With their unique photochemical properties, porphyrins have remained a central research topic for decades. Porphyrins can self-assemble into tubular structures at acidic pHs. However, the possibility of the disassembly of the aggregated porphyrin has not been investigated. Furthermore, quantitative understanding of the porphyrin optical activities is complicated by the complex interplay of the photon absorption, scattering, and fluorescence emission that can concurrently occur in porphyrin samples. Using meso- Tetrakis (4-sulfonato phenyl) porphyrin (TPPS) as the model molecule, discussed herein is combined UV-vis extinction, Stokes-shifted fluorescence, and polarized resonance synchronous spectroscopy (PRS2) study of porphyrin assembly and disassembly at acidic solutions. A series of optical constants, including photon absorption, scattering, and fluorescence emission cross-sections as well as its fluorescence and light scattering depolarizations has been quantified. Compared to UV-vis and SSF methods, the PRS2 is significantly more sensitivity in the detection of both concentration- and time-dependent porphyrin aggregation.

No description available.

Yes / The association constants for formation of 1 : 1 complexes between a H-bond donor, 1-naphthol, and a diverse range of charged and neutral H-bond acceptors have been measured using UV/vis absorption and fluorescence emission titrations. The performance of 1-naphthol as a dual colorimetric and fluorescent molecular recognition probe for determining the H-bond acceptor (HBA) parameters of charged and neutral solutes has been investigated in three solvents. The data were employed to establish self-consistent H-bond acceptor parameters (β) for benzoate, azide, chloride, thiocyanate anions, a series of phosphine oxides, phosphate ester, sulfoxide and a tertiary amide. The results demonstrate both the transferability of H-bond parameters between different solvents and the utility of the naphthol-based dual molecular recognition probe to exploit orthogonal spectroscopic techniques to determine the HBA properties of neutral and charged solutes. The benzoate anion is the strongest HBA studied with a β parameter of 15.4, and the neutral tertiary amide is the weakest H-bond acceptor investigated with a β parameter of 8.5. The H-bond acceptor strength of the azide anion is higher than that of chloride (12.8 and 12.2 respectively), and the thiocyanate anion has a β value of 10.8 and thus is a significantly weaker H-bond acceptor than both the azide and chloride anions. / Supported by the ESPRC.

Lyophilized pea chloroplasts were extracted in a stepwise manner in an organic solvent (petroleum ether) with increasing polarity which was obtained by addition of small amount of ETOH (0-1 %). Absorption and low temperature fluorescence emission spectra were measured on both the extracted thylakoids and on isolated chlorophylI-protein complexes. Extraction of chlorophyll from the membrane increased (and the ratio of chlorophyl l a/b decreased) with increasing polarity of the solvent. The gel scan revealed that after extraction with petroleum ether, CPa.. was lost from the gel and after extraction with petroleum ether +1 % ETOH only the CPa/b was left together with SDS-free chlorophyll. This shows that the chlorophyll in CPa/b are situated in a less hydrophobic environment than chlorophyll in CPa|| and CPa|. The long wavelength absorbing and emitting chlorophyll fraction associated to CPaj was found to be easily removed from the membrane. This caused a blue shift in the low temperature fluorescence emission peak and in the red absorption peak and it was also accompanied by a decrease in carote-noid absorption in isolated CPa|. It was found in different plant material lacking $-carotene in CPàj that a strong correlation between ß-carotene in CPa. and the existence of the long wavelength chlorophyll in isolated cPa. existed. Based on these data, it was suggested that excited chlorophyll can transfer energy in excess to ß-carotene by a triplet--triplet transfer.A method based on in vivo chlorophyll £ fluorescence was developed for studying photosynthetic capacity in unicellular algae. It was shown that DCMU-induced fluorescence increase was a good measure of photosynthetic capacity in four species of green algae tested.The effect of copper chloride on photosynthetic electron transport and chlorophyl1-protein complexes was studied in spinach chloroplasts. Copper(11) inhibited a PS I i reaction H2O—&gt; DPIP, a PS I reaction Asc/DPIP —&gt; NADP and the overall electron transport H2O —&gt; NAOP to different degrees. Chlorophyll protein complexes were only slightly affected by copper(ll) but with both copper and ascorbate in the reaction media, a rapid membrane destruction occurred. This was probably caused by a free radical reaction catalyzed by copper(ll). / <p>Härtill 4 delarbeten.</p> / digitalisering@umu

