Chemical characterization, in vivo safety profile and effects of the extract from unconventional food plant Acca sellowiana (O. BERG) Burret on endothelial cells under glucose overload.

"Guava" (Acca sellowiana) is an unconventional edible plant from Brazil. It is used in traditional medicine as an anti-diabetic; however, pharmacological studies on this plant are scarce. This study aimed to evaluate the chemical and safety profile of an aqueous A. sellowiana peel extract (ASPE) and its effects on endothelial EA.hy926 cells under glucose overload and in vivo (Artemia salina). An ethanolic extract from A. sellowiana peels (ASPEetOH) was also produced and characterized. Results showed that ASPE did not present in vivo toxicity, and it was found to contain high phenolic content and redox capacity. ASPE (50 µg/mL; 24 h) prevented oxidative stress and mitochondrial dysfunction, besides positively modulating Sirtuins 1 and 3, and prevented the increase of COX-2 and NF-kß expression levels in EA.hy926 cells under glucose overload. Chromatographic fractionation, metabolite profiling, spectroscopic and bioinformatics analyses revealed the presence of phenolic acids, flavan-3-ols, flavonols, flavones, flavanones, and anthocyanidins, displaying a diversity of compounds in the crude and fractionated ASPEetOH. This study provided evidence on the safety profile, chemical composition, and pharmacological activities of A. sellowiana.

Pre-clinical evidence for the therapeutic effect of Pitaya (Hylocereus lemairei) on diabetic intestinal microenvironment.

Intestinal glucose absorption plays a central role in the regulation of glucose plasmatic; however, current clinical management does not target the gut for treating diabetes. This study evaluated the effects of peel and pulp aqueous extract from Hylocereus lemairei on human enterocytes under high glucose concentration. Anti-hyperglycemic and antiobesity activities in vitro were also evaluated. Extracts did not cause cytotoxicity at 1 to 500 µg/mL. Moreover, they were effective in attenuating oxidative stress (DCFH-DA assay) and inflammation (•ON production) caused by high glucose. Intestinal enzymes (α- glucosidase and pancreatic lipase) were inhibited by pulp and peel extracts (>60% and >95%, respectively). Extracts exhibited a redox capacity superior to ascorbic and chlorogenic acids, presenting high phenolic content, mainly anthocyanins. The main compounds for both extracts were chlorogenic acid and naringin, and peel stood both qualitatively and quantitatively. Data suggest red Pitaya has potential as a new medicine for diabetes.

Dual effect of the herbal matcha green tea (Camellia sinensis L. kuntze) supplement in EA.hy926 endothelial cells and Artemia salina.

ETHNOPHARMACOLOGICAL RELEVANCE: Matcha green tea (Camellia sinensis) based-supplements have been widely used since they present a greater content of phenolic compounds than traditional green tea, which is popularly used in the treatment of diabetes. However, there are few studies on the effectiveness and safety of matcha supplements. AIM OF THE STUDY: This work aimed to evaluate the efficacy and safety of this supplement in endothelial cells (EA.hy926) in the hyperglycemic model and in vivo Artemia salina. MATERIALS AND METHODS: To assess the effect of Matcha herbal supplement (MHS), EA. hy926 endothelial cells were treated with 20 µg/mL of MHS for 24 h, in a hyperglycemic medium with 35 mM glucose. After treatment, cells were trypsinized and centrifuged at 4 °C and 47×g for 5 min. The pellet was used to determine the reaction products to thiobarbituric acid and the levels of nitric oxide. Electron transport chain activity and ATP levels were also evaluated. Intracellular pH, apoptosis, and mitochondrial membrane depolarization were evaluated by flow cytometry. MHS chemical characterization was performed by HPLC-UV and total phenolic content analysis. The evaluation of the antioxidant capacity of MHS was performed by 2,2-diphenyl-1-picrylhydrazyl radical scavenger assay. To determine the in vivo acute toxicity of MHS, an A. salina assay was conducted, using 0,2 mL of different concentrations of MHS (10, 50, 100, 250, 500, 750 and 1000 µg/mL). The LD50 values were obtained by interpolation of 50% (y = 50) of the dead individuals in the trend curves. RESULTS: Our data showed that MHS was able to avoid oxidative and nitrosative stress induced by hyperglycemia, demonstrating important antioxidant activity. However, it was observed that MHS reduced up to 90% the activity of the four-electron transport complexes, reducing the ATP production of the endothelial cells. In the toxicity assay performed in Artemia salina, MHS showed mild toxicity (LD50 = 0,4 mg/mL). The major compounds found in MHS were epigallocatechin gallate, epicatechin, rutin, kaempferol, and quercetin. CONCLUSIONS: This data draws attention to the fact that supplements with high content of phenolic compounds, capable of avoiding oxidative and nitrosative stress can have a dual effect and, simultaneously to antioxidant activity, can induce toxicity in different cell types.

