Fatty Acid Hydroxytyrosyl Esters of Olive Oils Are Bioaccessible According to Simulated In Vitro Gastrointestinal Digestion: Unraveling the Role of Digestive Enzymes on Their Stability

Recently, new bioactive compounds were identified in olive oil, lipophenols, which are composed of a fatty acid (FA) and a phenolic core, such as HT (HT-FA). However, their bioaccessibility remains unknown. Thus, the present study uncovers the impact of the separate phases of gastrointestinal digestion on the release and stability of HT-FAs from oily matrices under in vitro simulated conditions. Accordingly, it was found that the bioaccessibility of HT derivatives is largely dependent on the type of FA that esterifies HT, as well as the food matrix. Also, the generation of HT-FAs during intestinal digestion was observed, with pancreatin being the enzyme responsible, to a higher extent, for the de novo formation of lipophenolic derivatives. These findings prompt us to identify new applications to oily matrices and their byproducts as potential functional ingredients for the promotion of health, where the possible formation of new lipophenols during digestion should be taken into consideration

Chemical Characterization, Antioxidant Activity and GC-MS Analysis of the Extracted Oil from Seeds of Persea americana Mill. (Avocado)

From ancient times plants have provided a source of inspiration for novel drug compounds, as plant derived medicines have made large contribution to human health and well being. There is a long tradition of using the pits for various medicinal purposes. Medicinal plants have been of age long remedies for human diseases because they contain components of therapeutic value. In this research work, Avocado seed was selected for chemical analysis. Avocado (Htaw-Bat- Thee) was collected from Pyin-Oo-Lwin Township, Mandalay Region. Firstly, phytochemical screening for the Avocado seed was performed. According to phytochemical screening, alkaloid, glycoside, saponin, phenolic compound, polyphenol, reducing sugar, terpene, lipophenol and tannin compound were found to be present in the Avocado seed. The oil extracted from Avocado seed was done with petroleum ether by using Soxhlet extraction method. The physicochemical parameters, such as specific gravity, viscosity, acid value, saponification value, iodine value and unsaponifiable matter were determined by conventional method. Then, elemental analysis of Avocado seed was performed by using Energy Dispersive X-ray Fluorescence (EDXRF) method. Moreover, the fatty acid composition of Avocado seed oil was analyzed by Gas Chromatography Mass Spectrometry (GC-MS) method. Moreover, antioxidant activities of Avocado seed evaluated on the basis of their scavenging activity of the stable 1, 1-diphenyl 2-picrylhydrazyl (DPPH) free radical. IC50 value was 180.06 g/mL compare to that of ascorbic acid which was 0.65 g/mL

Quantitative Analysis of Protective Effect of Resveratrol Against Nanoplastic-Induced Oxidative Stress

Nanoplastics and microplastics are a growing ecological and health concern due to their widespread prevalence. The effects of these microscopic particles in biological systems primarily relate to their potential to produce reactive oxygen species and interactive effects with other cytotoxic agents. Of particular interest in this work is the effect on mammalian intestinal cells, due to the regular unintentional ingestion of micro- and nanoplastics in the normal western diet. In this study, Caco-2 cells were cultured and exposed to polystyrene particles in vitro as a simple model of the human digestive endothelium. Resveratrol, a naturally occurring polyphenolic compound, has been shown to protect against reactive oxygen species through a combination of direct neutralization or by enhancing the expression of genes responsible for cellular defense. This work sets out to demonstrate how resveratrol and other antioxidant compounds can be used to assuage the cellular damage caused by nanoplastics. Oxidative damage and apoptosis were measured as a function of nanoplastic size, concentration, and chemical modification. Significant cellular damage was detected in the presence of amine-modified nanoplastics, while cell health was recovered when the cells were pre-treated with resveratrol and a chemically-modified variant of resveratrol

Ex vivo Evaluation of a Liposome-Mediated Antioxidant Delivery System on Markers of Skin Photoaging and Skin Penetration.

