Exercise-induced heart mitochondrial cholesterol depletion influences the inhibition of mitochondrial swelling.

The significance of the reduction of the cholesterol pool in heart mitochondria after exercise is still unknown. Recently, published data have suggested that cholesterol may influence the components of mitochondrial contact site and affect mitochondrial swelling. Therefore, the aim of this study was to determine whether the decreased cholesterol content in heart mitochondria caused by prolonged swimming may provoke changes in their bioenergetics and result in an increased resistance to calcium chloride-induced mitochondrial swelling. Male Wistar rats were divided into a sedentary control group and an exercise group. The rats exercised for 3 h, burdened with an additional 3% of their body weight. Their hearts were removed immediately after completing the exercise. The left ventricle was divided and used for experiments. Mitochondrial cholesterol content, membrane fluidity and mitochondrial bioenergetics were measured in the control and exercised rat heart mitochondria. To assess whether mitochondrial modifications are linked to disruption of lipid microdomains, methyl-ß-cyclodextrin, a well-known lipid microdomain-disrupting agent and cholesterol chelator, was applied to the mitochondria of the control group. Cholesterol depletion, increased membrane fluidity and increased resistance to calcium chloride-induced swelling were observed in postexercise heart crude mitochondrial fraction. Similar results were achieved in control mitochondria treated with 2% methyl-ß-cyclodextrin. All of the mitochondrial bioenergetics parameters were similar between the groups. Therefore, the disruption of raft-like microdomains appears to be an adaptive change in the rat heart following exercise.

Imbalance in redox system of rat liver following permethrin treatment in adolescence and neonatal age.

The effect of different permethrin treatments on the redox system of rat liver, is presented. Two types of oral administration were chosen: (i) sub-chronic treatment (1/10 of LD50 for 60 days) during adolescence (5 weeks old) and (ii) sub-acute treatment (1/44 of LD50 for 15 days) during early life (from postnatal days 6-21). The results show that adolescent permethrin treatment induces damage to the liver redox system, increasing lipid and protein peroxidation and reducing membrane fluidity in the hydrophilic--hydrophobic region of the bilayer. In addition, glutathione peroxidase (GPx) and GSH levels resulted decreased, while glutathione transferase (GST) and catalase (CAT) levels increased. The rats treated in early life with permethrin and sacrificed in adult age, showed less signs of damage compared to those exposed during adolescence in which lipid peroxidation was increased by 32%, whereas for the first group the raise was only 11%. Moreover, fluidity improved in the deeper hydrophobic membrane region of the treated group, while the level of CAT was significantly lower compared to the control one. Although sub-chronic treatment increased CAT and GST and decreased GPx and GSH levels, the present data suggest that a shorter exposure to permethrin during neonatal age decreased CAT level and it could represent an important risk factor for the onset of long-term liver damage.

Early life permethrin exposure leads to hypervitaminosis D, nitric oxide and catecholamines impairment.

The aim of this study is to gain more knowledge on the impact of early life pesticide exposure on premature aging. The effect of a low dose of the insecticide permethrin administered to rats during early life (1/50 LD50, from 6th to 21st day of life) was analyzed by measuring some metabolites in plasma and urine of 500-day-old animals. Significant differences in early life treated rats compared to the control group were found in the plasma levels of Ca(++), Na(+), 25-hydroxy-vitamin D, adrenaline, noradrenaline, nitric oxide, cholesterol and urea while in urine only Na(+) content was different. These results add information on the impact of permethrin during the neonatal period, supporting the evidence that early life environmental exposure to xenobiotics has long-term effects, inducing modifications in adulthood that can be revealed by the analysis of some macroelements, metabolites and catecholamines in plasma, when rats are 500 days old.

Early-Life Exposure to Commercial Formulation Containing Deltamethrin and Cypermethrin Insecticides Impacts Redox System and Induces Unexpected Regional Effects in Rat Offspring Brain.

