Objectives: Type 1 Diabetes Mellitus (T1DM) is a chronic autoimmune disease that destroys insulin-producing pancreatic cells, affecting over 9 million people globally. The primary treatment involves insulin therapy, but improper delivery can lead to poor glucose control or hypoglycemia. Pancreatic islet transplantation offers a potential alternative, but long-term graft survival remains a challenge. This study aims to enhance cell-based therapies by developing a permeable, pro-angiogenic immune-isolation core- shell hydrogel using decellularized pancreatic extracellular matrix (dECM) for the encapsulation of beta cells. The hydrogel will be formed using Riboflavin (RB) a bio crosslinker also known as Vitamin B2, which is reported to ameliorate oxidative stress and inflammation, key factors in the pathophysiology of insulin resistance. Methods: Porcine pancreas was decellularized using a supercritical carbon dioxide (scCO2) protocol developed by our group, to effectively preserve the biochemical structure of the ECM, while removing the cellular components. After five cycles of decellularization (110-120 bar, 36-38ºC for 1h cycle), the tissue was thoroughly analyzed. The residual DNA content, a key indicator of the efficiency of decellularization, and the ECM components (glycosaminoglycans, collagen, and elastin) were quantified using Blyscan, Sircol and Fasin assay kits. The hydrogel was developed by digesting the dECM using pepsin/HCl and crosslinked with Riboflavin (RB) in combination with Horseradish peroxidase (HRP). Pancreatic islets from a pancreatic cell line were seeded into the hydrogel, by placing a beta cell line spheroid into the core of the dECM RB system. Also, cell viability will be assessed by live/dead assay, and insulin and PDX 1 expression will be characterized by Elisa and RT PCR. Results: The DNA quantification revealed a significant reduction in DNA content (?12.7 ng/mg) after five cycles, indicating effective removal of cellular material. Moreover, the results from the quantification of ECM components (GAGs, elastin and collagen) demonstrated that the decellularization process effectively preserved its ECM components. The crosslinked dECM-RB hydrogel exhibited mechanical stability, making it ideal for cell culture. The crosslinking process also helped to preserve the native structure of the ECM, which is crucial for maintaining the biological activity of the seeded cells. Ongoing studies are focused on regulating insulin secretion by stimulating the cells with glucose. The preliminary results revealed that beta cells within the hydrogel could produce insulin in response to glucose, showcasing its functional capability. Conclusion: The findings highlight the potential of the use of riboflavin as a bio crosslinker with an added activity to improve the stability and performance of pancreatic decellularized ECM. The dECM-RB hydrogel provided a favorable microenvironment for beta cell activity. Overall, a robust and functional cell delivery platform was created, and studies are presently ongoing to monitor cell longevity within the hydrogel and insulin production upon glucose stimulation.

Introduction: Cardiovascular diseases account for 32% of the global deaths, comprising different pathologies related to heart failure. [1] The heart self-regeneration is limited, which is associated to the highest mortality and morbidity rates of cardiovascular diseases. The incapacity of endogenous regeneration reduces therapeutic approaches to the relief of symptoms rather than restoring the proper function of the heart. The developed strategies to guide the regeneration of cardiac tissue lacks on biocompatibility, cell homing and engraftment. [2] The I-CARE project intends to develop an ECM-based implantable cardiac patch to promote in-situ the self-regeneration of myocardial tissue. Objectives: The I-CARE project proposes the use of Supercritical carbon dioxide (scCO2) technology to decellularize cardiac tissue. The decellularized matrix (dECM) will be further used to develop biocompatible patches with chemical and physical features able to support the recellularization with cardiomyocytes. The decellularized cardiac patch will stimulate in- situ cardiomyocyte contractility and support the regeneration of myocardial tissue. In addition, the dECM matrix will be manipulated to produce printable and injectable hydrogels, able to support the co- culture of cardiomyocytes (CMs) and mesenchymal stem cells (MSCs). Conclusions: The I-CARE project focuses on recovering damaged cardiac through the application of dECM-based patch, recellularized with viable cardiomyocytes. In addition, the dECM cardiac patch encapsulated with MSCs would benefit of its paracrine effects, including the anti-apoptosis, anti-inflammation effects, neovascularization features and the inhibition of fibrosis and; the perfectly aligned CMs capable of remuscularize the infarcted injury, electromechanical integration and synchronous activation with host myocardium. The proposed research is ambitious, and it is expected to generate relevant knowledge related to the behaviour of CMs on the developed cardiac ECM-mimic patches and, the crosstalk between CMs and MSCs towards cardiac tissue regeneration.

