Enzyme Micropump‐Based Inhibitor Assays

Rapid, easy‐to‐use, and portable devices that can provide a read‐out without the need for expensive equipment represent the future of sensing technology, with applications in areas like environmental, food, chemical, and biological safety. Enzymes immobilized on a surface function as micropumps in the presence of species (e.g., substrate, cofactor, or biomarker) that trigger the enzymatic reaction. The flow speed in these devices increases with increasing reaction rate. This allows the detection of substances that inhibit the enzymatic reaction. Using this principle, sensors for toxic substances, like mercury, cadmium, cyanide, and azide, were designed using urease and catalase‐powered pumps, respectively, with limits of detection well below the concentrations permitted by the Environmental Protection Agency. The study was also extended to other inhibitors for these enzymes. The sensing range of fluid flow‐based inhibitor assays depends on the type of inhibition, the enzyme concentration on the sensing platform, and, for competitive inhibition, the concentration of substrate used.     

Longitudinal Effects of Botulinum Toxin Injections on Voice-Related Quality of Life (V-RQOL) for Patients with Adductory Spasmodic Dysphonia

Adductory spasmodic dysphonia is a focal dystonia of laryngeal muscles. Patients with this disorder typically have severe vocal difficulties, with significant functional, social, and emotional consequences. There is no widely accepted cure for this condition, however, botulinum toxin injections of the thyroarytenoid muscles are considered by most voice clinicians to be the state of the art treatment. Based on extensive experience treating patients for adductory spasmodic dysphonia, we feel that traditional means of voice assessment do not adequately measure either the disease severity or the treatment outcomes. That is, listening to or acoustically analyzing limited phonatory samples does not capture the functional, social, and emotional consequences of this disorder. These consequences will be reflected in a patient's voice-related quality of life (V-RQOL). Using a validated voice outcomes instrument, the V-RQOL Measure, the purpose of this study was to quantify longitudinal changes in the V-RQOL of patients with adductory spasmodic dysphonia who are undergoing botulinum toxin injections. Twenty-seven consecutive new patients presenting with dysphonia to our institution during an 18-month period were diagnosed with adductory spasmodic dysphonia, and treated patients were evaluated prospectively using the V-RQOL Measure. Results indicated that (1) V-RQOL was initially very low for these patients, (2) botulinum toxin injections improved it significantly for each injection cycle studied, and (3) the magnitude of the treatment effect appears to change across injections.     

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.