Climatic and stratigraphic paleoindicators from the Canary Island

[EN] The last 5 Myr  are characterized by cliamatic variations globally and are reflected in ancient fossiliferous marine deposits visible in the Canary Islands. The fossils contained are identificated as paleoecological and paleoclimatic indicators. The Mio-Pliocene Transit is represented by the coral Siderastrea micoenica Osasco, 1897; the gastropods Rothpletzia rudista Simonelli, 1890; Ancilla glandiformis (Lamarck, 1822); Strombus coronatus Defrance, 1827 and Nerita emiliana Mayer, 1872 and the bivalve Gryphaea virleti Deshayes, 1832 as most characteristic fossils  and typical of a very warm climate and littoral zone. Associated  lava flows  have been dated radiometrically  and provides a range between  8.9 and about 4.2 Kyr. In the mid-Pleistocene, about 400,000 years ago, the called Marine Isotope Stage 11, a strong global warming that caused a sea level rise happens. Remains of the MIS 11  are preserved on the coast of Arucas (Gran Canaria), and associated with a tsunami in Piedra Alta (Lanzarote). These fossilifeorus  deposits contains the bivalve Saccostrea cucullata (Born, 1780), the gastropod Purpurellus gambiensis (Reeve, 1845) and the corals Madracis pharensis (Heller, 1868) and Dendrophyllia cornigera (Lamarck, 1816). Both sites have been dated by K-Ar on pillow lavas (approximately 420,000 years) and by Uranium Series on corals (about 481,000 years) respectively. The upper Pleistocene starts with another strong global warming known as the last interglacial or marine isotope  stage (MIS) 5.5, about 125,000 years ago, which also left marine  fossil deposits exposed in parallel to current in Igueste of San Andrés (Tenerife),  El Altillo, the  city of Las Palmas de Gran Canaria  and Maspalomas (Gran Canaria),  Matas Blancas, the Playitas and Morrojable (Fuerteventura ) and in Playa Blanca and Punta Penedo (Lanzarote ). The fossil coral Siderastrea radians (Pallas , 1766 ) currently living in the Cape Verde Islands , the Gulf of Guinea and the Caribbean has allowed Uranium series dating. The gastropods Strombus bubonius Lamarck, 1822 and Harpa doris (Röding , 1798 ) currently living in the Gulf of Guinea. Current biogeography using synoptic data obtained through satellites provided by the ISS Canary Seas provides data of Ocean Surface Temperature (SST) and Chlorophyll a (Chlor a) . This has allowed the estimation of these sea conditions during interglacials compared to today .

New synthetic bile acid analogue agonists of FXR and TGR5 receptors: Analytical methodologies for the study of their physico-chemical properties, pharmacokinetic activity and metabolism.

This thesis reports an integrated analytical approach for the study of physicochemical and biological properties of new synthetic bile acid (BA) analogues agonists of FXR and TGR5 receptors. Structure-activity data were compared with those previous obtained using the same experimental protocols on synthetic and natural occurring BA. The new synthetic BA analogues are classified in different groups according also to their potency as a FXR and TGR5 agonists: unconjugated and steroid modified BA and side chain modified BA including taurine or glycine conjugates and pseudo-conjugates (sulphonate and sulphate analogues). In order to investigate the relationship between structure and activity the synthetic analogues where admitted to a physicochemical characterization and to a preliminary screening for their pharmacokinetic and metabolism using a bile fistula rat model. Sensitive and accurate analytical methods have been developed for the quali-quantitative analysis of BA in biological fluids and sample used for physicochemical studies. Combined High Performance Liquid Chromatography Electrospray tandem mass spectrometry with efficient chromatographic separation of all studied BA and their metabolites have been optimized and validated. Analytical strategies for the identification of the BA and their minor metabolites have been developed. Taurine and glycine conjugates were identified in MS/MS by monitoring the specific ion transitions in multiple reaction monitoring (MRM) mode while all other metabolites (sulphate, glucuronic acid, dehydroxylated, decarboxylated or oxo) were monitored in a selected-ion reaction (SIR) mode with a negative ESI interface by the following ions. Accurate and precise data where achieved regarding the main physicochemical properties including solubility, detergency, lipophilicity and albumin binding . These studies have shown that minor structural modification greatly affect the pharmacokinetics and metabolism of the new analogues in respect to the natural BA and on turn their site of action, particularly where their receptor are located in the enterohepatic circulation.

