Identification and characterization of OmpT-like proteases in uropathogenic Escherichia coli clinical isolates.

Bacterial colonization of the urogenital tract is limited by innate defenses, including the production of antimicrobial peptides (AMPs). Uropathogenic Escherichia coli (UPEC) resist AMP-killing to cause a range of urinary tract infections (UTIs) including asymptomatic bacteriuria, cystitis, pyelonephritis, and sepsis. UPEC strains have high genomic diversity and encode numerous virulence factors that differentiate them from non-UTI-causing strains, including ompT. As OmpT homologs cleave and inactivate AMPs, we hypothesized that UPEC strains from patients with symptomatic UTIs have high OmpT protease activity. Therefore, we measured OmpT activity in 58 clinical E. coli isolates. While heterogeneous OmpT activities were observed, OmpT activity was significantly greater in UPEC strains isolated from patients with symptomatic infections. Unexpectedly, UPEC strains exhibiting the greatest protease activities harbored an additional ompT-like gene called arlC (ompTp). The presence of two OmpT-like proteases in some UPEC isolates led us to compare the substrate specificities of OmpT-like proteases found in E. coli. While all three cleaved AMPs, cleavage efficiency varied on the basis of AMP size and secondary structure. Our findings suggest the presence of ArlC and OmpT in the same UPEC isolate may confer a fitness advantage by expanding the range of target substrates.

Active bio-based food-packaging: Diffusion and release of active substances through and from cellulose nanofiber coating toward food-packaging design.

Cellulose nanofibers (CNFs) were recently investigated for the elaboration of new functional food-packaging materials. Their nanoporous network was especially of interest for controlling the release of active species. Qualitative release studies were conducted, but quantification of the diffusion phenomenon observed when the active species are released from and through CNF coating has not yet been studied. Therefore, this work aims to model CNF-coated paper substrates as controlled release system for food-packaging using release data obtained for two model molecules, namely caffeine and chlorhexidine digluconate. The applied mathematical model - derived from Fickian diffusion - was validated for caffeine only. When the active species chemically interacts with the release device, another model is required as a non-predominantly diffusion-controlled release was observed. From caffeine modeling data, a theoretical active food-packaging material was designed. The use of CNFs as barrier coating was proved to be the ideal material configuration that best meets specifications.

Antibacterial paperboard packaging using microfibrillated cellulose.

The industry and consumers are focusing more and more on the development of biodegradable and lightweight food-packaging materials, which could better preserve the quality of the food and improve its shelf-life. In an attempt to meet these requirements, this study presents a novel bio-substrate able to contain active bio-molecules for future food-packaging applications. Based on a paperboard substrate, the development of an antibacterial bio-packaging material is, therein, achieved using a chlorhexidine digluconate (CHX) solution as a model of an antibacterial molecule, mixed with microfibrillated cellulose (MFC) and used as coating onto paperboard samples. AFM and FE-SEM analyses were performed to underline the nanoporous MFC network able to trap and to progressively release the CHX molecules. The release study of CHX was conducted in an aqueous medium and showed a lower proportion (20 %) of CHX released when using MFC. This led to the constant release of low amounts of CHX over 40 h. Antibacterial tests were carried out to assess the preservation of the antibacterial activity of the samples after the release studies. Samples remained active against Bacillus subtilis, with better results being obtained when MFC was used. The preservation of the quality of a model food was finally evaluated paving the way for future promising applications in the food packaging industry.

Controlled release of chlorhexidine digluconate using ß-cyclodextrin and microfibrillated cellulose.