Sugarcane straw (Saccharum officinarum) is a valuable coproduct renowned for its abundant polyphenolic content. However, extracting these polyphenols for natural ingredients faces challenges due to their inherent variability, influenced by biotic stress factors and plant characteristics. We explored the impact of five crucial factors on sugarcane straw polyphenolic diversity: (i) production area (Guariba, Valparaíso), (ii) borer insect (Diatraea saccharalis) infestation, (iii) plant age (first to seventh harvest), (iv) harvest season, and (v) plant variety. Response surface methodology (RSM) and artificial neural networks (ANN) were used to optimize polyphenol extraction conditions. A second-order polynomial model guided us to predict ideal sugarcane straw harvesting conditions for polyphenol-rich extracts. The analysis identified CU0618-variety straw, harvested in Guariba during the dry season (October 2020), at the seventh harvest stage, with 13.81% borer insect infection, as the prime source for high hydroxybenzoic acid (1010 µg/g), hydroxycinnamic acid (3119 µg/g), and flavone (573 µg/g) content and consequently high antioxidant capacity. The ANN model surpasses the RSM model, demonstrating superior predictive capabilities with higher coefficients of determination and reduced mean absolute deviations for each polyphenol class. This underscores the potential of artificial neural networks in forecasting and enhancing polyphenol extraction conditions, setting the stage for AI-driven advancements in crop management.

The profile of polyphenol and carotenoid of peach were analysed on fresh fruits, immediately after freezing or after pasteurization, and during frozen storage at -20 ºC for 360 days. Freezing did not affect the concentration of polyphenols or carotenoids. Pasteurization of fresh and frozen samples induced an increase in the extractability of chlorogenic acid (38% and 24%) and of zeaxanthin (336% and 127%) and a decrease of lutein (22% and 56%) and b-cryptoxanthin (32 and 51%). Total antioxidant capacity and total phenolic content changed by less than 20% after frozen storage for 360 d. However, total carotenoid content decreased 41% and 48% in unpasteurized and pasteurized samples, after the same storage period. Long term storage induced an increase in the extractability of (+)-catechin (47%) and zeaxanthin (90%) and a decrease in neochlorogenic acid (59%), chlorogenic acid (39%), quercetin-3-glucoside (21%), lutein (24%) and b-cryptoxanthin (40%). Pasteurization of frozen samples promoted a decrease in the concentration of neochlorogenic acid (91%), chlorogenic acid (87%) and quercetin-3-rutinoside (71%) and increase in lutein (16%), b-cryptoxanthin (42%) and b-carotene (31%). Peach should be stored for periods no longer than 230 d, since pasteurization after that, induced a higher decrease in neochlorogenic, chlorogenic acids and quercetin-3-rutinoside content.

