No Effects of Decanoic Acid on Locomotor Activity and Antioxidant Defences in an Experimental Animal Model of Attention-Deficit/Hyperactivity Disorder.

BACKGROUND: Medium-chain triglycerides such as decanoic acid (C10), which is one of the fatty acids that constitute dietary fats, are of substantial interest for their potential therapeutic effects on neuropsychiatric disorders. However, the effects of C10 on attention-deficit/hyperactivity disorder (ADHD) remain to be studied. We explored the effects of C10 on behavioural activity and antioxidant defences in an experimental animal model of ADHD. METHODS: To establish an experimental animal model of ADHD, neonatal rats were subjected to unilateral striatal lesions using 6-hydroxydopamine (6-OHDA). The rats sequentially underwent open-field and Y-maze tests before treatment [postnatal day 25 (PN25)]. After the subcutaneous administration of either vehicle or C10 solution (250 mg/kg) for 14 days, the behavioural tests were repeated on PN39. Next, we examined the effects of C10 on the expression of the constitutive antioxidant enzymes catalase and glutathione peroxidase-1/2 and the phase II transcription factor nuclear factor erythroid 2-related factor 2 in four different regions of the rat brain. RESULTS: Injection of 6-OHDA unilaterally into the striatum resulted in elevated locomotor activity on PN39. The administration of C10 for a period of 14 days did not alter the locomotor hyperactivity. Moreover, the administration of C10 had no significant effects on the expression of proteins related to antioxidant defences in the hippocampus, prefrontal cortex, striatum or cerebellum of both control and lesioned rats. CONCLUSIONS: The lack of significant effects of C10 in our study may depend on the dose and duration of C10 administration. Further exhaustive studies are needed to verify the efficacy and effects of different doses and treatment durations of C10 and to explore the underlying mechanisms.

The Effect of Neuronal CoQ10 Deficiency and Mitochondrial Dysfunction on a Rotenone-Induced Neuronal Cell Model of Parkinson's Disease.

Parkinson's disease (PD) is the second most prevalent neurodegenerative disorder currently affecting the ageing population. Although the aetiology of PD has yet to be fully elucidated, environmental factors such as exposure to the naturally occurring neurotoxin rotenone has been associated with an increased risk of developing PD. Rotenone inhibits mitochondrial respiratory chain (MRC) complex I activity as well as induces dopaminergic neuronal death. The aim of the present study was to investigate the underlying mechanisms of rotenone-induced mitochondrial dysfunction and oxidative stress in an in vitro SH-SY5Y neuronal cell model of PD and to assess the ability of pre-treatment with Coenzyme Q10 (CoQ10) to ameliorate oxidative stress in this model. Spectrophotometric determination of the mitochondrial enzyme activities and fluorescence probe studies of reactive oxygen species (ROS) production was assessed. Significant inhibition of MRC complex I and II-III activities was observed, together with a significant loss of neuronal viability, CoQ10 status, and ATP synthesis. Additionally, significant increases were observed in intracellular and mitochondrial ROS production. Remarkably, CoQ10 supplementation was found to reduce ROS formation. These results have indicated mitochondrial dysfunction and increased oxidative stress in a rotenone-induced neuronal cell model of PD that was ameliorated by CoQ10 supplementation.

Bifurcaria bifurcata extract exerts antioxidant effects on human Caco-2 cells.