Exposure in the tobacco fields: Genetic damage and oxidative stress in tobacco farmers occupationally exposed during harvest and grading seasons.

Agricultural workers engaged in tobacco cultivation are constantly exposed to large amounts of harmful agents, such as pesticides and nicotine. Furthermore, most of the flue-cured tobacco leaves are manually graded exposing workers to agents such as tobacco-specific nitrosamines. This study aimed to evaluate genetic damage and oxidative stress in tobacco farmers occupationally exposed during the harvest and grading seasons. We obtained data on DNA damage detected in Comet assay in blood cells and micronucleus experiment with buccal cells from 241 individuals. The serum cotinine levels and nitrates were also evaluated. The Comet Assay results showed a showed an increased visual score for males and females during harvest time and tobacco grading. An increase of micronucleated and binucleated cells was observed in the grading group compared to the control and harvest groups. The oxidative stress measurements showed a clear increase of thiobarbituric acid reactive substances (TBARS) in tobacco farmers during harvest time, and trolox equivalent antioxidant capacity (TEAC) in individuals during harvest and grading time compared to the controls. Significant increases of the cotinine levels were observed during the harvest and grading period (harvest>grading), and nitrates for the grading period compared to the control. In this study, tobacco farmers presented compromised DNA integrity associated with enhanced oxidative stress levels.

[PARP Inhibitors: From the Mechanism of Action to Clinical Practice]. / Inibidores da PARP: do mecanismo de ação à prática clínica.

Repairing damage and errors that occur in the DNA molecule is essential to maintain the integrity of the genome and cell viability. Deficits in DNA repair mechanisms lead to an increased risk of genetic instability and contribute to neoplastic transformation. Poly (ADP-ribose) polymerases (PARP) are a group of enzymes that play a key role in signalling and repairing DNA errors. The inhibition of its activity is a therapeutic strategy that takes advantage of the mechanism of synthetic lethality and that can be used in the treatment of tumours with specific defects in DNA repair pathways, namely in tumours with mutations in the tumour suppressor genes BRCA1 and BRCA2. There are several PARP inhibitors (iPARP), already approved by the USA Food and Drug Administration and the European Medicines Agency used in the treatment of breast, ovarian, pancreatic and prostate cancer. However, as with other target therapies, despite being well tolerated and widely used in the clinical practice, iPARP resistance is common and can be developed through various molecular mechanisms. In this article, we intend to make an updated review on iPARP and its main role in tumour cells, highlighting the several resistance mechanisms that have been recently revealed, as well as the current clinical applications and toxicity associated with this target therapy.

A reparação dos danos que ocorrem na molécula de ADN é fundamental para manter a integridade do genoma e a viabilidade celular. Défices nos mecanismos de reparação desta molécula cursam com um aumento do risco para instabilidade genética e contribuem para a transformação neoplásica. As poly (ADP-ribose) polymerases (PARP) são um grupo de enzimas que apresentam um papel chave na sinalização e reparação dos erros no ADN. A inibição da sua atividade é uma estratégia terapêutica que tira partido do mecanismo de letalidade sintética e que pode ser usada no tratamento de tumores com defeitos específicos nas vias de reparação de ADN, nomeadamente em tumores com mutações nos genes supressores tumorais BRCA1 e BRCA2. Existem vários inibidores das PARP (iPARP) já aprovados pela Food and Drug Administration dos Estados Unidos da América e pela Agência Europeia do Medicamento e utilizados no tratamento do cancro da mama, ovário, pâncreas e próstata. No entanto, tal como acontece com outras terapias alvo, a resistência aos iPARP é comum apesar de bem tolerados e amplamente utilizados na prática clínica, e pode desenvolver-se através de vários mecanismos moleculares. Neste artigo, pretendemos realizar uma revisão atualizada sobre os iPARP e o seu principal modo de ação em células tumorais, dando a conhecer os vários mecanismos de resistência que têm sido recentemente revelados, assim como as atuais aplicações clínicas e a toxicidade associada a esta terapia alvo.

