Preparation and characterization of polyacrylic acid‐poly(vinyl alcohol)‐based interpenetrating hydrogels

Some structural features of hydrogels from poly(acrylic acid) (PAAc) of various crosslinking degrees have been investigated through mechanical and swelling measurements. Interpenetrating polymer hydrogels (IPHs) of poly(vinyl alcohol) (PVA) and PAAc have been prepared by a sequential method: crosslinked PAAc chains were formed in aqueous solution by crosslinking copolymerization of acrylic acid and N,N‐methylenebisacrylamide in the presence of PVA. The application of freeze–thaw (F–T) cycles leads to the formation of a PVA hydrogel within the synthesized PAAc hydrogel. The swelling and viscoelastic properties of the IPHs were evaluated as a function of the content of crosslinker and the application of one F–T cycle. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 102: 5789–5794, 2006     

Complexing the Oncolytic Adenoviruses Ad∆∆ and Ad-3∆-A20T with Cationic Nanoparticles Enhances Viral Infection and Spread in Prostate and Pancreatic Cancer Models

Oncolytic adenoviruses (OAd) can be employed to efficiently eliminate cancer cells through multiple mechanisms of action including cell lysis and immune activation. Our OAds, AdΔΔ and Ad-3∆-A20T, selectively infect, replicate in, and kill adenocarcinoma cells with the added benefit of re-sensitising drug-resistant cells in preclinical models. Further modifications are required to enable systemic delivery in patients due to the rapid hepatic elimination and neutralisation by blood factors and antibodies. Here, we show data that support the use of coating OAds with gold nanoparticles (AuNPs) as a possible new method of virus modification to help augment tumour uptake. The pre-incubation of cationic AuNPs with AdΔΔ, Ad-3∆-A20T and wild type adenovirus (Ad5wt) was performed prior to infection of prostate/pancreatic cancer cell lines (22Rv, PC3, Panc04.03, PT45) and a pancreatic stellate cell line (PS1). Levels of viral infection, replication and cell viability were quantified 24–72 h post-infection in the presence and absence of AuNPs. Viral spread was assessed in organotypic cultures. The presence of AuNPs significantly increased the uptake of Ad∆∆, Ad-3∆-A20T and Ad5wt in all the cell lines tested (ranging from 1.5-fold to 40-fold), compared to virus alone, with the greatest uptake observed in PS1, a usually adenovirus-resistant cell line. Pre-coating the AdΔΔ and Ad-3∆-A20T with AuNPs also increased viral replication, leading to enhanced cell killing, with maximal effect in the most virus-insensitive cells (from 1.4-fold to 5-fold). To conclude, the electrostatic association of virus with cationic agents provides a new avenue to increase the dose in tumour lesions and potentially protect the virus from detrimental blood factor binding. Such an approach warrants further investigation for clinical translation.     

3D Printable Composite Biomaterials Based on GelMA and Hydroxyapatite Powders Doped with Cerium Ions for Bone Tissue Regeneration

The main objective was to produce 3D printable hydrogels based on GelMA and hydroxyapatite doped with cerium ions with potential application in bone regeneration. The first part of the study regards the substitution of Ca²⁺ ions from hydroxyapatite structure with cerium ions (Ca_10-xCe_x(PO₄)₆(OH)₂, xCe = 0.1, 0.3, 0.5). The second part followed the selection of the optimal concentration of HAp doped, which will ensure GelMA-based scaffolds with good biocompatibility, viability and cell proliferation. The third part aimed to select the optimal concentrations of GelMA for the 3D printing process (20%, 30% and 35%). In vitro biological assessment presented the highest level of cell viability and proliferation potency of GelMA-HC5 composites, along with a low cytotoxic potential, highlighting the beneficial effects of cerium on cell growth, also supported by Live/Dead results. According to the 3D printing experiments, the 30% GelMA enriched with HC5 was able to generate 3D scaffolds with high structural integrity and homogeneity, showing the highest suitability for the 3D printing process. The osteogenic differentiation experiments confirmed the ability of 30% GelMA-3% HC5 scaffold to support and efficiently maintain the osteogenesis process. Based on the results, 30% GelMA-3% HC5 3D printed scaffolds could be considered as biomaterials with suitable characteristics for application in bone tissue engineering.     

Evaluation of bioprocesses to improve the antioxidant properties of chickpeas

Chickpea seeds (Cicer arietinum cv. Blanco lechoso) were submitted to different bioprocesses; i) germination of seeds for 2 and 3 days, ii) natural fermentation of flour or cracked seeds, and iii) induced fermentation of flour or cracked seeds with Lactobacillus plantarum, in order to improve the antioxidant properties. Determination of vitamins C and E, reduced glutathione (GSH) and total phenolic compounds (TPC) were carried out. Antioxidant capacity was measured by SOD-like activity, peroxyl radical-trapping capacity (PRTC), lipid peroxidation inhibition (LPI) and trolox equivalent antioxidant capacity (TEAC). Vitamin C and E increased after germination, whilst fermentation caused a decrease in vitamin E. Both bioprocesses caused an increment in TPC content while GSH decreased. Germination increased SOD-like activity (47-41%), PRTC (16-55%) and TEAC (12-23%) and a slight inhibition of LPI was observed. After fermentation, SOD-like activity suffer a drastic reduction. The LPI decreased between 11.3 and 21.3%, while an increase of TEAC (29-57%) and PRTC (27-44%) was observed, except in flour natural fermented seeds where decreased the PRTC. TPC contributed highly to total antioxidant capacity (TEAC). Results indicated that with the bioprocesses studied the antioxidant properties of chickpea flours are enhanced and they can be used as desired ingredients for new functional food formulations.     

