Characterization of soman-binding antibodies raised against soman analogs.

The production of antibodies against the highly toxic organophosphorus compound soman (GD) has been undertaken. Monoclonal antibodies were raised against two structural analogs of soman which served as haptens for immunization. In these soman analogs the chemically active P-F bond of the soman molecule was substituted by a P-OH group (which is ionized to P-O- under physiological conditions) or a P-H bond, creating compounds which we have named GDOH and GDH, respectively. These soman analogs were linked to carrier proteins through a short linker extending from the pinacolyl group. Monoclonal antibodies were selected according to their ability to bind to the immunizing hapten, and their specificities were determined by competitive inhibition assays. Out of total of 103 anti-GDOH antibodies 22 bound soman, whereas no binding was achieved with 62 anti-GDH antibodies. The two groups of monoclonal antibodies differed also in their structural specificity as demonstrated by different reactivities against a variety of soman analogs and substituted derivatives. These studies indicate that in order to achieve further improvement in anti-soman reactivity with protective potential, other groups (which resemble the OH group) have to be substituted for the F atom in the soman molecule.

Uranyl-selective electrode based on a new bifunctional derivative combining the synergistic properties of phosphine oxide and ester of phosphoric acid

Ion-selective electrodes based on the bifunctional chelating agent O-methyldihexylphosphine oxide O'-hexyl-2-ethylphosphoric acid (HL) incorporated into a poly(vinyl chloride) membrane were developed. This new derivative is proposed as a single molecular unit combining the overall properties of the synergistic single components, di-2-ethylhexylphosphoric acid and trioctylphosphine oxide. Two different ionophores, HL and its uranyl complex (UO2L2), were studied. The response of the electrodes to uranyl ion was Nernstian for UO2L2 and super-Nernstian for HL ionophores, with detection limits of 3.0 x 10(-6) and 2.0 x 10(-5) M, respectively. Results indicate a more effective interaction with the analyte in the case of having a unique molecule incorporating the two functional groups immobilized into a polymeric membrane, rather than the separated two synergistic ligands. Flow-through tubular electrodes based on both ionophores were also used as potentiometric detectors in flow injection techniques.

In vivo and in vitro toxicity of newly synthesized monofunctional sulfur mustard derivatives.

The toxicity of two new monofunctional sulfur mustard derivatives was tested. The compound (4-carboxybutyl 2-chloroethyl sulfide, CBCS; 10-carboxydecyl 2-chloroethyl sulfide, CDCS) possess the 2-chloroethyl sulfide moiety present in mustard gas. Exposure of guinea pig skin to CBCS resulted in a dose-related ulcerative effect. CDCS exhibited similar pathological effects. Dimethylsulfoxide (DMSO) exacerbated CBCS toxicity. Regeneration and healing were prominent six days after application. Concentration-related effects were found in in vitro systems, using human SH-SY5Y neuroblastoma cells for acute toxicity and Y79 retinoblastoma cells for colony forming assay. CBCS or derivatives may serve as models compounds for investigating the mechanism of action of alkylating agents.

Simple method for selecting catalytic monoclonal antibodies that exhibit turnover and specificity.

Monoclonal antibodies were raised against a mono-p-nitrophenyl phosphonate ester to elicit catalytic antibodies capable of hydrolyzing the analogous p-nitrophenyl ester or carbonate. Potential catalytic antibody producing clones were selected, by use of a competitive inhibition assay, on the basis of their affinity for a "short" transition-state analogue, a truncated hapten which maximizes the relative contribution of the transition-state structural elements to binding. Of 30-40 clones that would have been examined on the basis of hapten binding alone, 7 were selected and 4 of these catalyzed the hydrolysis of the relevant p-nitrophenyl ester. This competitive inhibition technique represents a general approach for selecting potential catalytic antibodies and significantly increases the probability of obtaining efficient catalytic monoclonal antibodies. Further study of the catalytic antibodies revealed significant rate enhancement (kcat/kuncat approximately 10(4)) and substrate specificity for the hydrolysis of the analogous ester and, for three of the antibodies, of the analogous carbonate. The antibodies displayed turnover, an essential feature of enzymes. Evidence that catalysis occurred at the antibody combining sites was provided by the identity of the binding and the catalysis-inhibition specificity patterns.

Protective effect of povidone iodine ointment against skin lesions induced by chemical and thermal stimuli.

Mustard gas (sulfur mustard, HD) is a powerful vesicant employed as a chemical weapon. The present study demonstrates the effect of povidone iodine (PI) ointment against skin toxicity caused by HD. Gross and histopathological examinations showed that application of PI 20 min or less following exposure to the vesicant resulted in marked skin protection. The shorter, interval between exposure and treatment, the better was the protection achieved. Povidone iodine was also effective against other mustards, such as carboxybutylchloroethyl sulfide (CBCS) and mechlorethamine. The fact that PI protected the skin against agents that cannot be oxidized, such as iodoacetic acid, divinylsulfone and cantharidine, indicated that the antidotal effect of PI was unrelated to oxidation of the nitrogen and sulfur atoms of the mustards. Furthermore, NMR spectroscopy of CBCS treated with iodine did not show oxidation of the sulfur atom. Clinical experience with patients after accidential heat burns (mostly of grade I) has shown that topical application of PI ointment immediately after the stimulus significantly reduced, and often prevented, skin lesions. Apart from being a safe and widely used disinfectant, PI ointment is recommended as an efficient protective agent against skin toxicity caused by hazardous chemicals and by heat stimuli.

Protective effect of povidone-iodine ointment against skin lesions induced by sulphur and nitrogen mustards and by non-mustard vesicants.

Mustard gas (sulphur mustard, SM) is a powerful vesicant employed as a chemical weapon. The present study demonstrates the effect of povidone iodine (PI) ointment against skin toxicity caused by SM. Gross and histopathological examinations showed that application of PI up to 20 min following exposure to the vesicant resulted in marked skin protection. The shorter the interval between exposure and treatment the better was the protection achieved. PI was also effective against other mustards such as carboxybutyl chloroethyl sulphide (CBCS) and mechlorethamine. The fact that PI protected the skin against agents which cannot be oxidized such as iodoacetic acid, divinylsulphone and cantharidine showed that the antidotal effect of PI was unrelated to oxidation of the nitrogen and sulphur atoms of the mustards. PI ointment is proposed as an efficient protective agent against skin toxicity caused by mustards and other alkylators.