Mutual Effects of Free and Nanoencapsulated Phenolic Compounds on Human Microbiota.

Phenolic compounds (PC) have many health benefits such as antioxidant, anticarcinogenic, neuroprotective, and anti-inflammatory activities. All of these activities depend on their chemical structures and their interaction with biological targets in the body. PC occur naturally in polymerized form, linked to glycosides and require metabolic transformation from their ingestion to their absorption. The gut microbiota can transform PC into more easily absorbed metabolites. PC, in turn, have prebiotic and antimicrobial actions on the microbiota. Despite this, their low oral bioavailability still compromises biological performance. Therefore, the use of nanocarriers has been demonstrated to be a useful strategy to improve PC absorption and, consequently, their health effects. Nanotechnology is an excellent alternative able to overcome the limits of oral bioavailability of PC, since it offers protection from degradation during their passage through the gastrointestinal tract. Moreover, nanotechnology is also capable of promoting controlled PC release and modulating the interaction between PC and the microbiota. However, little is known about the impact of nanotechnology on PC effects on the gut microbiota. This review highlights the use of nanotechnology for PC delivery on gut microbiota, focusing on the ability of such formulations to enhance oral bioavailability by applying nanocarriers (polymeric nanoparticles, nanostructured lipid carriers, solid lipid nanoparticles). In addition, the effects of free and nanocarried PC or nanocarriers per se on gut microbiota are also described.

Polyphenolic Content and Antioxidant Activity of Mycelia and Basidiomes of Oyster Mushrooms Pleurotus spp. (Agaricomycetes) from Brazil.

This study aimed to examine the antioxidant activity, total phenolic content, and phenolic profile of nine strains of Pleurotus spp. isolated in southern Brazil. Basidiomes were obtained from a solid-state culture in medium containing Pinus sp. sawdust (SCM-PSW), coffee grounds (CG), or organic grape waste (OGW). Mycelia were obtained from submerged culture-potato dextrose broth (MSC-PDB). Basidiomes had the highest total phenolic content (between 31.30 ± 0.26 and 47.00 ± 0.12 mg gallic acid equivalents [GAE]/g) compared with mycelia (between 8.15 ± 0.26 and 15.96 ± 0.82 mg GAE/g). Antioxidant activity of the basidiomes showed an IC50 value between 5.36 ± 0.27 (88F.13) and 10.68 ± 0.22 mg/mL (189H.3). Mushrooms produced in the OGW and CG media had higher total phenolic content than those from MSC, indicating that they can serve as sources of bioactive compounds on culture media. These findings show the potential of natural wastes to be used as a strategy for increasing secondary metabolites in edible mushrooms, proposing an interesting approach for the nutraceutical and pharmaceutical industries.

Impact of nicotine-induced green tobacco sickness on DNA damage and the relation with symptoms and alterations of redox status in tobacco farmers.

During the harvest period, tobacco workers are exposed to nicotine and it is known that absorption of the alkaloid via the leaves causes green tobacco sickness (GST). We investigated if GST and its symptoms are associated with DNA damage and alterations of the redox status. DNA damage was measured in lymphocytes of tobacco workers and controls (n = 40/group) in single cell gel electrophoresis assays. Exposure to nicotine was determined by plasma cotinine measurements, alterations of the redox status by quantification of the total antioxidant capacity (TEAC) and of thiobarbituric acid reactive substances (TBARS). The symptoms of GTS included nausea, abdominal cramps, headache, vomiting and dizziness, and 50% of the workers had more than one symptom. Cotinine levels were enhanced in the workers (111 ng/mL); furthermore, the extent of DNA damage was ca. 3-fold higher than in the controls. This effect was more pronounced in participants with GST compared to healthy nicotine exposed workers and increased in individuals with specific symptoms (range 22-36%). TBARS levels did not differ between workers and unexposed controls, while TEAC values were even increased (by 14.3%). Contact with nicotine present in tobacco leaves causes GTS and leads to damage of the DNA; this effect is more pronounced in workers with GTS symptoms and is associated with alterations of the redox status. Damage of the genetic material which was found in the workers may lead to adverse long-term effects that are caused by genomic instability such as cancer and accelerated ageing.