PURPOSE: The topical application of antioxidants has been shown to augment the skins innate antioxidant system and enhance photoprotection. A challenge of topical antioxidant formulation is stability and penetrability. The use of a targeted drug delivery system may improve the bioavailability and delivery of antioxidants. In this ex vivo study, we assessed the effects of the topical application of a liposome-encapsulated antioxidant complex versus a free antioxidant complex alone on skin photoaging parameters and penetrability in human skin explants. PATIENTS AND METHODS: Human organotypic skin explant cultures (hOSEC) were irradiated to mimic photoaging. The encapsulated antioxidant complex and free antioxidant complex were applied topically onto the irradiated hOSEC daily for 7 days. The two control groups were healthy untreated hOSEC and irradiated hOSEC. Photoprotective efficacy was measured with pro-inflammatory cytokine (IL-6 and IL-8) and matrix metalloproteinase 9 (MMP-9) secretion. Cell viability and metabolic activity were measured via resazurin assay. Tissue damage was evaluated via lactate dehydrogenase (LDH) cytotoxicity assay. Skin penetration of the encapsulated antioxidant complex was assessed via fluorescent dye and confocal microscopy. RESULTS: Compared to healthy skin, irradiated skin experienced increases in IL-6, IL-8 (p < 0.05), and MMP-9 (p < 0.05) secretion. After treatment with the encapsulated antioxidant complex, there was a 39.3% reduction in IL-6 secretion, 49.8% reduction in IL-8 (p < 0.05), and 38.5% reduction in MMP-9 (p < 0.05). After treatment with the free antioxidant complex, there were no significant differences in IL-6, IL-8, or MMP-9 secretion. Neither treatment group experienced significant LDH leakage or reductions in metabolic activity. Liposomes passed through the stratum corneum and into the epidermis. CONCLUSION: The topical application of a liposome-encapsulated antioxidant complex containing ectoin, astaxanthin-rich microalgae Haematococcus pluvialis extract, and THDA improves penetrability and restored IL-6, IL-8, and MMP-9 levels in irradiated human skin explants, which was not seen in the comparator free antioxidant complex group

Cellular Organelles Reorganization During Zika Virus Infection of Human Cells

Zika virus (ZIKV) is an enveloped positive stranded RNA virus belonging to the genus Flavivirus in the family Flaviviridae that emerged in recent decades causing pandemic outbreaks of human infections occasionally associated with severe neurological disorders in adults and newborns. The intracellular steps of flavivirus multiplication are associated to cellular membranes and their bound organelles leading to an extensive host cell reorganization. Importantly, the association of organelle dysfunction with diseases caused by several human viruses has been widely reported in recent studies. With the aim to increase the knowledge about the impact of ZIKV infection on the host cell functions, the present study was focused on the evaluation of the reorganization of three cell components, promyelocytic leukemia nuclear bodies (PML-NBs), mitochondria, and lipid droplets (LDs). Relevant human cell lines including neural progenitor cells (NPCs), hepatic Huh-7, and retinal pigment epithelial (RPE) cells were infected with the Argentina INEVH116141 ZIKV strain and the organelle alterations were studied by using fluorescent cell imaging analysis. Our results have shown that these three organelles are targeted and structurally modified during ZIKV infection. Considering the nuclear reorganization, the analysis by confocal microscopy of infected cells showed a significantly reduced number of PML-NBs in comparison to uninfected cells. Moreover, a mitochondrial morphodynamic perturbation with an increased fragmentation and the loss of mitochondrial membrane potential was observed in ZIKV infected RPE cells. Regarding lipid structures, a decrease in the number and volume of LDs was observed in ZIKV infected cells. Given the involvement of these organelles in host defense processes, the reported perturbations may be related to enhanced virus replication through protection from innate immunity. The understanding of the cellular remodeling will enable the design of new host-targeted antiviral strategies.Fil: Garcia, Cybele. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Vázquez, Cecilia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Giovannoni, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Russo, Constanza A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Cordo, Sandra Myriam. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Alaimo, Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Damonte, Elsa Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentin

Antisense Oligonucleotide-Based Rescue of Aberrant Splicing Defects Caused by 15 Pathogenic Variants in ABCA4

The discovery of novel intronic variants in the ABCA4 locus has contributed significantly to solving the missing heritability in Stargardt disease (STGD1). The increasing number of variants affecting pre-mRNA splicing makes ABCA4 a suitable candidate for antisense oligonucleotide (AON)-based splicing modulation therapies. In this study, AON-based splicing modulation was assessed for 15 recently described intronic variants (three near-exon and 12 deep-intronic variants). In total, 26 AONs were designed and tested in vitro using a midigene-based splice system. Overall, partial or complete splicing correction was observed for two variants causing exon elongation and all variants causing pseudoexon inclusion. Together, our results confirm the high potential of AONs for the development of future RNA therapies to correct splicing defects causing STGD1

Synthesis and characterization of a biopolymer pectin/ethanolic extract from olive mill wastewater. In vitro safety and efficacy tests on skin wound healing

