantatge de requerir baix consum de mostra, ser fiable, simple, de baix cost i de fàcil maneig, per la qual cosa no necessita de personal altament qualificat. A més, és una tècnica ràpida que proporciona els resultats en pocs minuts, i està basada en instrumentació portàtil, la qual cosa facilita la seua implantació en la línia de treball Els límits de detecció aconseguits en mel pels inmunosensors HFF-QCM per a carbaril, DDT i sulfatiazol van ser: 7, 24 i 0.10 µg/kg, respectivament. Per la seua banda, els límits de quantificació per als mateixos analits van ser: 17, 52 i 2 µg/kg, respectivament. Els valors obtinguts amb aquesta nova tecnologia són del mateix orde de magnitud que les tècniques de referència actuals (cromatografia, SPR i ELISA), i confirmen que complix amb els LMRs establits en el marc normatiu actual. Respecte a la validació d'esta nova tecnologia, l'inmunosensor HFF-QCM desenvolupat per a la detecció de sulfatiazol va complir els requisits de precisió i exactitud establits per la guia SANCO 12571/2013 per a concentracions de sulfatiazol en mel iguals o superiors a 10 µg/kg. Els inmunosensors HFF-QCM desenvolupats per a la detecció de carbaril i DDT es van aproximar als criteris de validació recomanats per la guia SANCO 12571/2013. Els resultats obtinguts amb la tècnica proposta per a l'anàlisi de mels reals han sigut corroborats per mitjà de cromatografia. En aquest sentit, cap de les mostres de mel reals analitzades per mitjà de cromatografia i HFF-QCM va revelar la presència de pesticides o d'antibiòtics en la mel, la qual cosa confirma la fiabilitat de la tècnica per a la detecció d'estos contaminants. Les troballes obtinguts en aquesta tesi són prometedors i estableixen un punt de partida per a futurs desenvolupaments encaminats a aconseguir la detecció simultània de diversos analits en un únic anàlisi i mostra. Açò podria aconseguir-se amb una matriu de sensors HFF-QCM. En definitiva, la investigació realitzada posa de manifest que els inmunosensors basats en la tecnologia HFF-QCM són una alternativa seriosa a les tècniques actuals, per a la quantificació in situ de pesticides i antibiòtics en la indústria d'envasament de la mel. / [EN] This thesis is part of one of the challenges demanded by society regarding food safety and quality, established in the current state and European plans for scientific, technical and innovation research. In particular, the research carried out in this doctoral thesis contributes to the validation of a novel analysis technique for the detection of pesticides and antibiotics of interest in honey. This technique is based on the combined use of high resolution acoustic sensor technology (High Fundamental Frequency Quartz Crystal Microbalance - HFF-QCM) functionalized with monoclonal antibodies, as a biological recognition element, for the detection of two pesticides (carbaryl and DDT ) and an antibiotic (sulfathiazole), in the honey food matrix. The developed methodology is presented as an analysis technique, alternative and / or complementary to those that already exist, which solves some of their drawbacks; with the advantage of requiring low sample consumption, being reliable, simple, low cost and easy to use, so it does not require highly qualified personnel. In addition, it is a fast technique that provides results in a few minutes, and is based on portable instrumentation, which facilitates its implementation in the industry. The detection limits reached in honey by the HFF-QCM immunosensors for carbaryl, DDT and sulfatiazole were: 7, 24 and 0.10 µg / kg, respectively. On the other hand, the quantification limits for the same analytes were: 17, 52 and 2 µg / kg, respectively. The values obtained with this new technology are of the same order of magnitude as the current reference techniques (chromatography, SPR and ELISA), and confirm that it meets the MRLs established in the current regulatory framework. Regarding the validation of this new technology, the HFF-QCM immunosensor developed for the detection of sulfathiazole meet the precision and accuracy requirements established by the SANCO 12571/2013 guideline for concentrations of sulfathiazole in honey equal to or greater than 10 ¿g / kg. The HFF-QCM immunosensors developed for the detection of carbaryl and DDT approached the validation criteria recommended by the SANCO guide 12571/2013. The results obtained with the proposed technique for the analysis of real honeys have been verified by chromatography. In this sense, none of the real honey samples analyzed by chromatography and HFF-QCM revealed the presence of pesticides or antibiotics in honey, confirming the reliability of this technique for the detection of these contaminants. The findings obtained in this thesis are promising and establish a starting point for