Targeting Inflammatory-Mitochondrial Response in Major Depression: Current Evidence and Further Challenges.

The prevalence of psychiatric disorders has increased in recent years. Among existing mental disorders, major depressive disorder (MDD) has emerged as one of the leading causes of disability worldwide, affecting individuals throughout their lives. Currently, MDD affects 15% of adults in the Americas. Over the past 50 years, pharmacotherapy, psychotherapy, and brain stimulation have been used to treat MDD. The most common approach is still pharmacotherapy; however, studies show that about 40% of patients are refractory to existing treatments. Although the monoamine hypothesis has been widely accepted as a molecular mechanism to explain the etiology of depression, its relationship with other biochemical phenomena remains only partially understood. This is the case of the link between MDD and inflammation, mitochondrial dysfunction, and oxidative stress. Studies have found that depressive patients usually exhibit altered inflammatory markers, mitochondrial membrane depolarization, oxidized mitochondrial DNA, and thus high levels of both central and peripheral reactive oxygen species (ROS). The effect of antidepressants on these events remains unclear. Nevertheless, the effects of ROS on the brain are well known, including lipid peroxidation of neuronal membranes, accumulation of peroxidation products in neurons, protein and DNA damage, reduced antioxidant defenses, apoptosis induction, and neuroinflammation. Antioxidants such as ascorbic acid, tocopherols, and coenzyme Q have shown promise in some depressive patients, but without consensus on their efficacy. Hence, this paper provides a review of MDD and its association with inflammation, mitochondrial dysfunction, and oxidative stress and is aimed at thoroughly discussing the putative links between these events, which may contribute to the design and development of new therapeutic approaches for patients.

Grape seed proanthocyanidins improves mitochondrial function and reduces oxidative stress through an increase in sirtuin 3 expression in EA.hy926 cells in high glucose condition.

Proanthocyanidins are phenolic compounds abundant in the diet, commonly found in grapes and derivatives, foods known for their health-promoting benefits. There is previous evidence showing the antidiabetic activity of proanthocyanidins, however, their mechanisms of action have not been fully elucidated. This study evaluated the capacity of grape seed proanthocyanidins extract (GSPE) to modulate oxidative stress, nitric oxide levels, mitochondrial dysfunction, apoptosis, and sirtuin expression in endothelial cells EA.hy926 under high glucose condition. In addition, the possible toxic effects of GSPE was evaluated in a zebrafish embryos model. The results showed that GSPE was able to enhance cell viability and avoid the disturbance in redox metabolism induced by high glucose. Moreover, GSPE was able to avoid mitochondria dysfunction and the increased in p53 and poly-(ADP-ribose) polymerase expression induced by high glucose exposition. These effects were attributed to the increase in expression of sirtuin 3, a protein able to regulate mitochondrial function. GSPE in an effective concentration did not show toxic effects in zebrafish embryos model. Taken together, these data elucidate the key molecular target of GSPE for future pharmacological interventions in diabetic patients.

Grape seed proanthocyanidins prevent H2 O2 -induced mitochondrial dysfunction and apoptosis via SIRT 1 activation in embryonic kidney cells.

Grape proanthocyanidins are compounds widely ingested in the diet. This study evaluated their effects on mitochondrial function, apoptosis, and sirtuin 1 and 3 expressions in HEK-293 cells exposed to H2 O2 . High-resolution mass spectrometry and high-performance liquid chromatography characterized the proanthocyanidins extract and the presence of procyanidins B and C was detected. The extract prevented H2 O2 -induced oxidative damage to proteins and lipids and depletion in superoxide dismutase activity. Moreover, it was able to regulate the expression of NADH: Ubiquinone oxidoreductase core subunit S7 and prevent mitochondrial electron transport chain dysfunction, ATP depletion, and apoptosis induced by H2 O2 . Finally, the extract was able to regulate sirtuin 1 and 3 expressions, thus maintaining cell viability. These data show that the grape seed proanthocyanidins can target mitochondrial proteins, which may represent an important approach for the management of numerous chronic illnesses associated with mitochondrial dysfunction. PRACTICAL APPLICATIONS: Proanthocyanidins are phenolic compounds abundant in regular diet, commonly found in grapes and derivatives, pomegranates, apples, and red fruits, all foods known for their beneficial effects on health. The current study highlights the role of proanthocyanidins as mitochondrial modulators that may explain the biological activity attributed to these compounds. This study brings evidence that proanthocyanidins might be considered as a value-added agent for the development of new nutraceutical and or pharmaceutical approaches.