Wound-healing delay is one of the major problems of type 2 diabetes, representing also a clinical emergency in non-healing chronic wounds. Natural antioxidants show interesting wound-healing properties, including those extracted from waste derived from olive oil production. Olive mill wastewater is one of the main by-products of the olive oil-making process, and it is rich in high-value secondary metabolites, mainly hydroxytyrosol. We proposed an eco-friendly extraction method, employing both ultrasound-assisted and Soxhlet techniques and ethanol as a solvent, to recover valuable molecules from Roggianella cv (Olea europea L.) olive mill wastewater, which was further entrapped in a pectin polymer via an enzymatic reaction using porcine pancreatic lipase. Pectin, in combination with other substances, promoted and accelerated wound healing and demonstrated good potential to produce a biomedical conjugate for wound treatment. The antioxidant activity of the extracts and conjugate were evaluated against lipophilic (IC50 equal to 0.152 mg mL−1) and hydrophilic (IC50 equal to 0.0371 mg mL−1) radical species as well as the in vitro cytotoxicity via NRU, h-CLAT, and a wound-healing scratch assay and assessment. The pectin conjugate did not exert hemolytic effects on the peripheral blood, demonstrating interesting wound-healing properties due to its ability to stimulate cell proliferation in a dose-dependent manner

Synthesis and Characterization of a Biopolymer Pectin/Ethanolic Extract from Olive Mill Wastewater: In Vitro Safety and Efficacy Tests on Skin Wound Healing

Wound-healing delay is one of the major problems of type 2 diabetes, representing also a clinical emergency in non-healing chronic wounds. Natural antioxidants show interesting wound-healing properties, including those extracted from waste derived from olive oil production. Olive mill wastewater is one of the main by-products of the olive oil-making process, and it is rich in high-value secondary metabolites, mainly hydroxytyrosol. We proposed an eco-friendly extraction method, employing both ultrasound-assisted and Soxhlet techniques and ethanol as a solvent, to recover valuable molecules from Roggianella cv (Olea europea L.) olive mill wastewater, which was further entrapped in a pectin polymer via an enzymatic reaction using porcine pancreatic lipase. Pectin, in combination with other substances, promoted and accelerated wound healing and demonstrated good potential to produce a biomedical conjugate for wound treatment. The antioxidant activity of the extracts and conjugate were evaluated against lipophilic (IC50 equal to 0.152 mg mL−1) and hydrophilic (IC50 equal to 0.0371 mg mL−1) radical species as well as the in vitro cytotoxicity via NRU, h-CLAT, and a wound-healing scratch assay and assessment. The pectin conjugate did not exert hemolytic effects on the peripheral blood, demonstrating interesting wound-healing properties due to its ability to stimulate cell proliferation in a dose-dependent manner

Influencia de la matriz alimentaria en la biodisponibilidad de metabolitos de hidroxitirosol y su capacidad moduladora de biomarcadores de estrés oxidativo e inflamación.

El hidroxitirosol (HT) es uno de los principales compuestos fenólicos del aceite de oliva. En los últimos años, de la mano de diversos estudios clínicos, se ha demostrado la asociación del consumo dietético de fuentes naturales de HT con beneficios para la salud, relacionados con su capacidad antioxidante y protectora frente a factores de riesgo cardiovascular (FRCV). Estos efectos beneficiosos dependen de su concentración en la matriz alimentaria, así como de su bioaccesibilidad y su biotransformación durante el proceso de digestión gastrointestinal en compuestos derivados que pueden contribuir a su biodisponibilidad total y al efecto biológico de esta molécula. Por tanto, resulta importante determinar la cantidad de HT y sus derivados como resultado de los diversos procesos fisiológicos (digestión) y reacciones metabólicas que darán lugar a metabolitos biológicamente activos. En este contexto, uno de los propósitos de este estudio fue analizar el efecto de la matriz alimentaria sobre la farmacocinética y biodisponibilidad del HT, destacando el aceite de oliva virgen extra como la matriz que proporcionó los mejores valores. Este trabajo ha contribuido a destacar la importancia de la matriz en la absorción final del HT. Sin embargo, recientemente se han identificado en el aceite de oliva derivados de HT esterificado con diferentes ácidos grasos (lipofenoles) que se caracterizan por un mayor carácter lipófilo. Esta propiedad redunda en una absorción celular y estabilidad metabólica mejoradas, de modo que ejercen su actividad biológica en tejidos u órganos diana de forma más eficiente. La presente Tesis Doctoral investigó mediante un modelo in vitro de simulación de digestión gastrointestinal el efecto de la matriz alimentaria y las enzimas digestivas sobre la bioaccesibilidad de lipofenoles de HT, concluyendo que su estabilidad depende en gran medida tanto de la matriz alimentaria como del ácido graso formando la esterificación. Asimismo, un enfoque lipidómico en un modelo de células mieloblásticas (línea monocítica THP-1) permitió investigar la actividad antioxidante y antiinflamatoria de las moléculas esterificadas en comparación con la molécula nativa, a través del estudio de la modulación de la síntesis de isoprostanos y prostaglandinas, marcadores de estrés oxidativo e inflamación, respectivamente. Los resultados obtenidos han permitido demostrar la función biológica de los lipofenoles de HT, siendo necesario la realización de estudios adicionales in vivo para la confirmación del potencial biológico de estos compuestos. Asimismo, seleccionar aquellos alimentos de origen vegetal que tengan un adecuado perfil de lipofenoles de HT en función de su actividad biológica, que permitan utilizarlos como coadyuvantes en la prevención de patologías cardiovasculares asociadas al estrés oxidativo y la inflamación.Ciencias de la AlimentaciónMedicin