Several studies have shown that the oxidative impact of pesticides is most prevalent in rural environments where they are intensively used. At different levels, pyrethroids are reported to promote neurodegeneration; they share the ability to promote oxidative stress, and to induce mitochondrial impairments, α-synuclein overexpression and neuronal cell loss. The present study evaluates the impact of early-life exposure to a commercial formulation containing deltamethrin (DM) and cypermethrin (CYP) at a dose of 1/100 LD50 (1.28 and 2.5 mg/kg, respectively). Rats aged 30 days old, treated from the 6th to the 21st day of life, were tested for brain antioxidant activity and α-synuclein levels. Four regions of the brain were analyzed: the striatum, cerebellum, cortex and hippocampus. Our data demonstrated a significant increase in catalase (CAT), superoxide dismutase (SOD) and glutathione (GSH) antioxidant levels in the brain regions compared to the controls. Pups exhibited no significant changes in protein carbonyl levels and lipid peroxidation. Striatal α-synuclein expression was significantly reduced in the rats exposed to DM + CYP, while the treatment resulted in a non-significant increase in the other brain areas. These findings indicate unexpected effects of postnatal treatment with the commercial formulation containing DM and CYP on brain redox state and α-synuclein expression, suggesting an adaptive response.

Leukocyte Nurr1 as peripheral biomarker of early-life environmental exposure to permethrin insecticide.

The effect of a low dose of the insecticide permethrin administered during early-life was evaluated on leukocytes inflammation mediators on 300- and 500-day-old rats. Nurr1, NF-κB-p65, Nrf2, lipid peroxidation and GSH levels increased with age but compared to the control group, treatment with permethrin induced a significant increase only of Nurr1 and lipid peroxidation in oldest rats. TNF-α and Rantes increased, while IL-1ß, IL-2, IL-13 decreased in oldest treated rats. The results propose Nurr1, TNF-α, Rantes, GSH and plasma lipid peroxidation as peripheral biomarkers for monitoring the impact of early-life environmental exposure to xenobiotics in old age.

Impact of JNK1, JNK2, and ligase Itch on reactive oxygen species formation and survival of prostate cancer cells treated with diallyl trisulfide.

PURPOSE: In our previous study, we demonstrated that diallyl trisulfide (DATS) induced iron-dependent G2-M arrest of prostate cancer cell cycle. Moreover, ferritin degradation and an increase of labile iron pool has been linked to the activation of the JNK signaling axis. In the present work, we extended this study to determine which of the c-jun kinases is responsible for ferritin degradation and the role of iron in DATS-induced cell death. We hypothesized that JNK1 activates Itch ligase which will lead to ferritin ubiquitination, an increase in iron-dependent ROS formation and cell death. METHODS: PC-3 prostate cancer cells were used in this study. Cell viability, concentration of ROS, labile iron pool, and changes in ferritin and P-Itch and DNA damage were determined. RESULTS: We observed that DATS induced ferritin degradation through JNK, Itch signaling axis. DATS did not induce neither ROS formation nor increase the LIP in JNK1-DN transfected cells. We also observed that DATS increased JNK-dependent activating phosphorylation of E3ligase Itch. The cells transfected with inactive form of Itch were more resistant against cytotoxicity of DATS and showed lower DATS-induced ferritin degradation. Desferrioxamine a specific iron chelator had no effect neither on cell viability nor DNA damage evaluated by comet assay. CONCLUSIONS: These results suggest that JNK1-dependent increase in LIP is mediated by Itch ubiquitin ligase.

Diallyl trisulfide-induced prostate cancer cell death is associated with Akt/PKB dephosphorylation mediated by P-p66shc.