Dietary total antioxidant capacity as a preventive factor against depression in climacteric women.

Depression is characterized by physical or psychological distress and in many cases can lead to suicide. OBJECTIVE: to assess the prevalence of depression and its possible relationship with dietary total antioxidant capacity (DTAC) and nutritional parameters in climacteric women participating in an extension university program in a Southern Brazilian city. METHODS: data were obtained through questionnaires and anthropometric measurements. Diet was assessed using a 24-hour dietary recall. The Beck Depression Inventory (BDI) was used to assess the intensity of symptoms of depression. RESULTS: DTAC of the population ranged from 435.60 to 4502.62 mg VCE/day. Among the most consumed antioxidant food/beverages, coffee ranked highest. Polyphenols were found to be directly linked to the antioxidant capacity of fresh foods (r=0.905; p=0.0001). Prevalence of depression in the population was 44%, and depressed women had lower intake levels of polyphenols (p=0.022; Cohen's d=0.80), and vitamin B6 (p=0.038; Cohen's d=0.65), vitamin A (p=0.044; Cohen's d =0.63), and vitamin C (p=0.050; Cohen's d =0.61). There was a significant negative correlation between BDI scores and polyphenol intake (r=-0.700; p=0.002). CONCLUSION: these results may contribute to a better understanding of the recommended dietary antioxidant intake as an adjuvant for preventing depression in women.

Depressão é caracterizada por causar sofrimento físico ou psicológico e em muitos casos pode levar ao suicídio. OBJETIVO: avaliar a prevalência de depressão e sua possível relação com a capacidade antioxidante total da dieta (DTAC) e parâmetros nutricionais em mulheres climatéricas participantes de um programa universitário de extensão em uma cidade do sul do Brasil. MÉTODOS: os dados foram obtidos por meio de questionários e medidas antropométricas. A dieta foi avaliada através de um recordatório alimentar de 24 horas. O Inventário de Depressão de Beck (BDI) foi usado para avaliar a intensidade dos sintomas de depressão. RESULTADOS: a DTAC da população variou de 435,60 a 4502,62 mg VCE/dia. Entre os alimentos/bebidas antioxidantes mais ingeridos, o café destacou-se em primeiro. Os polifenóis mostraram estar diretamente ligados à capacidade antioxidante dos alimentos in natura (r=0,905; p=0,0001). A prevalência de depressão na população foi de 44%, e as mulheres depressivas apresentaram menores níveis de ingestão de polifenóis (p=0,022; Cohen's d=0,80) e vitaminas B6 (p=0,038; Cohen's d=0,65), A (p=0,044; Cohen's d=0,63) e C (p=0,050; Cohen's d=0,61). Houve uma correlação negativa significativa entre os escores do BDI e a ingestão de polifenóis (r=-0,700; p=0,002). CONCLUSÃO: estes resultados podem contribuir para uma melhor compreensão da ingestão dietética recomendada de antioxidantes como adjuvante na prevenção da depressão feminina.

Dietary total antioxidant capacity as a preventive factor against depression in climacteric women / Capacidade antioxidante total dietética como um fator de prevenção contra a depressão em mulheres climatéricas

ABSTRACT Depression is characterized by physical or psychological distress and in many cases can lead to suicide. Objective: to assess the prevalence of depression and its possible relationship with dietary total antioxidant capacity (DTAC) and nutritional parameters in climacteric women participating in an extension university program in a Southern Brazilian city. Methods: data were obtained through questionnaires and anthropometric measurements. Diet was assessed using a 24-hour dietary recall. The Beck Depression Inventory (BDI) was used to assess the intensity of symptoms of depression. Results: DTAC of the population ranged from 435.60 to 4502.62 mg VCE/day. Among the most consumed antioxidant food/beverages, coffee ranked highest. Polyphenols were found to be directly linked to the antioxidant capacity of fresh foods (r=0.905; p=0.0001). Prevalence of depression in the population was 44%, and depressed women had lower intake levels of polyphenols (p=0.022; Cohen's d=0.80), and vitamin B6 (p=0.038; Cohen's d=0.65), vitamin A (p=0.044; Cohen's d =0.63), and vitamin C (p=0.050; Cohen's d =0.61). There was a significant negative correlation between BDI scores and polyphenol intake (r=-0.700; p=0.002). Conclusion: these results may contribute to a better understanding of the recommended dietary antioxidant intake as an adjuvant for preventing depression in women.