future developments aimed at achieving the simultaneous detection of several analytes in a single analysis and sample. This could be accomplished with an array of HFF-QCM sensors. Ultimately, the research carried out shows that immunosensors based on HFF-QCM technology are a serious alternative to current techniques for in situ quantification of pesticides and antibiotics in honey packaging industry. / Así mismo quiero agradecer a la Generalitat Valenciana y al Fondo Social Europeo por haberme proporcionado la financiación económica mediante el contrato “Ayudas para la contratación de personal investigador en formación de carácter predoctoral” (ACIF/2016/132), ya que sin este contrato la realización de esta tesis no habría sido posible. Además, quiero agradecer de nuevo a la Generalitat Valenciana y al Fondo Social Europeo por el apoyo financiero para llevar a cabo una estancia predoctoral en el Instituto de Ciencia y Tecnología de los Alimentos de la Universidad de León (ICTAL), España (BEFPI/2018/021). Y no menos importante es el agradecimiento al Ministerio Español de Economía y Competitividad, que mediante Fondos FEDER ha financiado esta investigación a través del proyecto titulado: “Diseño de una plataforma basada en matrices de sensores de cuarzo de alta resolución para HTS (High-Throughput Screening) de residuos en mieles” (PQHTS-HONEY, AGL 2013-48646-R). / Cervera Chiner, L. (2020). Aplicación y validación de la tecnología de inmunosensores piezoeléctricos de alta frecuencia (HFF-QCM) para la detección de pesticidas y antibióticos en miel [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/152488 / TESIS / Compendio

Metallic films are important in various applications, such as electric devices where it can act as contacts. In electrical devices, the substrate typically consists of silicon dioxide (SiO2) which is a temperature-sensitive substrate. Therefore, plasma enhanced chemical vapor deposition (PECVD) are better suited than thermally activated chemical vapor deposition (CVD). Depositing metallic films with PECVD demands co-reactants that act as reducing agents. However, these are not well-studied and do not always have the power enough to perform the reduction reaction for metals. Recently it has been concluded that electrons can act as reducing agents in the deposition of first row transition metallic films in a PECVD process. By supplying a positive bias to the substrate, the electrons got attracted to the surface of the substrate, which facilitated metal growth. The study concluded that metal growth only occurred at conductive -and semiconductive substrates and that the substrate bias and plasma power affected the metal growth. The process is however not well understood, which causes a knowledge gap, signifying that studies of the surface chemistry are needed. Here a new modified analytical method to study the surface chemistry in the newly developed process mentioned above is presented. The analytical method consists of an electrically modified quartz crystal microbalance (QCM) with gold electrodes as a conductive substrate. This allows the electron current to run through the QCM during the measurement. By supplying a DC-voltage to the front electrode it gets readily biased (negative and positive) and by placing a capacitor in the circuit, it connects the AC-circuit (oscillator circuit) and the DC-circuit (DC-voltage bias circuit). At the same time, it blocks the DC-current from going back to the oscillator but allows the high-frequency signal to pass from the QCM. The results in this thesis concluded that the QCM can be electrically modified to allow an electron flux to the QCM while using it as a substrate when electrons are used as reducing agents. Scanning electron microscopy (SEM) of a QCM crystal revealed that a 2 µm film had been deposited while SEM coupled with energy dispersive X-ray spectroscopy (EDS) showed that the film indeed contained iron. Further analysis was made by high-resolution X-ray photoelectron spectroscopy (HR-XPS) to find the elemental composition of the film, which revealed that the thin film contained 41 at.% iron. In addition, this study investigated if the QCM could be used to study CVD processes where electrons were used as reducing agents. The results indeed revealed that it is possible to study the surface chemistry where electrons are used as reducing agents with the electrically modified QCM to gain knowledge concerning film deposition. Initial results of the QCM showed that film growth could be studied when varying the plasma power between 5 W to 15 W and the QCM bias between -40 V to +40 V. The method generated easily accessible data concerning the process where electrons are used as reducing agents, which gained insight to the method that never has been disclosed before.