PURPOSE: P66Shc, an isoform of adaptor proteins, is known to mediate various signals including those leading to apoptosis or cell proliferation. Previously, we have shown that diallyl trisulfide (DATS)-induced prostate cancer cell death was mediated by increased ROS formation. In this study, we investigated the role of p66Shc protein and its serine 36 phosphorylation in DATS induced decrease in prostate cancer cell viability (PC-3). METHODS: PC-3 prostate cancer cells were used in this study. Stable cell lines expressing p66ShcS36A or an empty vector have been obtained. Cell viability, concentration of ROS, changes in P-p66Shc and P-Akt and DNA damage were determined. RESULTS: We observed that DATS treatment increased p66Shc phosphorylation at serine 36. Importantly, the phosphorylation was abolished by JNK inhibitor SP600125. Cells expressing plasmid-encoded variant of p66ShcS36A showed much higher resistance to DATS-induced cells death. In addition to that, we observed that DATS-induced ROS formation was completely abolished in cells expressing the p66ShcS36A variant. Interestingly, SP600125 proved to prevent DATS-induced Akt inactivation. In order to confirm that the observed effect is related to phosphorylation of p66Shc, we performed experiments on a stable cell line expressing p66ShcS36A. In such cells, DATS-induced Akt dephosphorylation was significantly reduced. On the other hand, hydrogen peroxide induced Akt activation in PC-3 cells, which was abrogated in cells expressing p66ShcS36A. CONCLUSIONS: Our results uncover a novel signaling pathway with p66Shc being indispensable for DATS-induced inactivation of Akt due to hypophosphorylation.

Gender-Related Differences in Trimethylamine and Oxidative Blood Biomarkers in Cardiovascular Disease Patients.

Gender differences in the burden of cardiovascular disease (CVD) have been observed worldwide. In this study, plasmatic levels of trimethylamine (TMA) and blood oxidative biomarkers have been evaluated in 358 men (89 controls and 269 CVD patients) and 189 women (64 control and 125 CVD patients). The fluorescence technique was applied to determine erythrocyte membrane fluidity using 1,6-diphenyl-1,3,5-hexatriene (DPH) and Laurdan, while lipid hydroperoxides were assessed by diphenyl-1-pyrenylphosphine (DPPP). Results show that levels of plasmatic TMA were higher in healthy men with respect to healthy women (p = 0.0001). Significantly lower TMA was observed in male CVD patients (0.609 ± 0.104 µM) compared to healthy male controls (0.680 ± 0.118 µM) (p < 0.001), while higher levels of TMA were measured in female CVD patients (0.595 ± 0.115 µM) with respect to female controls (0.529 ± 0.073 µM) (p < 0.001). DPPP was significantly higher in healthy control men than in women (p < 0.001). Male CVD patients displayed a lower value of DPPP (2777 ± 1924) compared to healthy controls (5528 ± 2222) (p < 0.001), while no significant changes were measured in females with or without CVD (p > 0.05). Membrane fluidity was significantly higher (p < 0.001) in the hydrophobic bilayer only in control male subjects. In conclusion, gender differences were observed in blood oxidative biomarkers, and DPPP value might be suggested as a biomarker predictive of CVD only in men.

Chemical and Sensory Profiling of Monovarietal Extra Virgin Olive Oils from the Italian Marche Region.

Chemical and sensory peculiarities of monovarietal extra virgin olive oils (MEVOOs) from the cultivars (cvs.) Ascolana tenera (ASC), Coroncina (COR), Mignola (MIG), Piantone di Mogliano (MOG), and Raggia (RAG) from Marche region (Italy) are investigated. Their polar phenolic substances and α-tocopherol are analysed through high performance liquid chromatography with different detectors. Volatile substances, fatty acid composition, and squalene are analysed by gas chromatography coupled to mass spectrometry (MS) and to the flame ionization detector, respectively. Total antioxidant activity and sensory analysis were also performed. MOG showed high squalene content (on average 0.88 ± 0.16 g/100 g), high relative amount of α-copaene among volatiles, and the highest oleic acid percentage. MIG had high α-tocopherol content (on average 350.0 ± 57.6 mg kg-1) and high α-farnesene in the volatile fraction. ASC showed the highest sensory quality and the lignan pinoresinol with higher concentration as compared to the other MEVOOs (p < 0.05), which resulted in a possible chemical marker for this cv. RAG was characterized by the sensory note of almond, which corresponds to its highest (E)-2-hexenal percentage. Sensory analysis and an antioxidant activity assay performed on a set of industrial extra virgin olive oils purchased in supermarkets, highlighted MEVOOs' superiority from these points of view. Principal component analysis displays the main characteristics of the cvs. investigated.