Ο Πατραϊκός Κόλπος βρίσκεται στη Δυτική Ελλάδα μεταξύ Στερεάς Ελλάδας και Πελοποννήσου. Είναι ένας σχετικά αβαθής θαλάσσιος κόλπος που συνδέει το βαθύ Κορινθιακό Κόλπο με το Ιόνιο Πέλαγος. Η περιοχή μελέτης εκτείνεται από την παραλία της Καλόγριας, Δ. Λαρισσού, μέχρι το Ακταίο του Δήμου Ρίου. Πραγματοποιηθήκαν 2 εποχικές δειγματοληψίες, για τη συλλογή θαλάσσιου νερού, μια χειμερινή το Μάρτιο του 2009 και μία θερινή το Σεπτέμβριο του ίδιου έτους. Εμβόλιμα έγινε δειγματοληψία λίγο μετά από ατύχημα πετρελαιοκηλίδας στο λιμάνι της Πάτρας. Στην εργασία αυτή με μία τροποποίηση της μεθόδου MARPOLMON-P, μετρήθηκε η εκπομπή φθορισμού των δειγμάτων μετά από την εκχύλιση τους με εξάνιο και διερευνήθηκε η παρουσία και η συγκέντρωση πετρελαϊκών ενώσεων καθώς και η διασπορά των πετρελαιοειδών στις παράκτιες περιοχές του Πατραϊκού Κόλπου. Σε κάθε δείγμα μετρήθηκαν φυσικές παράμετροι όπως η θερμοκρασία, το pH, η ηλεκτρική αγωγιμότητα και η θολερότητα του θαλάσσιου νερού. Η σύσταση των δειγμάτων της δεύτερης δειγματοληψίας διερευνήθηκε με τη χρήση GC-MS για τον εντοπισμό της πηγής των πετρελαϊκών ρύπων. Τα αποτελέσματα των δύο κύριων δειγματοληψιών παρουσίασαν μεγάλες διακυμάνσεις στη συγκέντρωση αλλά και στην κατανομή των πετρελαϊκών υδρογονανθράκων, με τα εύρη των τιμών των συγκεντρώσεων να κυμαίνονται για τη χειμερινή από 0 – 290 μg/L ισοδύναμων μονάδων χρυσενίου (Λιμάνι Προβλήτα Αγ. Νικολάου) και για τη θερινή δειγματοληψία από 0- 56 μg/L μονάδων χρυσενίου (Μαρίνα Τερψιθέας). Η κατανομή και η διασπορά των πετρελαιοειδών φαίνεται να επηρεάζεται άμεσα από τη διεύθυνση και ταχύτητα των κυμάτων και του επικρατούντος άνεμου στις περιοχές δειγματοληψίας, αλλά και από την ωκεανογραφική συμπεριφορά του Πατραϊκού Κόλπου. Στην ανάλυση GC/MS βρέθηκε μεγάλη αναλογία αλκάνιων σε όλα τα δείγματα, ενώ σε κάποια από αυτά βρέθηκαν πολυκυκλικοί αρωματικοί υδρογονάνθρακες και αλκένια, παράγωγα του πετρελαίου και των καυσίμων. Άλλες οργανικές ενώσεις όπως φθαλικοί εστέρες, σουλφίδια, αλκοόλες κ.α., ανιχνεύτηκαν σε χαμηλές συνήθως συγκεντρώσεις και πιθανόν προέρχονται από ρυπογόνες χερσαίες πηγές (επιφανειακή απορροή, αστικά και βιομηχανικά λύματα κ.α.). / The Gulf of Patras located between Continental Greece and Peloponnesus, in West Greece. The Gulf of Patras is relatively shallow and connects the deep Gulf of Corinth with Ionian Open Sea. The experimental area of our study extends from the beach of Kalogria, department of Municipality of Larissos, to Aktaion, department of Municipality of Rion. Two samplings, one in March of 2009 and one in September of the same year, were carried out in order to get sea water samples. After the occurrence of an oil slick accident in the port of Patras, an interstitial sampling was carried out. In this study, MARPOLMON-P method partially modified was used, in order to estimate the fluorescence emission of the hexane extracted sea water samples. The presence of Dissolved/Dispersed petroleum hydrocarbons (DDPH) was also investigated as well as their concentration and the distribution of those in the maritime areas of the Gulf of Patras. In every sample, physical parameters such as temperature, pH, electrical conductivity and turbidity of sea water were estimated. The identification of various substances within the samples of the second sampling (September of 2009) was conducted with GC-MS method, in order to find out the source of oil pollution. Considering our results of the two main samplings we found out that there were important ranges in concentration as well as in distribution of DDPH. At first sampling the range of values was between 0 to 290 μg/L of chrysene equivalents (Port of St. Nikolaos) and at second sampling was between 0 to 56 μg/L of chrysene equivalents (Port of Marina Terpsithea). The results showed that wave’s direction and speed, wind speed and oceanographical status of Gulf of Patras affected by the distribution and dispersion of petroleum hydrocarbons. During the GC/MS analysis high content of alkanes in all collected samples was identified, while some of them contained polycyclic aromatic hydrocarbons and alkenes, such as products of crude oil and mineral fuel. Other organic compounds such as pthallic esters, sulphides, alcoholes etc were found in low concentrations. These organic compounds were probably derived from pollutant ground sources (superficial rain flow or dilution drainage, urban and manufacturing wastesreams).

