Winery By-Products as Source of Bioactive Compounds for Pharmaceutical and Cosmetic Industries

It is well established in the scientific community that agro-food wastes represent economic advantages and contribute to circular economy. For instance, wine industries of Região Demarcada do Douro involve the production of large quantities of by-products, such as stem, pomace, trimmed vine shoots, or wine lees, presenting a remarkable valuable composition in phytochemicals with putative health-promoting qualities. Nevertheless, the bioactive compounds obtained from these natural sources depends on the extraction process employed. In order to reduce production costs and optimize processes, new technologies—such as ultrasound-assisted extraction (UAE)—have been employed to decrease energy consumption and increase the product or process safety/control and quality. This work aims to characterize the phenolic compounds extracted from winery by-products (WBPs), namely grape stems, grape pomace, and wine lees of two grape (Vitis vinifera L.) varieties (Sousão and Tinta Barroca) from the same geographical site, as well as the antioxidant capacity. Wine lees and grape stems presented the highest concentration of phenolic compounds and the highest antioxidant capacity for Tinta Barroca variety, while grape pomace presented the highest values of these parameters for Sousão variety, demonstrating the high influence of the variety studied. Furthermore, wine lees revealed to be the winery by-product with the lowest antioxidant capacity and content of phenolics. These by-products revealed to be a rich source of phenolic compounds with high antioxidant capacities reveling to be of interest for pharmaceutical and cosmetic industries

Honey as an Adjuvant in the Treatment of COVID-19 Infection: A Review

Since ancestor times, honey has been used to promote human health due to its medicinal, and nutritious properties, mainly due to bioactive compounds present, such as phenolic compounds. The emergence of COVID-19, caused by the SARS-CoV-2 virus, led to the pursuit of solutions for the treatment of symptoms and/or disease. Honey has proven to be effective against viral infections, principally due to its potential antioxidant and anti-inflammatory activities that attenuate oxidative damage induced by pathogens, and by improving the immune system. Therefore, the aim of this review is to overview the abilities of honey to attenuate different COVID-19 symptoms, highlighting the mechanisms associated with these actions and relating the with the different bioactive compounds present. A brief, detailed approach to SARS-CoV-2 mechanism of action is first overviewed to allow readers a deep understanding. Additionally, the compounds and beneficial properties of honey, and its previously application in other similar diseases, are detailed in depth. Despite the already reported efficacy of honey against different viruses and their complications, further studies are urgently needed to explain the molecular mechanisms of activity against COVID-19 and, most importantly, clinical trials enrolling COVID-19 patients.This research was funded by MTS/SAS/0077/2020—Honey+—New reasons to care honey from the Natural Park of Montesinho: A bioindicator of environmental quality & its therapeutic potential, and by the projects UIDB/50006/2020, UIDP/50006/2020, and LA/P/0008/2020, all supported by Fundação para a Ciência e a Tecnologia (FCT)/Ministério da Ciência, Tecnologia e Ensino Superior (MCTES). We are thankful to the Fundação para a Ciência e a Tecnologia (Portugal) through project MTS/SAS/0077/2020—Honey+—New reasons to care honey from the Natural Park of Montesinho: A bioindicator of environmental quality & its therapeutic potential. This work was also financially supported by Portuguese national funds through projects UIDB/50006/2020, UIDP/50006/2020, UIDB/04033/2020 and LA/P/0008/2020, from the Fundação para a Ciência e a Tecnologia (FCT)/Ministério da Ciência, Tecnologia e Ensino Superior (MCTES). Francisca Rodrigues and Clara Grosso are thankful for their contracts (CEECIND/01886/2020 and CEECIND/03436/2020) financed by FCT/MCTES—CEEC Individual 2020 Program Contract. Juliana Garcia is grateful to FCT and BPI La Caixa Foundation, within the project titled ‘AquaeVitae—Água Termal Como Fonte de Vida e Saúde”—“PROMOVE—O futuro do Interior” call 2020 and “AquaValor—Centro de Valorização e Transferência de Tecnologia da Água” (NORTE-01-0246-FEDER-000053), supported by Norte Portugal Regional Operational Programme (NORTE 2020) under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).info:eu-repo/semantics/publishedVersio