Extra Virgin Olive Oil and Nigella sativa Oil Produced in Central Italy: A Comparison of the Nutrigenomic Effects of Two Mediterranean Oils in a Low-Grade Inflammation Model.

Extra virgin olive (EVO) oil and Nigella sativa (NG) oil are two well-known Mediterranean foods whose consumption has been associated with beneficial effects on human health. This study investigates the nutrigenomic properties of two high quality EVO and NG oils in an in vitro model of low-grade inflammation of human macrophages (THP-1 cells). The aim was to assess whether these healthy foods could modulate inflammation through antioxidant and epigenetic mechanisms. When THP-1 cells were co-exposed to both lipopolysaccharides (LPS)-induced inflammation and oils, both EVO and NG oils displayed anti-inflammatory activity. Both oils were able to restore normal expression levels of DNMT3A and HDAC1 (but not DNMT3B), which were altered under inflammatory conditions. Moreover, EVO oil was able to prevent the increase in TET2 expression and reduce global DNA methylation that were measured in inflamed cells. Due to its antioxidant properties, EVO oil was particularly efficient in restoring normal levels of membrane fluidity, which, on the contrary, were reduced in the presence of inflammation. In conclusion, these data support the hypothesis that these Mediterranean oils could play a major role in the modulation of low-grade inflammation and metabolic syndrome prevention. However, NS oil seems to be more efficient in the control of proinflammatory cytokines, whereas EVO oil better helps to counteract redox imbalance. Further studies that elucidate the nutrigenomic properties of local produce might help to promote regional the production and consumption of high-quality food, which could also help the population to maintain and promote health.

Anti-Inflammatory, Anti-Arthritic and Anti-Nociceptive Activities of Nigella sativa Oil in a Rat Model of Arthritis.

This study investigated the preventive efficacy of the crude oil extracted from Nigella sativa seeds in a rat model of arthritis induced by using complete Freund's adjuvant (CFA). Nigella sativa oil at 1.82 mL/kg or 0.91 mL/kg (corresponding to 1596 and 798 mg/kg, respectively) was orally administered for 25 days from the day of immunization. One immunized group was treated orally with indomethacin (3 mg/kg) as a reference drug. Body weight growth rate, paw swelling, arthritis score, mechanical allodynia, locomotor activity and anxiety-like behavior were observed, and the levels of Interleukin 6 (IL-6), C-reactive protein, albumin and total cholesterol in plasma were measured on days 15 and 25. Nigella sativa oil showed anti-inflammatory, anti-arthritic and anti-nociceptive activities that were significant as compared to untreated arthritic rats but less than indomethacin. These results indicated that Nigella sativa oil significantly attenuated adjuvant-arthritis in rats and the higher dose (1.82 mL/kg) prevented the development of arthritis with an inhibition of 56%.

Positive effect of an electrolyzed reduced water on gut permeability, fecal microbiota and liver in an animal model of Parkinson's disease.

There is growing awareness within the scientific community of the strong connection between the inflammation in the intestine and the pathogenesis of Parkinson's disease (PD). In previous studies we developed a PD animal model exposing pup rats to permethrin (PERM) pesticide. Here, we intended to explore whether in our animal model there were changes in gut permeability, fecal microbiota and hepatic injury. Moreover, we tested if the co-treatment with an electrolyzed reduced (ERW) was effective to protect against alterations induced by PERM. Rats (from postnatal day 6 to 21) were gavaged daily with PERM, PERM+ERW or vehicle and gut, liver and feces were analyzed in 2-months-old rats. Increased gut permeability, measured by FITC-dextran assay, was detected in PERM group compared to control and PERM+ERW groups. In duodenum and ileum, concentration of occludin was higher in control group than those measured in PERM group, whereas only in duodenum ZO-1 was higher in control than those measured in PERM and PERM+ERW groups. Number of inflammatory focis and neutrophils as well as iNOS protein levels were higher in livers of PERM-treated rats than in those of PERM+ERW and control rats. Fecal microbiota analysis revealed that Lachnospira was less abundant and Defluviitaleaceae more abundant in the PERM group, whereas the co-treatment with ERW was protective against PERM treatment since the abundances in Lachnospira and Defluviitaleaceae were similar to those in the control group. Higher abundances of butyrate- producing bacteria such as Blautia, U.m. of Lachnospiraceae family, U.m. of Ruminococcaceae family, Papillibacter, Roseburia, Intestinimonas, Shuttleworthia together with higher butyric acid levels were detected in PERM+ERW group compared to the other groups. In conclusion, the PD animal model showed increased intestinal permeability together with hepatic inflammation correlated with altered gut microbiota. The positive effects of ERW co-treatment observed in gut, liver and brain of rats were linked to changes on gut microbiota.

