Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides

Despite evident regulatory roles of heparan sulfate (HS) saccharides in numerous biological processes, definitive information on the bioactive sequences of these polymers is lacking, with only a handful of natural structures sequenced to date. Here, we develop a “Shotgun” Ion Mobility Mass Spectrometry Sequencing (SIMMS2) method in which intact HS saccharides are dissociated in an ion mobility mass spectrometer and collision cross section values of fragments measured. Matching of data for intact and fragment ions against known values for 36 fully defined HS saccharide structures (from di- to decasaccharides) permits unambiguous sequence determination of validated standards and unknown natural saccharides, notably including variants with 3O-sulfate groups. SIMMS2 analysis of two fibroblast growth factor-inhibiting hexasaccharides identified from a HS oligosaccharide library screen demonstrates that the approach allows elucidation of structure-activity relationships. SIMMS2 thus overcomes the bottleneck for decoding the informational content of functional HS motifs which is crucial for their future biomedical exploitation

Herschel observations of extraordinary sources: Full Herschel/HIFI molecular line survey of Sagittarius B2(M)

We present a full analysis of a broadband spectral line survey of Sagittarius B2 (Main), one of the most chemically rich regions in the Galaxy located within the giant molecular cloud complex Sgr B2 in the Central Molecular Zone. Our goal is to derive the molecular abundances and temperatures of the high-mass star-forming region Sgr B2(M) and thus its physical and astrochemical conditions. Sgr B2(M) was observed using the Heterodyne Instrument for the Far-Infrared (HIFI) on board the Herschel Space Observatory in a spectral line survey from 480 to 1907 GHz at a spectral resolution of 1.1 MHz, which provides one of the largest spectral coverages ever obtained toward this high-mass star-forming region in the submillimeter with high spectral resolution and includes frequencies > 1 THz unobservable from the ground. We model the molecular emission from the submillimeter to the far-IR using the XCLASS program. For each molecule, a quantitative description was determined taking all emission and absorption features of that species across the entire spectral range into account. Additionally, we derive velocity resolved ortho / para ratios for those molecules for which ortho and para resolved molecular parameters are available. Finally, the temperature and velocity distributions are analyzed and the derived abundances are compared with those obtained for Sgr B2(N) from a similar HIFI survey. A total of 92 isotopologues were identified, arising from 49 different molecules, ranging from free ions to complex organic compounds and originating from a variety of environments from the cold envelope to hot and dense gas within the cores. Sulfur dioxide, methanol, and water are the dominant contributors. For the ortho / para ratios we find deviations from the high temperature values between 13 and 27 %. In total 14 % of all lines remain unidentified.Comment: 67 pages, 102 figures, submitted to A&

Alpine Glacier Reveals Ecosystem Impacts of Europe’s Prosperity and Peril Over the Last Millennium

Information about past ecosystem dynamics and human activities is stored in the ice of Colle Gnifetti glacier in the Swiss Alps. Adverse climatic intervals incurred crop failures and famines and triggered reestablishment of forest vegetation but also societal resilience through innovation. Historical documents and lake sediments record these changes at local—regional scales but often struggle to comprehensively document continental-scale impacts on ecosystems. Here, we provide unique multiproxy evidence of broad-scale ecosystem, land use, and climate dynamics over the past millennium from a Colle Gnifetti microfossil and oxygen isotope record. Microfossil data indicate that before 1750 CE forests and fallow land rapidly replaced crop cultivation during historically documented societal crises caused by climate shifts and epidemics. Subsequently, with technology and the introduction of more resilient crops, European societies adapted to the Little Ice Age cold period, but resource overexploitation and industrialization led to new regional to global-scale environmental challenges

