Pyrolysis of peptides and proteins. Analytical study and environmental applications

Analytical pyrolysis was used to investigate the formation of diketopiperazines (DKPs) which are cyclic dipeptides formed from the thermal degradation of proteins. A quali/quantitative procedure was developed combining microscale flash pyrolysis at 500 °C with gas chromatography-mass spectrometry (GC-MS) of DKPs trapped onto an adsorbent phase. Polar DKPs were silylated prior to GC-MS. Particular attention was paid to the identification of proline (Pro) containing DKPs due to their greater facility of formation. The GC-MS characteristics of more than 80 original and silylated DKPs were collected from the pyrolysis of sixteen linear dipeptides and four model proteins (e.g. bovine serum albumin, BSA). The structure of a novel DKP, cyclo(pyroglutamic-Pro) was established by NMR and ESI-MS analysis, while the structures of other novel DKPs remained tentative. DKPs resulted rather specific markers of amino acid sequence in proteins, even though the thermal degradation of DKPs should be taken into account. Structural information of DKPs gathered from the pyrolysis of model compounds was employed to the identification of these compounds in the pyrolysate of proteinaceous samples, including intrinsecally unfolded protein (IUP). Analysis of the liquid fraction (bio-oil) obtained from the pyrolysis of microalgae Nannochloropsis gaditana, Scenedesmus spp with a bench scale reactor showed that DKPs constituted an important pool of nitrogen-containing compounds. Conversely, the level of DKPs was rather low in the bio-oil of Botryococcus braunii. The developed micropyrolysis procedure was applied in combination with thermogravimetry (TGA) and infrared spectroscopy (FT-IR) to investigate surface interaction between BSA and synthetic chrysotile. The results showed that the thermal behavior of BSA (e.g. DKPs formation) was affected by the different form of doped synthetic chrysotile. The typical DKPs evolved from collagen were quantified in the pyrolysates of archaeological bones from Vicenne Necropolis in order to evaluate their conservation status in combination with TGA, FTIR and XRD analysis.

Biodiversità malacologica degli ecosistemi a coralli bianchi del Mediterraneo, dall'ultimo glaciale ad oggi

Nello studio di questa tesi è stata descritta una ricerca condotta allo scopo di fornire un contributo alla conoscenza della biodiversità analizzando la componente macrofaunistica subfossile e recente del Phylum Mollusca del Canale di Sicilia e del Golfo di Cadice. Tali organismi sono già da tempo noti in questi ambienti poiché associati a biocostruzioni a coralli bianchi presenti lungo i margini continentali atlantici e mediterranei i cui costituenti principali sono gli sclerattiniari coloniali Lophelia pertusa e Madrepora oculata. Tuttavia ben poco si conosce sull’ecologia di queste specie che si ritrovano spesso influenzate da scambi idrologici e biologici attraverso lo Stretto di Gibilterra. Nel corso di questo studio sono state analizzate 7 carote sedimentarie marine campionate durante diverse campagne oceanografiche: 6 carotaggi atlantici prelevati durante la campagna oceanografica Meteor nel Golfo di Cadice e 1 carota campionata nel Canale di Sicilia durante la campagna oceanografica MARCOS guidata dall’ISMAR-CNR di Bologna, sulla quale si concentra l’attenzione di questo lavoro. I dati raccolti sono stati analizzati in maniere quantitativa, per questo è stato utilizzato il software per analisi statistiche “Past3” e sono stati combinati con l’analisi qualitativa dei campioni in modo da ottenere il maggior numero di informazioni. Tra le analisi statistiche sono state effettuate il calcolo di tutti gli indici di dominanza, diversità, di equiripartizione e ricchezza specifica (Simpson, Shannon, Evenness e Margalef) e l’analisi multivariata NMDS (Non-metric Multi-Dimensional Scaling). Infine è stata effettuata l’analisi delle associazioni trofiche distribuite in una carota. Il risultato ha evidenziato un’elevata biodiversità per campioni prelevati in profondità e il ritrovamento di organismi appartenenti all’ultimo glaciale.

