Antigenic structure of rabies virus glycoprotein: ordering and immunological characterization of the large CNBr cleavage fragments.

After rabies virus glycoprotein was treated with CNBr, the peptide mixture was fractionated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. CNBr-cleaved peptide fragments were resolved into seven peptide bands under reducing conditions and six peptide bands under nonreducing conditions. The isolated nonreduced polypeptides were further analyzed by electrophoresis under reducing conditions. The N-terminal amino acid sequences were determined for the peptides in each of the isolated bands. The sequence data identified eight CNBr peptides and allowed the peptide fragments to be ordered within the deduced amino acid sequence of the glycoprotein. Analysis of the nonreduced CNBr peptides revealed two conformations of the glycoprotein. Two CNBr peptide fragments were specifically immunoprecipitated with a hyperimmune anti-rabies glycoprotein serum. These two and one other CNBr peptide induced the production of rabies virus-neutralizing antibodies, indicating the existence of at least three distinct antigenic sites on the rabies virus glycoprotein.     

S100A6 mediates nuclear translocation of Sgt1: a heat shock-regulated protein

Sgt1 was originally identified in yeast as a suppressor of the Skp1 protein. Later, it was found that Sgt1 is present in plant and mammalian organisms and that it binds other ligands such as S100A6, a calcium-binding protein. In this work we show that in HEp-2 cells Sgt1 translocates to the nucleus due to heat shock. We also found that in HEp-2 cells with diminished level of S100A6, due to stable transfection with siRNA against S100A6, such translocation occurred at a much smaller scale in comparison with cells expressing a normal level of S100A6. Moreover, translocation of Sgt1 was observed in HEp-2 cells treated with thapsigargin instead of heat shock. In contrast thapsigargin was ineffective in cells with diminished level of S100A6. Thus, our results suggest that increase in intracellular concentration of Ca²⁺, transduced by S100A6, is necessary for nuclear translocation of the Sgt1 protein.     

Reconstruction of non-uniformly sampled five-dimensional NMR spectra by signal separation algorithm

A method for five-dimensional spectral reconstruction of non-uniformly sampled NMR data sets is proposed. It is derived from the previously published signal separation algorithm, with major alterations to avoid unfeasible processing of an entire five-dimensional spectrum. The proposed method allows credible reconstruction of spectra from as little as a few hundred data points and enables sensitive resonance detection in experiments with a high dynamic range of peak intensities. The efficiency of the method is demonstrated on two high-resolution spectra for rapid sequential assignment of intrinsically disordered proteins, namely 5D HN(CA)CONH and 5D (HACA)CON(CO)CONH.     

A fluorescence in situ hybridization technique for retrospective cytogenetic analysis

Fluorescence in situ hybridization (FISH) is being used increasingly in clinical practice; however, current FISH techniques require fresh material, and there is considerable variation in hybridization efficiency between laboratories. We have modified a FISH technique described by Pinkel et al. (1986) that works not only on freshly G-banded material but also on cytogenetic preparations ranging in age from 2 wk to 12 yr. We have tested this technique on several centromeric alphoid satellite probes (D1Z5, D7Z1, D17Z1, DXZ1, and DYZ3) and one noncentromeric minisatellite probe (D1Z2). Our average hybridization efficiency on freshly banded preparations for these probes is consistently greater than 90%. The combination of higher efficiency and the ability to perform hybridization on previously G-banded material makes this a valuable technique for retrospective analyses.     

Die Feinstruktur des Lungengewebes nach dem Beginn der Atmung

Ohne Zusammenfassung     

The Physical and Linear Viscoelastic Properties of Fresh Wet Foams Based on Egg White Proteins and Selected Hydrocolloids

The aim of this work was to evaluate the physicochemical properties of fresh foams based on egg white proteins, xanthan gum and gum Arabic. The distributions of the size of gas bubbles suspended in liquid were determined, as well as density and volume fraction of gas phase of the generated foams. Additionally, the viscoelastic properties in the linear range were measured, and the results were analyzed with the use of the fractional Zener model. It was shown, that foam supplementation with hydrocolloids considerably decreased their volume fraction of gas phase in comparison to pure egg white protein-based foams. Application of gum Arabic did not cause an increase in the size of foam bubbles when compared to pure white egg foam, whereas application of xanthan gum significantly decreased the size of the bubbles. Application of the fractional Zener model allowed to determine the relaxation times, their intensity in analyzed suspensions and also equilibrium module (G _e ). The increase in the concentration of xanthan gum resulted in the prolongation of the relaxation time and increased its intensity. Gum Arabic, when added, weakened the viscoelastic properties of the mixture as a viscoelastic solid.