Toxicoproteomics and its application to human health risk assessment

Toxicoproteomics is the use of proteomic technologies to better understand environmental and genetic factors, toxic mechanisms, and modes of action in response to acute exposure to toxicants and in the long-term development of diseases caused or influenced by these exposures. Use of toxicoproteomic technologies to identify key biochemical pathways, mechanisms, and biomarkers of exposure and toxicity will decrease the uncertainties that are associated with human health risk assessments. This review provides an overview of toxicoproteomics from human health risk assessment perspectives. Key toxicoproteomic technologies such as 2-D gel-based proteomic methods and toxicoproteomic approaches are described, and examples of applications of these technologies and methodologies in the risk assessment context are presented. The discussion includes a focus on challenges and future directions.     

Chirality of synergistic sex pheromone components of the western hemlock looperLambdina fiscellaria lugubrosa (HULST) (Lepidoptera: Geometridae)

Bakers' yeast reduction of (2E)-3-(2-furanyl)-2-methyl-2-propenal yielded the synthetic intermediate, (2S)-3-(2-furanyl)-2-methylpropanol, of high chiral purity (>97% ee) for the synthesis of the enantiomers of 2,5-dimethylheptadecane and 7-methylheptadecane, two synergistic sex pheromone components of the western hemlock looper (WHL),Lambdina fiscellaria lugubrosa Hulst. In electrophysiological bioassays, (7S)- but not (7R)-7-methylheptadecane elicited strong antennal responses by male WHL antennae. In field trapping experiments, addition of (7S)- but not (7R)-7-methylheptadecane to (5R,11S)-5,11-dimethylheptadecane, the major sex pheromone component of WHL, increased attraction. Attraction to (5R,11S)-5,11-dimethylheptadecane in combination with (7S)-7-methyiheptadecane was further enhanced by the addition of (5S)- but not (5R)-2,5-dimethylheptadecane. Similarly, attraction to (5R,11S)-5,11-dimethylheptadecane combined with (5S)-2,5-dimethylheptadecane increased when 7S- but not (7R)-7-methylheptadecane was added as a third component. We conclude that (7S)-7-methylheptadecane and (5S)-2,5-dimethylheptadecane are the synergistic sex pheromone components of WHL. The synthetic methodology described is applicable to the synthesis of chiral methyl-branched pheromones in other orders of the Insecta, particularly Coleoptera, Diptera and Orthoptera.     

Effects of hydrogen on mechanical properties of vanadium-niobium alloys

Mechanical properties of V-Nb alloys were investigated from 78 to about 300 K. The effects of alloying composition on the yield and flow stress are consistent with a hardening model due to the “misfit”" of one element in the matrix of the other. Additions of up to about 0.6 at. pct of hydrogen have a moderate strengthening effect on the yield stress of high vanadium alloys, but relatively little effect on the high niobium alloys. Little or no effect of adding hydrogen to any of the alloys was observed for the strain hardening coefficient. High Nb alloys were found to be much less embrittled by small additions of hydrogen than were high V alloys. In addition, the failure of severely embrittled alloys was found to be initiated intergranularly by impingement of mechanical twins on grain boundaries. It is speculated that the grain boundaries may be enriched hydrogen.     

Directed evolution for the development of conformation-specific affinity reagents using yeast display

Yeast display is a powerful tool for increasing the affinity and thermal stability of scFv antibodies through directed evolution. Mammalian calmodulin (CaM) is a highly conserved signaling protein that undergoes structural changes upon Ca(2+) binding. In an attempt to generate conformation-specific antibodies for proteomic applications, a selection against CaM was undertaken. Flow cytometry-based screening strategies to isolate easily scFv recognizing CaM in either the Ca(2+)-bound (Ca(2+)-CaM) or Ca(2+)-free (apo-CaM) states are presented. Both full-length scFv and single-domain VH only clones were isolated. One scFv clone having very high affinity (K(d) = 0.8 nM) and specificity (>1000-fold) for Ca(2+)-CaM was obtained from de novo selections. Subsequent directed evolution allowed the development of antibodies with higher affinity (K(d) = 1 nM) and specificity (>300-fold) for apo-CaM from a parental single-domain clone with both a modest affinity and specificity for that particular isoform. CaM-binding activity was unexpectedly lost upon conversion of both conformation-specific clones into soluble fragments. However, these results demonstrate that conformation-specific antibodies can be quickly and easily isolated by directed evolution using the yeast display platform.     

Competitive Effects of Trichloroethylene on Cr(VI) Removal by Zero-Valent Iron

Eleven columns were set up under various groundwater geochemistry conditions to investigate the competitive effect of trichloroethylene (TCE) on hexavalent chromium [Cr(VI)] removal by zero-valent iron (Fe0). They were found to be electron competitors in the redox reactions. In the presence of TCE, the Cr(VI) removal capacities of Fe0 were decreased by about 40% when compared with their respective Cr(VI) removal capacities with identical groundwater geochemistry but without TCE. The specific reaction rate constant (kSA) of TCE was decreased by about 50% when Cr(VI) was singly applied. The kSA of TCE was further decreased by 75% in the presence of both Cr(VI) and carbonate. However, there was no apparent effect on the kSA of TCE when Cr(VI), hardness and carbonate were all present. It revealed that TCE was a stronger electron competitor of Cr(VI) and the degradation of TCE became more favorable when both hardness and carbonate were present. This suggests that the passivated precipitates formed on the Fe0 surface in the presence of both hardness and carbonate may significantly affect the Cr(VI) removal by Fe0 but has insignificant effect on the TCE removal.