Physico-chemical and microstructural properties of food dispersions

The macroscopic properties of oily food dispersions, such as rheology, mechanical strength, sensory attributes (e.g. mouth feel, texture and even flavour release) and as well as engineering properties are strongly determined by their microstructure, that is considered a key parameter in the understanding of the foods behaviour . In particular the rheological properties of these matrices are largely influenced by their processing techniques, particle size distribution and composition of ingredients. During chocolate manufacturing, mixtures of sugar, cocoa and fat are heated, cooled, pressurized and refined. These steps not only affect particle size reduction, but also break agglomerates and distribute lipid and lecithin-coated particles through the continuous phase, this considerably modify the microstructure of final chocolate. The interactions between the suspended particles and the continuous phase provide information about the existing network and consequently can be associated to the properties and characteristics of the final dispersions. Moreover since the macroscopic properties of food materials, are strongly determined by their microstructure, the evaluation and study of the microstructural characteristics, can be very important for a through understanding of the food matrices characteristics and to get detailed information on their complexity. The aim of this study was investigate the influence of formulation and each process step on the microstructural properties of: chocolate type model systems, dark milk and white chocolate types, and cocoa creams. At the same time the relationships between microstructural changes and the resulting physico-chemical properties of: chocolate type dispersions model systems dark milk and white chocolate were investigated.

Semi-synthetic bile acids as novel drug candidate in liver diseases: physico-chemical characterization and HPLC-ES-MS/MS methods for their quali-quantitative analysis in different experimental animal models

The physico-chemical characterization, structure-pharmacokinetic and metabolism studies of new semi synthetic analogues of natural bile acids (BAs) drug candidates have been performed. Recent studies discovered a role of BAs as agonists of FXR and TGR5 receptor, thus opening new therapeutic target for the treatment of liver diseases or metabolic disorders. Up to twenty new semisynthetic analogues have been synthesized and studied in order to find promising novel drugs candidates. In order to define the BAs structure-activity relationship, their main physico-chemical properties (solubility, detergency, lipophilicity and affinity with serum albumin) have been measured with validated analytical methodologies. Their metabolism and biodistribution has been studied in “bile fistula rat”, model where each BA is acutely administered through duodenal and femoral infusion and bile collected at different time interval allowing to define the relationship between structure and intestinal absorption and hepatic uptake ,metabolism and systemic spill-over. One of the studied analogues, 6α-ethyl-3α7α-dihydroxy-5β-cholanic acid, analogue of CDCA (INT 747, Obeticholic Acid (OCA)), recently under approval for the treatment of cholestatic liver diseases, requires additional studies to ensure its safety and lack of toxicity when administered to patients with a strong liver impairment. For this purpose, CCl4 inhalation to rat causing hepatic decompensation (cirrhosis) animal model has been developed and used to define the difference of OCA biodistribution in respect to control animals trying to define whether peripheral tissues might be also exposed as a result of toxic plasma levels of OCA, evaluating also the endogenous BAs biodistribution. An accurate and sensitive HPLC-ES-MS/MS method is developed to identify and quantify all BAs in biological matrices (bile, plasma, urine, liver, kidney, intestinal content and tissue) for which a sample pretreatment have been optimized.

Complementary mass spectrometric techniques for the characterization of the organic fraction in atmospheric aerosols