The present research study investigated the potential protective effect of Bifurcaria bifurcata extract on cell viability and antioxidant defences of cultured human Caco-2 cells submitted to oxidative stress induced by tert-butylhydroperoxide (tert-BOOH). Aqueous extracts were firstly characterized in terms of total phenolic contents. Concentrations of reduced glutathione (GSH) and malondialdehyde (MDA), generation of reactive oxygen species (ROS), nitric oxide (NO) production, antioxidant enzymes activities [NADPH quinone dehydrogenase 1 (NQO1) and glutathione S-transferase (GST)], caspase 3/7 activity and gene expression linked to apoptosis, proinflammation and oxidative stress signaling pathways were used as markers of cellular oxidative status. B. bifurcata extract prevented the cytotoxicity, the decrease of GSH, the increase of MDA levels and the ROS generation induced by tert-BOOH. B. bifurcata extract prevented the significant decrease of NQO1 and GST activities, and the significant increase of caspase 3/7 activity induced by tert-BOOH. B. bifurcata extract also caused an over-expression of GSTM2, Nrf2 and AKT1 transcriptors, as well as reduced ERK1, JNK1, Bax, BNIP3, NFκB1, IL-6 and HO-1 gene expressions induced by tert-BOOH suggesting an increase in cellular resistance against oxidative stress. The results of the biomarkers analyzed show that treatment of Caco-2 cells with B. bifurcata extract enhance antioxidant defences, which imply an improved cell response to an oxidative challenge. B. bifurcata extract possesses strong antioxidant properties and may be a potential effective alternative to oxidant agents in the functional food industry.

Early redox homeostasis disruption contributes to the differential cytotoxicity of imiquimod on transformed and normal endothelial cells.

The off-label use of imiquimod (IQ) for hemangioma treatment has shown clinical benefits. We have previously reported a selective direct IQ-cytotoxic effect on transformed (H5V) vs. normal (1G11) endothelial cells (EC). In the present study, we investigated the mechanism underlying this selective cytotoxicity in terms of TLR7/8 receptor expression, NF-κB signalling and time-dependent modifications of oxidative stress parameters (ROS: reactive oxygen species, catalase and superoxide dismutase activities, GSH/GSSG and lipid peroxidation). TLR7/8 level was extremely low in both cell lines, and IQ did not upregulate TLR7/8 expression or activate NF-κB signalling. IQ significantly induced ROS in H5V after 2 h and strongly affected antioxidant defenses. After 12 h, enzyme activities were restored to baseline levels but a robust drop in GSH/GSSG persisted together with increased lipid peroxidation levels and a marked mitochondrial dysfunction. Although in normal IQ-treated EC some oxidative stress parameters were affected after 4 h, mitochondrial health and GSH/GSSG ratio remained notably unaffected after 12 h. Therefore, the early alterations (0-2 h) in transformed EC breached redox homeostasis as strongly as to enhance their susceptibility to IQ. This interesting facet of IQ as redox disruptor could broaden its therapeutic potential for other skin malignancies, alone or in adjuvant schemes.

Circulating biomarkers of nitric oxide bioactivity and impaired muscle vasoreactivity to exercise in adults with uncomplicated type 1 diabetes.

AIMS/HYPOTHESIS: Early compromised endothelial function challenges the ability of individuals with type 1 diabetes to perform normal physical exercise. The exact mechanisms underlying this vascular limitation remain unknown, but may involve either formation or metabolism of nitric oxide (NO), a major vasodilator, whose activity is known to be compromised by oxidative stress. METHODS: Muscle microvascular reactivity (near-infrared spectroscopy) to an incremental exhaustive bout of exercise was assessed in 22 adults with uncomplicated type 1 diabetes (HbA1c 64.5 ± 15.7 mmol/mol; 8.0 ± 1.4%) and in 21 healthy individuals (18-40 years of age). NO-related substrates/metabolites were also measured in the blood along with other vasoactive compounds and oxidative stress markers; measurements were taken at rest, at peak exercise and after 15 min of recovery. Demographic characteristics, body composition, smoking status and diet were comparable in both groups. RESULTS: Maximal oxygen uptake was impaired in individuals with type 1 diabetes compared with in healthy participants (35.6 ± 7.7 vs 39.6 ± 6.8 ml min-1 kg-1, p < 0.01) despite comparable levels of habitual physical activity (moderate to vigorous physical activity by accelerometery, 234.9 ± 160.0 vs 280.1 ± 114.9 min/week). Compared with non-diabetic participants, individuals with type 1 diabetes also displayed a blunted exercise-induced vasoreactivity (muscle blood volume at peak exercise as reflected by ∆ total haemoglobin, 2.03 ± 5.82 vs 5.33 ± 5.54 µmol/l; interaction 'exercise' × 'group', p < 0.05); this was accompanied by lower K+ concentration (p < 0.05), reduced plasma L-arginine (p < 0.05)-in particular when HbA1c was high (mean estimation: -4.0, p < 0.05)-and lower plasma urate levels (p < 0.01). Nonetheless, exhaustive exercise did not worsen lipid peroxidation or other oxidative stress biomarkers, and erythrocytic enzymatic antioxidant resources were mobilised to a comparable extent in both groups. Nitrite and total nitrosation products, which are potential alternative NO sources, were similarly unaltered. Graphical abstract CONCLUSIONS/INTERPRETATION: Participants with uncomplicated type 1 diabetes displayed reduced availability of L-arginine, the essential substrate for enzymatic nitric oxide synthesis, as well as lower levels of the major plasma antioxidant, urate. Lower urate levels may reflect a defect in the activity of xanthine oxidase, an enzyme capable of producing NO from nitrite under hypoxic conditions. Thus, both canonical and non-canonical NO production may be reduced. However, neither of these changes exacerbated exercise-induced oxidative stress. TRIAL REGISTRATION: clinicaltrials.gov NCT02051504.

