Lower Reactive Oxygen Species Production Research Articles

Oxidative Stress in Cardiovascular Diseases.

Reactive oxygen species (ROS) are subcellular messengers in signal transductions pathways with both beneficial and deleterious roles. ROS are generated as a by-product of mitochondrial respiration or metabolism or by specific enzymes such as superoxide dismutases, glutathione peroxidase, catalase, peroxiredoxins, and myeloperoxidases. Under physiological conditions, the low levels of ROS production are equivalent to their detoxification, playing a major role in cellular signaling and function. In pathological situations, particularly atherosclerosis or hypertension, the release of ROS exceeds endogenous antioxidant capacity, leading to cell death. At cardiovascular levels, oxidative stress is highly implicated in myocardial infarction, ischemia/reperfusion, or heart failure. Here, we will first detail the physiological role of low ROS production in the heart and the vessels. Indeed, ROS are able to regulate multiple cardiovascular functions, such as cell proliferation, migration, and death. Second, we will investigate the implication of oxidative stress in cardiovascular diseases. Then, we will focus on ROS produced by NAPDH oxidase or during endothelial or mitochondrial dysfunction. Given the importance of oxidative stress at the cardiovascular level, antioxidant therapies could be a real benefit. In the last part of this review, we will detail the new therapeutic strategies potentially involved in cardiovascular protection and currently under study.

Copper-binding motifs Xxx-His or Xxx-Zzz-His (ATCUN) linked to an antimicrobial peptide: Cu-binding, antimicrobial activity and ROS production

Depending on the coordination, copper ions can have a very high activity in catalyzing the production of reactive oxygen species. Thus interest arose in increasing the activity of antimicrobial peptides (AMPs) by equipping them with a Cu-binding unit. Several examples, native and engineered, have been investigated with the motif Xxx-Zzz-His, called Amino Terminal Cu(II)- and Ni(II)-binding (ATCUN) motif. Here we investigate a short AMP that was equipped either with Xxx-Zzz-His or Xxx-His. Xxx-His is a shorter motif and yields a more redox active copper complex. The control AMP, Xxx-His-AMP and Xxx-Zzz-His-AMP were investigated toward Cu-binding, Reactive Oxygen Species (ROS) production and antimicrobial activity in E. coli. The data indicate that these Cu-binding motifs have very limited impact on antimicrobial activity and low ROS production capability.

Seed Priming Improved Antioxidant Defense System and Alleviated Ni-Induced Adversities in Rice Seedlings Under N, P, or K Deprivation.

Excess nickel (Ni) concentration in the growing medium severely hampers the plant growth by disturbing oxidative metabolism and nutrient status. The present study was carried out to investigate the individual and interactive effects of Ni toxicity (0.25 mM NiSO4.6H2O) and nutrient deprivation (no-N, no-P, or no-K) on growth, oxidative metabolism, and nutrient uptake in primed and non-primed rice seedlings. Rice seed was primed with distilled water (hydropriming), selenium (5 mg L-1), or salicylic acid (100 mg L-1). The Ni toxicity and deprivation of N, P, or K posed negative effects on the establishment of rice seedlings. The shoot length and fresh biomass were severely reduced by Ni toxicity and nutrient stresses; the minimum shoot growth was recorded for rice seedlings grown under Ni toxicity and no-N stress. The Ni toxicity reduced the root fresh biomass but did not significantly affect the root length of N-deprived seedlings. The rice seedlings with no-P or no-K recorded similar root fresh biomass compared with those grown with sufficient nutrient supply. The Ni toxicity alone or in combination with nutrient stresses triggered the production of reactive oxygen species (ROS) and caused lipid peroxidation in rice seedlings. Among antioxidants, only glutathione reductase and vitamin E were significantly increased by Ni toxicity under different nutrient stress treatments. The Ni toxicity also reduced the concentrations of N particularly in shoot of rice seedlings. The N-deprived (no-N) seedlings recorded maximum Ni concentration in shoot, while K-deprived (no-K) seedlings showed higher Ni concentrations in root. Seed priming with selenium or salicylic acid was effective to alleviate the detrimental effects of Ni toxicity and/or nutrient stresses on rice seedlings. The better growth and greater stress tolerance of primed seedlings was coordinately attributed to lower ROS production, higher membrane stability, strong antioxidative defense system, and maintenance of mineral nutrient status.

