Tyrosine phosphorylation of annexin II tetramer is stimulated by membrane binding

In the present article we have examined if the interaction of the Ca2+-binding protein, annexin II tetramer (AIIt) with the plasma membrane phospholipids or with the submembranous cytoskeleton, effects the accessibility of the tyrosine phosphorylation site of AIIt. In the presence of Ca2+, pp60(c-src) catalyzed the incorporation of 0.22 +/- 0.05 mol of phosphate/mol of AIIt (mean +/- S.D., n = 5). The Ca2+-dependent binding of AIIt to purified adrenal medulla plasma membrane or phosphatidylserine vesicles stimulated the pp60(c-src)-dependent phosphorylation of AIIt to 0.62 +/- 0.04 mol of phosphate/mol of AIIt (mean +/- S.D., n = 5) or 0.93 +/- 0.07 mol of phosphate/mol of AIIt (mean +/- S.D., n = 5), respectively. Phosphatidylserine- or phosphatidylinositol-containing vesicles but not vesicles composed of phosphatidylcholine or phosphatidylethanolamine, stimulated the phosphorylation of AIIt. In contrast, the binding of AIIt to F-actin resulted in the incorporation of only 0.04 +/- 0.04 mol of phosphate/mol of AIIt (mean +/- S.D., n = 5). These results suggest that the interaction of AIIt with plasma membrane and not the submembranous cytoskeleton, activates the tyrosine phosphorylation of AIIt by inducing a conformational change in the protein resulting in the enhanced exposure or accessibility of the tyrosine-phosphorylation site.     

Release and characterization of single side chains of white cabbage pectin and their complement‐fixing activity

A mixture of single side chains from white cabbage pectin were obtained by anion exchange chromatography after applying mild chemical conditions promoting β‐elimination. These pectin fragments were characterized by their molecular weight distribution, sugar composition, ¹³C‐NMR, and MALDI‐TOF‐MS analysis. These analyses revealed that the large oligosaccharides released by β‐eliminative treatment were composed of α‐1,5 linked arabinosyl residues with 2‐ and 3‐linked α‐arabinosyl side chains, and, or β‐1,4 linked galactosyl side chains. Fractions were tested for complement‐fixing activity in order to determine their interaction with the complement system. These results strongly indicated that there was a minimal unit size responsible for the complement‐fixing activity. Neutral pectin fragments (?8 kDa) obtained from β‐elimination were inactive in the complement system, although they contained a sugar composition previously shown to be highly active. Larger pectin fragments (?17 kDa) retained some activity, but much lower than polymers containing rhamnogalacturonan type 1 (RGI) structures isolated from the same source. This implied that structural elements containing multiple side chains is necessary for efficient complement‐fixing activity.     

On-line liquid chromatography-accurate radioisotope counting coupled with a radioactivity detector and mass spectrometer for metabolite identification in drug discovery and development

A novel detection method combining on-line liquid chromatography-accurate radioisotope counting (LC-ARC, advanced stop flow controller) coupled with a radioactivity detector and mass spectrometer has been developed. One of the major benefits of this method is that this system enhances the sensitivity of radioisotope measurement for metabolite identification in drug metabolism studies. Another advantage to this system is the easy interface with the mass spectrometer, which allows acquisition of mass spectrometric data on-line. For purposes of evaluating this system, in vitro microsomal incubations with [3Hlpropranolol were conducted. On-line separation and identification of [3H]propranolol metabolites was achieved without intensive sample preparation, concentration, or fraction collection. Mass spectrometric analysis showed the presence of propranolol major metabolites formed by hydroxylation, correlating with previously published results. Further evaluations of this system also were conducted using two 14C compounds, which are herein labeled X and Y. As our results show, 14C peaks were detected down to 6 cpm, which is approximately 20 times more sensitive than commercially available flow through radioactivity detectors. The overall results suggest that the combination of LC-ARC with radioactivity detection and mass spectrometry has great potential as a powerful tool for improving the sensitivity of radioisotope measurement in metabolite identification studies during drug discovery and development.     