Bisphenol S leads to cytotoxicity-induced antioxidant responses and oxidative stress in isolated rainbow trout (Oncorhyncus mykiss) hepatocytes.

BACKGROUND: Bisphenol S (BPS) is a chemical compound that is utilized in the plastic industry as an alternative to bisphenol A (BPA). The toxic effects of BPS in fish is less known and limited. Therefore, in the present study, the influence of BPS on rainbow trout (Oncorhyncus mykiss) hepatocytes in vitro was investigated. METHODS AND RESULTS: For this purpose the fish hepatocytes were isolated, and then the cultured cells were treated with increasing concentrations of BPS (0, 15.63, 31.25, 62.50, 125, 250, and 500 µM) for 24 h. The cytotoxic impact of BPS was determined in the culture media using lactate dehydrogenase assay and then, the antioxidant defence indicators were assayed. The results showed that concentration-dependent increases were observed in the percentage of cytotoxicity. The superoxide dismutase activity was reduced, while the catalase and glutathione peroxidase activity increased with all of the BPS concentrations. The glutathione S-transferase (GST) activity significantly increased after a BPS concentration of 31.25 µM or higher, while GST Theta 1-1 activity was decreased by the same concentrations of BPS. The reduced glutathione content significantly decreased with a BPS concentration of 31.25 µM or higher, and the malondialdehyde content increased after BPS concentrations of 125, 250, and 500 µM. CONCLUSIONS: The findings determined herein suggested that BPS causes cytotoxicity in fish hepatocytes and can lead to oxidative stress, resulting hepatotoxic in fish. Thus, the utilization of BPS instead of BPA as safe alternative in industry should be re-evaluated in the future for environmental health.

Selective dendritic susceptibility to bioenergetic, excitotoxic and redox perturbations in cortical neurons.

Neurodegenerative and neurological disorders are often characterised by pathological changes to dendrites, in advance of neuronal death. Oxidative stress, energy deficits and excitotoxicity are implicated in many such disorders, suggesting a potential vulnerability of dendrites to these situations. Here we have studied dendritic vs. somatic responses of primary cortical neurons to these types of challenges in real-time. Using a genetically encoded indicator of intracellular redox potential (Grx1-roGFP2) we found that, compared to the soma, dendritic regions exhibited more dramatic fluctuations in redox potential in response to sub-lethal ROS exposure, and existed in a basally more oxidised state. We also studied the responses of dendritic and somatic regions to excitotoxic NMDA receptor activity. Both dendritic and somatic regions experienced similar increases in cytoplasmic Ca²âº. Interestingly, while mitochondrial Ca²âº uptake and initial mitochondrial depolarisation were similar in both regions, secondary delayed mitochondrial depolarisation was far weaker in dendrites, potentially as a result of less NADH depletion. Despite this, ATP levels were found to fall faster in dendritic regions. Finally we studied the responses of dendritic and somatic regions to energetically demanding action potential burst activity. Burst activity triggered PDH dephosphorylation, increases in oxygen consumption and cellular NADH:NAD ratio. Compared to somatic regions, dendritic regions exhibited a smaller degree of mitochondrial Ca²âº uptake, lower fold-induction of NADH and larger reduction in ATP levels. Collectively, these data reveal that dendritic regions of primary neurons are vulnerable to greater energetic and redox fluctuations than the cell body, which may contribute to disease-associated dendritic damage. This article is part of a Special Issue entitled: 13th European Symposium on Calcium.