RESUMO Depressão é caracterizada por causar sofrimento físico ou psicológico e em muitos casos pode levar ao suicídio. Objetivo: avaliar a prevalência de depressão e sua possível relação com a capacidade antioxidante total da dieta (DTAC) e parâmetros nutricionais em mulheres climatéricas participantes de um programa universitário de extensão em uma cidade do sul do Brasil. Métodos: os dados foram obtidos por meio de questionários e medidas antropométricas. A dieta foi avaliada através de um recordatório alimentar de 24 horas. O Inventário de Depressão de Beck (BDI) foi usado para avaliar a intensidade dos sintomas de depressão. Resultados: a DTAC da população variou de 435,60 a 4502,62 mg VCE/dia. Entre os alimentos/bebidas antioxidantes mais ingeridos, o café destacou-se em primeiro. Os polifenóis mostraram estar diretamente ligados à capacidade antioxidante dos alimentos in natura (r=0,905; p=0,0001). A prevalência de depressão na população foi de 44%, e as mulheres depressivas apresentaram menores níveis de ingestão de polifenóis (p=0,022; Cohen's d=0,80) e vitaminas B6 (p=0,038; Cohen's d=0,65), A (p=0,044; Cohen's d=0,63) e C (p=0,050; Cohen's d=0,61). Houve uma correlação negativa significativa entre os escores do BDI e a ingestão de polifenóis (r=-0,700; p=0,002). Conclusão: estes resultados podem contribuir para uma melhor compreensão da ingestão dietética recomendada de antioxidantes como adjuvante na prevenção da depressão feminina.

Araucaria angustifolia (Bertol.) Kuntze has neuroprotective action through mitochondrial modulation in dopaminergic SH-SY5Y cells.

Brain disorders (BD) including neuropsychiatric and neurodegenerative diseases, are often associated with impairments in mitochondrial function and oxidative damage that can lead to neuronal injury. The mitochondrial complex I enzyme is one of the main sites of ROS generation and is implicated in many BD pathophysiologies. Despite advances in therapeutics for BD management, conventional pharmacotherapy still cannot efficiently control neuronal redox imbalance and mitochondrial dysfunction. Araucaria angustifolia is one of the main pine species in South America and presents a notable therapeutic history in folk medicine. A. angustifolia extract (AAE), obtained from the natural waste named bracts, is rich in flavonoids; molecules able to regulate cell redox metabolism. We examined the effects of AAE on rotenone-induced mitochondrial complex I dysfunction in human dopaminergic SH-SY5Y cells. AAE restored complex I assembly and activity mainly through overexpression of NDUFS7 protein and NDUFV2 gene levels. These findings were accompanied by a reduction in the generation of neuronal reactive oxygen species and lipid peroxidation. Our data demonstrates, for the first time, that AAE exerts in vitro neuroprotective effects, thus making it an interesting source for future drug development in BD-associated mitochondrial dysfunctions.

White grape juice increases high-density lipoprotein cholesterol levels and reduces body mass index and abdominal and waist circumference in women.

OBJECTIVES: Grape juice is a drink that is rich in phenolic compounds, which are important natural antioxidants that reduce the incidence of diseases linked to oxidative stress including cardiovascular disease. Studies on the effects of purple grape juice supplementation have already been conducted; however, there are no data on the effects of the consumption of white grape juice (WGJ) to date. The aim of this study was to evaluate the effects of WGJ supplementation on body mass index, waist and abdominal circumference, blood pressure and glucose, insulin, oxidative damage, and lipid profile in women. METHODS: A total of 25 women, ages 50 to 67 y, were included in this study. The volunteers were instructed to consume 7 mL/Kg/d of WGJ (Vitis labrusca) without other changes in diet energy consumption or habitual lifestyle. The supplementation occurred over 30 d. Anthropometric data and blood samples were collected before and after the intervention. RESULTS: Supplementation with WGJ reduced the women's body mass index and waist and abdominal circumference (P < 0.001). There were no significant differences in blood pressure before and after the consumption of WGJ. In addition, no changes in blood glucose, insulin, and levels of oxidative damage were found. Women who supplemented with WGJ showed an increase of 16% in high-density lipoprotein cholesterol levels. CONCLUSIONS: These results demonstrate that the consumption of WGJ can improve metabolic parameters in women, which may help reduce the risk of cardiovascular diseases.

