Ana R. Ferreira

Polysaccharide-Based Membranes in Food Packaging Applications

Plastic packaging is essential nowadays. However, the huge environmental problem caused by landfill disposal of non-biodegradable polymers in the end of life has to be minimized and preferentially eliminated. The solution may rely on the use of biopolymers, in particular polysaccharides. These macromolecules with film-forming properties are able to produce attracting biodegradable materials, possibly applicable in food packaging. Despite all advantages of using polysaccharides obtained from different sources, some drawbacks, mostly related to their low resistance to water, mechanical performance and price, have hindered their wider use and commercialization. Nevertheless, with increasing attention and research on this field, it has been possible to trace some strategies to overcome the problems and recognize solutions. This review summarizes some of the most used polysaccharides in food packaging applications.

Development and characterization of bilayer films of FucoPol and chitosan

Bilayer films of FucoPol and chitosan were prepared and characterized in terms of optical, morphologic, hygroscopic, mechanical and barrier properties, to evaluate their potential application in food packaging. Bilayer films have shown dense and homogeneous layers, and presented enhanced properties when comparing to monolayer FucoPol films. Though, a high swelling degree in contact with liquid water (263.3%) and a high water vapour permeability (0.75×10(-11)mol/msPa), typical of polysaccharide films, was still observed. However, they presented a low permeability to O2 and CO2 (0.47×10(-16)molm/m(2)sPa and 5.8×10(-16)molm/m(2)sPa, respectively). Tensile tests revealed a flexible and resistant film with an elongation at break of 38% and an elastic modulus of 137MPa. The studied properties, in particular the excellent barrier to gases, impart these bilayer films potential to be used in packaging of low moisture content products, as well as in multilayered hydrophobic/hydrophilic/hydrophobic barriers for food products with a broader range of water content.

Biodegradable films produced from the bacterial polysaccharide FucoPol

FucoPol, an exopolysaccharide produced by Enterobacter A47, grown in bioreactor with glycerol as carbon source, was used with citric acid to obtain biodegradable films by casting. The films were characterized in terms of optical, hygroscopic, mechanical and barrier properties. These films have shown to be transparent, but with a brown tone, imparting small colour changes when applied over coloured surfaces. They were hydrophilic, with high permeability to water vapour (1.01×10(-11)mol/msPa), but presented good barrier properties to oxygen and carbon dioxide (0.7×10(-16)molm/m(2)sPa and 42.7×10(-16)molm/m(2)sPa, respectively). Furthermore, films have shown mechanical properties under tensile tests characteristic of ductile films with high elongation at break, low tension at break and low elastic modulus. Although the obtained results are promising, films properties can be improved, namely by testing alternative plasticizers, crosslinking agents and blends with other biopolymers. Taking into account the observed ductile mechanical properties, good barrier properties to gases when low water content is used and their hydrophilic character, it is foreseen a good potential for FucoPol films to be incorporated as inner layer of a multilayer packaging material.

Peroxisomes are platforms for cytomegalovirus’ evasion from the cellular immune response

The human cytomegalovirus developed distinct evasion mechanisms from the cellular antiviral response involving vMIA, a virally-encoded protein that is not only able to prevent cellular apoptosis but also to inhibit signalling downstream from mitochondrial MAVS. vMIA has been shown to localize at mitochondria and to trigger their fragmentation, a phenomenon proven to be essential for the signalling inhibition. Here, we demonstrate that vMIA is also localized at peroxisomes, induces their fragmentation and inhibits the peroxisomal-dependent antiviral signalling pathway. Importantly, we demonstrate that peroxisomal fragmentation is not essential for vMIA to specifically inhibit signalling downstream the peroxisomal MAVS. We also show that vMIA interacts with the cytoplasmic chaperone Pex19, suggesting that the virus has developed a strategy to highjack the peroxisomal membrane proteins’ transport machinery. Furthermore, we show that vMIA is able to specifically interact with the peroxisomal MAVS. Our results demonstrate that peroxisomes constitute a platform for evasion of the cellular antiviral response and that the human cytomegalovirus has developed a mechanism by which it is able to specifically evade the peroxisomal MAVS-dependent antiviral signalling.

Hepatitis C virus NS3-4A inhibits the peroxisomal MAVS-dependent antiviral signalling response.

Hepatitis C virus (HCV) is the cause of one of the most prevalent viral infections worldwide. Upon infection, the HCV genome activates the RIG‐I‐MAVS signalling pathway leading to the production of direct antiviral effectors which prevent important steps in viral propagation.

Rheological studies of the fucose-rich exopolysaccharide FucoPol

In this work, the solution properties of the bacterial fucose-rich polysaccharide, FucoPol, were studied. The effect of pH (3.5-10.0) and ionic strength (0.02-1.0 M NaCl) on the intrinsic viscosity and steady shear flow were evaluated using a central composite rotatable design of experiments and surface response methodology. FucoPols intrinsic and apparent viscosities presented a quite low variation under a wide range of pH (3.5-8.0) and ionic strength (0.05-0.50 M NaCl) values. FucoPol produced viscous solutions with shear-thinning behavior at different polymer concentrations (0.2-1.2 wt.%). Flow curves were successfully described by the Cross model. The viscosity of the first Newtonian plateau varied from 0.01 to 2.47 Pas for polymer concentrations from 0.2 to 1.2 wt.%, and the dependence of the estimated relaxation time with polymer concentration suggests a large degree of interaction between FucoPol molecules. Given the results obtained, FucoPol is proposed as thickening agent for applications in which stability of the apparent viscosity under pH and ionic strength variations is required.

