Technological advancements for the detection of and protection against biological and chemical warfare agents

There is a growing need for technological advancements to combat agents of chemical and biological warfare, particularly in the context of the deliberate use of a chemical and/or biological warfare agent by a terrorist organization. In this tutorial review, we describe methods that have been developed both for the specific detection of biological and chemical warfare agents in a field setting, as well as potential therapeutic approaches for treating exposure to these toxic species. In particular, nerve agents are described as a typical chemical warfare agent, and the two potent biothreat agents, anthrax and botulinum neurotoxin, are used as illustrative examples of potent weapons for which countermeasures are urgently needed.     

Stability of nonautonomous second-order partial differential equations

Existence, uniqueness, and three sufficient stability criteria will be established for a class of differential equations of the form: $\text{d}{}^2\text{u(t)}\text{dt}{}^2\text{+ C(t)}\text{du(t)}\text{dt}\text{+ K(t) u(t) = 0}$.The stability criteria emphasize the situations where the nonautonomous portion of the linear operators C(t) and K(t): (a) have small magnitudes, (b) have small derivatives (slowly varying), and (c) have small integrals (rapidly varying). Although this paper directs attention only to the second-order equation, corresponding stability results for the first-order equation could be obtained utilizing similar arguments.     

Catalysis of the photo-Fries reaction: antibody-mediated stabilization of high energy states

A conformationally constrained hapten is presented that is capable of catalyzing the first antibody-mediated photo-Fries rearrangement. In this reaction, absorption of light energy by a diphenyl ether substrate results in homolytic C-O bond cleavage followed by recombination to yield biphenyl-derived products. The most proficient antibody studied converts 4-phenoxyaniline 15 into 2-hydroxy-5-aminobiphenyl 16 under high-intensity irradiation at a rate of 8.6 microM/min. These results support a recent hypothesis stating that immunization with conformationally constrained haptens provides higher titers for the acquisition of simple binding antibodies; however, in this case, conformational constraint does not ensure the development of more efficient catalysts. Using the obtained antibodies, the presence of products resulting from escape of free radicals from the solvent cage can be suppressed, altering the excited state energy surface such that free radicals are funneled into the formation of the desired biphenyl product. However, studies also show the inactivation of the antibodies as a result of photodecay of the biphenyl product. Using an isocyanate scavenging resin, the photodecay product could be removed and the inactivation of the antibody drastically reduced. Furthermore, despite the observed photodecay, turnover of the antibody was present; this represents the first case in which true turnover of a photochemical reaction using a catalytic antibody could be observed.     

Aqueous aldol catalysis by a nicotine metabolite

Nornicotine, an endogenous tobacco alkaloid and minor nicotine metabolite, can catalyze aldol reactions at physiological pH. Catalysis appears to be due to a covalent enamine mechanism, an unprecedented reaction with small organic molecule catalysts in aqueous buffer. Kinetic parameters for nornicotine as well as other related alkaloids were measured and demonstrate that both the pyrrolidine and pyridine rings are critical for optimal catalysis. Substrate compatibility of this catalyst and its implications in vivo are discussed.     

Size-selective poorly soluble particulate reference materials for evaluation of quantitative analytical methods

Owing to the absence of readily available certified particulate reference materials (RMs), most analytical methods used to determine particulate contaminant levels in workplace or other environments are validated using solution RMs, which do not assess the robustness of the digestion step for all forms and sizes of particles in a sample. A library of particulate RMs having a range of chemical forms and particle sizes is needed to support a shift in method evaluation strategies to include both solution and particulate RMs. In support of creating this library, we characterized bulk and physically size separated fractions of beryllium oxide (BeO) particles recovered from the machining fluid sludge of an industrial ceramic products grinding operation. Particles were large agglomerates of compact, crystalline BeO primary particles having diameters on the order of several micrometers. As expected, the particle surface area was independent of sieve size, with a range from 3.61 m(2)/g (53-63-microm fraction) to 4.82 m(2)/g (355-600-microm fraction). The density was near the theoretical value (3.01 g/cm(3)). The data support more detailed characterization of the sludge materials for use as size-selective RMs. This work illustrates an approach that can be used to develop RMs that are difficult to digest.     

Reaction of fluorogenic reagents with proteins I. Mass spectrometric characterization of the reaction with 3-(2-furoyl)quinoline-2-carboxaldehyde, Chromeo P465, and Chromeo P503

3-(2-Furoyl)quinoline-2-carboxaldehyde (FQ), Chromeo P465, and Chromeo P503 are weakly fluorescent reagents that react with primary amines to produce fluorescent products. We studied the reaction of these reagents with alpha-lactalbumin by mass spectrometry. The reaction generated a set of products by the addition of one or more labels to the protein. At room temperature, the reaction was an order of magnitude faster with the Chromeo reagents than with FQ; however, the steady-state labeling efficiency was a factor of two higher for FQ compared with the Chromeo reagents. The relative abundance of the products with FQ usually followed a binomial distribution, which suggests that the labeling sites were uniformly accessible to this reagent. In contrast, the distribution of reaction products with the Chromeo reagents did not follow a binomial distribution for reactions performed in the absence of sodium dodecyl sulfate (SDS); it appears that the protein labeled with the Chromeo reagents refolded into a relatively stable secondary structure that hid some reactive sites. The reaction with the Chromeo reagent did follow the binomial distribution if the protein underwent treatment with 1% SDS at 95 degrees C for 5 min, which apparently disrupts the protein's secondary structure and allowed uniform access to all labeling sites. Chromeo 503 labeled seven of the 13 primary amines in denatured alpha-lactalbumin.     

Superactivation of the botulinum neurotoxin serotype A light chain metalloprotease: a new wrinkle in botulinum neurotoxin

Small molecules based upon a 2-acylguanidine-5-phenyl thiophene scaffold that can activate the light chain metalloprotease of botulinum neurotoxin serotype A (BoNT LC/A) by an apparent reduction in Km are reported. On the basis of structure-activity relationships and the activation profile, one or more molecules of activator specifically bind to a defined site on the toxin, causing the observed rate enhancement. With the ever-growing clinical uses of BoNT, compounds such as those reported here may provide a method for combating the emerging adaptive immune responses to BoNT.