Differences in the oxidative balance of dispersing and non-dispersing individuals: an experimental approach in a passerine bird.

BACKGROUND: Dispersal is often associated with a suite of phenotypic traits that might reduce dispersal costs, but can be energetically costly themselves outside dispersal. Hence, dispersing and philopatric individuals might differ throughout their life cycle in their management of energy production. Because higher energy expenditure can lead to the production of highly reactive oxidative molecules that are deleterious to the organism if left uncontrolled, dispersing and philopatric individuals might differ in their management of oxidative balance. Here, we experimentally increased flight costs during reproduction via a wing load manipulation in female collared flycatchers (Ficedula albicollis) breeding in a patchy population. We measured the effects of the manipulation on plasmatic markers of oxidative balance and reproductive success in dispersing and philopatric females. RESULTS: The impact of the wing load manipulation on the oxidative balance differed according to dispersal status. The concentration of reactive oxygen metabolites (ROMs), a marker of pro-oxidant status, was higher in philopatric than dispersing females in the manipulated group only. Differences between dispersing and philopatric individuals also depended on habitat quality, as measured by local breeding density. In low quality habitats, ROMs as well as nestling body mass were higher in philopatric females compared to dispersing ones. Independently of the manipulation or of habitat quality, plasma antioxidant capacity differed according to dispersal status: philopatric females showed higher antioxidant capacity than dispersing ones. Nestlings raised by philopatric females also had a higher fledging success. CONCLUSIONS: Our results suggest that dispersing individuals maintain a stable oxidative balance when facing challenging environmental conditions, at the cost of lower reproductive success. Conversely, philopatric individuals increase their effort, and thus oxidative costs, in challenging conditions thereby maintaining their reproductive success. Our study sheds light on energetics and oxidative balance as possible processes underlying phenotypic differences between dispersing and philopatric individuals.

Propensity to metal accumulation and oxidative stress responses of two benthic species (Cerastoderma edule and Nephtys hombergii): are tolerance processes limiting their responsiveness?

The chronic exposure of benthic organisms to metals in sediments can lead to the development of tolerance mechanisms, thus diminishing their responsiveness. This study aims to evaluate the accumulation profiles of V, Cr, Co, Ni, As, Cd, Pb and Hg and antioxidant system responses of two benthic organisms (Cerastoderma edule, Bivalvia; Nephtys hombergii, Polychaeta). This approach will provide clarifications about the ability of each species to signalise metal contamination. Organisms of both species were collected at the Tagus estuary, in two sites with distinct contamination degrees (ALC, slightly contaminated; BAR, highly contaminated). Accordingly, C. edule accumulated higher concentrations of As, Pb and Hg at BAR compared to ALC. However, antioxidant responses of C. edule were almost unaltered at BAR and no peroxidative damage occurred, suggesting adjustment mechanisms to the presence of metals. In contrast, N. hombergii showed a minor propensity to metal accumulation, only signalising spatial differences for As and Pb and accumulating lower concentrations of metals than C. edule. The differences in metal accumulation observed between species might be due to their distinctive foraging behaviour and/or the ability of N. hombergii to minimise the metal uptake. Despite that, the accumulation of As and Pb was on the basis of the polychaete antioxidant defences inhibition at BAR, including CAT, SOD, GR and GPx. The integrated biomarker response index (IBRv2) confirmed that N. hombergii was more affected by metal exposure than C. edule. In the light of current findings, in field-based studies, the information of C. edule as a bioindicator should be complemented by that provided by another benthic species, since tolerance mechanisms to metals can hinder a correct diagnosis of sediment contamination and of the system's health. Overall, the present study contributed to improve the lack of fundamental knowledge of two widespread and common estuarine species, providing insights of the metal accumulation profiles under a scenario of chronic contamination. Finally, this work provided useful information that can be applied in the interpretation of future environmental monitoring studies.