Peach is a fruit with high content of phytochemical compounds with antioxidant activity, namely polyphenols and carotenes that make them a healthy fruit for consumers. Peach (Prunus persica (L.)), is an important commercial Mediterranean fruit during summer season not only for its high water content, fibre and minerals, but also for its high content in antioxidant molecules such as procyanidins, anthocyanins, catechins, phenolic acids and carotenoids (lutein, zeaxanthin, β-cryptoxanthin and β-carotene). These compounds in peach fruit vary strongly in relation to different environmental and agricultural factors, such as cultivar, rootstock, water and ripening stage at harvest. Besides the regular peach consumption as fresh fruit, processed products such as juices, jams, jellies, smoothies, canned fruit and others, are usually produced from fruit fresh market. Information about nutrient content of processed fruits is of interest to consumers as they are becoming more aware and interested about their related health benefits. Fruit processing and storage may lead to negative or positive effects concerning their nutritional value according the processing steps they are exposed to. There are occurrences during fruit processing where chemical reactions could release bound components leading to a promotion of nutritional quality. Among the phytochemicals, phenols, have received a great deal of attention because of their antioxidant activity, which help to neutralise free radicals. Thermal treatment is one of the most important factors affecting their stability. Previous studies have shown that concentrations of carotenoids often increase during heat treatment, probably because cell walls are softened, making the extraction of carotenoids easier during analysis. Extreme or prolonged heat treatment, however, enhances loss of carotenoids. Factors such as light, metals, oxygen, organic acids and heat can promote carotenoid isomerization and degradation. The aim of this chapter, therefore, is to review the impact of different industrial processing techniques of sterilization and associated storage conditions (e.g., time and temperature) on nutritional and functional characteristics of processed peach, and to establish their role on the nutritional and functional quality of fruit.

The sugarcane processing industry generates a large amount of straw, which has a negative environmental impact, and high costs are associated with their elimination, wasting their potential bioactive value attributed to their richness in polyphenols. In this study, an ethanolic extract produced from sugarcane straw was screened for its phenolic compounds content, and the potential use of this extract in the development of a food ingredient was further evaluated. Fifty different secondary metabolites belonging to the hydroxybenzoic acids, hydroxycinnamic acids, and flavonoids were identified by liquid chromatography–electrospray ionization–ultrahigh-resolution—quadrupole time of flight–mass spectrometry (LC-ESI-UHR-QqTOF-MS). The predominant phenolic compounds found were 4-hydroxybenzaldehyde, chlorogenic acid, and 5-O-feruloylquinic acid. The obtained extracts showed strong potential as food preservatives by exhibiting (a) antioxidant activity using both 2.2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt radical cation (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) methods; and (b) antimicrobial capacity, with a minimum inhibitory concentration of 50 mg/mL for Staphylococcus aureus, 74% inhibition for Bacillus cereus, and 44% for Salmonella enterica; and (c) the capacity to inhibit a food browning enzyme, tyrosinase (28–73% for 1–8 mg/ mL). Moreover, the extracts showed antidiabetic potential by inhibiting the enzymes α-glucosidase (15–38% for 1.25–5.00 mg/mL) and dipeptidyl peptidase-IV (DPP-IV) (62–114% for 0.31–5.00 mg/mL). The extract (0.625 mg/mL) also exhibited the capacity to reduce proinflammatory mediators (i.e., interleukins 6 and 8, and tumor necrosis factor alpha) when Caco-2 cells were stimulated with interleukin 1 beta. Thus, sugarcane straw extract, which is rich in phenolic compounds, showed high potential to be used in the development of food-preservative ingredients owing to its antioxidant and antimicrobial potential, and to be explored as a food supplement in diabetes prevention and as coadjuvant to reduce intestinal inflammation by reducing proinflammatory mediators.

Second-order polynomial models were used to relate the independent variables, temperature, relative humidity and storage time, to the quality attributes (electrolyte leakage, chlorophylls, weight loss, humidity and headspace gas composition inside packages before and after packaging) of readyto- eat spinach leaves. Results show that while waiting for processing the best conditions to assure high quality spinach leaves before packaging were 4 °C, 83 % RH and 4 h. The weight loss of spinach leaves was time-dependent: increasing the exposure time under these conditions decreased the amount of wet leaves before packaging. Under the same conditions, the electrolyte leakage was lower (4.63 %) and the chlorophyll content (45.44) was higher. After packaging and while waiting for dispatch, temperature most affected the package headspace atmosphere. Indeed, an increase in storage temperature from 3 to 7 °C led to a significant increase in respiration. The best combination of conditions in the highcare room was 3 °C, 74%relative humidity for a maximum of 1 day before dispatch. In this case, the moisture content (0.46 %) and electrolyte leakage (4.16 %) were lower and the chlorophyll content (43.37) and oxygen concentration inside the packages of ready-to-eat spinach leaves (19.55 kPa) were higher.