Aza-BODIPY Probe-Decorated Mesoporous Black TiO2 Nanoplatform for the Highly Efficient Synergistic Phototherapy

As an important noninvasive tumor treatment method, phototherapy has drawn extensive research interest. However, the requirements of separate excitation wavelengths, high degree of electron-hole recombination, and low reactive oxygen species (ROS) production capability are still the major barriers. This work reports the construction of a novel nanoplatform: design and synthesis of an aza-BODIPY (AB) probe-decorated mesoporous black titanium dioxide (TiO2) (MT) nanoparticles (NPs) for enhanced photodynamic therapy and photothermal therapy under single-wavelength near-infrared (NIR) laser irradiation for the first time. AB probe-decorated MT NPs (abbreviated as MTAB) were synthesized through the Al reduction of mesoporous anatase TiO2 NPs and subsequent adsorption of the AB probe. The mesoporous structure of MT ensured AB loading capacity and avoided the complicated modification and synthesis processes. Heterogeneous MTAB, which possessed staggered energy levels, were assessed for their capability for effective separation of photogenerated electrons and holes for the first time. Upon NIR laser light irradiation, MTAB exhibited sufficient ROS generation, resulting in distinct tumor cell killing and tumor tissue elimination. This unique heterogeneous nanoplatform with staggered energy levels provides a new strategy to enhance ROS generation and improve the therapeutic efficacy.

Effects of humic acid diet on the serum biochemistry and oxidative status markers in pheasants

This study investigated the effect of different concentrations of humic acids (HAs) on the selected serum biochemistry parameters and oxidative status markers in common pheasants (Phasianus colchicus). The control birds were fed a diet with no HA additives, while the birds in the experimental groups were fed diets containing HAs at the level of 0.5% (EG1), 0.75% (EG2) and 1.0% (EG3) from 1 to 90 days of age. The blood sodium concentration decreased (P < 0.01) in the group fed by 1.00% HAs compared to the control birds. The concentration of potassium in the EG2 group increased (P < 0.05) in comparison to the control group. The EG1 group showed a higher (P < 0.01) serum glucose than the EG2 group. Significant differences (P < 0.05; P < 0.01) were also found between the experimental groups (EG1 vs EG2, and EG2 vs EG3) in the cholesterol concentrations. The birds in the experimental groups showed lower ROS (reactive oxygen species) and MDA (malondialdehyde) production. An opposite effect was observed in the TAC (total antioxidant capacity), where its values significantly increased in the experimental groups. The diet supplementation affected the enzymatic antioxidant system of the fattened pheasants, and so the HAs exhibited an antioxidant potential in these birds.

Dendrimer Nanodevices and Gallic Acid as Novel Strategies to Fight Chemoresistance in Neuroblastoma Cells.

Human neuroblastoma (NB), a pediatric tumor inclined to relapse, after an initial response to therapy, usually develops resistance. Since several chemotherapeutics exert anticancer effect by increasing reactive oxygen species (ROS), NB cells overproduce antioxidant compounds becoming drugs-resistant. A strategy to sensitize NB cells to chemotherapy involves reducing their antioxidant defenses and inducing ROS overproduction. Concerning this, although affected by several issues that limit their clinical application, antioxidant/pro-oxidant polyphenols, such as gallic acid (GA), showed pro-oxidant anti-cancer effects and low toxicity for healthy cells, in several kind of tumors, not including NB. Herein, for the first time, free GA, two GA-dendrimers, and the dendrimer adopted as GA reservoir were tested on both sensitive and chemoresistant NB cells. The dendrimer device, administered at the dose previously found active versus sensitive NB cells, induced ROS-mediated death also in chemoresistant cells. Free GA proved a dose-dependent ROS-mediated cytotoxicity on both cell populations. Intriguingly, when administered in dendrimer formulations at a dose not cytotoxic for NB cells, GA nullified any pro-oxidant activity of dendrimer. Unfortunately, due to GA, nanoformulations were inactive on NB cells, but GA resized in nanoparticles showed considerable ability in counteracting, at low dose, ROS production and oxidative stress, herein induced by the dendrimer.