Antioxidant and Anti-Inflammatory Properties of Nigella sativa Oil in Human Pre-Adipocytes.

The oil obtained from the seeds of Nigella sativa L. (N. sativa), also known as black cumin, is frequently used in the Mediterranean area for its anti-inflammatory, anti-oxidant, and anti-cancer activities. The aim of the present study was to evaluate the antioxidant and anti-inflammatory properties of the oil extracted from seeds of a N. sativa cultivar produced in the Marche region of Italy, and to determine if the thymoquinone content, antioxidant properties, and biological activity would decay during storage. Cytotoxicity and anti-inflammatory properties of N. sativa oil were tested in an in vitro model of low-grade inflammation in Simpson⁻Golabi⁻Behmel syndrome human pre-adipocytes. The fresh extracted oil (FEO) contained 33% more thymoquinone than stored extracted oil (SEO), demonstrating that storage affects its overall quality. In addition, the thymoquinone content in the N. sativa oil from the Marche region cultivar was higher compared with other N. sativa oils produced in the Middle East and in other Mediterranean regions. Pro-inflammatory cytokines (e.g., Interleukin (IL)-1alpha, IL-1beta, IL-6) were differently modulated by fresh and stored extracts from N. sativa oils: FEO, containing more thymoquinone reduced IL-6 levels significantly, while SEO inhibited IL-1beta and had a higher antioxidant activity. Total antioxidant activity, reported as µM of Trolox, was 11.273 ± 0.935 and 6.103 ± 0.446 for SEO and FEO (p = 1.255 × 10-7), respectively, while mean values of 9.895 ± 0.817 (SEO) and 4.727 ± 0.324 (FEO) were obtained with the 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay (p = 2.891 × 10-14). In conclusion, the oil capacity to counteract proinflammatory cytokine production does not strictly depend on the thymoquinone content, but also on other antioxidant components of the oil.

Early impairment of epigenetic pattern in neurodegeneration: Additional mechanisms behind pyrethroid toxicity.

Permethrin is a synthetic pyrethroid extensively used as anti-woodworm agent and for indoor and outdoor pest control. The main route of human exposure is through fruit, vegetable and milk intake. Low dosage exposure to permethrin during neonatal brain development (from postnatal day 6 to postnatal day 21) leads to dopamine decrease in rat striatum nucleus, oxidative stress and behavioural changes linked to the development of Parkinson's like neurodegeneration later in life. The aim of this study was to evaluate the expression of genes involved in the dopaminergic pathway and epigenetic regulatory mechanisms in adolescent rats treated with permethrin during neonatal brain development. Furthermore, in order to shed light on the mechanisms associated with molecular impairments, in silico studies were performed. The outcomes show increased expression of genes related to the dopamine-synthesis pathway (Nurr1, Th, Snca), epigenetics (TET proteins and Mecp2) and exposure to toxicants (Pon1 and Pon2) in adolescent rats compared with control group. Furthermore, increased global 5mC and 5hmC levels were observed in the DNA extracted from striatum of early-life treated rats in comparison with controls. FAIRE-qPCR analysis shows that permethrin induces an enrichment of chromatin-free DNA at the level of Th and Nurr1 promoters, and ChIP-qPCR reveals a significant reduction in methylation levels at H3K9me3 position at both Th and Nurr1 promoter regions. In silico studies show that permethrin competes for the same two binding sites of known NURR1 agonists, with a lower binding free energy for permethrin, suggesting an important durable association of permethrin with the orphan receptor. Moreover, alpha-synuclein shows a strong affinity for NURR1, corroborating previous experimental outcomes on the interactions between them. This study focuses on an emerging role of early-life exposure to environmental pollutants in the regulation of late onset diseases through intriguing mechanisms that change crucial epigenetic patterns starting from adolescent age.

