Thrombolysis by chemically modified coagulation factor Xa.

UNLABELLED: Essentials Factor Xa (FXa) acquires cleavage-mediated tissue plasminogen activator (tPA) cofactor activity. Recombinant (r) tPA is the predominant thrombolytic drug, but it may cause systemic side effects. Chemically modified, non-enzymatic FXa was produced (Xai-K), which rapidly lysed thrombi in mice. Unlike rtPA, Xai-K had no systemic fibrinolysis activation markers, indicating improved safety. SUMMARY: Background Enzymatic thrombolysis carries the risk of hemorrhage and re-occlusion must be evaded by co-administration with an anticoagulant. Toward further improving these shortcomings, we report a novel dual-functioning molecule, Xai-K, which is both a non-enzymatic thrombolytic agent and an anticoagulant. Xai-K is based on clotting factor Xa, whose sequential plasmin-mediated fragments, FXaß and Xa33/13, accelerate the principal thrombolytic agent, tissue plasminogen activator (tPA), but only when localized to anionic phospholipid. Methods The effect of Xai-K on fibrinolysis was measured in vitro by turbidity, thromboelastography and chromogenic assays, and measured in a murine model of occlusive carotid thrombosis by Doppler ultrasound. The anticoagulant properties of Xai-K were evaluated by normal plasma clotting assays, and in murine liver laceration and tail amputation hemostatic models. Results Xa33/13, which participates in fibrinolysis of purified fibrin, was rapidly inhibited in plasma. Cleavage was blocked at FXaß by modifying residues at the active site. The resultant Xai-K (1 nm) enhanced plasma clot dissolution by ~7-fold in vitro and was dependent on tPA. Xai-K alone (2.0 µg g(-1) body weight) achieved therapeutic patency in mice. The minimum primary dose of the tPA variant, Tenecteplase (TNK; 17 µg g(-1) ), could be reduced by > 30-fold to restore blood flow with adjunctive Xai-K (0.5 µg g(-1) ). TNK-induced systemic markers of fibrinolysis were not detected with Xai-K (2.0 µg g(-1) ). Xai-K had anticoagulant activity that was somewhat attenuated compared with a previously reported analogue. Conclusion These results suggest that Xai-K may ameliorate the safety profile of therapeutic thrombolysis, either as a primary or tPA/TNK-adjunctive agent.

Comparison of mutagen accumulation in 3 estuarine species using the salmonella/microsome activation system.

3 Estuarine organisms- oysters (Crassostrea virginica), sea squirts (Mogulla sp.), and shrimp (Peneaus sp.)-were examined for Ames test detectable levels of mutagens. Whole-tissue extracts of these organisms were made and tested using S. Typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538, with and without S9 activation. Positive results were obtained with sea squirts and shrimp extracts. Activation was not necessary to show activity. Toxicity was encountered with oyster extracts. Histidine, a possible source of false positives, was eliminated from shrimp extracts using XAD-2 resin and thick-layer chromatography.

Auditory sensitivity, equal loudness, temporal resolving power, and vocalizations in the house finch (Carpodacus mexicanus).

Absolute thresholds for pure tones were measured in four house finches by use of avoidance conditioning and a modified method of limits. Response reaction time to each tone presentation served as a data base for generating a family of "equal loudness" contours. Temporal resolving power was measured in two additional birds and compared with similar measures in man. The results are discussed in relation to previous studies of vocalizations in the house finch, and the average power spectra of selected individual vocalizations are presented.