Assessment of adrenaline-induced DNA damage in whole blood cells with the comet assay

Harmful effects of elevated levels of catecholamines are mediated by various mechanisms, including gene transcription and formation of oxidation products. The aim of this study was to see whether the molecular mechanisms underlying the damaging action of adrenaline on DNA are mediated by reactive oxygen species (ROS). To do that, we exposed human whole blood cells to 10 mu mol L-1 adrenaline or 50 mu mol L-1 H2O2 (used as positive control) that were separately pre-treated or post-treated with 500 mu mol L-1 of quercetin, a scavenger of free radicals. Quercetin significantly reduced DNA damage in both pre- and post-treatment protocols, which suggests that adrenaline mainly acts via the production of ROS. This mechanism is also supported by gradual lowering of adrenaline and H2O2-induced DNA damage 15, 30, 45, and 60 min after treatment. Our results clearly show that DNA repair mechanisms are rather effective against ROS-mediated DNA damage induced by adrenaline

Nitric oxide alleviates cadmium- but not arsenic-induced damages in rice roots

Nitric oxide (NO) has signalling roles in plant stress responses. Cadmium (Cd) and arsenic (As) soil pollutants alter plant development, mainly the root-system, by increasing NO-content, triggering reactive oxygen species (ROS), and forming peroxynitrite by NO-reaction with the superoxide anion. Interactions of NO with ROS and peroxynitrite seem important for plant tolerance to heavy metal(oid)s, but the mechanisms underlying this process remain unclear. Our goal was to investigate NO-involvement in rice (Oryza sativa L.) root-system after exposure to Cd or As, to highlight possible differences in NO-behaviour between the two pollutants. To the aim, morpho-histological, chemical and epifluorescence analyses were carried out on roots of different origin in the root-system, under exposure to Cd or As, combined or not with sodium nitroprusside (SNP), a NO-donor compound. Results show that increased intracellular NO levels alleviate the root-system alterations induced by Cd, i.e., inhibition of adventitious root elongation and lateral root formation, increment in lignin deposition in the sclerenchyma/endodermis cell-walls, but, even if reducing As-induced endodermis lignification, do not recover the majority of the As-damages, i.e., enhancement of AR-elongation, reduction of LR-formation, anomalous tissue-proliferation. However, NO decreases both Cd and As uptake, without affecting the pollutants translocation-capability from roots to shoots. Moreover, NO reduces the Cd-induced, but not the As-induced, ROS levels by triggering peroxynitrite production. Altogether, results highlight a different behaviour of NO in modulating rice root-system response to the toxicity of the heavy metal Cd and the metalloid As, which depends by the NO-interaction with the specific pollutant

Oxidative Metabolism Genes Are Not Responsive to Oxidative Stress in Rodent Beta Cell Lines

Altered expression of oxidative metabolism genes has been described in the skeletal muscle of individuals with type 2 diabetes. Pancreatic beta cells contain low levels of antioxidant enzymes and are particularly susceptible to oxidative stress. In this study, we explored the effect of hyperglycemia-induced oxidative stress on a panel of oxidative metabolism genes in a rodent beta cell line. We exposed INS-1 rodent beta cells to low (5.6 mmol/L), ambient (11 mmol/L), and high (28 mmol/L) glucose conditions for 48 hours. Increases in oxidative stress were measured using the fluorescent probe dihydrorhodamine 123. We then measured the expression levels of a panel of 90 oxidative metabolism genes by real-time PCR. Elevated reactive oxygen species (ROS) production was evident in INS-1 cells after 48 hours (P < 0.05). TLDA analysis revealed a significant (P < 0.05) upregulation of 16 of the 90 genes under hyperglycemic conditions, although these expression differences did not reflect differences in ROS. We conclude that although altered glycemia may influence the expression of some oxidative metabolism genes, this effect is probably not mediated by increased ROS production. The alterations to the expression of oxidative metabolism genes previously observed in human diabetic skeletal muscle do not appear to be mirrored in rodent pancreatic beta cells

Platinacycles Containing a Primary Amine Platinum(II) Compounds for Treating Cisplatin-Resistant Cancers by Oxidant Therapy