The effect of methotrexate on neutrophil reactive oxygen species and CD177 expression in rheumatoid arthritis

Neutrophils are found in abundance in the synovial fluid of patients with rheumatoid arthritis (RA), where they are activated and show high reactive oxygen species (ROS) production. However, there is limited data on circulating neutrophils in peripheral blood of patients with RA in terms of ROS production, expression of activation markers and the effect of treatment with methotrexate (MTX) on ROS. This single-centre prospective study recruited patients of RA classified as per the 2010 ACR/EULAR criteria. In the cross-sectional arm, we included three groups, treatment-naïve RA (naïve-RA), MTX-treated RA (MTX-RA) and healthy controls, and compared ROS production and surface markers of neutrophil activation. In the longitudinal arm, we studied the change in neutrophil ROS production after 8 weeks of MTX treatment in naïve-RA patients. Neutrophil ROS production was measured by flow cytometry using dihydrorhodamine-123 (DHR) and by chemiluminescence using luminol. Surface expression of CD177, CD11b and CD64 was measured by flow cytometry. This study included 103 patients (50 naïve-RA, 53 MTX-RA) and 20 controls. Both naïve-RA and MTX-RA patients showed higher ROS production than healthy controls in unstimulated neutrophils in the DHR assay (p<0.001 and p=0.004). MTX-RA patients showed significantly lower ROS production than naive-RA, in both unstimulated (p=0.004) and PMA-stimulated neutrophils in the DHR assay (p=0.03). On longitudinal follow-up of 24 naïve-RA patients, there was a significant reduction of neutrophil ROS production (by 55% from baseline) (p<0.001) after 8 weeks of MTX. Neutrophil CD177 expression was higher in both naïve-RA and MTX-RA (trend) than controls (p=0.001 and p=0.09). MTX-RA neutrophils showed lower expression of CD177 than naïve-RA (p=0.01). CD11b expression was higher in MTX-RA compared to controls (p=0.01). Circulating neutrophils in RA showed higher ROS production and higher expression of CD177 and CD11b compared to controls. MTX treatment was associated with a reduction in ROS production and CD177 expression, which may be one of the mechanisms by which MTX works in RA.

Tumor Metabolism‐Engineered Composite Nanoplatforms Potentiate Sonodynamic Therapy via Reshaping Tumor Microenvironment and Facilitating Electron–Hole Pairs’ Separation

AbstractReactive oxygen species (ROS) depletion and low ROS production that result from the intratumoral redox metabolism equilibrium and low energy conversion efficiency from ultrasound mechanical energy to ROS‐represented chemical energy, respectively, are two vital inhibitory factors of sonodynamic therapy (SDT). To address the two concerns, a tumor metabolism‐engineered composite nanoplatform capable of intervening intratumoral ROS metabolism, breaking the redox equilibrium, and reshaping the tumor microenvironment is constructed to reinforce SDT against tumors. In this metabolism‐engineered nanoplatform, Nb2C nanosheets serve as the scaffold to accommodate TiO2 sonosensitizers and l‐buthionine‐sulfoximine. Systematic experiments show that such nanoplatforms can reduce ROS depletion via suppressing glutathione synthesis and simultaneously improving ROS production via the Nb2C‐enhanced production and separation of electron–hole pairs. Contributed by the combined effect, net ROS content can be significantly elevated, which results in the highly efficient anti‐tumor outcomes in vivo and in vitro. Moreover, the combined design principles, that is, tumor metabolism modulation for reducing ROS depletion and electron–hole pair separation for facilitating ROS production, can be extended to other ROS‐dependent therapeutic systems.