Modulation of Mitochondrial and Epigenetic Targets by Polyphenols-rich Extract from Araucaria angustifolia in Larynx Carcinoma.

BACKGROUND: Araucaria angustifolia extract (AAE) is a polyphenol-rich extract that has gained interest as a natural anticancer agent. Recent work suggests that AAE induces oxidative damage and apoptosis through its action on decreasing complex I activity of the mitochondrial Electron Transport Chain (ETC). AIMS AND METHODS: In the present study, we aimed to further examine the specific targets by which AAE exerts proapoptotic effects in HEp-2 cancer cells. Specifically, the effect of AAE on the: 1) levels of pyruvate dehydrogenase was assessed by ELISA assay; 2) levels of mitochondrial ETC complexes, focusing on complex I at the gene transcript and protein level relevant to ROS generation was evaluated by multiplex ELISA followed by qRT-PCR and immunoblotting; 3) mitochondrial network distribution analysis was assessed by MitoTracker Red CMXRos; and 4) chemical variations on DNA was evaluated by dot-blotting in HEp-2 cells. RESULTS: Results demonstrated that AAE increased protein levels of PDH, switching energy metabolism to oxidative metabolism. Protein expression levels of complex I and III were found decreased in AAE-treated HEp-2 cells. Analyzing the subunits of complex I, changes in protein and gene transcript levels of NDUFS7 and NDUFV2 were found. Mitochondria staining after AAE incubation revealed changes in the mitochondrial network distribution. AAE was able to induce DNA hypomethylation and decreased DNA (cytosine-5)-methyltransferase 1 activity. CONCLUSION: Our data demonstrate for the first time that AAE alters expression of NDUFS7 and NDUFV2 mitochondrial subunits and induce epigenetic changes in HEp-2 cancer cells leading to a possible suppression of oncogenes.

Role of PON1, SOD2, OGG1, XRCC1, and XRCC4 polymorphisms on modulation of DNA damage in workers occupationally exposed to pesticides.

Tobacco farming has been proving to induce poor health outcomes in agricultural workers, genomic instability being the triggering one. This study evaluated influence of PON1 (paraoxonase 1), SOD2 (superoxide dismutase), OGG1 (8-oxoguanine glycosylase), XRCC1 (X-ray repair cross-complementing protein 1), and XRCC4 (X-ray repair cross-complementing protein 4) genes polymorphisms on DNA damage in 121 subjects occupationally exposed to pesticides mixtures and nicotine at tobacco fields and 121 non-exposed individuals. Inorganic elements (Cl, P, S and Zn) and cotinine levels were found increased in farmers, confirming exposure. Results show higher frequencies of buccal micronucleus (MN), nuclear buds (NBUD), binucleated cells (BN) and damage index (comet assay), reduced telomere length (TL), and increased parameters of oxidative stress in farmers compared to non-exposed individuals. PON1 Gln/Gln genotype was associated with increased MN frequency. SOD2 Val/Val showed association with increased frequency of MN and NBUD and decreased antioxidant activity. The XRCC1 Arg/Arg showed protective effect for MN, BN and TL, which was also positively influenced by OGG1 -/Cys. MN was decreased in XRCC4 -/Ile farmers. These genotypes also showed a risk for antioxidant activity. Our study proposes that PON1 and SOD2 variants play a role in xenobiotic-metabolizing system in farmers, while base excision repair (BER) pathway could be the repair mechanism involved in genomic instability suffered by tobacco farmers.

Chronic occupational exposure endured by tobacco farmers from Brazil and association with DNA damage.