Signaling pathways in anchoring junctions of epithelial cells: cell-to-cell and cell-to-extracellular matrix interactions.

Abstract Epithelial cells form the epithelium, one of the basic tissues of the human body. These cells present specializations from tissue to tissue, determining different structures and functions. Tissues formed by epithelial cells are characterized by the few extracellular matrix found between adjacent cells. In this way, to preserve tissue integrity, cells have to stick to each other and have to maintain a strict communication with the environment via cell junctions. Signal transduction is the main way of cell communication, being vital for the regulation of cell survival and proliferation. In cell junctions, this communication occurs through cell adhesion molecules that promote cell-to-cell and cell-to-extracellular matrix adhesion, as well as, enable the flow of information to the inside and to the outside of the cell. These molecules include integrins and cadherins, among others. The impairment of cell signaling in epithelial junctions has been involved in several pathological processes that underlie the development of, for example, colorectal cancer. Thus, epithelial cell signaling mediators have been explored as potential therapeutic targets and efforts have been made to achieve a deeper understanding of molecular events that occur at cell junctions. In this review, we address the current knowledge on the main signaling events that take place in anchoring junctions of epithelial cells, focusing both on cell-to-cell and cell-to-matrix interactions. To conclude, we explore some relevant consequences from epithelial cell signaling impairment and demonstrate that the molecular mediators of the pathways analyzed may be putative therapeutic targets.

Removal of Pharmaceutical and Personal Care Products in Aquatic Plant-Based Systems

The use of pharmaceuticals and personal care products (PPCPs) is growing as they are used not only for treatment but also for prevention of illnesses. In wastewater treatment plants (WWTPs) many PPCPs are not completely degraded/removed, which contributes for their frequent detection in, e.g. salt marsh areas. These areas may be considered a sink, source and cycling centre of contaminants on the receiving water body flow rate. In this chapter, a study about the potential of salt marsh plants Halimione portulacoides and/or Spartina maritima for the removal of three PPCPs, caffeine (CAFF), oxybenzone (HMB) and triclosan (TCS), is summarized. Experiments, at microcosms level, had two main aims: (1) the simulation of natural estuarine environment and study of dispersion mechanisms and “natural” remediation potential of target PPCPs and (2) the optimization of biological treatment technologies in WWTPs by simulating constructed wetland composed by plants and light expanded clay aggregates (LECA). Results indicate that either in estuaries or in WWTP the presence of a physical support can help to decrease contaminant levels mainly through sorption processes. The tested salt marsh species also showed potential to directly or indirectly promote the remediation of contaminants. In any case the remediation potential and dispersion mechanism are highly dependent from the characteristics of contaminants, like octanol–water partition coefficient (Log Kow) and solubility.

Analysis of culture media screening data by projection to latent pathways: The case of Pichia pastoris X-33

Cell culture media formulations contain hundreds of individual components in water solutions which have complex interactions with metabolic pathways. The currently used statistical design methods are empirical and very limited to explore such a large design space. In a previous work we developed a computational method called projection to latent pathways (PLP), which was conceived to maximize covariance between envirome and fluxome data under the constraint of metabolic network elementary flux modes (EFM). More specifically, PLP identifies a minimal set of EFMs (i.e., pathways) with the highest possible correlation with envirome and fluxome measurements. In this paper we extend the concept for the analysis of culture media screening data to investigate how culture medium components up-regulate or down-regulate key metabolic pathways. A Pichia pastoris X-33 strain was cultivated in 26 shake flask experiments with variations in trace elements concentrations and basal medium dilution, based on the standard BSM+PTM1 medium. PLP identified 3 EFMs (growth, maintenance and by-product formation) describing 98.8% of the variance in observed fluxes. Furthermore, PLP presented an overall predictive power comparable to that of PLS regression. Our results show iron and manganese at concentrations close to the PTM1 standard inhibit overall metabolic activity, while the main salts concentration (BSM) affected mainly energy expenditures for cellular maintenance.

Design of Pathway-Level Bioprocess Monitoring and Control Strategies Supported by Metabolic Networks

In this chapter we explore the basic tools for the design of bioprocess monitoring, optimization, and control algorithms that incorporate a priori knowledge of metabolic networks. The main advantage is that this ultimately enables the targeting of intracellular control variables such as metabolic reactions or metabolic pathways directly linked with productivity and product quality. We analyze in particular design methods that target elementary modes of metabolic networks. The topics covered include the analysis of the structure of metabolic networks, computation and reduction of elementary modes, measurement methods for the envirome, envirome-guided metabolic reconstruction, and macroscopic dynamic modeling and control. These topics are illustrated with applications to a cultivation process of a recombinant Pichia pastoris X33 strain expressing a single-chain antibody fragment (scFv).