Cisplatin is an efficient anticancer drug, but its effects are often lost after several chemotherapy cycles, showing important secondary effects. For these reasons, new anticancer agents, with different coordination properties and mechanisms of action, are needed. Here we describe the reaction of 2-phenylaniline with cis-[PtCl2(dmso)(2)] and sodium acetate to afford a cycloplatinated compound 2 and the synthesis and some biological studies of 3-6 (two neutral and two ionic compounds): [PtCl(C-N)(L)], C-N cycloplatinated 2-phenylaniline with L = PPh3(3) or P(4-FC6H4)(3) (4) and [Pt(C-N)(L-L)]Cl with L-L = Ph2PCH2CH2Ph2(5) or (C6F5)(2)PCH2-CH2(C6F5)(2) (6). Ionic platinacycles 5 and 6 show a greater antiproliferative activity than that of cisplatin in human lung, breast, and colon cancer cell lines (A-549, MDA-MB-231 and MCF-7, and HCT-116), a remarkable result given the fact that they do not show covalent interaction with DNA. 5 and 6 have also been found able to oxidize NADH by a catalytic process prod- oducing H2O2 as ROS. The activity of these complexes to generate ROS seems to be the key factor to explain their potent anticancer activity; it should be noted that platinum(II) complexes showing biocatalytic activity for hydride transfer from NADH have not been described so far. Ionic complex 6 shows low affinity to some target proteins; the presence of perfluoroaromatic rings seems to hinder its interaction with some biomolecules

Aerobic Exercise Training Prevents the Onset of Endothelial Dysfunction via Increased Nitric Oxide Bioavailability and Reduced Reactive Oxygen Species in an Experimental Model of Menopause

ObjectivePrevious studies have shown that estrogen deficiency, arising in postmenopause, promotes endothelial dysfunction. This study evaluated the effects of aerobic exercise training on endothelial dependent vasodilation of aorta in ovariectomized rats, specifically investigating the role of nitric oxide (NO) and reactive oxygen species (ROS).MethodsFemale Wistar rats ovariectomized (OVX - n= 20) or with intact ovary (SHAM - n= 20) remained sedentary (OVX and SHAM) or performed aerobic exercise training on a treadmill 5 times a week for a period of 8 weeks (OVX-TRA and SHAM-TRA). in the thoracic aorta the endothelium-dependent and - independent vasodilation was assessed by acetylcholine (ACh) and sodium nitroprusside (SNP), respectively. Certain aortic rings were incubated with L-NAME to assess the NO modulation on the ACh-induced vasodilation. the fluorescence to dihydroethidium in aortic slices and plasma nitrite/nitrate concentrations were measured to evaluate ROS and NO bioavailability, respectively.ResultsACh-induced vasodilation was reduced in OVX rats as compared SHAM (Rmax: SHAM: 86 +/-3.3 vs. OVX: 57+/-3.0%, p<0.01). Training prevented this response in OVX-TRA (Rmax: OVX-TRA: 88+/-2.0%, p<0.01), while did not change it in SHAM-TRA (Rmax: SHAM-TRA: 80+/-2.2%, p<0.01). the L-NAME incubation abolished the differences in ACh-induced relaxation among groups. SNP-induced vasodilation was not different among groups. OVX reduced nitrite/nitrate plasma concentrations and increased ROS in aortic slices, training as effective to restore these parameters to the SHAM levels.ConclusionsExercise training, even in estrogen deficiency conditions, is able to improve endothelial dependent vasodilation in rat aorta via enhanced NO bioavailability and reduced ROS levels.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Biosci, São Paulo, BrazilUniv São Paulo, Inst Biomed Sci, Dept Physiol & Biophys, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biosci, São Paulo, BrazilFAPESP: 2012/17709-0FAPESP: 2010/50048-1Web of Scienc

Respostas de enzimas antioxidantes a bioestimulantes em plantas de milho e de soja sob estresse hídrico

Water stress is one of the most important environmental factors inducing physiological changes in plants, such as decrease in the water potential of the cells, the stomatal closure; and the development of oxidative processes mediated by reactive oxygen species (ROS). Antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) are efficient scavengers of ROS. The aim of this research was to examine how the application of biostimulant based on humic substances and aminoacids may affect activity levels of SOD, CAT, and APX of maize and soybean plants under well-watered or drought stress conditions. Pots (4.5 L) were filled with a Typic Hapludult soil where the biostimulants doses were applied. It was taken leaf samples in order to analyze SOD, CAT, and APX activities in plants. SOD and APX activity levels were increased by application of biostimulant 1 in maize subjected to stress. Catalase activity was not enhanced in plants by using the biostimulants. The composition of the biostimulants was not able to enhance stress tolerance in maize and soybean plants subjected to water stress.O estresse hídrico é um dos mais importantes fatores ambientais que induz mudanças fisiológicas, como diminuição do potencial de água na célula, o fechamento dos estômatos e o desenvolvimento de processos oxidativos mediante a formação das espécies reativas de oxigênio (ROS). As enzimas antioxidantes superóxido dismutase (SOD), catalase (CAT) e ascorbato peroxidase (APX) são eficientes eliminadores das ROS. O objetivo deste estudo foi examinar como a aplicação de bioestimulantes com substâncias húmicas e aminoácidos em sua composição afeta os níveis de SOD, CAT e APX nos tecidos das folhas de plantas de milho e de soja cultivadas com ou sem estresse hídrico. Amostras de um Argissolo foram colocadas em vasos (4,5 L) onde foram adicionadas as doses dos bioestimulantes. Foram retiradas amostras de folhas para análise da atividade da SOD, CAT e APX nas plantas. A SOD e APX aumentaram nas plantas de milho com a aplicação do bioestimulante 1. A atividade da CAT não aumentou nas plantas com a aplicação dos bioestimulantes. As composições dos bioestimulantes não possibilitaram aumento na resistência ao estresse hídrico em plantas de milho e de soja submetidas ao estresse hídricoCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Control of the Intracellular Redox State by Glucose Participates in the Insulin Secretion Mechanism