Gut morphology and hepatic oxidative status of European sea bass (Dicentrarchus labrax) juveniles fed plant feedstuffs or fishmeal-based diets supplemented with short-chain fructo-oligosaccharides and xylo-oligosaccharides.

The effects of short-chain fructo-oligosaccharides (scFOS) and xylo-oligosaccharides (XOS) on gut morphology and hepatic oxidative status were studied in European sea bass juveniles weighing 60 g. Fish were fed diets including fishmeal (FM diets) or plant feedstuffs (PF diets; 30 FM:70 PF) as main protein sources (control diets). Four other diets were formulated similar to the control diets but including 1 % scFOS or 1 % XOS. At the end of the trial, fish fed PF-based diets presented histomorphological alterations in the distal intestine, whereas only transient alterations were observed in the pyloric caeca. Comparatively to fish fed FM-based diets, fish fed PF diets had higher liver lipid peroxidation (LPO), superoxide dismutase (SOD) and catalase (CAT) activities, and lower glutathione peroxidase, glutathione reductase and glucose 6-phosphate dehydrogenase activities. In fish fed the PF diets, prebiotic supplementation decreased SOD activity and XOS supplementation further decreased CAT activity. In fish fed the FM diets, XOS supplementation promoted a reduction of all antioxidant enzyme activities. Overall, dietary XOS and scFOS supplementation had only minor effects on gut morphology or LPO levels. However, dietary XOS reduced antioxidant enzymatic activity in both PF and FM diets, which indicate a positive effect on reduction of hepatic reactive oxygen species production.

Systemic Oxidative Stress in Subacute Stroke Patients Undergoing Rehabilitation Treatment.

The imbalance in oxidative stress in acute stroke has been extensively studied; on the contrary, its investigation in the subacute phase is limited. The aim of this study was to analyse the variation in the systemic oxidative status in subacute post-stroke patients before (T0) and after a six-week rehabilitation treatment (T1) and to investigate the relationship between systemic oxidative status and rehabilitation outcomes. We enrolled 109 subjects in two different centres, and we analysed their serum hydroperoxide levels (d-ROMs), biological antioxidant power (BAP), thiol antioxidant components (-SHp), and relative antioxidant capacity (OSI and SH-OSI indices). Activity of Daily Living (ADL), hand grip strength, and walking endurance were evaluated using the modified Barthel Index, the Hand Grip test, and the 6-min walk test, respectively. At T0, most of the patients showed very high levels of d-ROMs and suboptimal levels of the BAP, OSI, and SH-OSI indices. Comparing the T1 and T0 data, we observed an improvement in the rehabilitation outcomes and a significant decrease in d-ROMs (549 ± 126 vs. 523 ± 148, p = 0.023), as well as an improvement in the OSI and SH-OSI indices (4.3 ± 1.3 vs. 4.7 ± 1.5, p = 0.001; 11.0 ± 0.4 vs. 1.2 ± 0.4, p < 0.001). In addition, significant correlations were seen between the oxidative stress parameters and the rehabilitation outcomes. These results suggest monitoring the systemic oxidative stress status in post-stroke patients in order to plan a tailored intervention, considering its relationship with functional recovery.

Chlorophyll fluorescence characteristics and H2O2 contents of Chinese tallow tree are dependent on population origin, nutrients and salinity.