This study aims to develop a high-performance delivery system using microfibrillated cellulose (MFC)-coated papers as a controlled release system combined with the well-known drug delivery agent, ß-cyclodextrin (ßCD). Chlorhexidine digluconate (CHX), an antibacterial molecule, was mixed with a suspension of MFC or a ßCD solution or mixed with both the substances, before coating onto a cellulosic substrate. The intermittent diffusion of CHX (i.e., diffusion interrupted by the renewal of the release medium periodically) was conducted in an aqueous medium, and the release mechanism of CHX was elucidated by field emission gun-scanning electron microscopy, SEM, NMR, and Fourier transform infrared analyses. According to the literature, both ßCD and MFC are efficient controlled delivery systems. This study indicated that ßCD releases CHX more gradually and over a longer period of time compared to MFC, which is mainly due to the ability of ßCD to form an inclusion complex with CHX. Furthermore from the release study, a complementary action when the two compounds were combined was deduced. MFC mainly affected the burst effect, while ßCD primarily controlled the amount of CHX released over time. In this paper, two different types of controlled release systems are proposed and compared. Depending on the final application, the use of ßCD alone would release low amounts of active molecules over time (slow delivery), whereas the combination of ß-cyclodextrin and MFC would be more suitable for the release of higher amounts of active molecules over time (rapid delivery).

Microfibrillated cellulose coatings as new release systems for active packaging.

In this work, a new use of microfibrillated cellulose (MFC) is highlighted for high-added-value applications. For the first time, a nanoporous network formed by MFC coated on paper is used for a controlled release of molecules. The release study was carried out in water with caffeine as a model molecule. The release process was studied by means of (i) continuous, and (ii) intermittent diffusion experiments (with renewal of the medium every 10 min). The effect of the MFC was first observed for the samples impregnated in the caffeine solution. These samples, coated with MFC (coat weight of about 7 g/m(2)), released the caffeine over a longer period (29 washings compared with 16), even if the continuous diffusions were similar for both samples (without and with MFC coating). The slowest release of caffeine was observed for samples coated with the mixture (MFC+caffeine). Moreover, the caffeine was only fully released 9h after the release from the other samples was completed. This study compared two techniques for the introduction of model molecules in MFC-coated papers. The latter offers a more controlled and gradual release. This new approach creates many opportunities especially in the food-packaging field. A similar study could be carried out with an active species.

Role of uropathogenic Escherichia coli OmpT in the resistance against human cathelicidin LL-37.

Uropathogenic Escherichia coli (UPEC) strains are among the most prevalent causative agents of urinary tract infections. To establish infection, UPEC must overcome the bactericidal action of host antimicrobial peptides. Previously, the enterohaemorrhagic E. coli outer membrane protease, OmpT, was shown to degrade and inactivate the human antimicrobial peptide LL-37. This study aims to investigate the involvement of UPEC OmpT in LL-37 degradation. An ompT deletion mutant was generated in the prototypical UPEC strain CFT073. Western blot analysis showed that the OmpT protein level is moderate in CFT073. In agreement, OmpT was shown to partially cleave LL-37. However, no difference in the minimum inhibitory concentration of LL-37 was observed between CFT073 and the ompT mutant. Plasmid complementation of ompT, which led to increased OmpT levels, resulted in complete cleavage of LL-37 and a fourfold increase in the minimum inhibitory concentration. The analysis of other UPEC isolates showed similar OmpT activity levels as CFT073. Although UPEC OmpT can cleave LL-37, we conclude that the low level of OmpT limits its contribution to LL-37 resistance. Collectively, these data suggest that UPEC OmpT is likely accompanied by other LL-37 resistance mechanisms.

Microfibrillated cellulose - its barrier properties and applications in cellulosic materials: a review.

Interest in microfibrillated cellulose (MFC) has been increasing exponentially. During the last decade, this bio-based nanomaterial was essentially used in nanocomposites for its reinforcement property. Its nano-scale dimensions and its ability to form a strong entangled nanoporous network, however, have encouraged the emergence of new high-value applications. In previous years, its mode of production has completely changed, as many forms of optimization have been developed. New sources, new mechanical processes, and new pre- and post-treatments are currently under development to reduce the high energy consumption and produce new types of MFC materials on an industrial scale. The nanoscale characterization possibilities of different MFC materials are thus increasing intensively. Therefore, it is critical to review such MFC materials and their properties. Moreover, very recent studies have proved the significant barrier properties of MFC. Hence, it is proposed to focus on the barrier properties of MFC used in films, in nanocomposites, or in paper coating.