Ice cave reveals environmental forcing of long-term Pyrenean tree line dynamics

1. Tree lines are supposed to react sensitively to the current global change. However, the lack of a long-term (millennial) perspective on tree line shifts in the Pyrenees prevents understanding the underlying ecosystem dynamics and processes. 2. We combine multiproxy palaeoecological analyses (fossil pollen, spores, conifer stomata, plant macrofossils, and ordination) from an outstanding ice cave deposit located in the alpine belt c. 200 m above current tree line (Armeña-A294 Ice Cave, 2, 238 m a.s.l.), to assess for the first time in the Pyrenees, tree line dynamics, and ecosystem resilience to climate changes 5, 700–2, 200 (cal.) years ago. 3. The tree line ecotone was located at the cave altitude from 5, 700 to 4, 650 cal year bp, when vegetation consisted of open Pinus uncinata Ramond ex DC and Betula spp. Woodlands and timberline were very close to the site. Subsequently, tree line slightly raised and timberline reached the ice cave altitude, exceeding its today''s uppermost limit by c. 300–400 m during more than four centuries (4, 650 and 4, 200 cal year bp) at the end of the Holocene Thermal Maximum. After 4, 200 cal year bp, alpine tundra communities dominated by Dryas octopetala L. expanded while tree line descended, most likely as a consequence of the Neoglacial cooling. Prehistoric livestock raising likely reinforced climate cooling impacts at 3, 450–3, 250 cal year bp. Finally, a tree line ecotone developed around the cave that was on its turn replaced by alpine communities during the past 2, 000 years. 4. Synthesis. The long-term Pyrenean tree line ecotone sensitivity suggests that rising temperatures will trigger future P. uncinata and Betula expansions to higher elevations, replacing arctic–alpine plant species. Climate change is causing the rapid melting of the cave ice; rescue investigations would be urgently needed to exploit its unique ecological information

Pantoprazole Does not Affect Serum Trough Levels of Tacrolimus and Everolimus in Liver Transplant Recipients

Background: Liver transplant recipients are frequently treated with proton pump inhibitors. Drug interactions have been described especially with respect to omeprazole. Due to the lower binding capacity of pantoprazole to CYP2C19 this drug became preferred and became the most used proton pump inhibitor in Germany. The data on the influence of pantoprazole on immunosuppressive drugs in liver transplant recipients a very scarce.Methods: The authors performed a single center analysis in liver transplant recipients on the effect of pantoprazole on the serum trough levels of different immunosuppressants. The trough levels were compared over a period of 1 year before and after start or stop of a continuous oral co-administration of 40 mg pantoprazole once daily.Results: The serum trough levels of tacrolimus (n = 30), everolimus (n = 7), or sirolimus (n = 3) remain constant during an observation period of at least 1 year before and after co-administration of pantoprazole. None of the included patients needed a change of dosage of the observed immunosuppressants during the observation period.Conclusions: The oral co-administration of pantoprazole is safe in immunosuppressed liver transplant recipients according to the serum trough levels of tacrolimus, everolimus, and sirolimus. This analysis provides first data on the influence of pantoprazole on immunosuppressive drugs in liver transplant recipients

Using Automated Glycan Assembly (AGA) for the Practical Synthesis of Heparan Sulfate Oligosaccharide Precursors

Herein we report synthesis of complex heparan sulfate oligosaccharide precursors by automated glycan assembly using disaccharide donor building blocks. Rapid access to a hexasaccharide was achieved through iterative solid phase glycosylations on a photolabile resin using Glyconeer™, an automated oligosaccharide synthesiser, followed by photochemical cleavage and glycan purification using simple flash column chromatography

Release of tryptophan and serotonin into the portal vein of the isolated perfused rat small intestine

To investigate the release of serotonin from intestinal enterochromaffin cells, we used an in vitro technique which allows studies excluding overlapping influences from outside the gut. The entire small intestine of rats fed a standard or tryptophan-enriched (3% of total) diet was totally isolated by ligatures with the exception of the superior mesentric artery and portal vein that supply and drain the intestine. Simultaneously to the vascular perfusion (Krebs-Ringer bicarbonate buffer, 0,4% human albumin, 5 m M glucose, 0.6 m M glutamine) the gut lumen was infused (buffer or 0.1 N HCL). Acidification of the gut lumen resulted in an increment of venously released tryptophan and serotonin. After feeding tryptophan-enriched food the release of tryptophan was increased. However, the total amount of released serotonin after tryptophan diet did not differ as compared to that after standard diet. Addition of a monoamino-oxidase inhibitor (pargyline) to the arterial perfusate enhanced the released amount of serotonin 3-fold in the portal venous effluent (at a concentration of 1 m M but not 0.1 m M ). Recovery studies done by arterial infusions of serotonin (1 µ M , 10µ M ) and evaluation of the amounts venously released revealed a high loss of infused serotonin (40%–70%). Our data suggest gut-born serotonin to more likely play a paracrine role than a role as a classical hormone.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47538/1/433_2005_Article_BF01852260.pd