Aerosol particles are strongly related to climate, air quality, visibility and human health issues. They contribute the largest uncertainty in the assessment of the Earth´s radiative budget, directly by scattering or absorbing solar radiation or indirectly by nucleating cloud droplets. The influence of aerosol particles on cloud related climatic effects essentially depends upon their number concentration, size and chemical composition. A major part of submicron aerosol consists of secondary organic aerosol (SOA) that is formed in the atmosphere by the oxidation of volatile organic compounds. SOA can comprise a highly diverse spectrum of compounds that undergo continuous chemical transformations in the atmosphere.rnThe aim of this work was to obtain insights into the complexity of ambient SOA by the application of advanced mass spectrometric techniques. Therefore, an atmospheric pressure chemical ionization ion trap mass spectrometer (APCI-IT-MS) was applied in the field, facilitating the measurement of ions of the intact molecular organic species. Furthermore, the high measurement frequency provided insights into SOA composition and chemical transformation processes on a high temporal resolution. Within different comprehensive field campaigns, online measurements of particular biogenic organic acids were achieved by combining an online aerosol concentrator with the APCI-IT-MS. A holistic picture of the ambient organic aerosol was obtained through the co-located application of other complementary MS techniques, such as aerosol mass spectrometry (AMS) or filter sampling for the analysis by liquid chromatography / ultrahigh resolution mass spectrometry (LC/UHRMS).rnIn particular, during a summertime field study at the pristine boreal forest station in Hyytiälä, Finland, the partitioning of organic acids between gas and particle phase was quantified, based on the online APCI-IT-MS and AMS measurements. It was found that low volatile compounds reside to a large extent in the gas phase. This observation can be interpreted as a consequence of large aerosol equilibration timescales, which build up due to the continuous production of low volatile compounds in the gas phase and/or a semi-solid phase state of the ambient aerosol. Furthermore, in-situ structural informations of particular compounds were achieved by using the MS/MS mode of the ion trap. The comparison to MS/MS spectra from laboratory generated SOA of specific monoterpene precursors indicated that laboratory SOA barely depicts the complexity of ambient SOA. Moreover, it was shown that the mass spectra of the laboratory SOA more closely resemble the ambient gas phase composition, indicating that the oxidation state of the ambient organic compounds in the particle phase is underestimated by the comparison to laboratory ozonolysis. These observations suggest that the micro-scale processes, such as the chemistry of aerosol aging or the gas-to-particle partitioning, need to be better understood in order to predict SOA concentrations more reliably.rnDuring a field study at the Mt. Kleiner Feldberg, Germany, a slightly different aerosol concentrator / APCI-IT-MS setup made the online analysis of new particle formation possible. During a particular nucleation event, the online mass spectra indicated that organic compounds of approximately 300 Da are main constituents of the bulk aerosol during ambient new particle formation. Co-located filter analysis by LC/UHRMS analysis supported these findings and furthermore allowed to determine the molecular formulas of the involved organic compounds. The unambiguous identification of several oxidized C 15 compounds indicated that oxidation products of sesquiterpenes can be important compounds for the initial formation and subsequent growth of atmospheric nanoparticles.rnThe LC/UHRMS analysis furthermore revealed that considerable amounts of organosulfates and nitrooxy organosulfates were detected on the filter samples. Indeed, it was found that several nitrooxy organosulfate related APCI-IT-MS mass traces were simultaneously enhanced. Concurrent particle phase ion chromatography and AMS measurements indicated a strong bias between inorganic sulfate and total sulfate concentrations, supporting the assumption that substantial amounts of sulfate was bonded to organic molecules.rnFinally, the comprehensive chemical analysis of the aerosol composition was compared to the hygroscopicity parameter kappa, which was derived from cloud condensation nuclei (CCN) measurements. Simultaneously, organic aerosol aging was observed by the evolution of a ratio between a second and a first generation biogenic oxidation product. It was found that this aging proxy positively correlates with increasing hygroscopicity. Moreover, it was observed that the bonding of sulfate to organic molecules leads to a significant reduction of kappa, compared to an internal mixture of the same mass fractions of purely inorganic sulfate and organic molecules. Concluding, it has been shown within this thesis that the application of modern mass spectrometric techniques allows for detailed insights into chemical and physico-chemical processes of atmospheric aerosols.rn

Millennial and sub-millennial scale climatic variations recorded in polar ice cores over the last glacial period

Since its discovery in Greenland ice cores, the millennial scale climatic variability of the last glacial period has been increasingly documented at all latitudes with studies focusing mainly on Marine Isotopic Stage 3 (MIS 3; 28–60 thousand of years before present, hereafter ka) and characterized by short Dansgaard-Oeschger (DO) events. Recent and new results obtained on the EPICA and NorthGRIP ice cores now precisely describe the rapid variations of Antarctic and Greenland temperature during MIS 5 (73.5–123 ka), a time period corresponding to relatively high sea level. The results display a succession of abrupt events associated with long Greenland InterStadial phases (GIS) enabling us to highlight a sub-millennial scale climatic variability depicted by (i) short-lived and abrupt warming events preceding some GIS (precursor-type events) and (ii) abrupt warming events at the end of some GIS (rebound-type events). The occurrence of these sub-millennial scale events is suggested to be driven by the insolation at high northern latitudes together with the internal forcing of ice sheets. Thanks to a recent NorthGRIP-EPICA Dronning Maud Land (EDML) common timescale over MIS 5, the bipolar sequence of climatic events can be established at millennial to sub-millennial timescale. This shows that for extraordinary long stadial durations the accompanying Antarctic warming amplitude cannot be described by a simple linear relationship between the two as expected from the bipolar seesaw concept. We also show that when ice sheets are extensive, Antarctica does not necessarily warm during the whole GS as the thermal bipolar seesaw model would predict, questioning the Greenland ice core temperature records as a proxy for AMOC changes throughout the glacial period.