Riprogettazione del processo di sviluppo nuovo prodotto - il caso Umbra Cuscinetti S.p.A

Questo lavoro di tesi nasce dall’opportunità di un tirocinio della durata di sei mesi presso Umbra Cuscinetti S.p.A., importante azienda leader nel settore aeronautico e industriale per la realizzazione di Cuscinetti a sfera, Viti a ricircolo di sfere e Attuatori Elettromeccanici (EMA). L’ambito di riferimento è quello del Processo di Sviluppo Nuovo Prodotto per la linea di Business EMA. La linea ha completato la fase “pionieristica” che ne ha segnato l’ingresso in un nuovo settore industriale e, oggi, ha la necessità di procedere ad una razionalizzazione delle proprie attività allo scopo di supportare al meglio i programmi di sviluppo. I prodotti della linea presentano un elevato grado di innovazione e non essendo standardizzati, richiedono una gestione oculata in tutti gli aspetti di costo, tempo e qualità. I progetti finora intrapresi non hanno soddisfatto appieno le aspettative per questa linea, le cui potenzialità sono molto elevate; da qui la necessità di una riprogettazione di tutto il processo. L’analisi è stata condotta mediante la metodologia DMAIC e prevede come obiettivo l’inserimento del Project Management come nuova metodologia di gestione del processo.

Analisi e confronto di sequenze di DNA mediante modelli Markoviani

Lo scopo di questa tesi è quello di evidenziare, attraverso varie analisi statistiche ed applicazione di modelli stocastici, il comportamento strutturale e funzionale dei dinucleotidi che compongono le sequenze di DNA di diversi organismi. Gli organismi che abbiamo scelto di prendere in considerazione sono l'uomo, il topo e l'Escherichia coli. Questa scelta non è stata casuale, ma oculata, al fine di mettere in risalto alcune differenze tra organismi eucarioti, quali l'uomo e il topo, ed organismi procarioti come il batterio E.coli. Nella prima parte del nostro studio, abbiamo computato le distanze che intercorrono tra occorrenze successive dello stesso dinucleotide lungo la sequenza, usando un metodo di non sovrapposizione, ed abbiamo iterato il calcolo per tutti i 16 dinucleotidi. Dopodiché ci siamo preoccupati di graficare le distribuzioni di distanza dei 16 dinucleotidi per l'E.Coli, il topo e l'uomo; gli istogrammi evidenziano un comportamento anomalo della distribuzione di CG che accomuna gli organismi eucarioti e di cui, invece, è esente l'organismo procariote esaminato. Questo dato statistico trova una spiegazione nei processi biologici di metilazione che possono innescarsi sul dinucleotide CG nelle sequenze eucariotiche. In seguito, per determinare quanto ciascuna delle 16 distribuzioni si discosti dalle altre abbiamo usato la divergenza di Jensen-Shannon. Per quantificare le differenze sostanziali tra le distribuzioni di CG dei 3 organismi considerati abbiamo deciso di verificare quale fosse il miglior fit per tali curve tra un esponenziale ed una power-law. L'esponenziale rappresenta un buon fit per le code delle distribuzioni di CG del topo e dell'uomo; ciò rivela la presenza di una lunghezza caratteristica per entrambi gli organismi. Nella seconda parte dello studio, i risultati vengono confrontati con modelli markoviani: sequenze random generate con catene di Markov di ordine zero (basate sulle frequenze relative dei nucleotidi) e uno (basate sulle probabilità di transizione tra diversi nucleotidi). Quest'ultima riproduce abbastanza fedelmente la sequenza biologica di partenza, per cui abbiamo scelto di utilizzare la catena Markov del 1° ordine per altre analisi statistiche riguardanti le distribuzioni dei nucleotidi, dinucleotidi, ed anche dei trinucleotidi con particolare interesse per quelli in cui è contenuto CG, in modo da verificare se l'anomalia si ripercuote anche in essi. Riteniamo pertanto che metodi basati su questo approccio potrebbero essere sfruttati per confermare le peculiarità biologiche e per migliorare l'individuazione delle aree di interesse, come le isole CpG, ed eventualmente promotori e Lamina Associated Domains (LAD), nel genoma di diversi organismi.