Effect of high-pressure processing and thermal treatment on quality attributes and nutritional compounds of "songold" plum purée

The application of hydrostatic high pressure on a "Songold" plum purée was assessed in comparison with heat pasteurization. To simulate industrial conditions, one-half of the total purée was manufactured with a pretreatment of thermal blanching (TB) and the other half without it (nonthermally blanched, NTB). Changes after thermal treatment and high-pressure processing (HPP: 400, 600 MPa) and after 20 d of refrigerated storage were evaluated. HPP maintained the microbial stability of the purées until the end of the storage period. Polyphenol oxidase activity was lower in TB purées than NTB purées. No treatment was completely effective to stop the enzyme activity, although a significant reduction was reached. Thermally treated purées showed more intense color changes after processing and storage than HP-treated purées. After processing, high-pressure (HP) purées treated at 600 MPa (TB and NTB) increased the extractability of carotenoids compared with initial untreated purée. Nevertheless, at the end of the storage, the highest carotenoid content was found in the TB purée treated at 400 MPa. After processing, total polyphenol levels were similar in all purées. TB and 600 MPa processing was more effective in the maintenance of the polyphenols than the other purées. TB increased the level of antioxidants after storage, compared to NTB purées. A previous TB step is necessary to inactivate browning enzymes before HPP to maintain the levels of bioactive compounds. HPP of plum purée could be a suitable alternative to traditional thermal processing, but more studies are necessary to ensure a major inactivation of polyphenol oxidase. Practical Application: High-pressure processing is one of the most successful technologies to obtain high-quality fruit purées without appreciable losses in taste, flavor, color, and nutritive value. However, for the introduction of a new technology, some advantages compared to the traditional thermal treatment need to be emphasized. The application of this technology could be reduced due to resistance of certain enzymes to pressurization. For this reason, in some cases the application of a previous thermal blanching can be necessary at industrial level. However, this can reduce the advantages of HPP application. This paper provides interesting information about the storage stability of plum purées after high-pressure treatments and assesses the need for applying heat pretreatments.     

Contribution of low‐molecular‐weight antioxidants to the antioxidant capacity of raw and processed lentil seeds

In this study four cultivars of lentil originating from Spain were examined: cv Paula, cv Agueda, cv Almar and cv Alcor. Since consumption of these seeds after heat treatment and as sprouts has been popularised, the impact of cooking (up to 30 min) and germination process (in dark, at 25°C, for up to 4 days) on peroxyl radical‐trapping capacity (PRTC) and Trolox‐equivalent antioxidant capacity (TEAC) of the processed seeds was addressed. Also, changes in the content of low‐molecular‐weight antioxidants (LMWA) and soluble proteins in the course of cooking and germination were studied. The analyzed LMWA were: total phenolics, tocopherols (α‐T, β‐T, γ‐T, δ‐T( reduced glutathione, and L ‐ascorbic acid. On the basis of the results obtained, the contribution of LMWA and soluble proteins to the PRTC and TEAC of raw, cooked, and germinated lentil seeds was calculated by multiple mean values for the content of investigated compounds and their relative potential with respect to Trolox. The results showed a very high molar percentage contribution of phenolic compounds and low contribution of tocopherols, glutathione, soluble proteins, and ascorbate (only in germinated seeds) to the total TEAC and total PRTC calculated as a sum of data provided for phosphate‐buffered and 80% methanolic extracts of raw and processed lentil seeds.     

Validation of a UHPLC-FLD method for the simultaneous quantification of aflatoxins,ochratoxin A and zearalenone in barley

A fast and simple UHPLC-FLD method has been developed for the simultaneous determination in barley of aflatoxins (B1, G1, B2 and G2), ochratoxin A (OTA) and zearalenone (ZEA), some of the most important mycotoxins due to their toxicity and occurrence. The procedure is based on the extraction of the six mycotoxins with a mixture of acetonitrile and water, and the purification of the extract with immunoaffinity columns before analysis. Detection of AFB1 and AFG1 is improved using a photochemical reaction. The method has been validated with satisfactory results. Limits of detection were 340 ng kg⁻¹ for ZEA, 13 ng kg⁻¹ for OTA and varied from 0.5 to 15 ng kg⁻¹ for aflatoxins. Recovery percentages were between 78.2% and 109.2%. After being validated, the method has been successfully applied to 20 barley samples cultivated in a region of northern Spain (Navarra).