Synthese C-glycosidischer und N-methylierter Glycosylaminosäuren für den Aufbau von Mimetika tumorassoziierter MUC1-Glycopeptidantigene unter Verwendung mikrostrukturierter Reaktoren

Zur Synthese hydrolysestabiler MUC1-Antitumorvakzine wurde im Rahmen dieser Arbeit zunächst ein Verfahren zur effizienten N Methylierung von Fmoc-Aminosäuren entwickelt. Die Synthese erfolgte in einer zweistufigen Umsetzung über Oxazolidinone unter Verwendung eines Tube-in-Tube-Durchflussreaktors mit einer semipermeablen Membran aus Teflon® AF 2400. In diesem Tube-in-Tube-Reaktor wurde in der ersten Stufe das Modellsubstrat Fmoc-Alanin bereits nach 2 h annähernd quantitativ in das entsprechende Oxazolidinon umgesetzt. In der zweiten Stufe wurde mit TFA erstmals eine Flüssigkeit durch eine solche Membran des Tube-in-Tube-Reaktors eingeleitet und lieferte innerhalb einer Stunde zahlreiche aliphatische, aromatische und funktionalisierte N-Methylaminosäuren in hohen Ausbeuten.rnDes Weiteren wurden erstmals sensible Glycosylaminosäuren, darunter auch TN Antigen-Strukturen, N-methyliert. Sie dienen als Bausteine für die Synthese von MUC1-Antitumorvakzinen. Neben Fmoc-N-Methyl-TN-Threonin konnten die Fmoc-geschützten N-Methyl-TN-Serin, N-Methyl-Sialyl-TN-Threonin sowie zwei N-Methyl-C Glycosylaminosäuren und in guten Ausbeuten erhalten werden. Anschließend wurde das N methylierte TN-Threonin gezielt in die tandem repeat-Sequenz des MUC1 in einer Festphasenpeptidsynthese eingebaut. Um einen direkten Vergleich bezüglich der N Methylierung im MUC1-Glycopeptide und dem darauf folgenden Einfluss auf die Tumorselektivität der resultierenden Vakzine erhalten zu können, wurde zudem ein Referenzpeptid aufgebaut. Zur Vollendung der Vakzinsynthese erfolgte die Konjugation beider Glycopeptidantigene an die jeweiligen BSA- und TTox-Proteine. rnEin alternativer Zugang zu hydrolysestabilen Glycopeptidbausteinen wurde im letzten Teil der Arbeit über die Synthese von α C Glycosylaminosäuren erarbeitet. Der entwickelte Syntheseweg basiert auf einer Ugi-Vier-Komponenten-Reaktion aus Aldehyd, Amin, Nitril und Carbonsäure. Als benötigte Aldehydkomponenten wurden ein einfaches Galactose- sowie ein Galactosamin-Derivat verwendet. Zum Aufbau des C-glycosidischen Grundgerüsts wurde eine Mikrowellen-unterstützte C-Allylierungsvariante im Durchfluss realisiert. Die Galactose- und Galactosaminaldehyde wurden danach mit chirale Glycosylaminen umgesetzt.

An investigation of phenolic glycoside and condensed tannin homeostasis in Populus by salicyl alcohol feeding to cell cultures and by transgenic manipulation of the sucrose transporter, PTSUT4, in planta