Sedimentology and stable oxygen isotope ratios of foraminifera from cores of the Porcupine Seabight, Northeast Atlantic

Large carbonate mound structures have been discovered in the northern Porcupine Seabight (Northeast Atlantic) at depths between 600 and 1000 m. These mounds are associated with the growth of deep-sea corals Lophelia pertusa and Madrepra oculata. In this study, three sediment cores have been analysed. They are from locations close to Propeller Mound, a 150 m high ridge-like feature covered with a cold-water coral ecosystem at its upper flanks. The investigations are concentrated on grain-size analyses, carbon measurements and on the visual description of the cores and computer tomographic images, to evaluate sediment content and structure. The cores portray the depositional history of the past ~31 kyr BP, mainly controlled by sea-level fluctuations and the climate regime with the advance and retreat of the Irish Ice Sheet onto the Irish Mainland Shelf. A first advance of glaciers is indicated by a turbiditic release slightly older than 31 kyr BP, coherent with Heinrich event 3 deposition. During Late Marine Isotope Stage 3 (MIS 3) and MIS 2 shelf erosion prevailed with abundant gravity flows and turbidity currents. A change from glaciomarine to hemipelagic contourite sedimentation during the onset of the Holocene indicates the establishment of the strong, present-day hydrodynamic regime at intermediate depths. The general decrease in accumulation of sediments with decreasing distance towards Propeller Mound suggests that currents (turbidity currents, gravity flows, bottom currents) had a generally stronger impact on the sediment accumulation at the mound base for the past ~31 kyr BP, respectively.

Lipids and nitrogen isotopes of deep-water corals

The lipid and organic nitrogen isotopic (delta15N) compositions of two common deep-water corals (Lophelia pertusa and Madrepora oculata) collected from selected locations of the NE Atlantic are compared to the composition of suspended particulate organic matter, in order to determine their principle food source. Initial results suggest that they may feed primarily on zooplankton. This is based on the increased abundances of mono-unsaturated fatty acids and alcohols and the different ratios of the polyunsaturated fatty acids, 22:6/20:5 of the corals when compared to those of the suspended particulate organic matter. There is enrichment in L. pertusa of mono-unsaturated fatty acids and of delta15N relative to M. oculata. It is unclear whether this reflects different feeding strategies or assimilation/storage efficiencies of zooplankton tissue or different metabolism in the two coral species.

Survival of Atlantic cod larvae from the Barents Sea and the Western Baltic sea due to ocean acidification from two experiments

The data show the survival data of Atlantic cod larvae from two different stocks, which were measured in two separate experiments in Kristineberg, Sweden in 2013 on the Western Baltic stock and in Tromsö, Norway in 2014 on the Barents Sea stock. Survival was measured as a response to ocean acidification, control tanks were kept at ambient CO2 concentrations. CO2 concentrations and feeding concentrations are also provided.

Vegetation in the coastal area of East Greenland

This paper deals with the syntaxonomy and ecology of debris, scree and alluvium vegetation of the Ammassalik district, Southeast Greenland, on more or less moist soil. The Oxyria digyna- and Chamaenerion latifoliumvegetation types are classified as Saxifrago-Oxyrietum digynae (Böcher 1933 ap. Nordh. 1943) Gjaerevoll 1950 respectively Chamaenerietum latifolii Böcher 1933 in the class Thlaspietea rotundifolii Br.-BI. ap. Br.-BI. et al. 1947. The chionophytic Saxifrago-Oxyrietum digynae and the Chamaenerietum latifolii occurring on river-banks are classified in the alliance Saxifrago stellaris-Oxyrion digynae Gjaerevoll 1950. This alliance belongs to the order Androsacetalia alpinae Br.-BI. ap. Br.-BI. & Jenny 1926, Thlaspietea rotundifolii Br.-BI. ap. Br.-BI. et al. 1947. The following syntaxa are described as new: Saxifrago-Oxyrietum digynae stellarietosum humifusae and typicum with two variants and one variant of the subassociation inops De Molenaar 1976, and the Chamaenerietum latifolii typicum with two variants and salicetosum herbaceae with three variants.

