The DNA aptamers that specifically recognize ricin toxin are selected by two in vitro selection methods

Aptamers which specifically recognize cytotoxin ricin were successfully selected using the two different in vitro selection methods. One selection method was used to isolate aptamers by affinity chromatography. Another selection method, named CE-SELEX, was carried out using CE as a separation approach. The high separation efficiency of CE evidently improved the rate of enrichment and obviously shortened the selection rounds, with near 87.2% binding just after the fourth round of selection. The aptamers A3, C1, and C5, derived from the two selection methods, were found to possess high affinity and specificity for ricin with the Kd values in the low nanomolar range, and did not recognize abrin toxin similar to ricin in the structures and properties, or BSA. Among the aptamers selected, A3 isolated by affinity chromatography shared extensive sequence similarity with C1 and C5 derived from CE-SELEX. They differed by only one base from each other. Their stable secondary structures predicted also had very similar structure motifs, and all folded a long and internal loop-embedded loop stem structure by base pairing. The ELISA and dot-blot analysis also proved that the selected DNA aptamers had the high specificity to ricin toxin.     

Synthesis of an Aminooxy Derivative of the Trisaccharide Globotriose Gb3

The synthesis of α-aminooxy trisaccharide moiety [α-d-Gal-(1,4)-β-d-Gal-(1,4)-β-d-Glc-α-aminooxy], related to the cell surface globotriaosylceramide (Gb3) receptor of the B subunit of the AB₅ Shiga toxin of Shigella dysenteriae, has been carried out for the first time in 11 steps with a 15% overall isolated yield. A highlight of this work entails utilizing chemically compatible synthetic transformations, including those related to glycosylation, incorporative of the succinimidyl moiety as a precursor to the aminooxy Gb3 derivative. The fully deprotected trisaccharide aminooxy compound was reacted with a carbonyl compound, leading to oxime formation in quantitative yield, which underscores the importance for future glyco-conjugations.     

Development of a new linker for the solid-phase synthesis of N-hydroxylated and N-methylated secondary amines

Merrifield resin was modified by the introduction of an ortho-nitrophenylethanal group that served as a linker moiety to attach amines to the resin by reductive amination. Resin-bound tertiary amines were shown to be readily transferred into the respective liberated N-hydroxylated or N-methylated derivatives by either an oxidation/Cope elimination or a permethylation/Hofmann elimination protocol. With these two divergent liberation/derivatization options, the new resin offers new flexibility in the solid phase synthesis of N-modified secondary amines, for instance in spider toxin synthesis.     

一类带毒的生化反应竞争模型的定性分析

有些微生物在连续培养中会产生毒素来抑制竞争者,同时竞争中也会产生一些振荡行为.本文研究两个微生物竞争同一营养,而其中一个竞争者会产生毒素抑制另一竞争者且产物系数为一般的形如δ1=A1+B1Sn,δ2=A2+B2Sm的函数时的生化反应模型.分析了系统平衡点的稳定性和当系统的某一微生物物种处于竞争劣势而趋于灭绝时另一微生物物种和营养的二维流形上极限环的存在性.     

Electrophoretic isolation of saponin fractions from Saponinum album and their evaluation in synergistically enhancing the receptor-specific cytotoxicity of targeted toxins

Saponinum album (SA) is a commercial mixture of saponins isolated from Gypsophila species. In the previously published work, we reported that SA dramatically improves the inhibition of tumor growth by targeted toxins in mice in a synergistic way. Here we report a simplified electrophoretic method for the isolation of a highly effective fraction of SA with a relative electrophoretic mobility to the dye front (R(f) ) of 0.63 from the mixture. In total, four different fractions were separated at a preparative scale, and evaluated by ESI-MS, HPLC and TLC analysis. Electrophoretic mobility and electrochemical properties of the different fractions of saponins from SA were set into relation to their ability to enhance the cytotoxicity of epidermal growth factor (EGF)-based targeted toxins. We here treated HER-14 cells, which are NIH-3T3 Swiss mouse embryo cells transfected with the human EGF receptor. Untransfected NIH-3T3 cells served as control. The major bulk of SA (72.3%) (R(f) =0.78) migrated the farthest and was found to be significantly ineffective (p<0.05) in enhancing the cytotoxicity of the targeted toxin, while the second fraction (R(f) =0.63) showed an enhancement of 9800-fold. The third (R(f) =0.56) had an enhancement factor of 3200, the fourth (R(f) =0.08) was again significantly ineffective (p<0.05) in exhibiting any enhancement of cytotoxicity.     

Current methods of analysis for the determination of trichothecene mycotoxins in food

This article describes the trends in analytical techniques for the determination of trichothecene mycotoxins, namely deoxynivalenol, and T-2 and HT-2 toxins in cereals and cereal products with particular emphasis on screening and rapid approaches. The driving force behind the changing methodologies is mainly attributed to legislative demands. However, for commercial and governmental testing laboratories, the need to use validated official methods is ever increasing to ensure quality assurance of results.Much research has been undertaken to improve screening assays, highlighted by the number of new methods using a variety of formats and platforms, including optical and electrochemical biosensors. Significant advances in the traditional reference methods have also been demonstrated in addition to the emergence of a variety of commercial immunoaffinity and solid-phase extraction columns for clean up. The use of liquid chromatography coupled to tandem mass spectrometry for mycotoxin detection is ever increasing, allowing simultaneous determination of many toxins in various sample matrices.