The production of postbiotics for food applications has been growing in recent years owing to their biological potential and superior technological performance over probiotics. Their production involves the use of synthetic culture media, and in this work, we propose using sugarcane straw as a source of sugar and biological components and a sustainable alternative. Thus, this by-product was used as a substrate to produce a postbiotic extract using Saccharomyces cerevisiae as a fermentative microorganism. Sugarcane straw underwent a precedent saccharification step to release the fermentable sugars. The final extracts were characterized for their total content of sugars, phenolic compounds, organic acids, and their respective chromatographic profiles. Seventeen different polyphenols were identified with the predominance of three classes, the hydroxybenzoic acids, hydroxycinnamic acids, and flavonoids, where ferulic acid, 3,4-dihydroxybenzaldehyde, and 2,5-dihydroxybenzoic acid were most prevalent. The current work evaluated the potential use of this postbiotic extract for food applications, its antioxidant activity, gut microbiota modulatory effect, and intestinal anti-inflammatory potential. The resultant extracts showed considerable antioxidant activity and the ability to lower the pro-inflammatory mediators (i.e., interleukin 6, 8, and tumor necrosis factor-alpha) in Caco-2 cells. During the fecal fermentability assay, no modulatory effect was observed on the main beneficial bacteria, such as Lactobacillus and Bifidobacterium. Nevertheless, a significant increase in short-chain fatty acids, namely, acetate, propionate, butyrate, and valerate was observed. Moreover, the extract also demonstrated capacity to inhibit the proliferation of putrefactive bacteria such as Enterobacteriaceae. Finally, sustainable postbiotic extracts produced by S. cerevisiae fermentation using sugarcane straw as a substrate exhibited relevant biological properties with potential use as food and nutraceutical ingredients.

The effect of calcium application and solution pH on ready-to-eat (RTE) baby-leaf spinach (Spinacia oleracea L.) texture and structure preservation was studied. Spinach leaves were treated with calcium chloride, calcium lactate and calcium propionate, at two different pH conditions (5 and 7), packaged and stored for 7 days at 5 C. After 24 h, the leaves crispness increased 49 and 29% for leaves treated with calcium chloride and lactate at pH 5 and the elasticity increased 100% after calcium propionate treatment at pH 7. During spinach shelf-life, the tissue flexibility decreased (20–60%) for all calcium treatments at pH 5 and 7 while tissue crispiness increased (7–40%) only for calcium treatments under pH 7. The electrolyte leakage increased throughout storage and was higher for all calcium treated samples when compared with controls. The chlorophyll content slightly decreased (12%) by the end of product shelf-life and did not differ according the calcium treatments. Total vitamin C content was lower in leaves treated with calcium chloride at pH 5 (31%) and pH 7 (19%) while the remaining treatments did not affect vitamin C content. The different calcium additives tested for potential texture quality maintenance did not provide the expected benefits on baby spinach leaves but increasing solution pH from 5 to 7 lead to an increased firmness by the end of shelf-life.

The interactions effect between (+)-catechin, β-lactoglobulin and two types of polysaccharides (pectin and chitosan) were studied at pH 4. Dynamic light scattering (DLS) revealed that pectin formed smaller complexes in the presence of (+)-catechin and β-lg, while chitosan presented a reduced number of populations with higher particle size. Dynamic interfacial data (obtained by a drop tensiometer) revealed that the complexes formed between pectin (+)-catechin and β-lg slowed down the β-lg migration to the oil/water interface; contrarily to the chitosan mixtures where protein migrated faster. The surface dilatational modulus of β-lg was ∼45 mN/m and increased to ∼60 mN/m when mixing with pectin and (+)-catechin, while the values of the surface dilatational modulus for the chitosan mixed interfacial films decreased to ∼35 mN/m. The free (+)-catechin content decreased 12% and 10% when interacting with pectin and chitosan, respectively, as compared to pure (+)-catechin. However, the (+)-catechin antioxidant activity were not affected by the interactions. Complexes formed between polyphenols, proteins and polysaccharides could be used as a good alternative to understand and consequently improve the phytochemicals stability in food matrices.