Urolithin a Partially Protects against Oxidative Damage Induced for Microcistyn-lr in C6 Cells.

Microcystin-LR (MIC-LR) is a toxin which the mechanism of intoxication involves oxidative stress. Urolithin A (URO-A) is a metabolic product from the colonic fermentation of ellagic acid with antioxidant potential. This study aimed to evaluate the putative protective effect of URO-A against MIC-LR toxicity in C6 cells. C6 cells were incubated with MIC-LR (1 and 10 µM) and/or URO-A (3, 30, 60 and 100 µM) for 24 h. MIC-LR induced reactive species (RS) generation, depletion of total thiol (SH) groups, and survival loss when compared with the control group. Also, at 10 µM, MIC-LR induced CAT activity inhibition. URO-A caused CAT activity inhibition and showed a trend to increase RS generation (60 and 100 µM) per se. URO-A at 3 µM completely attenuated the RS generation and the impairment in SH groups caused by MIC-LR. Our results demonstrated that URO-A might offer a protective effect against toxicity caused by MIC-LR in glial cells by restoring the levels of RS and thiol groups.

Bioavailable Phenolic Compounds from Olive Pomace Present Anti-Neuroinflammatory Potential on Microglia Cells.

The neuroinflammatory process is considered one of the main characteristics of central nervous system diseases, where a pro-inflammatory response results in oxidative stress through the generation of reactive oxygen and nitrogen species (ROS and RNS). Olive (Olea europaea L.) pomace is a by-product of olive oil production that is rich in phenolic compounds (PCs), known for their antioxidant and anti-inflammatory properties. This work looked at the antioxidant and anti-neuroinflammatory effects of the bioavailable PC from olive pomace in cell-free models and microglia cells. The bioavailable PC of olive pomace was obtained through the process of in vitro gastrointestinal digestion of fractionated olive pomace (OPF, particles size < 2 mm) and micronized olive pomace (OPM, particles size < 20 µm). The profile of the PC that is present in the bioavailable fraction as well as its in vitro antioxidant capacity were determined. The anti-neuroinflammatory capacity of the bioavailable PC from olive pomace (0.03-3 mg L-1) was evaluated in BV-2 cells activated by lipopolysaccharide (LPS) for 24 h. The total bioavailable PC concentration and antioxidant activity against peroxyl radical were higher in the OPM than those observed in the OPF sample. The activation of BV-2 cells by LPS resulted in increased levels of ROS and nitric oxide (NO). The bioavailable PCs from both OPF and OPM, at their lowest concentrations, were able to reduce the ROS generation in activated BV-2 cells. In contrast, the highest PC concentration of OPF and OPM was able to reduce the NO levels in activated microglial cells. Our results demonstrate that bioavailable PCs from olive pomace can act as anti-neuroinflammatory agents in vitro, independent of particle size. Moreover, studies approaching ways to increase the bioavailability of PCs from olive pomace, as well as any possible toxic effects, are needed before a final statement on its nutritional use is made.

Phenolic compounds and antioxidant activity in vitro and in vivo of Butia and Opuntia fruits.

Despite the consumption recommendations and the potential health benefits, Brazilian biodiversity has a large number of fruit species that are still unexplored, such as Butia catarinensis (Butiá da Praia), Butia eriospatha (Butiá da Serra) and Opuntia elata (Arumbeva). The phenolic compounds of these fruits were determined by HPLC-DAD-MS/MS. Morever, in vitro assays of antioxidant capacity on hydroethanolic extracts against hydrogen peroxide (H2O2), hydroxyl (OH), peroxyl (ROO) and ABTS radicals were evaluated. In vivo assays evaluating the survival of worms and reactive oxygen species (ROS) generation were performed using the nematode Caenorhabditis elegans. Eighteen, twenty-eight and seventeen phenolic compounds were identified in Butiá da Praia, Butiá da Serra and Arumbeva, respectively. The main groups of phenolic compounds found in the fruits were hydroxybenzoic acids (60.5, 26.5 and 96.1% of the total phenolic compounds for Butiá da Praia, Butiá da Serra and Arumbeva, respectively), flavan-3-ols (23.6 and 61.2% of the total phenolic compounds for Butiá da Praia and Butiá da Serra) and flavonols (2.6% of the total phenolic compounds for Arumbeva). The hydroethanolic extracts of these fruits were free radical scavenger, sources of phenolic compounds and did not cause toxic effects in vivo. In hydroethanolic extracts of Butiá da Praia and Arumbeva, the total phenolic content increased by around 67% and 35%, respectively. Besides the health benefits, these proved to be promising sources of natural antioxidants, with phenolic composition variating among species and collection site. The obtained results enable future applications of studied fruits extracts in food and/or pharmaceutical products, encouraging and valuing the sustainable use of biodiversity.

New insights into the phenolic compounds and antioxidant capacity of feijoa and cherry fruits cultivated in Brazil.