Benthos fauna in the North Sea from 2 surveys in 1986 and 1987

During two surveys in the North Sea, in summer 1986 and in winter 1987, larger epibenthos was collected with a 2 m beam trawl. The distributions of the species were checked for average linkage by means of the JACCARD-index cluster analysis. In summer two main clusters can be recognized. These are situated to the north and to the south of the Dogger Bank. In winter two main clusters may be recognized as well, but these clusters divide the North Sea into a western and an eastern part. We conclude, that these differences of epibenthos characteristics are correlated with seasonal changes in water body distributions.

Neogene diatoms of the Japan Sea

A series of excellent upper Miocene through Quaternary diatomaceous sequences recovered at four sites during Leg 127 was examined for diatoms. The diagenetic transition from opal-A to opal-CT is a diachronic horizon from the uppermost part of the Denticulopsis katayamae Zone (8.5 Ma) at Hole 797B to the uppermost part of the Neodenticula kamtschatica Zone (5.73 Ma) at Hole 795A. The diatom zonation of Koizumi (1985) best divides the upper Miocene to Quaternary sequences above the opal-A/opal-CT boundary and also is useful to date carbonate concretions including diatoms below the boundary. Forty diatom datum levels were evaluated biostratigraphically based on the sediment accumulation rate curve, and several isochronous datum levels are newly proposed for the Japan Sea area. A warm-water current did not penetrated into the Japan Sea through the Tsushima strait during the late Miocene and Pliocene time, because subtropical warm-water diatoms are essentially not present in such sediment samples. The occurrences of diatom are cyclic throughout the Quaternary sediments and are affected by eustatic sea level changes.

Diatom deposits in the ostpreußischen Haffe

Als man nach dem ersten Weltkrieg im verkleinerten Deutschland nach der Möglichkeit von Neulandgewinnung suchte, dachte man auch an eineTrockenlegung der ostpreußischen Haffe. Aus diesem Anlaß wurden umfangreiche Bohrungen ausgeführt, um ein möglichst genaues Bild vom Untergrunde der Haffe zu bekommen. Auf Veranlassung der Preußischen Geologischen Landesanstalt wurde ich mit der Untersuchung der Diatomeen in den Bohrproben beauftragt. Die Arbeit wurde 1934 begonnen und Ende 1937 wurde der letzte Arbeitsbericht abgeliefert. Die beabsichtigte Veröffentlichung ist bisher unterblieben, weil die Druckvorlagen später verloren gegangen sind. Seitdem sind über die Haffuntersuchungen mehrere Teilergebnisse veröffentlicht worden, von denen hier schon wegen der Terminologie die pollenanalytischen Arbeiten von L. HEIN (1941) und HUGO GROSS (1941) erwähnt seien, auf die im Abschnitt Il 2e näher eingegangen wird. Bei der geologischen Auswertung war Zurückhaltung geboten; denn es wäre gewagt, allein aus der Perspektive der Diatomeenforschung endgültige Aussagen machen zu wollen. Darum habe ich mich bemüht, das Material so weit aufzuschließen, daß es Geologen später auch bei veränderter Fragestellung auswerten können. "Die Theorien wechseln, aber die Tatsachen bleiben." Der Initiative des Herrn Prof. Dr. K. GRIPP und der finanziellen Hilfe der Deutschen Forschungsgemeinschaft ist es zu verdanken, daß die vorliegende Arbeit im Druck erscheinen kann. Zusammenfassung 1. Nur in den alluvialen Schichten des Kurischen Haffs wurden Diatomeen gefunden. 2. Die Diatomeenflora des Kurischen Haffs besteht zur Hauptsache aus Süßwasserformen. 3. Salzwasserformen finden sich in allen Schichten verstreut unter der Süßwasserflora. Wenn sie auch nach Zahl der Arten in manchen Proben einen erheblichen Prozentsatz der Flora ausmachen, so ist doch die Zahl der Individuen stets so gering, daß man nirgends von einer Brackwasserflora sprechen kann. 4. Die Süßwasserflora besteht in den unteren Schichten vorwiegend aus Grundformen; und zwar machen die epiphytischen Bewohner flacher Sumpfgewässer einen großen Teil der Flora aus. 5. In einzelnen Bohrungen kommt in den untersten alluvialen Schichten eine Grundflora mit zahlreichen Mastogloien vor. Dies sind die ältesten diatomeenführenden Schichten, entstanden in isolierten Sumpfgewässern. 6. Die übrigen Schichten mit überwiegender Grundflora sind vermutlich Ablagerungen der Ancyluszeit. 7. Die oberen Schichten, in denen die Planktondiatomeen überwiegen, dürften größtenteils der Litorina-Transgressionszeit angehören, jedoch ist der Transgressions-Kontakt nicht klar zu erkennen. 8. Das Ende der Litorinazeit ist noch weniger erkennbar, da eine grundsätzliche Veränderung der Flora nach oben nicht zu beobachten ist. 9. Die ostbaltischen Charakterformen sind in allen Schichten vertreten.

