Modulation of Diverse Procoagulant Venom Activities by Combinations of Platinoid Compounds

Procoagulant snake venoms have been inhibited by the ruthenium containing compounds CORM-2 and RuCl3 separately, presumably by interacting with critical histidine or other sulfur-containing amino acids on key venom enzymes. However, combinations of these and other platinoid containing compounds could potentially increase, decrease or not affect the procoagulant enzyme function of venom. Thus, the purpose of this investigation was to determine if formulations of platinoid compounds could inhibit venom procoagulant activity and if the formulated compounds interacted to enhance inhibition. Using a human plasma coagulation kinetic model to assess venom activity, six diverse venoms were exposed to various combinations and concentrations of CORM-2, CORM-3, RuCl3 and carboplatin (a platinum containing compound), with changes in venom activity determined with thrombelastography. The combinations of CORM-2 or CORM-3 with RuCl3 were found to enhance inhibition significantly, but not in all venoms nor to the same extent. In sharp contrast, carboplatin-antagonized CORM-2 mediated the inhibition of venom activity. These preliminary results support the concept that platinoid compounds may inhibit venom enzymatic activity at the same or different molecular sites and may antagonize inhibition at the same or different sites. Further investigation is warranted to determine if platinoid formulations may serve as potential antivenoms.

Effects of Heme Modulation on Ovophis and Trimeresurus Venom Activity in Human Plasma

Geographic isolation and other factors result in evolution-driven diversity of the enzymatic composition of venom of pit vipers in the same genus. The present investigation sought to characterize venoms obtained from such genetically diverse Ovophis and Trimeresurus pit vipers utilizing thrombelastographic coagulation kinetic analyses. The coagulation kinetics of human plasma were assessed after exposure to venom obtained from two Ovophis and three Trimeresurus species. The potency of each venom was defined (µg/mL required to equivalently change coagulation); additionally, venoms were exposed to carbon monoxide (CO) or a metheme-inducing agent to modulate any enzyme-associated heme. All venoms had fibrinogenolytic activity, with four being CO-inhibitable. While Ovophis venoms had similar potency, one demonstrated the presence of a thrombin-like activity, whereas the other demonstrated a thrombin-generating activity. There was a 10-fold difference in potency and 10-fold different vulnerability to CO inhibition between the Trimeresurus species. Metheme formation enhanced fibrinogenolytic-like activity in both Ovophis species venoms, whereas the three Trimeresurus species venoms had fibrinogenolytic-like activity enhanced, inhibited, or not changed. This novel “venom kinetomic” approach has potential to identify clinically relevant enzymatic activity and assess efficacy of antivenoms between genetically and geographically diverse species.

Kinetic Modeling and Optimization of Milk Coagulation Affected by Several Prevalent Cheesemaking Factors and Essence Addition

The effect of Tarragon essence concentration (0–1,000 ppm), whey protein concentration (0.7–2.8 % milk), calcium chloride (0.15–0.35 g/kg), rennet concentration (0.01–0.03 g/kg), starter content (0.01–0.03 g/kg), and renneting temperature (25–45 °C) on kinetic parameters of milk coagulation was investigated using statistical approach. A central composite design was used to investigate the effect of studied factors on coagulation parameters such as coagulation rate, lag time and inflection time. In general, with a decrease in whey protein concentration or an increase in starter level, rennet content and renneting temperature, coagulation rate increased while the lag time and time at inflection point decreased. Essence addition and calcium chloride did not have significant effects on rennet coagulation kinetic. The optimum conditions of milk coagulation were: rennet content, 0.025 g/kg of milk; whey protein concentration, 0.93 %; and renneting temperature, 44.9 °C.