Plants from invasive populations often have higher growth rates than conspecifics from native populations due to better environmental adaptability. However, the roles of improved chlorophyll fluorescence or antioxidant defenses in helping them to grow better under adverse situations are insufficient, even though this is a key physiological question for elucidating mechanisms of plant invasion. Here, we conducted experiments with eight native (China) and eight introduced (USA) populations of Chinese tallow tree (Triadica sebifera). We tested how salinity, nutrients (overall amount or N:P in two separate experiments) and their interaction affected T. sebifera aboveground biomass, leaf area, chlorophyll fluorescence and antioxidant defenses. Plants from introduced populations were larger than those from native populations, but salinity and nutrient shortage (low nutrients or high N:P) reduced this advantage, possibly reflecting differences in chlorophyll fluorescence based on their higher PSII maximum photochemical efficiency (F v/F m) and PSI maximum photo-oxidizable P700 in higher nutrient conditions. Native population plants had lower F v/F m with saline. Except in high nutrients/N:P with salinity, introduced population plants had lower electron transfer rate and photochemical quantum yield. There were no differences in antioxidant defenses between introduced and native populations except accumulation of hydrogen peroxide (H2O2), which was lower for introduced populations. Low nutrients and higher N:P or salinity increased total antioxidant capacity and H2O2. Our results indicate that nutrients and salinity induce differences in H2O2 contents and chlorophyll fluorescence characteristics between introduced and native populations of an invasive plant, illuminating adaptive mechanisms using photosynthetic physiological descriptors in order to predict invasions.

Developmental post-natal stress can alter the effects of pre-natal stress on the adult redox balance.

Across diverse vertebrate taxa, stressful environmental conditions during development can shape phenotypic trajectories of developing individuals, which, while adaptive in the short-term, may impair health and survival in adulthood. Regardless, the long-lasting benefits or costs of early life stress are likely to depend on the conditions experienced across differing stages of development. Here, we used the Japanese quail (Coturnix coturnix japonica) to experimentally manipulate exposure to stress hormones in developing individuals. We tested the hypothesis that interactions occurring between pre- and post-natal developmental periods can induce long-term shifts on the adult oxidant phenotype in non-breeding sexually mature individuals. We showed that early life stress can induce long-term alterations in the basal antioxidant defences. The magnitude of these effects depended upon the timing of glucocorticoid exposure and upon interactions between the pre- and post-natal stressful stimuli. We also found differences among tissues with stronger effects in the erythrocytes than in the brain in which the long-term effects of glucocorticoids on antioxidant biomarkers appeared to be region-specific. Recent experimental work has demonstrated that early life exposure to stress hormones can markedly reduce adult survival (Monaghan et al., 2012). Our results suggest that long-term shifts in basal antioxidant defences might be one of the potential mechanisms driving such accelerated ageing processes and that post-natal interventions during development may be a potential tool to shape the effects induced by pre-natally glucococorticoid-exposed phenotypes.

Counteraction of oxidative damage by pomegranate juice: influence of the cultivar.

BACKGROUND: Pomegranate (Punica granatum) has gained widespread popularity as a potential functional food due to its high phenolics content. Although in different studies pomegranate juice has been shown to exert anti-atherogenic, anti-inflammatory and antioxidant effects, the cultivar-related differences in phenolics content and antioxidant activity must be carefully taken into account when evaluating the health effects. The first aim of this study was to rank the juices of 15 different varieties of pomegranate according to their total phenolic content and antioxidant capacity. Then three juices were selected, better characterised and added to HepG2 cells as a supplement to evaluate the protective effect against induced oxidative stress. RESULTS: The cultivars Wonderful, Hicaz and G2 were chosen according to the ranking and the corresponding juices used at two different concentrations as a supplement to HepG2 cells. Overall, all juices were able to protect cells from oxidative damage, but differences that could be related to the different phenolics content and pattern were detected among the three juices. CONCLUSION: Our results show the advantage of screening cultivars prior to efficacy studies. This approach can be useful for food companies that focus on the development of food that has added nutritional and health value.