Les fonctions organiques du corps humain sont liées à des constants biologiques. Variations de ces constantes induisent divers états pathologiques. Parmi ces constantes, le pH constitue le cœur de ces travaux de thèse. Chez les êtres vivants, les fonctions biologiques dépendent de constant acides ou alcalines. En fait, l’action d’une protéine dépend du pH du milieu environnant. Une valeur inadéquate du pH rend les protéines inactives ce qui est délétère pour l’organisme. Il existe donc un besoin pour des capteurs de pH qui puissant être utilisés dans le corps humain pour des applications cliniques (échelle macroscopique), sur des cellules en culture pour des recherches en biologie (échelle mesoscopique) et pour étudier les échanges ioniques au niveau des membranes cellulaires pour des travaux plus fondamentaux (échelle microscopique). Parmi le large éventail de technologies potentiellement candidates pour ces applications, la mesure de pH par fibre optique exploitant la fluorescence permet d’être adaptée aux trois échelles dimensionnelles susnommées. Ce manuscrit de thèse adresse des contraintes par l’étude de capteurs fluorescents à fibre optique utilisant deux types d’indicateur de pH: les SNARF et la fluorescéine. En parallèle de ces développements expérimentaux, des descriptions mathématiques des propriétés de fluorescence de ces deux indicateurs sont proposées. Ces descriptions permettent de progresser vers une mesure du pH sans calibration / Organic functions of the human body are linked to biological constants. Variations of these constants induce pathological troubles. Among these constants, the pH is the central subject of this PhD work. In living beings, biological functions are related to either acid or alkaline constants. Indeed, the action of a protein depends on the surrounding pH. An inadequate value of the pH makes the proteins non active which is deleterious for the organism. There exist a need for pH sensors which can be used in the human body for clinical applications (macroscopic scale), on cells in culture for biology researches (mesoscopic scale) or at a cell membrane level for more fundamental studies (microscopic scale). Among the wide range of technologies potentially useful for these applications, fiber optic fluorescence pH sensing offers the possibility to be adapted to the three above mentioned dimensional scales. This PhD dissertation addresses these constraints by studying fluorescence fiber optics pH sensors using two kind of pH indicators: SNARF and fluorescein. Together with these experimental studies, mathematical descriptions of the fluorescence properties of these indicators are proposed. They allow progressing towards calibration free pH sensing.