Sinapine, but not sinapic acid, counteracts mitochondrial oxidative stress in cardiomyocytes

IntroductionWhen confronted to stress or pathological conditions, the mitochondria overproduce reactive species that participate in the cellular dysfunction. These organelles are however difficult to target with antioxidants. A feature of mitochondria that can be used for this is the negatively charged compartments they form. Most of mitochondrion-targeting antioxidants are therefore cationic synthetic molecules. Our hypothesis is that such mitochondriotropic traits might also exists in natural molecules. AimWe tested here whether sinapine, a natural phenolic antioxidant-bearing a permanent positive charge, can target mitochondria to modulate mitochondrial oxidative stress. MethodsExperiments were performed in-vitro, in-cellulo, ex-vivo, and in-vivo, using cardiac tissue. The sinapic acid -lacking the positively-charged-choline-moiety present in sinapine-was used as a control. Sinapine entry into mitochondria was investigated in-vivo and in cardiomyocytes. We used fluorescent probes to detect cytosolic (H2DCFDA) and mitochondrial (DHR123) oxidative stress on cardiomyocytes induced with either hydrogen peroxide (H2O2) or antimycin A, respectively. Finally, ROS production was measured with DHE 10 min after ischemia-reperfusion (IR) on isolated heart, treated or not with sinapine, sinapic acid or with a known synthetic mitochondrion-targeted antioxidant (mitoTempo). ResultsWe detected the presence of sinapine within mitochondria in-vitro, after incubation of isolated cardiomyocytes, and in-vivo, after oral treatment. The presence of sinapic acid was not detected in the mitochondria. Both the sinapine and the sinapic acid limited cytosolic oxidative stress in response to H2O2. Only sinapine was able to blunt oxidative stress resulting from antimycin A-induced mtROS. Both mitoTempo and sinapine improved cardiac functional recovery following IR. This was associated with lower ROS production within the cardiac tissue. ConclusionSinapine, a natural cationic hydrophilic phenol, commonly and substantially found in rapeseed species, effectively (i) enters within the mitochondria, (ii) selectively decreases the level of mitochondrial oxidative stress and, (iii) efficiently limits ROS production during cardiac ischemia-reperfusion.

Exo/endogenous dual-augmented chemodynamic therapy based on bio-reducible and bio-breakable copper (Ⅱ)-based truncated octahedron

Recently, chemodynamic therapy (CDT) has become an outstanding cancer treatment because of its high specificity for tumor microenvironment (TME). Nevertheless, lower reactive oxygen species production efficiency and cellular antioxidant defense system are still the main obstacles in CDT. To this end, we propose a highly efficient nanoplatform for exogenous and endogenous dual-augmented mitochondrial targeted CDT based on bio-reducible and bio-breakable copper (II) truncated octahedron (CuO@AuCu-TPP). In vitro, copper exists in the form of bio-reducible Cu (II) without Fenton-like reaction. After entering the tumor cells, however, Cu (II) was converted into Cu (I) to trigger rapid Fenton-like reaction because of the special exposed surface and morphology of truncated octahedron in the presence of endogenous glutathione, thus generating ROS as well as consuming GSH to disrupt the redox homeostasis. Moreover, exogenous near-infrared light induces hyperthermia of tumor site, which significantly improves the production efficiency of hydroxyl radicals, thus achieving the effect of dual-augmented CDT (4.6 times). Importantly, using triphenylphosphonium (TPP) as a targeted component, Fenton catalyst is localized into the mitochondrial site of tumor, which plays an important role in cell apoptosis through cytochrome c pathway. In addition, the truncated octahedron framework could be collapsed to small-sized nanoparticles under overexpressed GSH and acidic environment, thereby avoiding accumulation of nanocarriers in the body. Overall, both cellular and animal experiments show that CuO@AuCu-TPP nanocarriers exhibit excellent therapeutic efficacy and low systemic toxicity, which may hold great application prospects in dual-augmented chemodynamic cancer treatment.