食用核苷酸

<正> 以前,人们认为核苷酸并非人体所需的营养物质。人们普遍认为生物,包括人类在内,有能力在体内合成对于正常生长和发育所需的核苷酸。如今这个观点发生了改变。有证据表明食用核苷酸对人体起着至关重要的作用。核苷酸是一种低分子量的生物化合物,它几乎在所有的生化反应中部起着非常重要的作用。核苷酸由嘌呤或嘧啶(一种含氮碱基)组成。它们是组成脱氧核糖核酸和核糖核酸的基本物质,在细胞运动和新陈代谢过程中起着很多重要的作用,尤其在许多辅酶中它们也是ATP和其它核苷三磷酸的组成成分。     

Performance assessment of electrospun nanofibers for filter applications

The aim of this study was to evaluate the use of nanofiber microfiltration membranes, spun by an innovative electrospinning technique, in water filtration applications. As such, this study bridges between developments in electrospinning techniques for the production of flat sheet membranes and the application of these membranes in water filtration. Three different applications were examined. First, the use of the membrane (functionalized or non-functionalized) for the removal of pathogens was investigated. Second the electrospun flat sheet membranes were applied in a lab scale submerged membrane bioreactor (MBR). Third, the electrospun membranes were applied as stand-alone filter for water treatment. Next to these applications, physical properties such as clean water permeability (CWP) and strength were also examined. The test showed that the electrospun membranes can be used for water filtration applications, but that further research is necessary towards irreversible fouling properties and level of functionalisation.     

Pectin isolated from white cabbage--structure and complement-fixing activity

This study was done to investigate whether white cabbage contained polysaccharides with immunostimulatory activity using the complement-fixing test as an indicator. The main polysaccharide isolated was of pectin nature. Methanolysis and (13)C-NMR showed that the polymers consisted of highly esterified alpha-galactopyranoside (alpha-GalpA), significant amounts of alpha-arabinose furanoside (alpha-Araf), beta-Galp and lesser amounts of rhamnose in the pyranose form (Rhap) and xylose in the pyranose form (Xylp). Linkage analyses showed that the alpha-GalpA residues were mainly 1,4-linked with small amounts of 1,3,4-linkages. The alpha-Araf residues were mainly terminally (t)- and 1,5-linked, whereas beta-Galp was t-, 1,3-, 1,6-, and 1,3,6-linked. Positive Yariv reaction indicated polymers with arabinogalactan type 2 like structures. alpha-Rhap was mainly present as 1,2- and 1,2,4-linked residues and Xylp was t- and 1,4-linked. The molecular weight varied greatly and was from 10 to 150 kDa. Cabbage polymers had biological activity and this complement-fixing activity was greatly affected by hydrolytic removal of Araf from pectic side chains.     

Screening of steam explosion conditions for glucose production from non-impregnated wheat straw

The conversion of wheat straw to fermentable sugar for bioethanol production typically involves a thermal pretreatment step, followed by enzymatic hydrolysis. In this study we have investigated the effect of steam explosion parameters on wheat straw digestibility using a newly designed steam explosion unit and a process without acid impregnation. The wheat straw was steam pretreated using 18 different conditions in the temperature range of 170-220 °C and the resulting material was used directly (i.e. without washing) for enzymatic hydrolysis and fermentation in either a separate hydrolysis and fermentation (SHF)-type or a simultaneous saccharification and fermentation (SSF)-type set-up. Maximum glucose yields upon enzymatic hydrolysis were obtained after pretreatment at 210 °C for 10 min and yields were similar at harsher conditions. Xylose yields increased with temperature and residence time up to 190 °C, but decreased at harsher pretreatment conditions since these led to xylan degradation and concomitant production of furfural. In an SHF-type set-up ethanol formation did not follow enzymatic glucose release and was inversely correlated with furfural levels. An SFF-type set-up displayed a straightforward correlation between the expected amount of released glucose and the ethanol yields. The highest saccharification yields corresponded to about 90% of the cellulose in the substrate. Overall, this study shows that steam explosion without an acid catalyst is a good pretreatment method for saccharification of wheat straw. Optimal steam explosion conditions need to be a compromise between sugar accessibility and sugar degradation.