Plasma membrane calcium ATPase powered by glycolysis is the main mechanism for calcium clearance in the hippocampal pyramidal neuron.

AIMS: This study sought to elucidate the primary ATP-dependent mechanisms involved in clearing cytosolic Ca2+ in neurons and determine the predominant ATP-generating pathway-glycolysis or tricarboxylic acid cycle/oxidative phosphorylation (TCA/OxPhos)-associated with these mechanisms in hippocampal pyramidal neurons. MAIN METHODS: Our investigation involved evaluating basal Ca2+ levels and analyzing the kinetic characteristics of evoked neuronal Ca2+ transients after selectively combined the inhibition/blockade of key ATP-dependent mechanisms with the suppression of either TCA/OxPhos or glycolytic ATP sources. KEY FINDINGS: Our findings unveiled that the plasma membrane Ca2+ ATPase (PMCA) serves as the principal ATP-dependent mechanism for clearance cytosolic Ca2+ in hippocampal pyramidal neurons, both during rest and neuronal activity. Remarkably, during cellular activity, PMCA relies on ATP derived from glycolysis, challenging the traditional notion of neuronal reliance on TCA/OxPhos for ATP. Other mechanisms for Ca2+ clearance in pyramidal neurons, such as SERCA and NCX, appear to be dependent on TCA/OxPhos. Interestingly, at rest, the ATP required to fuel PMCA and SERCA, the two main mechanisms to keep resting Ca2+, seems to originate from a source other than glycolysis or the TCA/OxPhos. SIGNIFICANCE: These findings underscore the vital role of glycolysis in bolstering PMCA neuronal function to uphold Ca2+ homeostasis. Moreover, they elucidate the varying dependencies of cytoplasmic Ca2+ clearance mechanisms on distinct energy sources for their operation.

Interpenetrating alginate network as drug delivery matrix: Effects on protein stability and release.

The study investigates the rheological properties and protein release capacity of a uniform hydrogel composed of sodium alginate (SA) and poloxamer (P407). The hydrogel is prepared through the sustained release of calcium ions, resulting in a reinforced and homogeneous interpenetrating networks (IPNs) of SA and P407 polymeric chains. By adjusting the amount of crosslink agent, the hydrogel exhibites an adjustable dissolution ratio and adaptable gelling time. Moreover, the composite showed a well-structured network and superior mechanical strength, enabling the sustained release of both calcium ions and Soybean Trypsin Inhibitor (STI) protein, a model of Bone Morphogenic Protein (BMP). Importantly, the protein release kinetic can be tuned based on the SA content in the polymeric blend, highlighting the versatile nature of this hydrogel for drug delivery purposes.

Influence of the mRNA initial region on protein production: a case study using recombinant detoxified pneumolysin as a model

Recombinant proteins are of great importance in modern society, mostly as biopharmaceutical products. However, challenging and complex processes with low production yield are major drawbacks. Normally, the optimization to overcome these obstacles is focused on bioreactor and purification processes, and the biomolecular aspects are neglected, seen as less important. In this work, we present how the 5′ mRNA secondary structure region can be relevant for translation and, therefore, protein production. For this, Escherichia coli BL21(DE3) clones, producing recombinant detoxified pneumolysin (PdT) with and without the N-terminal His-tag, were cultivated in 10-L bioreactors. Another version of the pdt gene (version 2) with synonymous changes in the 5′-end nucleotide sequence was also obtained. Protein production, plasmid stability, carbon sources, and acetic acid were quantified during the cultures. Furthermore, in silico mRNA analyses were performed using TIsigner and RNAfold. The results showed that the His-tag presence at the N-terminus generated a minimum 1.5-fold increase in target protein synthesis, which was explained by the in silico mRNA analyses that returned an mRNA secondary structure easier to translate and, therefore, higher protein production than without the His-tag. The pdt gene version 2 showed lower 5′ mRNA opening energy than version 1, allowing higher PdT production even without a tag. This work reveals that simple mRNA analyses during heterologous gene design and production steps can help reach high-recombinant protein titers in a shorter time than using only traditional bioprocess optimization strategies.

Fully Automated Production of [68Ga]GaFAPI-46 with Gallium-68 from Cyclotron Using Liquid Targets.

