In vivo and in vivo biological activity of cola beverages and different additives ; Actividad biológica in vivo e in vitro de bebidas de cola y sus aditivos

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Titel: In vivo and in vivo biological activity of cola beverages and different additives ; Actividad biológica in vivo e in vitro de bebidas de cola y sus aditivos

Beteiligte: Mateo-Fernández, Marcos [VerfasserIn]

Erschienen: [Erscheinungsort nicht ermittelbar]: Universidad de Córdoba, UCOPress, 2019

Sprache: Englisch

Identifikator:

Entstehung:

Hochschulschrift: Dissertation, Universidad de Córdoba, UCOPress, 2019

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Beschreibung: Environmental factors such as diet may modify cancer risk and tumor behavior since they could be involved in gene expression changes. One of the worldwide consumed products is Coca-Cola. Originally developed as medical supplements, Cola-based drinks and several beverages such as beer and wine were proposed as medicinal substances. However, a relationship between the consumption of these beverages and an increase in the prevalence of several diseases such as child obesity, diabetes, hypertension and dental diseases was also demonstrated. In spite of the worldwide importance and spread of cola beverages, scientific studies assessing their effects on health and wellbeing, are quite scarce and controversial. Therefore, assays are needed to research about their food safety and nutraceutic potential. Genotoxicological screening tests have been extensively used over time for assessing the health properties of compounds prior to being considered as safe substances. Nowadays, the list of foods with documented health-benefit activities is endless, and scientific evidence supporting the concept of health-promoting food ingredients is steadily growing. The aim of this work was to assess the biological and food safety of different colabased beverages and theirs distinctive constituents, adding to science, a new data corpus by providing more information about the promising properties that cola beverages could provide. In order to analyse the safety and protective effects against an oxidative toxin in the in vivo Drosophila model organism, toxicity, antitoxicity, genotoxicity, antigenotoxicity and lifespan assays were carried out. In addition, the chemopreventive potential and the induction of DNA damage ability in the in vitro tumour cells (human leukaemia HL-60) were conducted using cytotoxicity, internucleosomal DNA fragmentation, single cell gel electrophoresis and methylation status assays. All these studies consist of a set of biological screening assays where several points related to degenerative processes can be checked. Neither toxic nor genotoxic activity was found according to the results obtained. Classic Coca-Cola, Caffeine-Free Coca-Cola, Zero Coca-Cola, energy drinks, Glucose, Phosphoric acid and Citric acid behaved as antioxidant against hydrogen peroxide. In addition, all tested substances and the constituents, except for Zero Coca-Cola and Fructose showed to be antigenotoxic exerting a protective ability against hydrogen peroxide in the genome of Drosophila melanogaster. However, promising results were not found in the lifespan assay on D. melanogaster. Only 2 out of 8 lyophilised beverages (Classic Coca-Cola and Diet Coca-Cola) resulted in increasing the life expectancy of this fly. Caffeine-Free Coca-Cola, Zero Coca-Cola, Pepsi-Cola and RedBull Sugar Free (4 out of 8 complex mixtures) significantly decreased the lifespan at some tested concentration at least, being Pepsi-Cola and RedBull Sugar Free the worst ones. According to the constituents, Caffeine, Fructose, Aspartame and Citric Acid increased the life expectancy in some extent and only Phosphoric acid decreased it. All lyophilised beverages and constituents were cytotoxic in HL-60 cell line, except Caffeine, Cyclamate and Taurine. Classic Coca-Cola, Fructose and Classic RedBull induced DNA fragmentation in HL-60 cell line when treated with the second-highest tested concentrations. However, despite all the tested compounds and beverages, except Citric acid, Caramel colour IV, Pepsi-Cola, and RedBull Sugar Free, induced DNA damage when single cell level is analysed using SCGE test, none of the tested compounds produced significant apoptotic TM values. Therefore, the cell death showed by most of the lyophilised beverages and biological compounds is not produced by apoptotic way but, according to SCGE, it is due to necrotic activity. Considering hypermethylation status as genomic stability marker in the HL-60 cancer line, Diet Coca-Cola, Zero Coca-Cola and Caffeine-Free Diet Coca-Cola produced an increased level of methylation at each tested concentration. At least, one concentration of Classic Coca- Cola, Caffeine-Free Coca-Cola, Pepsi-Cola