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- Biopreservation strategies in combination with mild high pressure treatments in traditional Portuguese ready-to-eat meat sausagePublication . Castro, S. M.; Kolomeytseva, M.; Casquete, R.; Silva, J.; Queirós, R.; Saraiva, J. A.; Teixeira, P.Thrombomodulin (TM) is an endothelial cell membrane protein that acts as a major cofactor in the protein C anticoagulant pathway. The EGF-like domains 4-6 of TM (TM456) are essential for PC activation. In this study, we proposed a liposomal recombinant TM conjugate to mimic the membrane TM structure and its anticoagulant activity. First, a DSPE-PEG2000-TM456 was successfully synthesized by site-specific conjugation of azido-TM456 with DSPE-PEG2000-DBCO via copper-free click chemistry quantitatively. Then, liposome-TM456 was fabricated via direct liposome formation with the DSPE-PEG2000-TM456 and other lipids. This liposomal formulation of TM456 retained protein C activation activity as that of TM456. Also, liposome-TM456 was much more stable and had a longer plasma half-life than TM456 and DSPE-PEG2000-TM456, respectively. Moreover, liposome-TM456 showed in vivo anticoagulant effect by decreasing the mortality from 80% to 20% in a thrombin-induced thromboembolism mouse model. The reported liposome-TM456 conjugate mimics the endothelial TM anticoagulation activity and may serve as an effective anticoagulant agent candidate for future development.
- Combined effect of pediocin bacHA-6111-2 and high hydrostatic pressure to control listeria innocua in fermented meat sausagePublication . Castro, S. M.; Silva, J.; Casquete, R.; Queirós, Rui; Saraiva, J. A.; Teixeira, P.In and ex situ production of bacteriocin (bacHA-6111-2) combined with high hydrostatic pressure (300 MPa, 5 min, 10ºC) were assessed on the survival of Listeria innocua reviously inoculated in Alheira, a traditional Portuguese fermented meat product. The effect was evaluated immediately after each treatment, alone and combined, and during 60 days of storage (4ºC). For higher concentrations of L. innocua, a bacteriostatic effect was verified: i) during the first days of storage for the pressure treatment alone and in combination with ex situ production of bacteriocin; and ii) for longer periods, when pressure was combined with in situ production of bacteriocin. After this effect, the first treatments revealed rapid growth up to the initial values; while the latest treatment, and for lower initial cells, resulted in a decrease > 2 log CFU g-1, from day 3 of storage until the end of the study. A similar result was obtained for analogous pressurized L. innocua samples with ex situ production of bacteriocin. The developed work has demonstrated the potential of using the combination of natural antimicrobial compounds (pediocin bacHA-6111-2, produced either in or ex situ) with mild pressure (300 MPa, 5 min) treatments to control effectively L. innocua in fermented meat products such as Alheiras, when contaminated with values closed to the ones that might potentially arise by natural postcontamination.