68Ga-based radiopharmaceuticals are routinely used for PET imaging of multiple types of tumors. Gallium-68 is commonly obtained from 68Ge/68Ga generators, which are limited in the quantity of activity produced. Alternatively, gallium-68 can easily be produced on a cyclotron using liquid targets. In this study, we optimized the GMP production of [68Ga]GaFAPI-46 using gallium-68 produced via a standard medical cyclotron using liquid targets. Starting from the published synthesis and quality control procedures described for other 68Ga-based radiopharmaceuticals, we have validated the synthesis process and the analytical methods to test the quality parameters of the final product to be used for routine clinical studies. [68Ga]GaFAPI-46 was successfully produced with high radiochemical purity and yield using an IBA Synthera® Extension module. Gallium chloride was produced on a medical cyclotron using a liquid target with activity of 4.31 ± 0.36 GBq at the end of purification (EOP). Analytical methods were established and validated, meeting Ph. Eur. standards. Full GMP production was also validated in three consecutive batches, producing 2.50 ± 0.46 GBq of [68Ga]GaFAPI-46 at the end of synthesis (EOS), with 98.94 ± 0.72% radiochemical purity measured via radio-HPLC. Quality was maintained for up to 3 h after the EOS. Production of [68Ga]GaFAPI-46 was performed and validated using a standard medical cyclotron with liquid targets. The quality control parameters (e.g., sterility, purity, and residual solvents) conformed to Ph. Eur. and a shelf life of 3 h was established. The activity of [68Ga]GaFAPI-46 produced was substantially higher than the one obtained with generators, enabling a better response to the clinical need for this radiopharmaceutical.

Novel Hydrogel Membranes Based on the Bacterial Polysaccharide FucoPol: Design, Characterization and Biological Properties.

FucoPol, a fucose-rich polyanionic polysaccharide, was used for the first time for the preparation of hydrogel membranes (HMs) using Fe3+ as a crosslinking agent. This study evaluated the impact of Fe3+ and FucoPol concentrations on the HMs' strength. The results show that, above 1.5 g/L, Fe3+ concentration had a limited influence on the HMs' strength, and varying the FucoPol concentration had a more significant effect. Three different FucoPol concentrations (1.0, 1.75 and 2.5 wt.%) were combined with Fe3+ (1.5 g/L), resulting in HMs with a water content above 97 wt.% and an Fe3+ content up to 0.16 wt.%. HMs with lower FucoPol content exhibited a denser porous microstructure as the polymer concentration increased. Moreover, the low polymer content HM presented the highest swelling ratio (22.3 ± 1.8 g/g) and a lower hardness value (32.4 ± 5.8 kPa). However, improved mechanical properties (221.9 ± 10.2 kPa) along with a decrease in the swelling ratio (11.9 ± 1.6 g/g) were obtained for HMs with a higher polymer content. Furthermore, all HMs were non-cytotoxic and revealed anti-inflammatory activity. The incorporation of FucoPol as a structuring agent and bioactive ingredient in the development of HMs opens up new possibilities for its use in tissue engineering, drug delivery and wound care management.

Effect of Olive Pomace Extract Application and Packaging Material on the Preservation of Fresh-Cut Royal Gala Apples.

The efficiency of natural olive pomace extracts for enhancing the quality of fresh-cut apples was compared with commercial ascorbic acid and two different packaging films (biodegradable polylactic acid (PLA) and oriented polypropylene (OPP)) were tested. The composition of atmosphere inside the packages, the physicochemical parameters (firmness, weight loss and color), the microbial load, total phenolic content and antioxidant activity of fresh-cut apples were evaluated throughout 12 days of storage at 4 °C. After 12 days of refrigerated storage, a significant decrease in O2 was promoted in PLA films, and the weight loss of the whole packaging was higher in PLA films (5.4%) than in OPP films (0.2%). Natural olive pomace extracts reduced the load of mesophilic bacteria (3.4 ± 0.1 log CFU/g and 2.4 ± 0.1 log CFU/g for OPP and PLA films, respectively) and filamentous fungi (3.3 ± 0.1 log CFU/g and 2.44 ± 0.05 log CFU/g for OPP and PLA films, respectively) growth in fresh-cut apples after five days of storage at 4 °C, and no detection of coliforms was verified throughout the 12 days of storage. In general, the olive pomace extract preserved or improved the total phenolic index and antioxidant potential of the fruit, without significant changes in their firmness. Moreover, this extract seemed to be more effective when combined with the biodegradable PLA film packaging. This work can contribute to the availability of effective natural food additives, the sustainability of the olive oil industries and the reduction of environmental impact. It can also be useful in meeting the food industries requirements to develop new functional food products.