Bioactive-rich Sideritis scardica tea (mountain tea) is as potent as Camellia sinensis tea at inducing cellular antioxidant defences and preventing oxidative stress.

BACKGROUND: In several countries, tea (hot-water infusions of dried Camellia sinensis (CS) leaves) is a major source of antioxidant flavonoids, and its consumption has been associated with several favourable outcomes. Other plants used for the preparation of herbal teas are sources of phenolic antioxidant compounds; among them Sideritis scardica (SS) is used for the preparation of a popular drink throughout Eastern and Central Europe. We have compared the effects of an SS extract to a CS extract in HepG2 cells to set the scientific basis for the exploitation of other herbal teas in counteraction of oxidative stress. RESULTS: Although SS extract had a lower phenolic concentration and total antioxidant capacity than CS extract, their cellular antioxidant effects were similar. The different phenolic pattern of the extracts suggests that the protective activity is not limited to catechins. CONCLUSION: Although further research is needed, our data represent a first contribution for the evaluation of the potential effect of SS in increasing antioxidant defences. © 2013 Society of Chemical Industry.

Atomoxetine Decreases Mitochondrial Biogenesis, Fission and Fusion In Human Neuron-like Cells But Does Not Alter Antioxidant Defences.

Atomoxetine (ATX) is a presynaptic norepinephrine transporter (NET) inhibitor widely prescribed for attention-deficit/hyperactivity disorder (ADHD) due to its low abuse potential and absence of psychostimulant effects. While NET inhibition is implicated in the clinical response, several additional pharmacoactivities may contribute to clinical efficacy or unwanted side effects. We recently reported that ATX can dose-dependently alter mitochondrial function and cellular redox status. Here, we assessed potential alterations in mitochondrial biogenesis, mitochondrial dynamics and cellular antioxidant capacity following high- and low-dose ATX treatment of differentiated human neuroblastoma cells. Human SH-SY5Y neuroblastoma cells were treated with ATX (1, 5, 10, 20 and 50 µM) for 7 days under differentiation culture conditions. Changes in the expression levels of protein markers for mitochondrial biogenesis, fusion and fission as well as of antioxidant proteins were analysed by Western blot. High-dose ATX (50 µM) reduced while low-dose ATX (10 µM) increased mitochondrial biogenesis as evidenced by parallel changes in SDHA, COX-I, PGC1α and TFAM expression. High-dose ATX also reduced mitochondrial fusion as evidenced by OPA1 and MFN2 downregulation, and mitochondrial fission as indicated by DRP1 and Fis1 downregulation. In contrast, ATX did not alter expression of the antioxidant enzymes SOD1 and catalase, the phase II transcription factor Nfr2, or the Nfr2-regulated antioxidant enzyme NQO1. Clinical responses and side effects of ATX may be mediated by dose-dependent modulation of mitochondrial biogenesis and dynamics as well as NET inhibition.

Impact of carob (Ceratonia siliqua L.) pulp inclusion and warm season on gastrointestinal morphological parameters, immune-redox defences and coccidiosis in concentrate-fed light lambs.

This study aimed to evaluate the effects of dietary carob (Ceratonia siliqua L.) pulp and warm season on gastrointestinal morphological parameters, immune-redox defences and coccidiosis in concentrate-fed light lambs. Weaned lambs were assigned to one of three concentrate-based diets: C0 (without carob pulp), C15 (150 g/kg of carob pulp) and C30 (300 g/kg of carob pulp) from 40 to 80 days of age in two consecutive cold and warm batches. Blood samples were collected at Day 80 to determine the metabolic status. Rectal faeces were sampled at Days 50, 65 and 80 to determine consistency and oocyst count per gram. Inclusion of carob pulp in lamb diets did not affect lamb growth but reduced coccidia oocyst excretion, improved faecal consistency and gastrointestinal morphological parameters, enhancing the ruminal thickness of the papilla living strata and reducing the darkness of the epithelium colour. Moreover, carob condensed tannins in the lambs' diet enhanced the expression of antioxidant SOD2 in rumen, while down-regulated NRF2, SOD1, CAT and PPARG in ileum. There was no interaction between the treatments and season in the evaluated variables. Lambs from the warm season exhibited reduced growth performance, altered ruminal epithelium, lower circulating iron levels, increased protein concentrations and higher coccidiosis susceptibility. In addition, regulatory immune and antioxidant mechanisms to counterbalance reactive oxygen species production in gastrointestinal tissues were evident. Dietary inclusion of carob pulp (150 and 300 g/kg) in lamb diets improved gastrointestinal health and homeostasis but did not ameliorate the deleterious effects of warm season.