Acca sellowiana (feijoa) and Eugenia involucrata (cherry) are fruits species of Brazilian biodiversity (Myrtaceae family). In this study, a sampling process was used with three different harvesting sites. The composition of phenolic compounds of these fruits was determined by HPLC-DAD-MS/MS. Moreover, the antioxidant capacity of hydroethanolic extracts against hydrogen peroxide (H2O2), hydroxyl (OH), peroxyl (ROO-) and ABTS radicals was evaluated. Thirty and twenty-seven phenolic compounds were identified in feijoa and cherry, respectively. The major phenolic compounds found were pedunculagin isomer (5040.87, 3443.66 and 1324.95 µg·g-1) in feijoa and procyanidin (1406.54, 1888.00 and 1380.64 µg·g-1) in cherry. Hydroethanolic extract of these fruits was a potent scavenger of free radicals and excellent source of phenolic compounds. In hydroethanolic extracts of feijoa, the phenolic content increased by around 50%, while in the cherry the content was similar to that found in the fruit. For ORAC method, sample 2 of feijoa and cherry showed values of 383 and 126 µM·TE·g-1, respectively, featuring the highest antioxidant capacity. This study is the first to report the identification of castalagin, catechin and epicatechin in feijoa, and rutin in cherry. Besides, the health benefits, these fruits can contribute to biodiversity conservation.

Ochratoxin A presence in Cabernet Sauvignon wine changes antioxidant activity in vitro and oxidative stress markers in vivo.

Ochratoxin A (OTA) is a mycotoxin found in grape products and oxidative stress has been reported as an important mechanism involved in its toxicity, classified as possible carcinogenic to humans. Conversely, phenolics are known bioactive compounds in grapes and display great antioxidant properties. However, the biological effects of the concomitant presence of phenolic compounds and OTA remains unclear. The aim of this study was to evaluate, for the first time, the effect of OTA presence in Cabernet Sauvignon wine on antioxidant activity in vitro and on oxidative stress markers in vivo. In addition, the phenolic composition of wine was evaluated by LC-DAD-MS/MS. In vitro assays were based on spectrophotometric methods, while in vivo assays were performed evaluating oxidative stress markers in the nematode Caenorhabditis elegans, an alternative model to animal testing. A total of 23 phenolic compounds were identified in the Cabernet sauvignon red wine, including the anthocyanins delphinidin-3-O-glicoside and malvidin-3-O-glicoside, the flavonol quercetin-3-O-glucuronide and the phenolic acids caffeic, verbascoside and caftaric. Trans-resveratrol and trans-piceid were the only stilbenes found in the samples. OTA presence in the red wine was accompanied by reduction in GSH content and increase in hydroxyl radical generation in vitro. The presence of OTA in wine also increased lipoperoxidation and induced overexpression of the antioxidant enzymes superoxide dismutase and catalase in vivo. This study demonstrates that OTA presence in red wine can reduce its antioxidant potential in vitro and induces oxidative stress in vivo, without affecting the phenolic compounds levels in the samples. Thus, this work provides insights into the negative effects of the presence of OTA in wine, not only by its known toxicity, but also by prejudicing the antioxidant potential of wine. It is important to be aware of these effects when developing a complete description of OTA toxicity in humans.

Effect of Aspergillus carbonarius on ochratoxin a levels, volatile profile and antioxidant activity of the grapes and respective wines.

Aspergillus carbonarius can produce a possibly carcinogenic mycotoxin named ochratoxin A (OTA). The metabolism of this fungus can also impact grape and wine quality as it influences the volatile and phenolic profiles, which are related to aroma and antioxidant activity, respectively. The objective of this study was to evaluate the effect of A. carbonarius on OTA levels and for the first time on volatile profile and antioxidant activity of grapes and their respective wines. Cabernet Sauvignon (CS, red) grapes presented higher susceptibility to A. carbonarius than Moscato Italico (MI, white) grapes and OTA levels in their respective musts were in accordance with this same trend. However, vinification of red grapes resulted in 67% reduction of OTA, while the reduction observed with white wines was 45%. The presence of acids (hexanoic, octanoic, nonanoic and decanoic, fatty odor) was found to be an indicative of the fungus incidence in grapes. These acids were precursors of esters that might impart negative aroma (methyl nonanoate and isoamyl octanoate, fatty odor) or provide desirable fruity characteristics (ethyl hexanoate, ethyl octanoate and methyl octanoate) for wine. In addition, terpenes were detected only in wines produced with grapes (CS and MI) inoculated with A. carbonarius. The presence of A. carbonarius increased the antioxidant activity of CS grapes. For MI grapes and both wines (CS and MI) no differences were verified in the antioxidant activity of the samples affected or not affected by this fungus. Although A. carbonarius occurrence has shown no influence on the antioxidant activity of wines, it produced OTA and has negatively influenced the wine odor profile, due to the production of some volatiles that impart a deleterious effect on wine aroma.