Secondary metabolites play an important role in plant protection against biotic and abiotic stress. In Populus, phenolic glycosides (PGs) and condensed tannins (CTs) are two such groups of compounds derived from the common phenylpropanoid pathway. The basal levels and the inducibility of PGs and CTs depend on genetic as well as environmental factors, such as soil nitrogen (N) level. Carbohydrate allocation, transport and sink strength also affect PG and CT levels. A negative correlation between the levels of PGs and CTs was observed in several studies. However, the molecular mechanism underlying such relation is not known. We used a cell culture system to understand negative correlation of PGs and CTs. Under normal culture conditions, neither salicin nor higher-order PGs accumulated in cell cultures. Several factors, such as hormones, light, organelles and precursors were discussed in the context of aspen suspension cells’ inability to synthesize PGs. Salicin and its isomer, isosalicin, were detected in cell cultures fed with salicyl alcohol, salicylaldehyde and helicin. At higher levels (5 mM) of salicyl alcohol feeding, accumulation of salicins led to reduced CT production in the cells. Based on metabolic and gene expression data, the CT reduction in salicin-accumulating cells is partly a result of regulatory changes at the transcriptional level affecting carbon partitioning between growth processes, and phenylpropanoid CT biosynthesis. Based on molecular studies, the glycosyltransferases, GT1-2 and GT1-246, may function in glycosylation of simple phenolics, such as salicyl alcohol in cell cultures. The uptake of such glycosides into vacuole may be mediated to some extent by tonoplast localized multidrug-resistance associated protein transporters, PtMRP1 and PtMRP6. In Populus, sucrose is the common transported carbohydrate and its transport is possibly regulated by sucrose transporters (SUTs). SUTs are also capable of transporting simple PGs, such as salicin. Therefore, we characterized the SUT gene family in Populus and investigated, by transgenic analysis, the possible role of the most abundantly expressed member, PtSUT4, in PG-CT homeostasis using plants grown under varying nitrogen regimes. PtSUT4 transgenic plants were phenotypically similar to the wildtype plants except that the leaf area-to-stem volume ratio was higher for transgenic plants. In SUT4 transgenics, levels of non-structural carbohydrates, such as sucrose and starch, were altered in mature leaves. The levels of PGs and CTs were lower in green tissues of transgenic plants under N-replete, but were higher under N-depleted conditions, compared to the levels in wildtype plants. Based on our results, SUT4 partly regulates N-level dependent PG-CT homeostasis by differential carbohydrate allocation.

Synthesis of oligosaccharides corresponding to the polysaccharides of Lactobacillus and Thermophlilus strains

Synthesis of a branched trisaccharide and a tetrasaccharide repeating units corresponding to the polysaccharides of Lactobacillus spp. G-77 and Thermus thermophilus Samu-SA1 as their methyl glycosides has been achieved in excellent yield. Most of the glycosyl linkages are 1,2-cis in these oligosaccharide fragments

Role of two UDP-Glycosyltransferases from the L group of arabidopsis in resistance against pseudomonas syringae

The role of the salicylic acid (SA) glycosides SA 2-O-β-D-glucose (SAG), SA glucose ester (SGE) and the glycosyl transferases UGT74F1 and UGT74F2 in the establishment of basal resistance of Arabidopsis against Pseudomonas syringae pv tomato DC3000 (Pst) was investigated. Both mutants altered in the corresponding glycosyl transferases (ugt74f1 and ugt74f2) were affected in their basal resistance against Pst. The mutant ugt74f1 showed enhanced susceptibility, while ugt74f2 showed enhanced resistance against the same pathogen. Both mutants have to some extent, altered levels of SAG and SGE compared to wild type plants, however, in response to the infection, ugt74f2 accumulated higher levels of free SA until 24 hpi compared to wild type plants while ugt74f1 accumulated lower SA levels. These SA levels correlated well with reduced expression in PR1 and EDS1 in ugt74f1. In contrast, ugt74f2 has enhanced expression of Enhanced Disease Susceptibility 1 (EDS1) but a strong reduction in the expression of several jasmonate (JA)-dependent genes. Bacterial infection interfered with the expression of Fatty Acid Desaturase (FAD), Lipoxygenase2 (LOX2), carboxyl methyltransferase1 (BSMT1) and 9-cis-epoxycarotenoid dioxygenase (NCED3) genes in ugt74f1, thus promoting an antagonistic effect with SA-signalling and leading to enhanced bacterial growth. UGT74F2 might be a target for bacterial effectors since bacterial mutants affected in effector synthesis were impaired in inducing UGT74F2 expression. These results suggest that UGT74F2 negatively influences the accumulation of free SA, hence leading to an increased susceptibility due to reduced SA levels and increased expression of the JA and ABA markers LOX-2, FAD and NCED-3.

Changes in calcareous nannofossil assemblages across the Paleocene/Eocene transition from the paleo-equatorial Pacific Ocean