Cadmium effects on the freshwater teleost Prochilodus lineatus: Accumulation and biochemical, genotoxic, and behavioural biomarkers.

In order to evaluate the effects of Cd, juveniles of the Neotropical fish Prochilodus lineatus were exposed to 1 and 10 µg L-1 Cd, for 24 and 96 h. Fish exposed to Cd showed metal accumulation in the gills, kidney, and liver, an increase in DNA damage in erythrocytes, and an increase in lipid peroxidation (LPO) in the kidney. Cd exposure also caused a reduction in catalase activity, metallothionein induction, and LPO in the liver. Cd stimulated the swimming activity of exposed fish, resulting in longer swimming times and distances travelled, especially for the shortest exposure time. Changes in acetylcholinesterase activity (AChE) in the muscle and brain are probably related to these behavioural responses. These results show that Cd affects the functioning of several organs in P. lineatus, which is indicated by the genotoxic damage and changes in the AChE and swimming pattern of the exposed fish.

Oxidative damages and antioxidant defences after feeding a single meal in rainbow trout.

Feeding and digestion are metabolically demanding causing a rise on metabolic rate called Specific Dynamic Action (SDA). Although SDA has been vastly reported in fish, its potential consequences on the oxidative-antioxidant balance has not been evaluated to date in fish, a model with a long alkaline tide associated with feeding as well. Using rainbow trout (Oncorhynchus mykiss) as a model species, the aims of the present study were to: (1) assess potential oxidative damages and changes in oxidative defences after feeding on a single meal, and (2) identify the timescale of such changes over a 96 h post-feeding period. Oxidative damage in proteins and lipids and the activities of four enzymatic antioxidant defences: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) were measured in gill, stomach, intestine and liver. DNA damage was measured in red blood cells. Fish were sampled before and after 1.5, 6, 24, 48, 72 and 96 h of ingestion of a 3% body mass ration. Trends of post-prandial damage were present in all tissues, but only protein oxidation varied significatively during digestion in the stomach. The intestine and stomach presented the highest enzymatic activities, likely due to the high metabolic action that these tissues have during digestion, with peaks during post-feeding: at 24 h of SOD in stomach and at 48 h of CAT in intestine. Observed GPx peaks during post-feeding in gills are likely due to the exacerbated demands for ion fluxes and/or oxygen during feeding. The differential response of the antioxidant system observed in tissues of rainbow trout during digestion indicates a coordinated and tissue-specific antioxidant defence.

Stress response to trace elements mixture of different embryo-larval stages of Paracentrotus lividus.

This study investigated for the first time the oxidative biomarkers responses in all larval stages of sea urchin. The contamination effects were reproduced by using contaminated seawater to concentrations measured in the area adjacent to an old asbestos mine at factors of 5 and 10. The results suggested that the concentrations were not sufficiently high to induce a major oxidative stress. The biometric differences make this method a more sensitive approach for assessing the effects on sea urchin larvae. Measurements of specific activities of antioxidant enzymes at each stage suggested a high capacity of the larvae to respond to oxidative stress. This normal activity of the organism must be considered in future research. This work also highlighted the importance of spawners provenance in ecotoxicological studies. These data are essential to better understand the stress responses of sea urchin larvae and provide baseline information for later environmental assessment research.