Development of a functional prebiotic strawberry preparation by in situ enzymatic conversion of sucrose into fructo-oligosaccharides.

Food industry has been pressed to develop products with reduced sugar and low caloric value, while maintaining unchanged their rheological and physicochemical properties. The development of a strawberry preparation for the dairy industry, with prebiotic functionality, was herein investigated by in situ conversion of its intrinsic sucrose content into prebiotic fructo-oligosaccharides (FOS). Two commercial enzymatic complexes, Viscozyme® L and Pectinex® Ultra SP-L, were evaluated for the synthesis of FOS. Operational parameters such as temperature, pH, and enzyme:substrate ratio (E:S) were optimized to maximize FOS yield. The rheological and physicochemical properties of the obtained strawberry preparation were evaluated. For functional analysis, the resistance of FOS to the harsh conditions of the gastro-intestinal digestion was evaluated by applying the standardized INFOGEST static protocol. At optimal conditions (60 ℃, pH 5.0), Pectinex® produced 265 ± 3 g·L-1 FOS, yielding 0.57 ± 0.01 gFOS·ginitial sucrose-1 after 7 h reaction (E:S:1:40); and Viscozyme® produced 295 ± 1 g·L-1 FOS, yielding 0.66 ± 0.00 gFOS·ginitial sucrose-1 after 5 h (E:S:1:30). The obtained strawberry preparations contained more than 50 %(w/w) prebiotic FOS incorporated (DP 3-5), with 80 % reduction of its sucrose content. The caloric value was therefore reduced by 26-31 %. FOS showed resistance to gastrointestinal digestion being only slightly hydrolysed (<10 %). 1F-Fructofuranosylnystose was not digested at any phase of the digestion. Although the physicochemical properties of the prebiotic preparations were different from the original one, parameters such as the lower °Brix, water activity, consistency and viscosity, and its different color, may be easily adjusted. Results indicate that in situ synthesis strategies are efficient alternatives in the manufacture of reduced sugar and low-caloric food products with prebiotic potential.

Recovery and Purification of Cutin from Tomato By-Products for Application in Hydrophobic Films.

Tomato pomace is a low-cost, renewable resource that has been studied for the extraction of the biopolyester cutin, which is mainly composed of long-chain hydroxy fatty acids. These are excellent building blocks to produce new hydrophobic biopolymers. In this work, the monomers of cutin were extracted and isolated from tomato pomace and utilized to produce cutin-based films. Several strategies for the depolymerization and isolation of monomeric cutin were explored. Strategies differed in the state of the raw material at the beginning of the extraction process, the existence of a tomato peel dewaxing step, the type of solvent used, the type of alkaline hydrolysis, and the isolation method of cutin monomers. These strategies enabled the production of extracts enriched in fatty acids (16-hydroxyhexadecanoic, hexadecanedioic, stearic, and linoleic, among others). Cutin and chitosan-based films were successfully cast from cutin extracts and commercial chitosan. Films were characterized regarding their thickness (0.103 ± 0.004 mm and 0.106 ± 0.005 mm), color, surface morphology, water contact angle (93.37 ± 0.31° and 95.15 ± 0.53°), and water vapor permeability ((3.84 ± 0.39) × 10-11 mol·m/m2·s·Pa and (4.91 ± 1.33) × 10-11 mol·m/m2·s·Pa). Cutin and chitosan-based films showed great potential to be used in food packaging and provide an application for tomato processing waste.

Chitin-Glucan Complex Hydrogels: Physical-Chemical Characterization, Stability, In Vitro Drug Permeation, and Biological Assessment in Primary Cells.