The in origin intertidal marine brown alga Fucus vesiculosus L. grow permanently sublittoral in the brackish Bothnian Sea, side by side with the recently discovered F. radicans L. Bergström et L. Kautsky. Environmental conditions like salinity, light and temperature are clearly different between F. vesiculosus growth sites in the Bothnian Sea (4-5 practical salinity units, psu; part of the Baltic Sea) and the tidal Norwegian Sea (34-35 psu; part of the Atlantic Ocean). The general aims of this thesis were to compare physiological aspects between the marine ecotype and the brackish ecotype of F. vesiculosus as well as between the two Bothnian Sea species F. vesiculosus and F. radicans. The result in the study indicates a higher number of water soluble organic compounds in the marine ecotype of F. vesiculosus compared to the brackish ecotype. These compounds are suggested to be compatible solutes and be due to an intertidal and sublittoral adaptation, respectively; where the intertidal ecotype needs the compounds as a protection from oxygen radicals produced during high irradiation at low tide. The sublittoral ecotype might have lost the ability to synthesize these compound/compounds due to its habitat adaptation. The mannitol content is also higher in the marine ecotype compared to the brackish ecotype of F. vesiculosus and this is suggested to be due to both higher level of irradiance and higher salinity at the growth site. 77 K fluorescence emission spectra and immunoblotting of D1 and PsaA proteins indicate that both ecotypes of F. vesiculosus as well as F. radicans have an uneven ratio of photosystem II/photosystem I (PSII/PSI) with an overweight of PSI. The fluorescence emission spectrum of the Bothnian Sea ecotype of F. vesiculosus however, indicates a larger light-harvesting antenna of PSII compared to the marine ecotype of F. vesiculosus and F. radicans. Distinct differences in 77 K fluorescence emission spectra between the Bothnian Sea ecotype of F. vesiculosus and F. radicans confirm that this is a reliable method to use to separate these species. The marine ecotype of F. vesiculosus has a higher photosynthetic maximum (Pmax) compared to the brackish ecotype of F. vesiculosus and F. radicans whereas both the brackish species have similar Pmax. A reason for higher Pmax in the marine ecotype of F. vesiculosus compared to F. radicans is the greater relative amount of ribulose-1.5-bisphosphate carboxylase/oxygenase (Rubisco). The reason for higher Pmax in marine ecotype of F. vesiculosus compare to the brackish ecotype however is not due to the relative amount of Rubisco and further studies of the rate of CO2 fixation by Rubisco is recommended. Treatments of the brackish ecotype of F. vesiculosus in higher salinity than the Bothnian Sea natural water indicate that the most favourable salinity for high Pmax is 10 psu, followed by 20 psu. One part of the explanation to a high Pmax in 10 psu is a greater relative amount of PsaA protein in algae treated in 10 psu. The reason for greater amount of PsaA might be that the algae need to produce more ATP, and are able to have a higher flow of cyclic electron transport around PSI to serve a higher rate of CO2 fixation by Rubisco. However, studies of the rate of CO2 fixation by Rubisco in algae treated in similar salinities as in present study are recommended to confirm this theory. / Fucus vesiculosus L. (Blåstång) är en brunalg som i huvudsak växer i tidvattenzonen i marint vatten men arten klarar också att växa konstant under ytan i det bräckta Bottenhavet. Norska havet och den del av Bottenhavet, där algerna är insamlade i denna studie, har salthalterna 34-35 psu (praktisk salthaltsenhet) respektive 4-5 psu. F. radicans L. Bergström et L. Kautsky (Smaltång) är en nyligen upptäckt art (2005) som har utvecklats i Bottenhavet. F. radicans och Bottenhavets ekotyp av F. vesiculosus växer sida vid sida och har tidigare ansetts vara samma art. Sett till hela Östersjön, så ändras ytans salthalt från 25 till 1-2 psu mellan Östersjöns gräns mot Kattegatt och norra Bottenviken. Den låga salthalten i Östersjön beror på det höga flödet av sötvatten från älvarna och på ett litet inflödet av saltvatten i inloppet vid Kattegatt. Salthaltsgradienten är korrelerad med antalet arter som minskar med minskad salthalt. Östersjön är ett artfattigt hav och de arter som finns är till stor del en blandning av söt- och saltvattenarter. Det finns bara ett fåtal arter som är helt anpassade till bräckt vatten och F. radicans är en av dem. Exempel på miljöskillnader för F. vesiculosus i Norska havet och i Bottenhavet är salthalten, tidvattnet, ljuset och temperaturen. Tidvattnet i Norska havet gör att algerna växlar mellan att vara i vattnet och på land, vilket utsätter algerna för stora ljusskillnader, snabba och stora temperaturväxlingar samt även torka. De alger som växer i Bottenhavet har däremot en jämnare och lägre temperatur, istäcke på vintern och mindre tillgång på ljus eftersom de alltid lever under vattenytan. Skillnaderna i miljön mellan växtplatserna leder till