Acorn flour from holm oak (Quercus rotundifolia): Assessment of nutritional, phenolic, and technological profile

Acorn is the fruit of holm oak (Quercus rotundifolia), being mainly used nowadays to feed animals, however a substantial part remains in the fields without any valorization. Underexploited crops are gaining new interest, driven by food security concerns and health benefits potential as well. In the present work, it was studied the physicochemical characteristics and functional perspective of acorn flour, as an ingredient for human diet. The study included nutritional composition analysis, phenolic compounds profile through HPLC, starch content and its microstructure, fibre, and pasting properties assessment. Acorn flour presented a high content in fat, particularly monounsaturated and polyunsaturated (oleic and linoleic acids), and high minerals content in particular K. Concerning phenolic profile, rutin, catechin, ellagic acid, gallic acid, and syringic acid were identified. In regards to technological profile, fibre was mainly insoluble, with around 11%, and starch content was 50%. Its pasting behaviour revealed a high gelatinization temperature (85 ◦C), with low breakdown, and higher retrogradation consistency. These results show acorn flour potential as a valuable and sustainable multipurpose food ingredientinfo:eu-repo/semantics/publishedVersio

Apple Flour in a Sweet Gluten-Free Bread Formulation: Impact on Nutritional Value, Glycemic Index, Structure and Sensory Profile

Baking bread without gluten presents many challenges generally related with poor sensorial and nutritional characteristics, and strategies to overcome this issue are needed. Despite many glutenfree (GF) bread studies, to the best of our knowledge, few are dedicated to sweet GF bread. Sweet breads have traditionally been an important type of food and are still frequently consumed worldwide. Apple flour is naturally GF, and is obtained from apples which do not accomplish market quality requirements and are being wasted. Apple flour was, therefore, characterized in terms of nutritional profile, bioactive compounds, and antioxidant capacity. The aim of this work was to develop a GF bread with incorporation of apple flour, in order to study its effect on nutritional, technological, and sensory characteristics of sweet GF bread. Additionally, in vitro starch hydrolysis and glycemic index (GI) were also analyzed. Results demonstrated the influence of apple flour in dough’s viscoelastic behavior, increasing G’ and G”. Regarding bread characteristics, apple flour led to better acceptance by the consumer, with firmness increasing (21.01; 26.34; 23.88 N), and consequently specific volume decreasing (1.38; 1.18; 1.13 cm3/g). In addition, an increase of bioactive compounds content and antioxidant capacity of the breads were revealed. As expected, the starch hydrolysis index increased, as well as GI. Nevertheless the values were really close to low eGI (56), which is a relevant result for a sweet bread. Apple flour showed good technological and sensory properties as a sustainable and healthy food ingredient for GF breadinfo:eu-repo/semantics/publishedVersio

Acorn flour as a source of bioactive compounds in gluten-free bread

Polyphenols are important bioactive compounds whose regular ingestion has shown di erent positive impacts in health. Celiac patients have nutritional deficiencies, bringing many problems to their health. Thus, it is important to develop gluten-free (GF) products, such as bread, with nutritional benefits. The acorn is the fruit of holm oak and cork oak, being an underexploited resource nowadays. Its nutritional and functional characteristics are remarkable: rich in unsaturated fatty acids and fiber, vitamin E, chlorophylls, carotenoids, phenolic compounds, and antioxidant properties. The purpose of this study was to assess the use of acorn flour as a bioactive compounds source and natural GF ingredient for baking GF bread. Bread loaves were prepared with buckwheat, rice, acorn flour, and potato starch. Two levels of acorn flour (23% and 35% of the flour mixture) were tested. The physical, nutritional, and sensory characteristics of the bread were analysed, as well as the composition of phenolic compounds: total phenols, ortho-diphenols, and flavonoids. The phenolic profile was assessed by Reverse Phase–High-Performance Liquid Chromatography–Diode Array Detector (RP-HPLC-DAD). The antioxidant activity of the bread extracts was determined by 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), diphenyl-1-picrylhidrazyl radical (DPPH), and ferric reducing antioxidant power (FRAP) methodologies. Acorn flour can be considered a good source of bioactive compounds and antioxidants in GF bread. Acorn flour showed good technological properties in GF baking, improving bread nutritional and sensory characteristicsinfo:eu-repo/semantics/publishedVersio