Artificial lipid bilayers formed on solid surface supports are widespread model systems to study physical, chemical, as well as biological aspects of cell membranes and fundamental interfacial interactions. The approach to use a thin polymer film representing a cushion for lipid bilayers prevents incorporated membrane proteins from pinning to the support and mimics the native environment of a lipid bilayer in certain aspects of the extracellular matrix and intracellular structures. A key component for cell anchorage to extracellular fibronectin is the transmembrane adhesion receptor alpha(5)beta(1) integrin. Its transport dynamics and clustering behavior plays a major role in the assembly of focal adhesions, which mediate mechanical forces and biochemical signals of cells with their surrounding. The system investigated herein is envisioned to use extrinsically controlled stimuli-responsive polymer cushions to tune the frictional drag between polymer cushion and mobile membranes with incorporated integrins to actively regulate lipid membrane characteristics. To attain this goal, a temperature- and pH-responsive polymer based on poly(N-isopropylacrylamide) copolymers containing varying amounts of carboxyl-group-terminated comonomers at different aliphatic spacer lengths (PNIPAAm-co-carboxyAAM) was surface-grafted to a poly(glycidyl methacrylate) anchorage layer. The swelling transitions were characterized using atomic force microscopy, ellipsometry and quartz crystal microbalance with dissipation monitoring (QCM-D) and found to be tunable over a wide range of temperature and pH. In agreement with the behavior of the polymers in solution, longer alkyl spacers decreased the phase transition temperature T(P) and higher contents of carboxylic acid terminated comonomers increased T(P) at alkaline conditions and decreased T(P) at acidic conditions. Remarkably, the point where the degree of carboxyl group deprotonation balances the T(P)-lowering effect of the alkyl spacer was distinctive for each alkyl spacer length. These findings illustrate how the local and global balance of hydrophilic and hydrophobic interactions along the copolymer chain allows to adjust the swelling transition to temperatures below, comparable, or above those observed for PNIPAAm homopolymers. Additionally, it could be shown that surface-grafting leads to a decrease in T(P) for PNIPAAm homopolymers (7°C) and copolymers (5°C - 10°C). The main reason is the increase in local polymer concentration of the swollen film constrained by dense surface anchorage in comparison to the behavior of dilute free chains in solution. In accordance with the Flory-Huggins theory, T(P) decreases with increasing concentration up to the critical concentration. Biological functionalization of the PNIPAAm-co-carboxyAAm thin films was demonstrated for the cell adhesion ligand peptide cRGD via carbodiimide chemistry to mimic extracellular binding sites for the cell adhesion receptors integrin. The outcome of QCM-D measurements of cRGD-functionalized surfaces showed a maintained stimuli-responsiveness with slight reduction in T(P). A drying/rehydration procedure of a 9:1 lipid mixture of the cationic lipid dioleoyl-trimethylammoniumpropane (DOTAP) and the zwitterionic dioleoyl-phosphatidylcholine (DOPC) was utilized to form lipid bilayer membranes on PNIPAAm-co-carboxyAAM cushions. Fluorescence recovery after photobleaching (FRAP) revealed that lipid mobility was distinctively higher (6.3 - 9.6) µm2 s-1 in comparison to solid glass support ((3.0 - 5.9) µm2 s-1). In contradiction to the initial expectations, modulation of temperature and pH led to poor variations in lipid mobility that did not correlate with the PNIPAAm cushion swelling state. The results suggested a weak coupling of the lipid bilayer with PNIPAAm polymer cushions that can be slightly tuned by electrostatic interactions. The transmembrane adhesion receptor alpha(5)beta(1) integrin was reconstituted into liposomes consisting of DOPC/sphingomyelin/cholesterol 2:2:1 for the formation of polymer cushioned bilayers. PNIPAAm- co-carboxyAAM and maleic acid (MA) copolymers were used as cushions, both with the option for cRGD functionalization. On the MA copolymer cushions, fusion of proteoliposomes resulted in supported bilayers with mobile lipids as confirmed by FRAP. However, incorporated integrins were immobile. In an attempt to explain this observation, the medium-sized cytoplasmic integrin domain was accounted to hamper the movement by steric interactions with the underlying polymer chains in conjunction with electrostatic interactions of the cationic cytoplasmic domain with the oppositely charged MA copolymer. On the PNIPAAm-co-carboxyAAM cushion only a drying/rehydration procedure lead to bilayer formation. However, again the integrins were immobile, presumably due to the harsh treatment during preparation. Nevertheless, the results of the investigated set of PNIPAAm copolymer films suggest their application as temperature- and pH-responsive switchable layers to control interfacial phenomena in bio-systems at different physiological conditions. The PNIPAAm-co-carboxyAAm cushioned bilayer system represents a promising step towards extrinsically controlled membrane – substrate interactions.

