Wheat (Triticum spp. L.), rye (Secale cereal L.), and barley (Hordeum vulgare L.) seeds contain peptides toxic to celiac patients. Maize (Zea mays L.) and rice (Oryza sativa L.) are distant relatives of wheat as well as sorghum (Sorghum bicolor (L.) Moench) and are known to be safe for celiacs. Both immunochemical studies and in vitro and in vivo challenge of wheat-free sorghum food products support this conclusion, although molecular evidence is missing. The goal of the present study was to provide biochemical and genetic evidence that sorghum is safe for celiac patients. In silico analysis of the recently published sorghum genome predicts that sorghum does not contain peptides that are toxic for celiac patients. Aqueous/alcohol-soluble prolamins (kafirins) from different sorghum varieties, including pure lines and hybrids, were evaluated by SDS-PAGE and HPLC analyses as well as an established enzyme-linked immunosorbent assay (ELISA) based on the R5 antibody. These analyses provide molecular evidence for the absence of toxic gliadin-like peptides in sorghum, confirming that sorghum can be definitively considered safe for consumption by people with celiac disease.
One of the main objectives of wheat glutenin subunit (GS) analysis is the identification of protein components linked to wheat quality. The proteomic characterization of glutenin has to consider the relatively low levels of arginine and lysine residues and the close sequence similarity among the different groups of these subunits, which hinders or even prevents the identification of the GS. In this study, a proteomic approach has been applied to resolve the heterogeneity of wheat glutenin components. Proteins extracted from Triticum durum flour were first analyzed by two-dimensional gel electrophoresis, which greatly reduced glutenin complexity. The identity of each spot was confirmed by nano liquid chromatography tandem mass spectrometry analysis of tryptic peptides. In parallel, measurements of the high mass range by matrix-assisted laser desorption/ionization time-of-flight analysis allowed detection of the large tryptic peptides. Gathering all data from search engine interrogation, very high sequence coverage was obtained for high molecular weight GS, including Bx7 and By8, in agreement with the known genetic profile of durum wheat. In addition, a truncated form of By8, never detected before, was also found. Low molecular weight GS (LMW-GS) B-type was identified with reasonable sequence coverage, while a clear identification of LWM-GS C- and D-type was hindered by the incompleteness of the wheat DNA databases. This study represents the first comprehensive analysis of the glutenin proteome and provides a reliable method for classifying wheat varieties according to their glutenin profile.
We report the results of x-ray diffraction and small angle scattering studies of Ar and Kr confined in sol-gel and Vycor glasses. The confined liquid crystallizes in a disordered hcp structure on freezing. Upon further cooling a sharp transition occurs at a reduced temperature of T/T(m) approximately 0.65, where the crystalline structure disappears and the total scattering decreases. This behavior marks the onset of a well-defined mobility transition, where the confined sample migrates out of the pore space.
We have established a primary cell culture of the marine demosponge Ircinia muscarum. The culture was started from a cell suspension obtained by a combination of mechanical chemical means. Microbial contamination was controlled by the use of a pool of antibiotics. Optical density, rather than hemocytometer count, is suggested to monitor the cellular growth. Analysis of the chemical composition of I. muscarum cells revealed absence of sterols, showing that the cells were unable to biosynthesize sterols. When the medium was supplemented with cholesterol an increase of about 70% in the number of cells was observed. These results suggest that the classic mammalian nutrient medium was not satisfactory for I. muscarum cell growth, and sterols were needed to satisfy the membrane requirements.
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