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UHEP prolonged the intrinsic pathway by APTT model (Mourao et al.
2004), but the relationship between structure and anticoagulation still lacks in-depth details since the existence of complexity and heterogeneity of the algae glycans make comparison difficult with UHEP at molecular level and bioactivity (Athukorala et al.
The anticoagulant dynamic of the fractions was monitored for 60 min at 37[degrees]C in parallel with the control curve of UHEP used (Figure 2) (Rodrigues et al.
As expected, it was observed that a single injection of UHEP in a bolus, given 5min before the thrombogenic stimulus, produced a dose-dependent inhibition of thrombus formation (~0.
1] (better antithrombotic dose) could not produce haemorrhagic events in comparison to UHEP (data not shown).
The major results showed that the administration of fragments of 5kDa are capable of extinguishing the effect on factor XII activation and a similar effect is obtained compared to UHEP Therefore, a similar study also suggest to C.
8-fold higher of SPs than UHEP for totally abolish the contact-activated pathway since the TG curve (positive control) indicated high values of absorbance until reaching a plateau (~ 24 min) (Figure 3).
This marked difference of Cc-SP1 on both experimental coagulation protocols could be possibly explained by its lack or less specificity of interaction with the plasmatic proteases or regulators (antithrombin and/or heparin cofactor II), displaying inhibition of the tenase and prothrombinase systems and/or the formation of a direct complex with thrombin, as revealed for other algal SPs that showed very distinct structures and effects than that of UHEP (a SP that contains a specific pentasaccharide sequence with high antithrombin affinity) on the coagulation (Mourao, 2015).
Agarose gel (A and B) and polyacrylamide gel (C and D) electrophoreses of Cc-SP1 and standards chondroitin-6-sulfate (C-6-S, ~ 60 kDa), chondroitin-4-sulfate (C-4-S ~ 40 kDa), dextran sulfate (DexS, ~ 8 kDa) and/or UHEP (~ 14 kDa).
Since the end product of the acid reaction was similar in size to UHEP (Figure 2D), a glycosaminoglycan that has the highest negative charge density of any known natural biomaterial found in vertebrate tissues thus far (Mourao, 2015), it was speculated that the depolymerization process of Am-2 would, perhaps, lead to the elimination of sulfate of the structure in detriment of the reduced molecular size; therefore, possibly differing among the fragments in the pattern of sulfation (Leite et al.
muscoides, using the chromogenic method by a continuous detection system parallel with UHEP testing as a function of its in vitro inhibitory reference, as shown in Figure 3.
In addition, the evident difference in molecular size between different classes of SPs makes comparison with UHEP difficult (Melo & Mourao, 2008; Mourao et al.