HTR2C Gene Variant and Salivary Cortisol Levels after Endurance Physical Activity: A Pilot Study.

INTRODUCTION: The 5-hydroxytryptamine 2C receptor (HTR2C) rs6318 polymorphism has been associated with increased sensitivity to stress. This study investigated whether the rs6318 genotype modified the cortisol response to endurance physical activity. METHODS: The HTR2C SNP was genotyped in a population of agonistic cyclists, and salivary cortisol levels were measured before and after an endurance competition. RESULTS AND CONCLUSION: Salivary cortisol levels increased after the competition (from 20.72 ± 12.36 ng/mL to 33.80 ± 21.53 ng/mL; p = 3.189 × 10-5). rs6318 C carriers displayed higher baseline cortisol levels compared to G carriers (26.60 ± 9.35 ng/mL vs. 19.50 ± 12.63 ng/mL; p = 0.04). Baseline cortisol levels were able to predict the cortisol response to exercise (ß = -0.846; p = 1.2 × 10-5). Although regression analysis did not identify an association between HTR2C genotype and change in cortisol levels, a secondary analysis in which the population was classified by median cortisol changes suggested that they might be weakly associated, thus warranting further investigation.

Chemical and sensory differences between high price and low price extra virgin olive oils.

The aim of the study was to identify new potential chemical markers of extra virgin olive oil (EVOO) quality by using a multicomponent analysis approach. Sixty-six EVOOs were purchased from the Italian market and classified according to their price as low price EVOOs (LEVOOs) and high price EVOOs (HEVOOs) costing 3.60-5.90euro/L and 7.49-29.80euro/L respectively. Sensory and chemical parameters strictly related to olive oil quality have been investigated, like volatile substances, polar phenolic substances, antioxidant activity, fatty acid composition, and α-tocopherol. Significant differences in terms of chemical composition and sensory features have been highlighted between the two EVOOs classes investigated, proving a generally lower level of quality of LEVOOs, clearly showed also by means of principal component analysis. Among the most interesting outcomes, R ratio (free tyrosol and hydroxytyrosol over total free and bound forms), measuring the extent of secoiridoids hydrolysis, resulted to be significantly higher in LEVOOs than in HEVOOs. Other key differences were found in the volatile substances composition, in the stearic acid percentage and in p-coumaric acid content.

Lutein, zeaxanthin and astaxanthin protect against DNA damage in SK-N-SH human neuroblastoma cells induced by reactive nitrogen species.

The purpose of this study was to evaluate the ability of the predominant carotenoids (lutein and zeaxanthin) of the macular pigment of the human retina, to protect SK-N-SH human neuroblastoma cells against DNA damage induced by different RNOS donors. Although astaxanthin has never been isolated from the human eye, it was included in this study because its structure is very close to that of lutein and zeaxanthin and because it affords protection from UV-light. DNA damage was induced by GSNO-MEE, a nitric oxide donor, by Na(2)N(2)O(3), a nitroxyl anion donor and by SIN-1, a peroxynitrite-generating agent. DNA damage was assessed using the comet assay, a rapid and sensitive single cell gel electrophoresis technique able to detect primary DNA damage in individual cells. The tail moment parameter was used as an index of DNA damage. The values of tail moment increased in all the samples incubated with the RNOS donors, indicating DNA impairment. Data obtained show that the ability of zeaxanthin, lutein, and astaxanthin to reduce the DNA damage depends on the type of RNOS donor and the carotenoid concentration used. All the carotenoids studied were capable of protecting against DNA damage in neuroblastoma cells when the cells were exposed to GSNO-MEE. However, a different behaviour was present when the other two RNOS donors were used. The presence of a carotenoid alone (without an RNOS donor) did not cause DNA damage. Spectrophotometric studies showed that the order with which tested carotenoids reacted with RNOS was not always in agreement with the DNA protection results. The data from this study provides additional information on the activities of the macular pigment carotenoids of the human retina.