Impact of genetic background, maturation, and infection by Colletotrichum acutatum on the phytochemical composition of olives and olive oils: implementation of faster infrared screening tools

Tese de Doutoramento em Ciências Químicas e BiológicasA oliveira (Olea europaea L.) é uma das culturas mais importantes dos países do Mediterrâneo, cujo cultivo tem vindo a sofrer um incremento ao longo dos últimos anos, particularmente devido às diversas propriedades benéficas e sensoriais que os seus produtos (azeitona e azeite) apresentam. Estas propriedades relacionam-se com a composição nutricional e não nutricional das azeitonas, e dependem de vários fatores de pré-colheita (condições agro-ambientais, cultivar e estado de maturação, entre outros) e pós-colheita (condições de armazenamento). Uma vez que durante o processo de extração de azeite não são adicionados quaisquer químicos, a composição nutricional e fitoquímica mantém-se praticamente inalterável. Para além dos fatores anteriormente referidos, a oliveira é ainda por vezes afetada por diferentes doenças, que podem reduzir o rendimento e a qualidade final dos seus produtos, nomeadamente em termos de metabolitos secundários. A antracnose da oliveira (também denominada por doença da gafa) é provocada pelos fungos Colletotrichum acutatum e C. gloeosporioides e, em condições propícias de humidade e temperatura, pode levar a elevadas perdas de produção. Além disso, a incidência da antracnose depende das condições ambientais, da virulência dos isolados e da suscetibilidade das cultivares. Os objetivos deste trabalho consistiram em determinar a composição fenólica de diferentes cultivares de azeitona que diferem em termos de suscetibilidade ao C. acutatum (‘Galega Vulgar’ (altamente suscetível), ‘Cobrançosa’ (moderadamente suscetível) e ‘Picual’ tolerante), quer durante o processo de maturação, quer durante o processo de infeção pelo fungo. Para além da determinação da composição fenólica por métodos clássicos, e uma vez que estes requerem o uso de reagentes e solventes dispendiosos, e o pré-tratamento das amostras, foram ainda aplicados os métodos vibracionais espectroscópicos, por transformada de Fourier e Raman. Além disso, a expressão do gene OePAL foi determinada a fim de tentar relacioná-la com a atividade da enzima fenilalanina amónia-liase (PAL) e o conteúdo em compostos fenólicos. Para atingir estes objetivos, as três cultivares acima referidas foram avaliadas ao longo dos processos de maturação e infeção em termos de composição fenólica e atividade antioxidante dos frutos e azeites. Estas análises, juntamente com a determinação do conteúdo em metais e dos parâmetros de qualidade dos azeites (acidez livre e índice de peróxidos) e a aplicação de métodos espectroscópicos e análise multivariada foram utilizados de forma a discriminar os azeites das diferentes cultivares. Para além deste estudo, foi ainda avaliado o efeito do ataque do fungo C. acutatum na expressão do gene OePAL, na atividade da enzima PAL e no conteúdo e perfil fenólico das azeitonas. Para tal, azeitonas não infetadas (controlo) e infetadas às 0, 16, 48, 144 e 360 horas após a infeção foram colhidas e analisadas. Além disso, sabendo que a cutícula é o tecido da azeitona com o qual o fungo estabelece o primeiro contacto durante o processo de infeção, a mesma foi analisada através da técnica de espectroscopia de infravermelho por transformada de Fourier (FTIR) a fim de discriminar não só as cultivares, mas também os diversos tempos de infeção, e adicionalmente avaliar esta abordagem como ferramenta auxiliar para aferição das diferentes suscetibilidades ao fungo, apresentadas pelas distintas cultivares. Os resultados referentes ao estudo da maturação demonstraram uma diminuição do conteúdo em compostos fenólicos, independentemente da cultivar estudada. Além disso, as condições climáticas demonstraram ter um papel importante nesta composição. Relativamente aos métodos espectroscópicos (FTIR e Raman) e clássicos utilizados para a determinação do conteúdo fenólico, juntamente com análise multivariada, foi possível, não só criar modelos de previsão para a determinação da composição fenólica e dos parâmetros de qualidade dos azeites, mas também discriminar as diferentes cultivares e estados de maturação. O mesmo foi conseguido através da análise do conteúdo em metais das amostras de azeites. Relativamente ao estudo do efeito do ataque do C. acutatum nas azeitonas, verificou-se um aumento da produção do conteúdo em compostos fenólicos, principalmente nas azeitonas da cultivar Galega Vulgar, descrita como suscetível a este fungo. Além disso, a expressão do gene OePAL e a atividade da enzima PAL foram induzidas pelo ataque do fungo, nomeadamente para as cultivares Galega Vulgar e Picual, que demonstraram expressão e atividade máxima às 16 horas de infeção. Em suma, este trabalho permitiu obter um maior conhecimento àcerca do efeito da maturação e da infeção pelo C. acutatum no conteúdo e perfil fenólico de azeites e azeitonas. Além disso, o uso de métodos espectroscópicos vibracionais, tais como o FTIR e Raman, articulados com a aplicação de análises quimiométricas, permitiu descrever de uma forma mais rápida e eficiente a discriminação de diferentes cultivares, estados de maturação e infeção.The olive tree (Olea europaea L.) is one of the