Red seaweeds Grateloupia turuturu and Porphyra umbilicalis as nutraceuticals and functional food: nutritional/chemical composition and immunostimulatory activity

1st Natural Products Application: health, cosmetic and food - International online conference, 4-5 February 2021The number of nutraceuticals and functional foodsfrom marine origin is increasing in the worldwide market. Specifically, the red seaweeds Grareloupia turuturu and Porphyra umbilicalis have shown their potential considering relevant bioactive compounds and nutritional value [1,2]. Nevertheless, it is pertinent to explore knowledge gaps regarding their nutritional/chemical composition and bioactivities. Hence firstly G. turuturu and P. umbilicalis proximate composition and lipid classes were determined; secondly, after preparing hydroethanolic and aqueous infusion and decoction) seaweed extracts, mycosporine-like ami no acids (MAAs) and carbohydrates were quantified. and the immunomodulatory potential was assessed using RAW 264.7 cell line. Results show that dietary fibre was the most abundant nutritional parameter for both seaweed species, followed by relevant ash and protein contents. Free fatty acids were the main lipid class in both seaweed species, whereas tocopherols exhibited residual contentsby RP-HPLC-FLD. For hydroethanolic and aqueous extracts, several MAAs were quantified by RP-HPLC-DAD and LC-DAD ESI-MS. P. umbilicalis extracts demonstrated greater MAAs content than G. turuturu, and porphyra-334 was the main MAA. The aqueous extracts of both seaweeds showed higher carbohydrate contents than the hydroethanolic, with P. umbilicalis decoction exhibiting the highest level. Concerning the immunomodulatory activity, for 0.02 mg/mL, the exposure to P. umbilicalis aqueous extracts resulted in the most accentuated 'NO production by RAW 264.7 cells, showing immunostimulatory potential. MAAs and carbohydrates are believed to be responsible for the immunostimulatory activity [3,4]. G. turuturu and P. umbilicalis potential as functional food and nutraceuticals, relying on their relevance as sources of bioactive compounds to the food, pharmaceutical and cosmetic industries, is thus reinforcedThis work was financed by European Funds FEDER/COMPETE/POCI under project POCI-01-0145-FEDER-006958 (CITAB), and by FCT to CECAV (UIDB/CVT/00772/2020), CITAB (UlDB/04033/2020) and CESAM (UIDP/5001712020+UIDB/50017/2020). FCT is also acknowledged for a doctoral grant to Joao Ferreira (SFRH/BD/144882/2019)N

An optimized comet-based in vitro DNA repair assay to assess base and nucleotide excision repair activity

This optimized protocol (including links to instruction videos) describes a comet-based in vitro DNA repair assay that is relatively simple, versatile, and inexpensive, enabling the detection of base and nucleotide excision repair activity. Protein extracts from samples are incubated with agarose-embedded substrate nucleoids (‘naked’ supercoiled DNA) containing specifically induced DNA lesions (e.g., resulting from oxidation, UVC radiation or benzo[a]pyrene-diol epoxide treatment). DNA incisions produced during the incubation reaction are quantified as strand breaks after electrophoresis, reflecting the extract’s incision activity. The method has been applied in cell culture model systems, human biomonitoring and clinical investigations, and animal studies, using isolated blood cells and various solid tissues. Once extracts and substrates are prepared, the assay can be completed within 2 d

Cytotoxic effects of euterpe oleraceae fruit oil (acai) in rat liver and thyroid tissues

Euterpe oleracea Mart., Arecaceae, fruit (acai) presents considerable potential for the development of new medicines due to its phytochemical composition and antioxidant activity. More recently, special attention has been given to the pharmacological potential of the fruit's oil. This study analysed the histological and histochemical effects of different dosages of acai oil on rat's liver and thyroid cells, in order to evaluate its cytotoxic potential after administration for consecutive days. Male Wistar rats were treated with the acai oil by gavage at doses of 30, 100 and 300 mg/kg, for 14 days, within a 24 h interval. Liver and thyroid fragments were collected for histology (hematoxylin and eosin) and histochemistry analysis (blue of Nilo (lipids), Baker (lipids), bromophenol blue (protein), PAS (polysaccharides)). The results showed that animals exposed to acai oil presented alterations in the liver cells, where the integrity of the liver tissue was increasingly lost as the acai oil doses increased. Nuclear pyknosis was observed in several hepatocytes, evidencing the occurrence of cell death. Alteration in the amount of lipids, polysaccharides, vacuoles in the cytoplasm, and proliferation of Kupffer cells were observed in histochemical analyzes. As for the thyroid of the treated rats, alterations were observed in the size of the follicular lumen and also in the connective tissue found between the follicles. Under the experimental conditions employed in the present study, the cytotoxicity observed in this work is worrying, specially considering the liver, when frequent or continuous damage could lead pathological disorders in this organ2915461CAPES - Coordenação de Aperfeiçoamento de Pessoal e Nível SuperiorFAPESP – Fundação de Amparo à Pesquisa Do Estado De São PauloSem informação2012/17241-

Measuring DNA modifications with the comet assay: a compendium of protocols

Supplementary information The online version contains supplementary material available at https://doi.org/10.1038/s41596-022-00754-yThe comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA-DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.The comet assay is commonly used to assess DNA damage. This collection of consensus protocols includes adaptations for a wide range of species and sample types, assay formats and detection of different types of DNA lesions.We thank the hCOMET project (COST Action, CA 15132) for support. A. Azqueta thanks the Ministry of Science and Innovation (AGL2015-70640-R and PID2020-115348RB-I00) of the Spanish Government. S.G. thanks the national funds (OE), through FCT- Fundacao para a Ciencia e a Tecnologia (IP, in the scope of the framework contract foreseen in the numbers 4,5 and 6 of the article 23, of the Decree-Law 57/2016, of 29 August, changed by Law 57/2017, of 19 July) for personal support. V.M.d.A. thanks the National Council of Technological and Scientific Development (CNPq-304203/2018-1) for personal support. D.M. thanks the program Ayudas para la formacion de profesorado universitario (FPU) of the Spanish Government for the predoctoral grant received . N.O. thanks the NIEHS Superfund Research Program ES ES027707 for personal support . J.S.-S. thanks the Government of Navarra for the predoctoral grant received. V.V. thanks the Ministerio de Educacion, Cultura y Deporte (Beatriz Galindo' program, BEAGAL18/00142) of the Spanish Government for personal support . M.S.C. acknowledges personal support from the National Institute of Environmental Health Sciences of the National Institutes of Health under award number: 1R41ES030274-01. This paper reflects the views of the authors and does not necessarily reflect those of the US Food and Drug Administration or the National Institutes of Health