Quantitative analyses of selected calcareous nannofossils in deep-sea sections recovered from the paleo-equatorial Pacific (ODP Leg 199) provide new information about biostratigraphy, biochronology and the evolutionary history of calcareous nannofossils across the Paleocene/Eocene transition interval. The sediment cores from ODP Leg 199 represent the first continuous Paleocene/Eocene boundary sections ever to be sampled in the central equatorial Pacific Ocean. Calcareous nannofossil assemblages are studied to document the distribution of biostratigraphically useful taxa such as Ericsonia, Discoaster, Fasciculithus, Rhomboaster and Tribrachiatus. Focus is given to the evolution of the Rhomboaster-Tribrachiatus lineage in the lower Eocene interval at Site 1215, and on the stratigraphic relationship of these taxa relative to species in the genus Fasciculithus. Critical intervals of North Atlantic DSDP Site 550 have also been re-examined. The Tribrachiatus digitalis morphotype was described at Site 550 from an interval affected by down-hole contamination, partly originating from within the Tribrachiatus orthostylus range. The T. digitalis morphotype represents an evolutionary transitional form between T. contortus and T. orthostylus, entering the stratigraphic record within the range of the former species and disappearing within the lower part of the range of the latter species. The subzonal subdivision of Zone NP10 hence collapses. Lithological and colour variability reflecting orbital cyclicity occur in the lower Eocene of Site 1215, permitting a relative astronomical age calibration of the Tribrachiatus taxa. The distinct Rhomboaster spp.-Discoaster araneus association also occurs in the paleo-equatorial Pacific Ocean, together with a marked decrease in diversity of Fasciculithus spp. Site 1220 reveals a short peak abundance of Thoracosphaera spp. just above the P/E boundary interval, which probably reflects a stressed surface water environment.

The hydropolyp fauna of the Red Sea area

46 hydropolyp species of 28 genera and 10 families were sampled during the "Meteor" passage 1964/65 (IIOE) through the Red Sea and its northern and southern exits and on the occasion of several ecological investigations of 29 selected coral reef sections of the central Red Sea and the Gulf of Aqaba. These collections comprise 128 single records of hydropolyp species. Three species and two genera each with one species are doubtful. 25 species, seven genera, one family and one subfamily, together from 49 records have not previously been found in the Red Sea and its exits. Including these newly reported species, the total list increases from 64 species and 112 records to 89 species and 240 single records and 51 additional ones. Scanning microscopical photos, made for the first time for the illustration of the hydropolyps, have been shown to be suitable for a better characterization and diagnosis of the species. Qualified results on the reasons for the horizontal distribution of the species known from the Red Sea area cannot be given because of the low number of samples sporadically distributed through the whole area. In contrast with this fact, the vertical spread of the species sampled seems primarily to be regulated by water exchange and light intensity. For example, four species of hydropolyps are excellent indicators of certain abiotic factors or combinations of them: Gymnangium eximium reacts extremely stenophote-photophobe-rheophil, Eudendrium ramosum moderately stenophote-photophobe-rheophobe, Lytocarpus philippinus moderately stenophote-photophil-rheophil, and Halocordyle disticha var. australis extremely stenophote-photophil but moderately rheophil. Other species have been found throughout all the light zones. Combined with the small size of their colonies their euryphotic behaviour does not allow their use as indicator species.

Jurassic microfossils of DSDP Hole 79-547B

The first marine incursion of the incipient North Atlantic Ocean is recorded in the uppermost Triassic to Lower Jurassic sequence of DSDP Site 547 off central Morocco. A lithologic change from continental red beds below to slope breccias and hemipelagic carbonates above indicates that a carbonate ramp was probably established by Sinemurian time along the Moroccan continental margin and that subsidence in the adjacent basin was rapid in the early phases of continental rift. Foraminifers recovered from the Liassic (Sinemurian-Pliensbachian) basinal deposits are diverse and well preserved. The faunas are compositionally similar to contemporaneous neritic assemblages of Europe and the Grand Banks of Newfoundland. The Middle Jurassic in Hole 547B is characterized by regressive deposits that are poor in foraminifers. The major Late Jurassic "Atlantic" transgression is again represented by basinal deposits consisting of limestone breccias and pelagic carbonates. Foraminifers recovered from this interval are transitional between Late Jurassic assemblages reported from deep-sea deposits in the North Atlantic and typical Late Jurassic neritic assemblages of Europe. The Late Jurassic assemblages of Hole 547B are primarily dominated by nodosariids and spirillinids with moderate abundances of simple arenaceous forms. Nonreticulate epistominids occur very rarely in the Upper Jurassic of Hole 547B. It is tentatively suggested that these represent upper bathyal assemblages.

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.

Palynological records of sediments from the Odderade type locality, north Germany

Reinvestigation of the Odderade Interstadial in its type locality led to an augmentation of the flora list and correction of some misinterpretations (e.g. Omorica, Frangula). The Eemian, stadials FW 1 and 3, and the interstadials Amersfoort/Broerup and Odderade have been manifested by pollen analyses. FW 1 and FW 3 are probably not completely free from forested areas. The new pollen diagrams considered with older data from Odderade, and in comparison with other regions in Central Europe, fit essentially with the classification and development of Vegetation during the Early Weichselian in Oerel by Behre & Lade (1986).