Label‐Free and Ultra‐Low Level Detection of Salmonella enterica Serovar Typhimurium Using Electrochemical Impedance Spectroscopy

An immunosensor for rapid and low level detection of the bacterial pathogen Salmonella enterica Serovar Typhimurium was designed and developed based upon label‐free electrochemical impedance spectroscopy and correlated to viable cell counts. The immunosensor was fabricated by electroplating gold onto a disposable printed circuit board (PCB) electrode by immobilizing monoclonal antibody (MAb) specific against Salmonella typhimurium cell surface lipopolysaccharide (LPS) onto the surface of the electrode. Use of mass‐fabricated and electroplated PCB electrodes allowed for disposable, highly sensitive, and rapid detection of Salmonella in an aqueous environment. Results demonstrate that in purified solution, Salmonella can be detected as low as 10 CFU in a 100 μL volume and label‐free and rapid manner in fewer than 90 s. The cost effective approach described here can be used for detection of pathogens with relevance for healthcare, food, and environmental applications.     

Suicide Inactivation of Dioldehydrase by 2‐Chloroacetaldehyde: Formation of the ‘cis‐Ethanesemidione’ Radical,and the Role of a Monovalent Cation

Dioldehydrase is an adenosylcobalamin‐dependent enzyme that catalyzes the dehydration of (R)‐ or (S)‐propane‐1,2‐diol to propanal. The reaction proceeds by a radical mechanism initiated by the homolytic scission of the covalent Co? C(5′) bond in the coenzyme to form cob(II)alamin and the 5‐deoxyadenosyl radical as transient intermediates. Dioldehydrase is subject to ‘suicide inactivation’ by substrate/product analogs. Inactivation by 2‐chloroacetaldehyde converts the inactivator into the ‘cis‐ethanesemidione’ radical. A mechanism for this process includes reaction of chloroacetaldehyde in the reverse of the normal catalytic process to a rearranged radical that eliminates HCl. K⁺ and other monovalent cations of similar size, including Tl⁺, are required for dioldehydrase activity and for suicide inactivation by glycolaldehyde or 2‐chloroacetaldehyde. A K⁺ ion is bound to propane‐1,2‐diol in dioldehydrase. Both EPR and pulsed‐EPR experiments show that the magnetic nuclei of thallous ions (²⁰³Tl⁺, ²⁰⁵Tl⁺) do not interact with the unpaired electron in the cis‐ethanesemidione radical at the active site of dioldehydrase. Pulsed‐EPR experiments implicate a ¹⁴NH group, possibly of His¹⁴³, interacting with the radical at the active site.     

Oligomeric carbocation-like species from protonation of chloroalkanes

The protonation of chloroethane by the strongest known solid superacid, the carborane acid H(CHB(11)Cl(11)), has been studied by quantitative IR spectroscopic methods to track mass balance and uncover previously unobserved chemistry. In the first step, an intermediate EtCl·H(CHB(11)Cl(11)) species without full proton transfer to EtCl can be observed when d(5)-deuterated chloroethane is used. It rapidly eliminates HCl (but not DCl) to form ethyl carborane, Et(CHB(11)Cl(11)), which binds a second molecule of chloroethane to form the Et(2)Cl(+) chloronium ion. This undergoes a slower, previously unrecognized HCl elimination reaction to form a butyl carborane, Bu(CHB(11)Cl(11)), beginning an oligomerization process whereby unsymmetrical dialkylchloronium ions decompose to alkyl carboranes of formula Bu(C(2)H(4))(n)(CHB(11)Cl(11)) up to n = 4. Over time, a parallel competing process of de-oligomerization take place in the presence of free carborane acid that finishes with the formation of hexyl or butyl carboranes. Upon heating to 150 C, the final products are all converted to the remarkably stable tert-butyl cation carborane salt.     

Various aspects of enantiomeric recognition of (S,S)-dimethylpyridino-18-crown-6 by several organic ammonium salts

Abstract

Factors responsible for complex stability and enantiomeric recognition for the interactions of (S,S)-dimethylpyridino-18-crown-6 with several organic ammonium salts were examined using an ¹H NMR technique. The results indicate that cation structures have a significant effect on enantiomeric recognition; solvents play a very important role in the stability of the complexes, and anions can compete with ligands for the ammonium cations.     