most important crops in the Mediterranean Basin, which cultivation has increased over the last decades, encouraged by the diverse health benefits associated to olive oils consumption and the sensorial characteristics. However, the health benefits associated to dietary olive oils are closely dependent on the nutritional and non-nutritional composition of olive fruits, which are influenced by several pre-harvest (agro-environmental conditions, cultivar, and ripening stage) and post-harvest (storage conditions) factors. Once initialized the olive fruits processing towards obtaining olive oils, the nutritional and phytochemical compounds remain almost unaltered, keeping the final products all the characteristics and constituents described in olives. In addition to the above mentioned factors, the olive tree can also be affected by diverse diseases, conditioning yield and final product quality. This fact is of special relevance regarding the content in (poly)phenols. Among the currently identified diseases affecting olive tree, olive antrachnose, caused by the fungus Colletotrichum acutatum and C. gloeosporioides, is one of the most relevant constraint, which severity is dependent on the weather conditions, pathogen virulence, and cultivar susceptibility. The objective of this work was to evaluate the phenolic composition in diverse maturation stages and infection degree in three different olive cultivars characterized by distinct susceptibility to C. acutatum (‘Galega Vulgar’ (highly susceptible), ‘Cobrançosa’ (moderately susceptible), and ‘Picual’ tolerant). The assessment of olives and olive oils on the phenolic composition was developed by classical spectrophotometric and chromatographic methods, which were complemented by spectroscopical approaches, such as Fourier- Transform Infrared spectroscopy (FTIR) and Raman. Furthermore, the expression of OePAL gene, was explored to understand the relationship between OePAL expression, phenylalanine ammonia-lyase (PAL) activity, and the (poly)phenolic content. To achieve these goals, the three olive cultivars referred above were investigated during the fruit maturation and infection process, regarding the phenolic composition and radical scavenging capacity of olive fruits and oils. The information recorded on these characteristics together with metal content, free acidity, and the peroxide value, resorting to spectroscopical methods combined with multivariate analysis was aimed to discriminate olive oils obtained from different cultivars. Besides the basic information on olives and olive oils composition, olive fruits were also assessed on the susceptibility to the infection by C. acutatum. For this purpose, the olives were infected in the olive grove. Non-infected controls and olives infected for 0, 16, 48, 144, and 360 hours after infection (hai) were harvested and assessed concerning the expression level of OePAL gene, PAL activity, and phenolic profile and content. These determinations allowed to study the correlation between OePAL gene expression, PAL activity, and phenolic content. Since cuticle constitutes the first contact point of the fungus during infection, this tissue was analysed by FTIR combined with Attenuated Total Reflectance (ATR) accessory to discriminate cultivars and infection times, besides the susceptibility/response to the infection. The major results obtained allowed to state that during the maturation process, the content on phenolic compounds decreased independently of the cultivar, whilst the climatic conditions appeared as a critical factor for the (poly)phenolic composition. The use of spectroscopical (FTIR and Raman) and classical methods for determining phenolic compounds and quality parameters of olive oils, combined with Partial Least Square regression allowed to discriminate the cultivar and the distinct maturation stages, and to create models for predicting the chemical characteristics of this food matrix during the maturation process. Besides the discrimination based in these analyses, the evaluation of the metal content was also applied alongside Partial Least Square regression, allowing to discriminate the campaign and the cultivar. Apart from the evaluation of the basic chemical and phytochemical composition and the mineral content of olives and olive oils in different cultivars and throughout the ripening stage, it was determined the effect of C. acutatum on the level of secondary metabolites (phenolics). These increased throughout the infection as a response to the fungal attack, especially concerning ‘Galega Vulgar’, which has been described as the most susceptible cultivar. Furthermore, OePAL gene expression and PAL activity were boosted by the fungal infection, mainly in cultivars Galega Vulgar and Picual, which showed the highest gene expression level and PAL activity at 16 hours after infection. As overall conclusion, this work allowed to gain a further insight in the effect of maturation and C. acutatum infection processes on the phenolic profile and (poly)phenolic content of olives and olive oils. In addition, the application of FTIR methods combined with chemometric analyses allowed to describe a faster spectroscopical method for the discrimination of different olive cultivars, maturation stages and infection incidence.FCT (Fundação para a Ciência e Tecnologia