Byl studován kontakt mezi koulí a deskou pod tangenciálním zatížením ve vodě. Jako zdroj střižných kmitů byl použit akustický rezonátor (křemíkové mikrováhy – QCM). Kontakt koule s povrchem resonátoru indukuje změnu resonanční frekvence a šířky pásma. Byla měřena změna frekvence f a změna šířky pásma v závislosti na amplitudě oscilací. S rostoucí amplitudou docházelo k poklesu f a růstem , což je chování typické pro parciální skluz. Díky aplikaci Cattaneo-Mindlinova modelu byl vypočítán kontaktní poloměr a třecí koeficient. Kontaktní poloměr při nízké amplitudě stoupal při zvětšujícím se normálovým zatížením. Tato závislost se dobře shodovala s JKR modelem. Třecí koeficient se nacházel v odpovídajícím rozsahu. Při zvyšování externí normálové síly, docházelo k nepatrnému snižování hodnoty třecího koeficientu. Toto chování je vysvětleno příspěvkem adhezivních sil k totální normálové síle. Výpočtem byly získány dva typy třecích koeficientů, první ze změny frekvence f a druhý ze změny šířky pásma . Tyto dvě hodnoty se spolu shodovaly z ± 20 % pro měření prováděná ve vodě, zatímco pro dvě měření prováděných na hydrofilním povrchu ve vzduchu se lišila. Tento nesoulad poukazuje na nedostatek Cattaneo Mindlinovy teorie a mohl by být vysvětlen přítomností kapilárních sil.

Artificial lipid bilayers formed on solid surface supports are widespread model systems to study physical, chemical, as well as biological aspects of cell membranes and fundamental interfacial interactions. The approach to use a thin polymer film representing a cushion for lipid bilayers prevents incorporated membrane proteins from pinning to the support and mimics the native environment of a lipid bilayer in certain aspects of the extracellular matrix and intracellular structures. A key component for cell anchorage to extracellular fibronectin is the transmembrane adhesion receptor alpha(5)beta(1) integrin. Its transport dynamics and clustering behavior plays a major role in the assembly of focal adhesions, which mediate mechanical forces and biochemical signals of cells with their surrounding. The system investigated herein is envisioned to use extrinsically controlled stimuli-responsive polymer cushions to tune the frictional drag between polymer cushion and mobile membranes with incorporated integrins to actively regulate lipid membrane characteristics. To attain this goal, a temperature- and pH-responsive polymer based on poly(N-isopropylacrylamide) copolymers containing varying amounts of carboxyl-group-terminated comonomers at different aliphatic spacer lengths (PNIPAAm-co-carboxyAAM) was surface-grafted to a poly(glycidyl methacrylate) anchorage layer. The swelling transitions were characterized using atomic force microscopy, ellipsometry and quartz crystal microbalance with dissipation monitoring (QCM-D) and found to be tunable over a wide range of temperature and pH. In agreement with the behavior of the polymers in solution, longer alkyl spacers decreased the phase transition temperature T(P) and higher contents of carboxylic acid terminated comonomers increased T(P) at alkaline conditions and decreased T(P) at acidic conditions. Remarkably, the point where the degree of carboxyl group deprotonation balances the T(P)-lowering effect of the alkyl spacer was distinctive for each alkyl spacer length. These findings illustrate how the local and global balance of hydrophilic and hydrophobic interactions along the copolymer chain allows to adjust the swelling transition to temperatures below, comparable, or above those observed for PNIPAAm homopolymers. Additionally, it could be shown that surface-grafting leads to a decrease in T(P) for PNIPAAm homopolymers (7°C) and copolymers (5°C - 10°C). The main reason is the increase in local polymer concentration of the swollen film constrained by dense surface anchorage in comparison to the behavior of dilute free chains in solution. In accordance with the Flory-Huggins theory, T(P) decreases with increasing concentration up to the critical concentration. Biological functionalization of the PNIPAAm-co-carboxyAAm thin films was demonstrated for the cell adhesion ligand peptide cRGD via carbodiimide chemistry to mimic extracellular binding sites for the cell adhesion receptors integrin. The