Background: Production of reactive oxygen species (ROS) due to chronic exposure to glucose has been associated with impaired beta cell function and diabetes. However, physiologically, beta cells are well equipped to deal with episodic glucose loads, to which they respond with a fine tuned glucose-stimulated insulin secretion (GSIS). In the present study, a systematic investigation in rat pancreatic islets about the changes in the redox environment induced by acute exposure to glucose was carried out. Methodology/Principal Findings: Short term incubations were performed in isolated rat pancreatic islets. Glucose dose- and time-dependently reduced the intracellular ROS content in pancreatic islets as assayed by fluorescence in a confocal microscope. This decrease was due to activation of pentose-phosphate pathway (PPP). Inhibition of PPP blunted the redox control as well as GSIS in a dose-dependent manner. The addition of low doses of ROS scavengers at high glucose concentration acutely improved beta cell function. The ROS scavenger N-acetyl-L-cysteine increased the intracellular calcium response to glucose that was associated with a small decrease in ROS content. Additionally, the presence of the hydrogen peroxide-specific scavenger catalase, in its membrane-permeable form, nearly doubled glucose metabolism. Interestingly, though an increase in GSIS was also observed, this did not match the effect on glucose metabolism. Conclusions: The control of ROS content via PPP activation by glucose importantly contributes to the mechanisms that couple the glucose stimulus to insulin secretion. Moreover, we identified intracellular hydrogen peroxide as an inhibitor of glucose metabolism intrinsic to rat pancreatic islets. These findings suggest that the intracellular adjustment of the redox environment by glucose plays an important role in the mechanism of GSIS.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)(CAPES) Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior, Brazi

Efficiency of antimicrobial photodynamic therapy with photodithazine® on mssa and mrsa strains

Staphylococccus aureus is a ubiquitous and opportunistic bacteria associated with high mortality rates. Antimicrobial photodynamic therapy (aPDT) is based on the application of a light source and a photosensitizer that can interact with molecular oxygen, forming Reactive Oxygen Species (ROS) that result in bacterial inactivation. This study aimed to analyze, in vitro, the action of aPDT with Photodithazine® (PDZ) in methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) strains. The strains were incubated with PDZ at 25, 50, 75, and 100 mg/L for 15 min and irradiated with fluences of 25, 50, and 100 J/cm2 . The internalization of PDZ was evaluated by confocal microscopy, the bacterial growth by counting the number of colony-forming units, as well as the bacterial metabolic activity post-aPDT and the production of ROS. In both strains, the photosensitizer was internalized; the production of ROS increased when the aPDT was applied; there was a bacterial reduction compared to the control at all the evaluated fluences and concentrations; and, in most parameters, it was obtained complete inactivation with significant difference (p < 0.05). The implementation of aPDT with PDZ in clinical strains of S. aureus has resulted in its complete inactivation, including the MRSA strains.Fil: Nunes Souza, Beatriz Müller. University of Vale do Paraíba; BrasilFil: Pinto, Juliana Guerra. University of Vale do Paraíba; BrasilFil: Correia Pereira, André Henrique. University of Vale do Paraíba; BrasilFil: Miñan, Alejandro Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas; ArgentinaFil: Ferreira Strixino, Juliana. University of Vale do Paraíba; Brasi

A quantitative proof of the H\"older regularity of stable solutions to semilinear elliptic equations

This article provides quantitative proofs of the results obtained in [Cabre, Figalli, Ros-Oton, and Serra, Acta Math. 224 (2020)], a work that proved the H\"older regularity of stable solutions to semilinear elliptic equations in the optimal range of dimensions $n \leq 9$. Some results in that paper were established through compactness proofs, arguing by contradiction -- the boundary regularity arguments being specially delicate. Here we provide proofs that do not rely on compactness and, in particular, allow to quantify the H\"older regularity exponent. For this, we establish two new estimates which control the solution in $L^1$ by the $L^1$ norm of its radial derivative. For expository purposes, we provide self-contained proofs of the full H\"older regularity results. They only involve basic Analysis tools