Chitin-glucan complex (CGC) hydrogels were fabricated by coagulation of the biopolymer from an aqueous alkaline solution, and their morphology, swelling behavior, mechanical, rheological, and biological properties were studied. In addition, their in vitro drug loading/release ability and permeation through mimic-skin artificial membranes (Strat-M) were assessed. The CGC hydrogels prepared from 4 and 6 wt% CGC suspensions (Na51*4 and Na51*6 hydrogels, respectively) had polymer contents of 2.40 ± 0.15 and 3.09 ± 0.22 wt%, respectively, and displayed a highly porous microstructure, characterized by compressive moduli of 39.36 and 47.30 kPa and storage moduli of 523.20 and 7012.25 Pa, respectively. Both hydrogels had a spontaneous and almost immediate swelling in aqueous media, and a high-water retention capacity (>80%), after 30 min incubation at 37 °C. Nevertheless, the Na51*4 hydrogels had higher fatigue resistance and slightly higher-water retention capacity. These hydrogels were loaded with caffeine, ibuprofen, diclofenac, or salicylic acid, reaching entrapment efficiency values ranging between 13.11 ± 0.49% for caffeine, and 15.15 ± 1.54% for salicylic acid. Similar release profiles in PBS were observed for all tested APIs, comprising an initial fast release followed by a steady slower release. In vitro permeation experiments through Strat-M membranes using Franz diffusion cells showed considerably higher permeation fluxes for caffeine (33.09 µg/cm2/h) and salicylic acid (19.53 µg/cm2/h), compared to ibuprofen sodium and diclofenac sodium (4.26 and 0.44 µg/cm2/h, respectively). Analysis in normal human dermal fibroblasts revealed that CGC hydrogels have no major effects on the viability, migration ability, and morphology of the cells. Given their demonstrated features, CGC hydrogels are very promising structures, displaying tunable physical properties, which support their future development into novel transdermal drug delivery platforms.

Peptide linker increased the stability of pneumococcal fusion protein vaccine candidate.

Streptococcus pneumoniae is a bacterial pathogen exclusive to humans, responsible for respiratory and systemic diseases. Pneumococcal protein vaccines have been proposed as serotype-independent alternatives to currently used conjugated polysaccharide vaccines, which have presented limitations regarding their coverage. Previously in our group, pneumococcal surface protein A (PspA) and detoxified pneumolysin (PdT) were genetically fused and the hybrid protein protected mice against pneumococcal challenge, offered higher cross-protection against different strains and showed greater opsonophagocytosis rate than co-administered proteins. As juxtaposed fusion was unstable to upscale production of the protein, flexible (PspA-FL-PdT) and rigid (PspA-RL-PdT) molecular linkers were inserted between the antigens to increase stability. This work aimed to produce recombinant fusion proteins, evaluate their stability after linker insertion, both in silico and experimentally, and enable the production of two antigens in a single process. The two constructs with linkers were cloned into Escherichia coli and hybrid proteins were purified using chromatography; purity was evaluated by SDS-PAGE and stability by Western blot and high performance size exclusion chromatography. PspA-FL-PdT showed higher stability at -20°C and 4°C, without additional preservatives. In silico analyses also showed differences regarding stability of the fusion proteins, with molecule without linker presenting disallowed amino acid positions in Ramachandran plot and PspA-FL-PdT showing the best scores, in agreement with experimental results. Mice were immunized with three doses and different amounts of each protein. Both fusion proteins protected all groups of mice against intranasal lethal challenge. The results show the importance of hybrid protein structure on the stability of the products, which is essential for a successful bioprocess development.

Trends and Application of Chemometric Pattern Recognition Techniques in Medicinal Plants Analysis.

Medicinal plants have been used and studied for ages, from very old registers to modern ethnopharmacology, which encompasses analytical chemistry, foods, and pharmacy. Based on international norms and governmental organizations of health, phytomedicine-for example, herbal drugs-needs to guarantee the quality control of products and identify contaminants, biomarkers, and chemical profiles, among other issues. In this sense, is necessary to develop advanced analytical methods that show interesting possibilities and obtain a great amount of data. In order to treat the data, a set of mathematical and statistical procedures named chemometrics is necessary. In terms of herbal drugs, chemometric tools may be used to identify the following in plants: parts, development stages, processing, geographic origin, authentication, and chemical markers. This review describes applications of chemometric pattern recognition tools to analyze herbal drugs in different conditions associated with analytical methods in the last six years (2015-2020).

Rheological characterization of the exopolysaccharide produced by Alteromonas macleodii Mo 169.