Assessment of Phenolic Content, Antioxidant and Anti-Aging Activities of Honey from <i>Pittosporum undulatum</i> Vent. Naturalized in the Azores Archipelago (Portugal)

Pittosporum undulatum Vent. is an invasive species scattered across all of the Azores’s Islands. Identifying processes to obtain economic returns from the plant is of great interest. This work aims to evaluate honey from P. undulatum from the Azores by determining its phenolic content and biological activities, to enhance its value and equate its potential applications in the food, cosmetic, and/or pharmaceutical industries. Herein, the phenolic content and antioxidant capacity were evaluated by spectrophotometric methods. Furthermore, and for the first time, anti-aging capacity was determined in honey samples. The total phenols content revealed values from 20.82 to 112.13 mg GA/100 g, from 10.25 to 103.26 mg GA/100 g for ortho-diphenols, and from 2.94 to 40.96 mg CAT/100 g for flavonoids content. Regarding the antioxidant capacity, the values ranged from 0.05 to 2.27 mmol Trolox/100g. Concerning the anti-aging capacity, promising results were obtained, namely for tyrosinase inhibitory capacity, with values ranging between 4.36% and 9.37%, while the values of elastase inhibitory capacity ranged from 37.52% to 45.88%. This study allowed us to understand the phytochemicals and biological activities of honey from P. undulatum, enhancing the possible health benefits, namely for potential anti-aging treatment, valorizing, at the same time, a national food product

Optimization of the Extraction Methodology of Grape Pomace Polyphenols for Food Applications