Working on a baseline for the Kongsfjorden food web: production and properties of macroalgal particulate organic matter (POM), supplementary material

Macroalgae, in particular kelps, produce a large amount of biomass in Kongsfjorden, which is to a great extent released into the water in an annual cycle. As an example, the brown alga Alaria esculenta loses its blade gradually, 3 ± 0.8 % of the blade area per day (August 2012), thereby adding to the pool of particulate organic matter (POM) in the fjord. Upon release small thallus pieces are "aging" in that they are prone to leaching and serving as substrate for microorganisms, thus turning into palatable food for suspension and bottom feeders. In order to define a macroalgal baseline for the Kongsfjorden food web, stable isotopes d14C and d15N were measured in individuals of A. esculenta, Saccharina latissima and Laminaria digitata directly sampled after collection and in artificially produced POM (aPOM) of A. esculenta that was allowed to age under experimental conditions. In aPOM from this species sampled in August 2012 the C/N ratios decreased between d1 and d8 of a 14-day culture period in parallel to the fading photosynthetic activity of the algal fragments as demonstrated by use of an Imaging-PAM. Microscopic observations of the aPOM in August 2012 and 2013 revealed the frequent occurrence of small brown algal endo- and epiphytes. First feeding experiments with Mysis oculata (Mysids) and Hiatella arctica (Bivalves) showed that these species can ingest macroalgal POM. The importance of kelp-derived POM for the food web is subject of the current research.

Pollen and diatom analysis on varved sediments in Lake Jues (Harz Mountains, Germany)

Studies combining sedimentological and biological evidence to reconstruct Holocene climate beyond the major changes, and especially seasonality, are rare in Europe, and are nearly completely absent in Germany. The present study tries to reconstruct changes of seasonality from evidence of annual algal successions within the framework of well-established pollen zonation and 14C-AMS dates from terrestrial plants. Laminated Holocene sediments in Lake Jues (10°20.70' E, 51°39.30' N, 241 m a.s.l.), located at the SW margin of the Harz Mountains, central Germany, were studied for sediment characteristics, pollen, diatoms and coccal green algae. An age model is based on 21 calibrated AMS radiocarbon dates from terrestrial plants. The sedimentary record covers the entire Holocene period. Trophic status and circulation/stagnation patterns of the lake were inferred from algal assemblages, the subannual structure of varves and the physico-chemical properties of the sediment. During the Holocene, mixing conditions alternated between di-, oligo- and meromictic depending on length and variability of spring and fall periods, and the stability of winter and summer weather. The trophic state was controlled by nutrient input, circulation patterns and the temperature-dependent rates of organic production and mineralization. Climate shifts, mainly in phase with those recorded from other European regions, are inferred from changing limnological conditions and terrestrial vegetation. Significant changes occurred at 11,600 cal. yr. BP (Preboreal warming), between 10,600 and 10,100 cal. yr. BP (Boreal cooling), and between 8,400 and 4,550 cal. yr. BP (warm and dry interval of the Atlantic). Since 4,550 cal. yr. BP the climate became gradually cooler, wetter and more oceanic. This trend was interrupted by warmer and dryer phases between 3,440 and 2,850 cal. yr. BP and, likely, between 2,500 and 2,250 cal. yr. BP.