Low abundance of NDUFV2 and NDUFS4 subunits of the hydrophilic complex I domain and VDAC1 predicts mammalian longevity

Mitochondrial reactive oxygen species (ROS) production, specifically at complex I (Cx I), has been widely suggested to be one of the determinants of species longevity. The present study follows a comparative approach to analyse complex I in heart tissue from 8 mammalian species with a longevity ranging from 3.5 to 46 years. Gene expression and protein content of selected Cx I subunits were analysed using droplet digital PCR (ddPCR) and western blot, respectively. Our results demonstrate: 1) the existence of species-specific differences in gene expression and protein content of Cx I in relation to longevity; 2) the achievement of a longevity phenotype is associated with low protein abundance of subunits NDUFV2 and NDUFS4 from the matrix hydrophilic domain of Cx I; and 3) long-lived mammals show also lower levels of VDAC (voltage-dependent anion channel) amount. These differences could be associated with the lower mitochondrial ROS production and slower aging rate of long-lived animals and, unexpectedly, with a low content of the mitochondrial permeability transition pore in these species.

Calorie Restriction Delays the Aging of Saccharomyces cerevisiae by Improving Complex III Activity via Glutathione Peroxidase 2 (Gpx2)

Comparative studies among organisms of different species of the same genus have found that long ‐ lived species contains polymorphisms in the sequence of the cytochrome b (Cyt b) subunit of the complex III leading to lower ROS production, which has been suggested to contribute for longer lifespans. Cyt b is a highly hydrophobic protein that depends on cardiolipin for its function. Cardiolipin peroxidation impairs electron transfer at the cytochrome b and enhances ROS by inducing electron leak. Cardiolipin peroxidation is repaired in mitochondria by Gpx2. Calorie restriction, the only non‐genetic manipulation increasing the lifespan, induces a hormetic response enhancing the activity of antioxidant systems like Gpx. Thus, it is feasible to hypothesize that calorie restriction increases longevity in yeast by preserving complex III activity and electron transfer in the Cyt b in a Gpx2 – dependent manner. Calorie restriction was induced in yeast with 0.05% glucose in YPD medium. YPD 0.05% extended the chronological lifespan of yeast in comparison with YPD medium with standard 2% glucose. Calorie restriction also improved complex III activity in aged yeasts and enhanced the reduction of Cyt b. The beneficial effects of calorie restriction on chronological lifespan, complex III activity and Cyt b reduction were lost in a GPX2Δ yeast mutant. These results suggest that preservation of complex III activity by enhanced electron transfer at Cyt b might be another mechanism by which calorie restriction extends longevity, which might be related with lower ROS generation and protection of cardiolipin against oxidative damage induced by aging.Support or Funding InformationThis work was supported by a grant from Programa de Investigación 2019, Coordinación de la Investigación Científica ‐ UMSNH (CCR).

In situ experiment to evaluate biochemical responses in the freshwater mussel Diplodon chilensis under anthropogenic eutrophication conditions.

An in-situ experiment was performed to study metabolic responses of the freshwater mussel Diplodon chilensis to water contaminated by leachates from an open dump and cattle activity, in order to analyze both the effects of those contaminants on aquatic environments and the potential use of a native bivalve to evaluate the effects of anthropic influence and eutrophication. Bivalves from a reference site were cage-transplanted to a control site (site A) and to a temporal water pond (site B) over 30 and 60 periods. Water quality analyses revealed that the site B was affected by anthropogenic influence. Mussel's hemocytes from site B showed 50% lower reactive oxygen species production and 130% higher lysosomal membrane stability in the site B mussels. In addition, no oxidative stress was evident in gills, despite the elevated copper and iron concentrations recorded in the site B water samples (CuB=0.3350±0.0636mg. L-1vs. CuA=0.0045±0.0007mg. L-1; FeB=3.8650±0.4031mg. L-1vs. FeA=0.0365±0.0049mg. L-1). In contrast, the adductor muscle accumulated more Fe (~10-20-fold) than the gills and showed signs of oxidative stress, e.g. superoxide dismutase activity and TBARS levels were increased by 10% were 34%, respectively, in the site B compared with the site A after 60 days of exposure. Additionally, the adductor muscle showed signs of anaerobic metabolism activation. Cu is accumulated in gills from both sites' individuals, at 60 days, in concordance with the increase in the activity of the cu-containing enzyme cytochrome-c-oxidase. There was a reduction in the overall condition and digestive gland index in bivalves exposed at site B, associated with diminished levels of lipid and protein contents. Metal-pollution and eutrophication affects D. chilensis metabolism and is associated to tissue-specific exposure, anaerobic metabolism and general energetic condition depletion.