Rheology modifiers are essential additives in numerous products in a variety of industries. Due to environmental awareness, consumer-oriented industries are interested in novel natural rheological agents that can replace synthetic chemicals. In this study, the chemical composition and rheological properties of a novel exopolysaccharide (EPS) produced by Alteromonas macleodii Mo 169 were investigated. It was mainly composed of uronic acids (50 mol%) and total carbohydrates were 17 % sulfated. The EPS viscosity increased with concentration, and a non-Newtonian shear thinning behavior was found for concentrations above 0.1 wt%. The elastic and viscous moduli indicated a weak gel-like structure above 0.4 wt%. It maintained its shear thinning behavior and viscoelastic properties in the presence of NaCl and CaCl2 for pH range 5-7 and temperatures up to 55 °C. Though the apparent viscosity decreased at pH 3 and 9 and temperatures above 65 °C, the shear thinning behavior was retained. The viscous and viscoelastic properties were recovered after heating (95 °C) and cooling (0 °C), indicating a good thermal stability and recoverability. After high shear force, the solution recovered original rheological properties within few seconds, demonstrating self-healing properties.

Recombinant T-cell epitope conjugation: A new approach for Dermatophagoides hypoallergen design.

BACKGROUND: Allergen-specific immunotherapy (AIT) is the only clinical approach that can potentially cure some allergic diseases by inducing immunological tolerance. Dermatophagoides pteronyssinus is considered as the most important source of mite allergens worldwide, with high sensitization rates for the major allergens Der p 1, Der p 2 and Der p 23. The aim of this work is to generate a hypoallergenic hybrid molecule containing T-cell epitopes from these three major allergens. METHODS: The hybrid protein termed Der p 2231 containing T-cell epitopes was purified by affinity chromatography. The human IgE reactivity was verified by comparing those with the parental allergens. The hybrid was also characterized immunologically through an in vivo mice model. RESULTS: The hybrid rDer p 2231 stimulated in peripheral blood mononuclear cells (PBMCs) isolated from allergic patients with higher levels of IL- 2, IL-10, IL-15 and IFN-γ, as well as lower levels of IL-4, IL-5, IL-13, TNF-α and GM-CSF. The use of hybrid molecules as a therapeutic model in D. pteronyssinus allergic mice led to the reduction of IgE production and lower eosinophilic peroxidase activity in the airways. We found increased levels of IgG antibodies that blocked the IgE binding to the parental allergens in the serum of allergic patients. Furthermore, the stimulation of splenocytes from mice treated with rDer p 2231 induced higher levels of IL-10 and IFN-γ and decreased the secretion of IL-4 and IL-5, when compared with parental allergens and D. pteronyssinus extract. CONCLUSIONS: rDer p 2231 has the potential to be used in AIT in patients co-sensitized with D. pteronyssinus major allergens, once it was able to reduce IgE production, inducing allergen-specific blocking antibodies, restoring and balancing Th1/Th2 immune responses, and inducing regulatory T-cells.

Influence of the mRNA initial region on protein production: a case study using recombinant detoxified pneumolysin as a model.

Recombinant proteins are of great importance in modern society, mostly as biopharmaceutical products. However, challenging and complex processes with low production yield are major drawbacks. Normally, the optimization to overcome these obstacles is focused on bioreactor and purification processes, and the biomolecular aspects are neglected, seen as less important. In this work, we present how the 5' mRNA secondary structure region can be relevant for translation and, therefore, protein production. For this, Escherichia coli BL21(DE3) clones, producing recombinant detoxified pneumolysin (PdT) with and without the N-terminal His-tag, were cultivated in 10-L bioreactors. Another version of the pdt gene (version 2) with synonymous changes in the 5'-end nucleotide sequence was also obtained. Protein production, plasmid stability, carbon sources, and acetic acid were quantified during the cultures. Furthermore, in silico mRNA analyses were performed using TIsigner and RNAfold. The results showed that the His-tag presence at the N-terminus generated a minimum 1.5-fold increase in target protein synthesis, which was explained by the in silico mRNA analyses that returned an mRNA secondary structure easier to translate and, therefore, higher protein production than without the His-tag. The pdt gene version 2 showed lower 5' mRNA opening energy than version 1, allowing higher PdT production even without a tag. This work reveals that simple mRNA analyses during heterologous gene design and production steps can help reach high-recombinant protein titers in a shorter time than using only traditional bioprocess optimization strategies.

