Utilization of Polyspecific Antiserum for Specific Radioimmunoassays: Radioimmunoassays for Rat Fetuin and Bikunin Were Developed by Using Antiserum Against Total Rat Serum Proteins

Polyspecific antiserum against total rat serum proteins was used to develop specific and sensitive radioimmunoassays for fetuin and bikunin, two minor protein components of rat plasma. The radioimmunoassays proved to be highly useful to trace bikunin and fetuin in the course of developing isolation procedures, since neither specific functional assays nor monospecific antisera were available. The two examples demonstrate that, in general, it will be possible to develop a specific and sensitive radioimmunoassay with antiserum raised against a crude antigen preparation, such as a body fluid or a tissue extract, provided that a minute amount of pure antigen is available for preparing the radioiodinated antigen

The nucleotide and partial amino acid sequences of rat fetuin

Fetuins are among the major plasma proteins, yet their biological role has remained elusive. Here we report the molecular cloning of rat fetuin and the sequence analysis of a full-length clone, RF619 of 1456 bp with an open reading frame of 1056 bp encoding 352 amino acid residues. The coding part of RF619 was identical with the cDNA sequence of the natural inhibitor of the insulin receptor tyrosine kinase from rat (pp63) except for four substitutions and a single base insertion causing divergence of the predicted protein sequences. Partial amino acid sequences of rat plasma fetuin were in agreement with the predictions based on the RF619 cDNA. Purified rat fetuin inhibited the insulin receptor tyrosine kinase in vitro. Therefore, we conclude that RF619 and pp63 cDNA encode the same protein, i.e. authentic rat fetuin which is a functional tyrosine kinase inhibitor

A Hepatic Protein, Fetuin-A, Occupies a Protective Role in Lethal Systemic Inflammation

A liver-derived protein, fetuin-A, was first purified from calf fetal serum in 1944, but its potential role in lethal systemic inflammation was previously unknown. This study aims to delineate the molecular mechanisms underlying the regulation of hepatic fetuin-A expression during lethal systemic inflammation (LSI), and investigated whether alterations of fetuin-A levels affect animal survival, and influence systemic accumulation of a late mediator, HMGB1.LSI was induced by endotoxemia or cecal ligation and puncture (CLP) in fetuin-A knock-out or wild-type mice, and animal survival rates were compared. Murine peritoneal macrophages were challenged with exogenous (endotoxin) or endogenous (IFN-γ) stimuli in the absence or presence of fetuin-A, and HMGB1 expression and release was assessed. Circulating fetuin-A levels were decreased in a time-dependent manner, starting between 26 h, reaching a nadir around 24-48 h, and returning towards base-line approximately 72 h post onset of endotoxemia or sepsis. These dynamic changes were mirrored by an early cytokine IFN-γ-mediated inhibition (up to 50-70%) of hepatic fetuin-A expression. Disruption of fetuin-A expression rendered animals more susceptible to LSI, whereas supplementation of fetuin-A (20-100 mg/kg) dose-dependently increased animal survival rates. The protection was associated with a significant reduction in systemic HMGB1 accumulation in vivo, and parallel inhibition of IFN-γ- or LPS-induced HMGB1 release in vitro.These experimental data suggest that fetuin-A is protective against lethal systemic inflammation partly by inhibiting active HMGB1 release

Progress towards assessing the contemporary evolution of the Greenland ice sheet.

A more accurate assessment of the contemporary evolution of the Greenland ice sheet and its major drainage basins requires a close interaction between observational data and modeling. The main challenge when interpreting satellite and observational data is to separate the ice mass contribution from the contribution of postglacial isostatic rebound, to separate ice-sheet dynamic changes from interannual surface mass balance changes, and to separate long-term ice-dynamic changes from short-term flow fluctuations. Here we report from recent progress towards these goals within the DFG SPP 1257 project 'Assessing the current evolution of the Greenland ice sheet' from studies combining observational data with glaciological modeling. This comprises studies to reconstruct the surface mass balance of the Greenland ice sheet between 1866 and 2006, optical satellite data from ASTER to obtain surface velocities, modelled balance velocities, and simulations with a three-dimensional thermomechanical ice-sheet model. In combination with GRACE data, these studies are expected to contribute to an improved estimate of the present-day contribution of the Greenland ice sheet to global sea-level change and a better understanding of the various contributions to current ice mass changes and their associated uncertainties

S-type lectins occur also in invertebrates: high conservation of the carbohydrate recognition domain in the lectin genes from the marine sponge Geodia cydonium

The marine sponge Geodia cydonium contains several lectins. The main component, called lectin-1, is composed of three to four identical subunits. The subunits of the lectins were cloned from a cDNA library; two clones were obtained. From the deduced aa sequence of one clone, LECT-1, a mol. wt of 15,313 Da is calculated; this value is in good agreement with mass spectrometric analysis of 15,453 ± 25 Da. The sequence of another clone, LECT-2, was analysed and the aa sequence was deduced (15,433 Da). The two subunits have a framework sequence of 38 conserved aa which are characteristic for the carbohydrate-binding site of vertebrate S-type lectins. Clustering of lectin sequences of various species following their pairwise comparison establishes a dendrogram, which reveals that the sponge lectin could be considered as the ancestor for vertebrate S-type lectins