Composição fenólica e perfil antioxidante do suco de uva Concord / Phenolic composition and antioxidant profile of Concord grape juice

O suco de uva apresenta na constituição química uma diversidade de substâncias com ações benéficas, especialmente compostos fenólicos. O trabalho teve como objetivo analisar a composição fenólica e a capacidade antioxidante in vitro do suco de uva Concord. A atividade antioxidante foi avaliada pelos métodos de oxidação da GSH, geração do radical ABTS e ensaio da desoxirribose. A análise da composição fenólica foi realizada utilizando HPLC-DAD-MS. O suco reduziu a geração dos radicais ABTS e hidroxil bem como elevou o conteúdo de GSH. Os compostos fenólicos majoritários no suco de uva Concord foram antocianinas, ácidos fenólicos e flavonols. Os resultados obtidos demonstram o potencial antioxidante do suco da variedade Concord, possivelmente associado à sua composição fenólica.

Efeito da ocratoxina A em suco de uva tinto sobre sobrevivência e geração de EROs em Caenoaerbiditis elegans / Effects of ochratoxin A in red grape juice on survival rate and ROs generation in Caenoaerbiditis elegans

O consumo do suco de uva vem crescendo ao longo dos anos em função das suas propriedades funcionais. No entanto, este produto não está livre de contaminantes como a ocratoxina A (OTA). O presente trabalho teve por objetivo avaliar o efeito da presença de OTA em suco de uva Concord sobre geração de espécies reativas de oxigênio (EROs) e taxa de sobrevivência em Caenoarbiditis elegans. Os vermes foram expostos por 30 minutos ao suco na presença de OTA (0, 1, 2 e 4 µg∙L-1). O suco em presença de OTA não afetou a sobrevivência do C. Elegans. A adição de suco livre de OTA reduziu a sobrevivência dos nematoides, embora não tenha influenciado a geração de EROs. Contudo o suco com a concentração mais elevada de OTA reduziu a geração de EROs. Assim, são necessários maiores estudos para entender os mecanismos de toxicidade da OTA neste modelo vivo.

Exposure risk to carbonyl compounds and furfuryl alcohol through the consumption of sparkling wines / Risco da exposição a compostos carbonílicos e álcool furfurílico através do consumo de espumantes

ABSTRACT: The goals of this study were to verify the occurrence of furfuryl alcohol (FA) and carbonyl compounds (acetaldehyde, acrolein, ethyl carbamate (EC), formaldehyde and furfural) in sparkling wines and to evaluate, for the first time, whether the consumption of the samples under study could represent risk to consumers health. These compounds are electrophilic; and therefore, may covalently bind to DNA, which may result in mutagenicity. EC and formaldehyde were present at low levels (<1μg L-1) in all samples. Acetaldehyde, furfural and acrolein were also found in low levels (<1.5, 1.4 and 1.0μg L-1, respectively) in 57, 71 and 76% of samples. In the other samples, levels of acetaldehyde, furfural and acrolein ranged from 5.2 to 54.8, 10.5 to 41.0 and 20.3 to 36.7μg L-1, respectively. Furfuryl alcohol was also reported in all samples in levels from 10.4 to 33.5μg L-1. Acrolein was the only compound reported at levels sufficient to represent risk to health, which occurred in 24% of the samples. A study focused on the origin of acrolein deserves attention, investigating the influence of the concentration of precursors and the role of fermentation in the formation of this aldehyde, besides the evaluation of possible environmental contamination of grapes during cultivation.

RESUMO: Os objetivos deste estudo foram verificar a ocorrência de álcool furfurílico (FA) e compostos carbonílicos (acetaldeído, acroleína, carbamato de etila (CE), formaldeído e furfural) em espumantes e avaliar, pela primeira vez, se o consumo das amostras em estudo poderia representar risco para a saúde do consumidor. Esses compostos são eletrofílicos e, portanto, podem se ligar covalentemente ao DNA, o que pode resultar em mutagenicidade. CE e formaldeído foram encontrados em baixos níveis (<1μg/L) em todas as amostras. Acetaldeído, furfural e acroleína também foram encontrados em baixos níveis (<1,5; 1,4 e 1,0μg L-1, respectivamente) em 57, 71 e 76% das amostras. Nas demais amostras, os níveis de acetaldeído, furfural e acroleína variaram de 5,2 a 54,8, 10,5 a 41,0 e 20,3 a 36,7μg L-1, respectivamente. O álcool furfurílico também foi encontrado em todas as amostras em níveis de 10,4 a 33,5μg L-1. A acroleína foi o único composto encontrado em níveis suficientes para representar risco à saúde, que ocorreu em 24% das amostras. Uma avaliação focada na origem da acroleína merece atenção, investigando a influência da concentração dos precursores e o papel da fermentação na formação do aldeído, além da avaliação da possível contaminação ambiental das uvas durante o cultivo.