Peptide linker increased the stability of pneumococcal fusion protein vaccine candidate

Streptococcus pneumoniae is a bacterial pathogen exclusive to humans, responsible for respiratory and systemic diseases. Pneumococcal protein vaccines have been proposed as serotype-independent alternatives to currently used conjugated polysaccharide vaccines, which have presented limitations regarding their coverage. Previously in our group, pneumococcal surface protein A (PspA) and detoxified pneumolysin (PdT) were genetically fused and the hybrid protein protected mice against pneumococcal challenge, offered higher cross-protection against different strains and showed greater opsonophagocytosis rate than co-administered proteins. As juxtaposed fusion was unstable to upscale production of the protein, flexible (PspA-FL-PdT) and rigid (PspA-RL-PdT) molecular linkers were inserted between the antigens to increase stability. This work aimed to produce recombinant fusion proteins, evaluate their stability after linker insertion, both in silico and experimentally, and enable the production of two antigens in a single process. The two constructs with linkers were cloned into Escherichia coli and hybrid proteins were purified using chromatography; purity was evaluated by SDS-PAGE and stability by Western blot and high performance size exclusion chromatography. PspA-FL-PdT showed higher stability at −20°C and 4°C, without additional preservatives. In silico analyses also showed differences regarding stability of the fusion proteins, with molecule without linker presenting disallowed amino acid positions in Ramachandran plot and PspA-FL-PdT showing the best scores, in agreement with experimental results. Mice were immunized with three doses and different amounts of each protein. Both fusion proteins protected all groups of mice against intranasal lethal challenge. The results show the importance of hybrid protein structure on the stability of the products, which is essential for a successful bioprocess development.

Deacetylation and Desuccinylation of the Fucose-Rich Polysaccharide Fucopol: Impact on Biopolymer Physical and Chemical Properties.

FucoPol is an acylated polysaccharide with demonstrated valuable functional properties that include a shear thinning fluid behaviour, a film-forming capacity, and an emulsion forming and stabilizing capacity. In this study, the different conditions (concentration, temperature, and time) for alkaline treatment were investigated to deacylate FucoPol. Complete deacetylation and desuccinylation was achieved with 0.02 M NaOH, at 60 °C for 15 min, with no significant impact on the biopolymer's sugar composition, pyruvate content, and molecular mass distribution. FucoPol depyruvylation by acid hydrolysis was attempted, but it resulted in a very low polymer recovery. The effect of the ionic strength, pH, and temperature on the deacetylated/desuccinylated polysaccharide, d-FucoPol, was evaluated, as well as its emulsion and film-forming capacity. d-FucoPol aqueous solutions maintained the shear thinning behaviour characteristic of FucoPol, but the apparent viscosity decreased significantly. Moreover, contrary to FucoPol, whose solutions were not affected by the media's ionic strength, the d-FucoPol solutions had a significantly higher apparent viscosity for a higher ionic strength. On the other hand, the d-FucoPol solutions were not affected by the pH in the range of 3.6-11.5, while FucoPol had a decreased viscosity for acidic pH values and for a pH above 10.5. Although d-FucoPol displayed an emulsification activity for olive oil similar to that of FucoPol (98 ± 0%) for an oil-to-water ratio of 2:3, the emulsions were less viscous. The d-FucoPol films were flexible, with a higher Young's modulus (798 ± 152 MPa), a stress at the break (22.5 ± 2.5 MPa), and an elongation at the break (9.3 ± 0.7%) than FucoPol (458 ± 32 MPa, 15.5 ± 0.3 MPa and 8.1 ± 1.0%, respectively). Given these findings, d-FucoPol arises as a promising novel biopolymer, with distinctive properties that may render it useful for utilization as a suspending or emulsifier agent, and as a barrier in coatings and packaging films.

Characterisation of Films Based on Exopolysaccharides from Alteromonas Strains Isolated from French Polynesia Marine Environments.

This work assessed the film-forming capacity of exopolysaccharides (EPS) produced by six Alteromonas strains recently isolated from different marine environments in French Polynesia atolls. The films were transparent and resulted in small colour alterations when applied over a coloured surface (ΔEab below 12.6 in the five different colours tested). Moreover, scanning electron microscopy showed that the EPS films were dense and compact, with a smooth surface. High water vapour permeabilities were observed (2.7-6.1 × 10-11 mol m-1 s-1 Pa-1), which are characteristic of hydrophilic polysaccharide films. The films were also characterised in terms of barrier properties to oxygen and carbon dioxide. Interestingly, different behaviours in terms of their mechanical properties under tensile tests were observed: three of the EPS films were ductile with high elongation at break (ε) (35.6-47.0%), low tensile strength at break (Ꞇ) (4.55-11.7 MPa) and low Young's modulus (εm) (10-93 MPa), whereas the other three were stiffer and more resistant with a higher Ꞇ (16.6-23.6 MPa), lower ε (2.80-5.58%), and higher εm (597-1100 MPa). These properties demonstrate the potential of Alteromonas sp. EPS films to be applied in different areas such as biomedicine, pharmaceuticals, or food packaging.