Tobacco farming is an important economic income in Brazil, although it has been challenged as regard the occupational exposure to both pesticides and nicotine endured by farmers. Chronic occupational exposure to complex mixtures can lead to health hazardous. We examined genomic instability and epigenetic changes in tobacco farmers occupationally exposed to pesticide mixtures and nicotine at tobacco fields. DNA damage was assessed by alkaline comet assay in blood cells. Genomic DNA was isolated, and telomere length was measured using quantitative polymerase chain reaction assay. We measured 5-methyl-2'-deoxycytidine, a marker of global DNA methylation, and p16 promoter methylation. The oxidative profile was evaluated by trolox equivalent antioxidant capacity and lipid peroxidation (thiobarbituric acid reactive substances) in serum. Exposure parameters, plasma cotinine and inorganic element levels, were also measured. DNA damage was significantly elevated for farmers in relation to unexposed group (P < 0.001; Mann-Whitney test) and positively associated with years of exposure. Inverse relationship between DNA damage and total equivalent antioxidant activity was demonstrated for exposed and unexposed groups. Exposed group showed significantly shorter telomeres (P < 0.001; unpaired t-test) and DNA hypomethylation (P < 0.001; unpaired t-test), as well as p16 hypermethylation (P = 0.003; Mann-Whitney test). Lipid peroxidation was increased for exposed group in relation to unexposed one (P = 0.02; Mann-Whitney test) and presented a positive correlation with global DNA methylation (P = 0.0264). Farmers have increased plasma cotinine levels (P < 0.001) and inorganic elements (phosphorus, sulphur and chlorine) in relation to unexposed group. Elevated oxidative stress levels due to chronic occupational pesticide mixtures and nicotine exposure in tobacco farmers were associated with higher DNA damage, shorter telomeres and altered DNA methylation. Telomere-accelerated attrition due to exposure may be potential intermediate step before a disease state.

Antioxidant defences and haemocyte internalization in Limnoperna fortunei exposed to TiO2 nanoparticles.

TiO2 nanoparticles (TiO2-NP) have been incorporated into a large range of materials for different applications in the last decades and are very likely to appear in wastewater and effluents, eventually reaching the aquatic environment. Therefore, the assessment of the biological impact of TiO2-NP on aquatic ecosystem is of a major concern. The mussels represent a target group for TiO2-NP toxicity, as they are filter feeders and are capable of bioaccumulating toxic compounds. Furthermore, the exotic organism Limnoperna fortunei, golden mussel, is a freshwater bivalve that has been used in biomonitoring environmental conditions. In this work, the TiO2-NP's ability to interact with haemocytes of golden mussel was assessed by transmission electron microscopy. The enzymatic and non-enzymatic antioxidant defenses were evaluated by superoxide dismutase (Sod) and catalase (Cat) activities and protein sulfhydryl content, which were measured after the golden mussel was exposed to TiO2-NP (1, 5, 10 and 50µgmL(-1)). Results demonstrate that TiO2-NP was internalized by cells, causing alterations in haemocytes membrane. Antioxidant activity of Sod and Cat decreased after 2h TiO2-NP exposure. After 4h exposure, the enzymatic antioxidant activity was restored. Notably, the protein sulfhydryl content decreased after 2h to all the TiO2-NP concentrations and no alterations were observed after 4h of TiO2-NP exposure. These results demonstrate the potential of golden mussel as sentinel organism to TiO2-NP exposure.

Pharmacological perspectives from Brazilian Salvia officinalis (Lamiaceae): antioxidant, and antitumor in mammalian cells.

Salvia officinalis (Lamiaceae) has been used in south of Brazil as a diary homemade, in food condiment and tea-beverage used for the treatment of several disorders. The objective of this study was to characterize chemical compounds in the hydroalcoholic (ExtHS) and aqueous (ExtAS) extract from Salvia officinalis (L.) by gas chromatography-mass spectrometry (GC-MS) and by high-resolution electrospray ionization mass spectrometry (ESI-QTOF MS/MS), evaluate in vitro ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), catalase (CAT-like) and superoxide dismutase (SOD-like) activity, moreover cytotoxic by MTT assay, alterations on cell morphology by giemsa and apoptotic-induced mechanism for annexin V/propidium iodide. Chemical identification sage extracts revealed the presence of acids and phenolic compounds. In vitro antioxidant analysis for both extracts indicated promising activities. The cytotoxic assays using tumor (Hep-2, HeLa, A-549, HT-29 and A-375) and in non-tumor (HEK-293 and MRC-5), showed selectivity for tumor cell lines. Immunocytochemistry presenting a majority of tumor cells at late stages of the apoptotic process and necrosis. Given the results presented here, Brazilian Salvia officinalis (L.) used as condiment and tea, may protect the body against some disease, in particularly those where oxidative stress is involved, like neurodegenerative disorders, inflammation and cancer.