outcome of QCM-D measurements of cRGD-functionalized surfaces showed a maintained stimuli-responsiveness with slight reduction in T(P). A drying/rehydration procedure of a 9:1 lipid mixture of the cationic lipid dioleoyl-trimethylammoniumpropane (DOTAP) and the zwitterionic dioleoyl-phosphatidylcholine (DOPC) was utilized to form lipid bilayer membranes on PNIPAAm-co-carboxyAAM cushions. Fluorescence recovery after photobleaching (FRAP) revealed that lipid mobility was distinctively higher (6.3 - 9.6) µm2 s-1 in comparison to solid glass support ((3.0 - 5.9) µm2 s-1). In contradiction to the initial expectations, modulation of temperature and pH led to poor variations in lipid mobility that did not correlate with the PNIPAAm cushion swelling state. The results suggested a weak coupling of the lipid bilayer with PNIPAAm polymer cushions that can be slightly tuned by electrostatic interactions. The transmembrane adhesion receptor alpha(5)beta(1) integrin was reconstituted into liposomes consisting of DOPC/sphingomyelin/cholesterol 2:2:1 for the formation of polymer cushioned bilayers. PNIPAAm- co-carboxyAAM and maleic acid (MA) copolymers were used as cushions, both with the option for cRGD functionalization. On the MA copolymer cushions, fusion of proteoliposomes resulted in supported bilayers with mobile lipids as confirmed by FRAP. However, incorporated integrins were immobile. In an attempt to explain this observation, the medium-sized cytoplasmic integrin domain was accounted to hamper the movement by steric interactions with the underlying polymer chains in conjunction with electrostatic interactions of the cationic cytoplasmic domain with the oppositely charged MA copolymer. On the PNIPAAm-co-carboxyAAM cushion only a drying/rehydration procedure lead to bilayer formation. However, again the integrins were immobile, presumably due to the harsh treatment during preparation. Nevertheless, the results of the investigated set of PNIPAAm copolymer films suggest their application as temperature- and pH-responsive switchable layers to control interfacial phenomena in bio-systems at different physiological conditions. The PNIPAAm-co-carboxyAAm cushioned bilayer system represents a promising step towards extrinsically controlled membrane – substrate interactions.

I värme- och kraftvärmeverk förbränns olika typer av bränslen för produktion av el och fjärrvärme. Vid förbränningen bildas rökgaser som innehåller föroreningar, till exempel tungmetaller, från bränslet. Anläggningarna har ofta krav på utsläpp både via rökgaserna och avloppsvatten. Rökgaserna renas därmed genom olika tekniker var av en vanlig teknik är rökgaskondensering. Vid rökgaskondenseringen bildas en vätska, kallad rökgaskondensat, som delvis innehåller tungmetaller från bränslet. Rökgaskondensatet måste renas innan det kan lämna anläggningen och det görs bland annat med tungmetalljonbytare. Jonbytarmassan i tungmetalljonbytarkolonnerna behöver bytas ungefär två gånger per driftsäsong då den inte längre kan binda mer tungmetaller. Detta är en kostnad för värme- och kraftvärmeverken som de vill minimera. I denna studie undersöktes om lokal provtagning och analys på ett kraftvärmeverk av ett antal utvalda tungmetaller i rökgaskondensat är en bra metod för att optimering av reningssteget med tungmetalljonbytare. Samt om detta kan säkerställa att miljökraven för tungmetaller i det renade rökgaskondensatet uppfylls. Med optimering avses att jonbytarmassornas fulla kapacitet utnyttjas, d.v.s. att byten av jonbytarmassor kan reduceras utan att riskera otillåtna halter av tungmetaller i de renade rökgaskondensatet till följd av att jonbytarmassorna använts för länge. Även tiden som behöver avsättas för lokal provtagning och analys dokumenterades. I dagsläget sker analyser hos ackrediterade laboratorium där det tar drygt två veckor att få resultatet och under väntetiden kan mycket på anläggningen förändras. En verifiering av resultaten från studien gjordes mot resultat från ett sådant. I denna studie undersöktes lokal provtagning och analys med mätinstrumentet FREEDD som bygger på tekniken kvartskristall mikrobalans (QCM-teknik). Andra alternativ för lokal analys har inte undersökts här.  