The Use of the PREPTM System for Handling of Body Fluids

Abstract

The Du Pont PREP^TM automated sample processor is a centrifugally based, microprocessor controlled instrument that was designed for extraction of samples from biological fluids. Extraction takes place in cartridges containing either organic resins or bonded silica packings as extraction sorbants. This paper will discuss the application of several lipophilic and ion exchange sorbants to the extraction of biological samples from body fluids. The advantages of these different types of sorbants will be compared and their performance with automated sample preparation will be shown. A variety of applications including the extraction of benzodiazepine, barbiturate, aminoglycoside and anticonvulsant drugs and their metabolites from serum, urine, and tissue homogenates will be discussed.     

Coupling of NMR to ACOMP for Terpolymerization Monitoring and Control

Monitoring and control of free radical copolymerization using automatic continuous online monitoring of polymerization reactions (ACOMP) with UV detection has been recently achieved. It is difficult, however, to achieve spectral separation of similar monomers with UV alone. Here, nuclear magnetic resonance (NMR) is coupled to ACOMP for the first time and terpolymerization reactions involving acrylamide (Am), sodium acrylate (Ac), and styrene sulfonate (SS) are monitored, and a first attempt at active composition control is made. The NMR resolves the concentrations of Am and Ac, while the UV resolves SS and the sum of Am and Ac. NMR analysis is performed in water, using signal suppression, to eliminate the need for deuterated solvents. From this, instantaneous fractions of each comonomer in the terpolymer are continuously known, along with weight average molecular weight and intrinsic viscosity, IV. Am and Ac have similar reactivities, whereas the reactivity ratio of SS is much larger. Hence, there is high composition drift in batch polymerization and SS is rapidly consumed, leaving a final copolymer of Am and SS to form. Maintaining constant terpolymer composition is taken as a first step toward active control.     

Azacrown ethers containing oximic and Schiff base sidearms - potential heteronuclear metal ion receptors

A simple synthetic pathway for the preparation of oxime- and Schiff base-containing aza- and diazacrown ethers is reported. N-Methoxymethyl-substituted aza-15-crown-5 and aza-18-crown-6 as well as N,N′-bis(methoxymethyl)-substituted diaza-18-crown-6 were treated with 5-bromosalicylaldehyde to produce the N-(2′-hydroxy-3′-carbonyl-5′-bromobenzyl)-substituted aza-15-crown-5 (8), aza-18-crown-6 (9) and N,N′-bis(2′-hydroxy-3′-carbonyl-5′-bromobenzyl)-substituted diaza-18-crown-6 (10) compounds. Compounds 8 and 10 were treated with hydroxylamine to give oxime-substituted ligands 12 and 13. A series of bis-Schiff base-containing diaza-18-crown-6 ligands were prepared by reacting 10 with 2-hydroxyaniline (to form 14), 5-nitro-2-hydroxyaniline (15), 2-aminopyridine (16), 2-hydrazinopyridine (17) and N-aminomorpholine (18). Compounds 12–18 are potential complexing agents for simultaneous binding of soft transition and hard alkali or alkaline earth metal ions in one molecule. These new oxime- and Schiff base-containing ligands interacted strongly with Na⁺ and K⁺ in methanol. The interaction of the aromatic portions of 9, 10, and 12–15 with transition metal ions was shown by the UV spectra of the metal ion complexes in 50% aqueous DMF. The X-ray structure of 10 is reported.     

Recognizing Planar Strict Quasi-Parity Graphs

 A graph is a strict-quasi parity (SQP) graph if every induced subgraph that is not a clique contains a pair of vertices with no odd chordless path between them (an “even pair”). We present an O(n ³) algorithm for recognizing planar strict quasi-parity graphs, based on Wen-Lian Hsu's decomposition of planar (perfect) graphs and on the (non-algorithmic) characterization of planar minimal non-SQP graphs given in [9]. Received: September 21, 1998 Final version received: May 9, 2000     

The “least flexible job first” rule in scheduling and in queueing

We consider an environment with m$m$ machines in parallel operating at different speeds. The processing requirements of all jobs are independent and have the same exponential distribution. Job j$j$ may only be processed on a specific subset of the m$m$ machines, referred to as its restricted set. The restricted sets are nested and preemptions are allowed. We show that the Least Flexible Job to the Fastest Machine (LFJ-FM) minimizes the expected makespan and the total expected completion time.     

Critical random graphs and the structure of a minimum spanning tree

We consider the complete graph on n vertices whose edges are weighted by independent and identically distributed edge weights and build the associated minimum weight spanning tree. We show that if the random weights are all distinct, then the expected diameter of such a tree is Θ(n^1/3). This settles a question of Frieze and Mc‐Diarmid (Random Struct Algorithm 10 (1997), 5–42). The proofs are based on a precise analysis of the behavior of random graphs around the critical point. © 2008 Wiley Periodicals, Inc. Random Struct. Alg., 2009