Membrane Technology for Valorization of Mango Peel Extracts.

Mango peel is rich in nutritional and functional compounds, such as carbohydrates, dietary fibers, proteins, and phenolic compounds, with high potential to be applied in the food industry. Most of the investigation about recovery of bioactive compounds from fruit bioproducts involves extraction techniques and further separation of target compounds. There is still a lack of information about the potential of membrane processes to recover the nutritive/functional compounds present in aqueous extracts of those bioproducts. This research is addressed to study the performance of ultrafiltration (UF), followed by nanofiltration (NF) of UF permeates, to fractionate the compounds present in aqueous extracts of mango peel. Both UF and NF concentration processes were carried up to a volume concentration factor of 2.0. Membranes with molecular weight cut-offs of 25 kDa and 130 Da were used in the UF and NF steps, respectively. UF and NF concentrates showed antioxidant activity, attributed to the presence of phenolic compounds, with rejections of about 75% and 98.8%, respectively. UF membranes totally rejected the higher molecular weight compounds, and NF membranes almost totally concentrated the fermentable monosaccharides and disaccharides. Therefore, it is envisaged that NF concentrates can be utilized by the food industry or for bioenergy production.

Storage Stability and In Vitro Bioaccessibility of Microencapsulated Tomato (Solanum Lycopersicum L.) Pomace Extract.

Tomato pomace is rich in carotenoids (mainly lycopene), which are related to important bioactive properties. In general, carotenoids are known to react easily under environmental conditions, which may create a barrier in producing stable functional components for food. This work intended to evaluate the storage stability and in vitro release of lycopene from encapsulated tomato pomace extract, and its bioaccessibility when encapsulates were incorporated in yogurt. Microencapsulation assays were carried out with tomato pomace extract as the core material and arabic gum or inulin (10 and 20 wt%) as wall materials by spray drying (160 and 200 °C). The storage stability results indicate that lycopene degradation was highly influenced by the presence of oxygen and light, even when encapsulated. In vitro release studies revealed that 63% of encapsulated lycopene was released from the arabic gum particles in simulated gastric fluid, whereas for the inulin particles, the release was only around 13%. The feed composition with 20% inulin showed the best protective ability and the one that enabled releasing the bioactives preferentially in the intestine. The bioaccessibility of the microencapsulated lycopene added to yogurt increased during simulated gastrointestinal digestion as compared to the microencapsulated lycopene alone. We anticipate a high potential for the inulin microparticles containing lycopene to be used in functional food formulations.

Production of GMP-Compliant Clinical Amounts of Copper-61 Radiopharmaceuticals from Liquid Targets.

PET imaging has gained significant momentum in the last few years, especially in the area of oncology, with an increasing focus on metal radioisotopes owing to their versatile chemistry and favourable physical properties. Copper-61 (t1/2 = 3.33 h, 61% ß+, Emax = 1.216 MeV) provides unique advantages versus the current clinical standard (i.e., gallium-68) even though, until now, no clinical amounts of 61Cu-based radiopharmaceuticals, other than thiosemicarbazone-based molecules, have been produced. This study aimed to establish a routine production, using a standard medical cyclotron, for a series of widely used somatostatin analogues, currently labelled with gallium-68, that could benefit from the improved characteristics of copper-61. We describe two possible routes to produce the radiopharmaceutical precursor, either from natural zinc or enriched zinc-64 liquid targets and further synthesis of [61Cu]Cu-DOTA-NOC, [61Cu]Cu-DOTA-TOC and [61Cu]Cu-DOTA-TATE with a fully automated GMP-compliant process. The production from enriched targets leads to twice the amount of activity (3.28 ± 0.41 GBq vs. 1.84 ± 0.24 GBq at EOB) and higher radionuclidic purity (99.97% vs. 98.49% at EOB). Our results demonstrate, for the first time, that clinical doses of 61Cu-based radiopharmaceuticals can easily be obtained in centres with a typical biomedical cyclotron optimised to produce 18F-based radiopharmaceuticals.