Resultatet visade att det i dagsläget är svårt att med lokal provtagning optimera reningssteget med jonbytarmassor samt kontrollera utsläppen av tungmetaller via det renade rökgaskondensatet. Korrigeringar hos mätinstrumentet och provpunkterna behöver göras för att få pålitligt resultat. Tiden som behöver avsättas för provtagning och analys beror på vilken metall som ska analyseras då tiden för preparering av prov varierar. Men om det kan möjliggöra att anläggningarna kan använda jonbytarmassorna längre samt får kontroll på utsläppen via det renade rökgaskondensatet kan det vara lönsamt att avvara den tiden. / In heating and combined heat and power plants, different types of fuels are burned to produce electricity and district heating. During the combustion flue gases containing pollutants, such as heavy metals, are formed from the flue. The plants have requirements for low emissions, both from the flue gases and the wastewater. The flue gases are purified by various techniques and a common technique is flue gas condensation. During the flue gas condensation, a liquid called flue gas condensate, is formed, which partly contains heavy metals from the flue. The flue gas condensate must be cleaned before it can leave the plant. A step in the purification of the flue gas condensate is usually heavy metal ion-exchanger. The ion-exchange mass in the heavy metal ion-exchange columns needs to be changed approximately twice per operating season as it no longer has room to bind more heavy metals. This is an expensive cost for the heating and combined heat and power plants that they want to minimize. This study investigated whether local sampling and analysis at a cogeneration plant of a number selected heavy metals in flue gas condensate is a good method for optimizing the purifications step with heavy metal ion-exchangers. And if this can ensure that the environmental requirements for the heavy metals in the purified flue gas condensate are met. Optimization means that the full capacity of the ion-exchange masses is utilized, i.e. that the exchange of ion-exchange masses can be reduced without risking unauthorized levels of heavy metals in the purified flue gas condensate as a result of the ion exchange masses being used for too long.  The time needed for local sampling and analysis was also documented. At present, analyzes are done at accredited laboratories where it takes over two weeks to get the result and during that time much can be changes at the plant. A verification of the result of the study was also made against the result of an accredited laboratory. In this study, local analysis was made with the measuring instrument FREEDD which is based on quartz crystal microbalance (QCM-technology). Other options for local sampling and analysis have not been investigated. The result showed that, in the present, it is difficult to optimize the purification step with ion-exchange masses and check emissions of heavy metals with the purified flue gas condensate. To obtain reliable result, corrections to the measuring instrument and test points need to be made. The time that needs to be set aside for sampling and analysis depends on the metal, as the time for sample preparation varies.  But if it can enable the plants to use the ion-exchange masses longer and gain control of the emissions of heavy metals with the purified flue gas condensate, it can be profitable to save that time.

Characterizing nanoparticles to be able to understand how they functions in the body is important for development of drugs. Furthermore with increasing number of nanoparticle product the nanotoxicity of nanoparticles is important to understand. This report is a part of the EU-project Nanoclassifier which purpose is to “develop a cost effective, high throughput screening platform for characterization of the bionanointerface and its cell-binding partners”. Single-cycle kinetic was used to determine the number of binding epitopes on polystyrene nanoparticle with transferrin corona. The number of available epitopes describes how active the Nanoparticle will be in the body. For this purpose Single-cycle kinetic methodology was successfully used on nanoparticles. Single-cycle kinetic methodology has great potential to become the standard method for high throughput nanoparticle epitope characterization.