Confronto tra l’impatto ambientale dell’analisi tradizionale e mediante spettroscopia NIR sull’oliva

Obiettivi Le analisi che tradizionalmente si effettuano sulle olive richiedono l\u2019utilizzo di molteplici strumentazioni, lunghi tempi di analisi, l\u2019utilizzo di solventi potenzialmente dannosi, sia per la salute umana che per l\u2019ecosistema, e la distruzione del campione. Le stesse analisi effettuate con una tecnologia ottica, non distruttiva (spettroscopia nella regione del visibile e vicino infrarosso, Vis-NIR), permettono la stima dei parametri di interesse, in modo molto rapido, con una sola lettura, anche condotta dagli addetti alla raccolta. L\u2019unico aspetto laborioso \ue8 la costruzione di robuste calibrazioni. Il grande vantaggio sarebbe un ridotto impatto ambientale della tecnologia ottica, la cui valutazione, a confronto con i metodi tradizionali, \ue8 lo scopo di questo lavoro. Esso \ue8 parte di un progetto AGER (Progetto Agroalimentare e Ricerca), denominato \u201cSustainability of the Olive-Oil System - S.O.S.\u201d, il cui obiettivo \ue8 quello di migliorare e rafforzare la sostenibilit\ue0 della filiera dell\u2019olio extravergine di oliva in Italia. Metodologia Le analisi tradizionali per la valutazione qualitativa delle olive individuate come riferimento sono state: contenuto di acqua, olio e fenoli totali. Il confronto \ue8 stato effettuato per le stesse determinazioni mediante analisi ottica. Il metodo Life Cycle Assessment (ISO 14040:2006, ISO 14044:2006), \ue8 stato applicato per valutare l\u2019impatto ambientale mediante un preciso schema di esecuzione. L\u2019approccio utilizzato (from cradle to grave), ha considerato tutti gli input e output di ogni analisi, tenendo conto della strumentazione necessaria per l\u2019esecuzione (estrazione materie prime, costruzione, utilizzo e smaltimento), dei solventi utilizzati (ottenimento solvente, utilizzo e smaltimento), delle risorse energetiche necessarie. Inoltre, per la strumentazione Vis-NIR sono state considerate le analisi necessarie per la calibrazione dello strumento. I dati raccolti sono stati elaborati con il software SimaPro. Risultati Il confronto tra l\u2019impatto ambientale delle analisi tradizionali e quello delle analisi ottiche ha evidenziato un netto vantaggio nell\u2019applicazione della strumentazione ottica la quale, non solo garantisce la non distruzione del campione e un risultato in tempi brevi ma, ha un impatto sull\u2019ambiente 36 volte inferiore rispetto alle analisi tradizionali di riferimento. Se le maggiori voci di impatto ambientale per le analisi tradizionali sono la corrente elettrica, la strumentazione e le sostanze chimiche, per l\u2019analisi non distruttiva, l\u2019impatto ambientale \ue8 dettato maggiormente dalla necessit\ue0 di calibrare lo strumento, attivit\ue0 che richiede di effettuare su un relativamente elevato numero di campioni (500 per la calibrazione iniziale, 200 per il mantenimento) sia le analisi tradizionali che le letture ottiche (Vis-NIR)