This study aims to take advantage of the wine industry by-products and extract bioactive compounds from grape pomace by applying methodologies susceptible to be integrated easily into industrial workflows because of the association with standard instrumentation and facilities, while the main factors affecting the efficiency of the process have been optimized. The sampling consisted of two grape varieties: ‘Touriga Nacional’ and ‘Sousão’. A response surface methodology (RSM) method was used to optimize the extraction conditions based on three independent variables according to the chemical characteristics and stability/lability traits associated with polyphenols; the main bioactive phytochemical in grape pomace: solvent (50%, 70%, and 90% ethanol); temperature (20 °C, 40 °C, and 60 °C); and pH (0.5% HCl, 2% HCl, and 3.5% HCl). The phytochemical profile, as well as the radical scavenging and reducing powers were determined on 27 different samples. The highest yield and antioxidant activity corresponded to extracts obtained at 60 °C using 3.5% HCl and 70% ethanol. The values for total phenols and flavonoids were 44.93 mg of gallic acid equivalents (GAE) and 22.95 mg of catechins equivalents (CE) per gram, respectively. Concerning the evaluation of antioxidant capacity using various assays such as ABTS, DPPH, and FRAP, the results obtained were 0.30, 0.43, and 0.36 mmol of Trolox equivalent antioxidant capacity (TEAC) per gram, correspondingly. The analysis of the extract obtained with the best extraction performance using these parameters via High-Performance Liquid Chromatography–Mass Spectrometry has been also performed, allowing us to identify fourteen (14) compounds, including phenolic acids (n = 3), flavonols (n = 7), and anthocyanins (n = 4). As a result of this process, the best conditions for the production of a natural and environmentally friendly dye, not only avoiding waste but also reusing these by-products, were achieved

Impact of Colletotrichum acutatum Pathogen on Olive Phenylpropanoid Metabolism

Olive anthracnose caused by the hemibiotrophic fungal pathogen Colletotrichum acutatum is a serious threat to the olive sector. Olive oil and fruit production is severely constrained by Colletotrichum spp. infection, being C. acutatum the most distributed pathogen in Portuguese olive orchards. To understand the impact of C. acutatum on phenylpropanoids biosynthesis, the enzyme activity, phenolic compounds, ortho-diphenols, and flavonoids content were determined and correlated with the expression of gene encoding key enzymes within phenylpropanoids metabolism in susceptible and tolerant olive fruits, during maturation and when infected with C. acutatum. Differences between cultivars was observed, the tolerant olive cv. Picual presented a higher basal value and a stable phenolic content throughout the infection process, supporting its high C. acutatum tolerance, whereas in the susceptible olive cv. Galega these secondary metabolites were significantly increased only after the elicitation with C. acutatum

Seed phytochemical profiling of three olive cultivars, antioxidant capacity, enzymatic inhibition, and effects on human neuroblastoma cells (SH-SY5Y)

This work evaluated the phytochemical composition of olive seed extracts from different cultivars (‘Cobrançosa’, ‘Galega’, and ’Picual’) and their antioxidant capacity. In addition, it also appraised their potential antineurodegenerative properties on the basis of their ability to inhibit enzymes associated with neurodegenerative diseases: acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase (TYR). To achieve this goal, the phenolic composition of the extracts was deter-mined through high-performance liquid chromatography coupled with photodiode-array detection and electrospray ionization/ion trap mass spectrometry (HPLC-DAD-ESI/MSn ).The antioxidant capacity was assessed by two different methods (ABTS•+ and DPPH•), and the antineurodegenerative potential by the capacity of these extracts to inhibit the aforementioned related enzymes. The results showed that seed extracts presented a high content of phenolic compounds and a remarkable ability to scavenge ABTS•+ and DPPH•. Tyrosol, rutin, luteolin-7-glucoside, nüzhenide, oleuropein, and ligstroside were the main phenolic compounds identified in the extracts. ‘Galega’ was the most promising cultivar due to its high concentration of phenolic compounds, high antioxidant capacity, and remarkable inhibition of AChE, BChE, and TYR. It can be concluded that olive seed extracts may provide a sustainable source of bioactive compounds for medical and industrial applications.</p