skillnader i fysiologiska anpassningar. Anledningen till att F. vesiculosus och F. radicans valdes som studieobjekt i denna avhandling är att de är viktiga nyckelarter i Bottenhavet. F. vesiculosus och F. radicans är de enda större bältesbildande alger som finns i det artfattiga ekosystemet och de används därför flitigt som mat, gömställe, parningsplats och barnkammare för t.ex. fisk. Att de är nyckelarter gör det angeläget att försöka förstå hur algerna är anpassade och hur de reagerar på miljöförändringar för att få veta hur de kan skyddas och bevaras. F. radicans inkluderades även för att se hur en naturlig art i Bottenhavet är anpassad i jämförelse med den invandrade F. vesiculosus. Marin F. vesiculosus inkluderades för att vara en artreferens från artens naturliga växtplats. Studien visar att det finns fler vattenlösliga organiska substanser (finns vissa organiska substanser som har en proteinskyddande funktion) i den marina ekotypen av of F. vesiculosus än i Bottenhavets ekotyp. Anledningen till detta föreslås vara en anpassning till att växa i tidvattenzonen. Vid lågvatten utsätts F. vesiculosus från Norska havet för starkt ljus, uttorkning, och snabba temperatur- växlingar vilket gör att den kan behöva dessa organiska substanser som skydd mot fria syreradikaler som bildas under lågvattenexponeringarna. F. vesiculosus från Bottenhavet har troligen mist förmågan att syntetisera dessa substanser på grund av anpassning till att hela tiden växa under ytan. Mängden mannitol (socker) är högre i den marina ekotypen av of F. vesiculosus än i Bottenhavets ekotyp. Detta föreslås bero på högre fotosyntetiskt maximum i F. vesiculosus från Norska havet jämfört med ekotypen från Bottenhavet. Skillnaden i fotssyntetiskt maximum är bland annat kopplat till ljus- och salthaltskillnaden på algernas växtplatser. Denna teori styrks av att både fotosyntesen och halten av mannitol ökar i Bottenhavets ekotyp när den behandlas i högre salthalt. Studien visar även att båda ekotyperna av F. vesiculosus samt F. radicans har ett ojämnt förhållande mellan fotosystem II och I (PSII och PSI) med en dominans av PSI. Denna slutsats är baserad på fluorescens emissions mätningar vid 77 K (-196 °C) och mätning av den relativa mängden D1 protein (motsvarar PSII) och PsaA protein (motsvarar PSI). F. vesiculosus från Bottenhavet visar ett emission spektrum som pekar mot en jämnare fördelning av PSII och PSI jämfört med den marina ekotypen och F. radicans. Detta stämmer dock inte med förhållandet mellan D1/PsaA som indikerar att alla tre har mer PSI än PSII. Förklaringen till avvikelsen mellan metoderna antas vara att F. vesiculosus från Bottenhavet har större ljus-infångande antennpigment än marin F. vesiculosus och F. radicans. De tydliga skillnaderna i 77 K fluorescens emission spektra mellan Bottenhavets F. vesiculosus och F. radicans visar att denna metod kan användas som säker artidentifiering. Den marina ekotypen av F. vesiculosus har högre fotosyntetiskt maximum än de båda arterna från Bottenhavet. Mätningar av den relativa mängden av enzymet Rubisco, viktigt för upptaget av koldioxid hos växter och alger, visar att mängden enzym är en sannolik förklaring till skillnaden i fotosyntetiskt maximum mellan den marina ekotypen av F. vesiculosus och F. radicans och detta är troligen en normal artskillnad. Mängden Rubisco kan dock inte förklara skillnaden i fotosyntetiskt maximum mellan de båda ekotyperna av F. vesiculosus. För att undersöka vad skillnaden mellan dessa två beror på så föreslås istället mätningar av Rubisco’s koldioxidfixeringshastighet. Det är en ökning av fotosyntetiskt maximum i Bottenhavets ekotyp av F. vesiculosus när den behandlas i högre salthalt (10, 20 och 35 psu) och det högsta fotosyntetiska maximumet uppmättes i alger som behandlats i 10 psu. Denna ökning beror inte på ökning i den relativa mängden av Rubisco. Ökningen i fotosyntesen speglas dock av en ökning av den relativa mängden PsaA. Detta antas bero på att det behövs mer energi i form av ATP och att en ökning av detta kan ske på grund av att mer PsaA kan driva den cykliska elektrontransporten i fotosyntesreaktionen. Ökat behov av ATP antas bero på en ökning av Rubisco aktiviteten men mätning av aktiviteten krävs för att bekräfta detta.

Chlorophyll fluorescence imaging is noninvasive technique often used in plant physiology, molecular biology and precision farming. Captured sequences of images record the dynamic of chlorophyll fluorescence emission which contain the information about spatial and time changes of photosynthetic activity of plant. The goal of this Ph.D. thesis is to contribute to the development of chlorophyll fluorescence imaging by application of advanced statistical techniques. Methods of statistical pattern recognition allow to identify images in the captured sequence that are reach for information about observed biotic stress and to find small subsets of fluorescence images suitable for following analysis. I utilized only methods for identification of small sets of images providing high performance with realistic time consumptions.