and RedBull Sugar Free hypermethylated transposable element on HL-60 cell line. Acesulphame-K and Phosphoric acid increased the level of methylation at each concentration. The methylation status of HL-60 cell line decreased when they are treated with Cyclamate, Citric acid, Caramel colour IV and Classic RedBull in one tested concentration at least and none of the tested concentrations hypermethylated them. As for the quantitative approach results, ACK and CAR could be the responsable compounds for the low scores obtained in PEP and diet beverages, especially in DCFCC. The mark obtained in CRB, SFRB and TAU was the same hence TAU may possess an influence on CRB and SFRB and the other compounds could be disguised one another in a synergic effect. In summary, lyophilised cola beverages and energy drinks showed to be safe to D. melanogaster. Further studies are needed to elucidate the biological activities and nutraceutic potential of cola beverages to solve the controversial information found in scientific databases due to the promising results obtained in flies and HL-60 cell line in some extent. For instance, non-tumoural cell must be used to be compared with our in vitro studies. When appropriate, clinic assays should also be performed. ; Existen factores medioambientales, tales como la dieta, que pueden modificar el riesgo de padecer cáncer y el comportamiento tumoral cambiando la regulación de la expresión génica. Una de las bebidas más consumidas en nuestra dieta a lo largo del mundo es la Coca-Cola y se desarrolló originalmente como un fármaco al igual que la cerveza o el vino y otras bebidas. Sin embargo, también se demostró una relación entre el consumo de estas bebidas y un aumento en la prevalencia de varias enfermedades como la obesidad infantil, diabetes, hipertensión y enfermedades dentales. A pesar de la importancia mundial y la propagación de las bebidas de cola, los estudios científicos que evalúan sus efectos en la salud y el bienestar son bastante escasos y controvertidos. Por lo tanto, se necesitan ensayos para investigar sobre su inocuidad alimentaria y potencial nutracéutico. Las pruebas de detección genotoxicológica se han utilizado ampliamente a lo largo del tiempo para evaluar las propiedades saludables de los compuestos antes de ser considerados como sustancias seguras. Hoy en día, la lista de alimentos con propiedades beneficiosas para la salud es interminable, y la evidencia científica que respalda el concepto de ingredientes alimentarios que promueven la salud está en constante crecimiento. El objetivo de este trabajo fue evaluar la seguridad biológica y alimentaria de diferentes bebidas a base de cola y sus constituyentes distintivos, agregando a la ciencia, un nuevo cuerpo de datos al aportar más información sobre las propiedades prometedoras de las bebidas de cola. Con el fin de analizar los efectos de seguridad y protección contra una toxina oxidativa en el organismo in vivo Drosophila, se llevaron a cabo ensayos de toxicidad, antitoxicidad, genotoxicidad, antigenotoxicidad y ensayos de longevidad. Además, el potencial quimiopreventivo y la capacidad de inducir daño en el ADN en las células tumorales (leucemia humana HL-60) se evalúo mediante ensayos in vitro de citotoxicidad, fragmentación del ADN internucleosómica, ensayo del cometa y ensayos de estado de metilación. Todos estos estudios consisten en un conjunto de ensayos de detección biológica donde se pueden verificar varios puntos relacionados con los procesos degenerativos. Los resultados obtenidos mostraron que no se encontraron compuestos tóxicos ni genotóxicos para ninguna sustancia probada. La Coca-Cola Clásica, Coca-Cola Sin Cafeína, Coca-Cola Zero, bebidas energéticas, glucosa, ácido fosfórico y ácido cítrico se comportaron como antioxidantes contra el peróxido de hidrógeno. Además, todas las sustancias analizadas y los constituyentes, excepto Coca-Cola Zero y fructosa, mostraron ser antigenotóxicos y ejercen una capacidad protectora contra el peróxido de hidrógeno en el genoma de Drosophila melanogaster. Sin embargo, no se encontraron resultados prometedores en el ensayo longevidad de D. melanogaster. Solo 2 de 8 bebidas liofilizadas (Coca-Cola Clásica y Coca-Cola Light) aumentaron la esperanza de vida de esta mosca. La Coca-Cola Sin Cafeína, Coca-Cola Zero, Pepsi-Cola y RedBull Sin Azúcar (4 de 8 mezclas complejas) redujeron significativamente la longevidad en algunas concentraciones probadas, siendo la Pepsi-Cola y el RedBull Sin azúcar las peores. En cuanto a los constituyentes, la cafeína, la fructosa, el aspartamo y el ácido cítrico aumentaron la esperanza de vida en cierta medida y solo el ácido fosfórico la disminuyó. Todas las bebidas liofilizadas y los compuestos simples fueron citotóxicos en la línea celular HL-60, excepto la caf...

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