Permethrin pesticide induces NURR1 up-regulation in dopaminergic cell line: Is the pro-oxidant effect involved in toxicant-neuronal damage?

The mechanisms associated to the development of neurodegeneration due to pesticide exposure are not clear yet. In this study we evaluated how permethrin pesticide (PERM) can influence the Nurr1 gene and protein expression, and if a pro-oxidant activity of the pesticide contributes to up-regulation of Nurr1 in a dopaminergic cell line. Incubation of PC12 cells with 1µM PERM for 72h, leads to over expression of Nurr1 gene. This effect occurs with both corn oil and extra virgin olive oil (EVO) used to solubilize the toxicant. In order to investigate if the Nurr1 up-regulation induced by PERM, was associated to the pro-oxidant activity of the pesticide, anti-oxidants as glutathione (GSH), tocotrienols (TOC) and Electrolyzed Reduced Water (ERW) were tested. RT-PCR of Nurr1 showed that its up-regulation was significantly reduced in the presence of antioxidants, especially by addition of ERW. Western-blot analysis reveals that ERW was able to counterbalance the up-regulation of Nurr1 protein induced by permethrin exposure.

In vivo and in silico studies to identify mechanisms associated with Nurr1 modulation following early life exposure to permethrin in rats.

The present work was designed to study the mechanisms associated with Nurr1 modulation following early life permethrin (PERM) treatment during rat's life span. Here we demonstrate that PERM exposure in rats, at a dose close to No Observed Adverse Effect Level (NOAEL) for 15days during neonatal brain development leads to its accumulation long after exposure. In striatum from adolescent rats we detected an increase in DNA methyltransferases (DNMTs) such as DNMT1, DNMT3a, Tyrosine hydroxylase, monomeric and aggregated α-synuclein protein levels. Adult rats showed enhanced DNMT3b and α-synuclein aggregation compared to the control group, while with aging a significant decrease in all biomarkers studied was observed. No changes in Nurr1 promoter methylation in adolescent, adult and old rats were found. In silico studies showed clear evidence of a strong binding interaction between PERM and its metabolite 3-phenoxybenzoic acid with the nuclear orphan receptor Nurr1. These findings suggest that an additional interference with the dopaminergic neuron pathway could occur in situ during PERM accumulation in brain. Therefore, Nurr1 modulation in early life PERM-treated rats, depends on age-related adaptive responses in animals.

Flunisolide attenuates nitric oxide-induced DNA damage in rat trachea epithelial cells.

In asthma the bronchial epithelium is highly abnormal, with various structural changes. As a consequence, the epithelium becomes an important source of inflammatory mediators that contribute to the ongoing inflammation and remodeling responses occurring in asthma. Compared with normal individuals, the fraction of exhaled nitric oxide (NO) is elevated in patients with asthma, and these levels have been shown to vary with disease activity. Thus, in asthma, epithelial cells may be exposed to large amounts of NO. Increased NO production is associated with the formation of various nitrosating species capable of promoting DNA damage. In this study we investigated the effect of NO on DNA of rat trachea epithelial cells in the presence or absence of flunisolide. Rat airway epithelial cells were prepared and incubated with the NO donor S-nitroso-L-glutathione monoethyl ester (GSNO-MEE). DNA damage was evaluated using single cell gel electrophoresis 'comet assay.' The parameters used as an index of DNA damage were tail length, tail intensity, and tail moment. Results of our study demonstrated that NO induced significant DNA damage in rat airway epithelial cells. Flunisolide in amounts of 11-110 mumol/L significantly reduced all the considered parameters indicating DNA damage. These data indicate that flunisolide may protect epithelial cells from the NO-mediated DNA damage. NO overproduction could contribute to epithelial injury in asthma, and flunisolide seems to attenuate this damage.