Antibiotic Resistance Profile, Outer Membrane Proteins, Virulence Factors and Genome Sequence Analysis Reveal Clinical Isolates of Enterobacter Are Potential Pathogens Compared to Environmental Isolates.

Outer membrane proteins (OMPs) of gram-negative bacteria play an important role in mediating antibacterial resistance, bacterial virulence and thus affect pathogenic ability of the bacteria. Over the years, prevalence of environmental antibiotic resistant organisms, their transmission to clinics and ability to transfer resistance genes, have been studied extensively. Nevertheless, how successful environmental bacteria can be in establishing as pathogenic bacteria under clinical setting, is less addressed. In the present study, we utilized an integrated approach of investigating the antibiotic resistance profile, presence of outer membrane proteins and virulence factors to understand extent of threat posed due to multidrug resistant environmental Enterobacter isolates. Also, we investigated clinical Enterobacter isolates and compared the results thereof. Results of the study showed that multidrug resistant environmental Enterobacter isolates lacked OmpC, lacked cell invasion abilities and exhibited low reactive oxygen species (ROS) production in neutrophils. In contrast, clinical isolates possessed OmpF, exhibited high invasive and adhesive property and produced higher amounts of ROS in neutrophils. These attributes indicated limited pathogenic potential of environmental Enterobacter isolates. Informations obtained from whole genome sequence of two representative bacterial isolates from environment (DL4.3) and clinical sources (EspIMS6) corroborated well with the observed results. Findings of the present study are significant as it highlights limited fitness of multidrug resistant environmental Enterobacter isolates.

Cytochrome c phosphorylation: Control of mitochondrial electron transport chain flux and apoptosis.

Cytochrome c (Cytc)1is a cellular life and death decision molecule that regulates cellular energy supply and apoptosis through tissue specific post-translational modifications. Cytc is an electron carrier in the mitochondrial electron transport chain (ETC) and thus central for aerobic energy production. Under conditions of cellular stress, Cytc release from the mitochondria is a committing step for apoptosis, leading to apoptosome formation, caspase activation, and cell death. Recently, Cytc was shown to be a target of cellular signaling pathways that regulate the functions of Cytc by tissue-specific phosphorylations. So far five phosphorylation sites of Cytc have been mapped and functionally characterized, Tyr97, Tyr48, Thr28, Ser47, and Thr58. All five phosphorylations partially inhibit respiration, which we propose results in optimal intermediate mitochondrial membrane potentials and low ROS production under normal conditions. Four of the phosphorylations result in inhibition of the apoptotic functions of Cytc, suggesting a cytoprotective role for phosphorylated Cytc. Interestingly, these phosphorylations are lost during stress conditions such as ischemia. This results in maximal ETC flux during reperfusion, mitochondrial membrane potential hyperpolarization, excessive ROS generation, and apoptosis. We here present a new model proposing that the electron transfer from Cytc to cytochrome c oxidase is the rate-limiting step of the ETC, which is regulated via post-translational modifications of Cytc. This regulation may be dysfunctional in disease conditions such as ischemia-reperfusion injury and neurodegenerative disorders through increased ROS, or cancer, where post-translational modifications on Cytc may provide a mechanism to evade apoptosis.

Flow-cytometric Detection of Low-level Reactive Oxygen Species in Cell Lines and Primary Immune Cells.

Depending on its concentration and cellular origin the production of reactive oxygen species (ROS) in the organism serves a variety of functions. While high concentrations during an oxidative burst are used to fight pathogens, low to moderate amounts of ROS act as signaling molecules important for several physiological processes such as regulation of immune responses. The ROS-sensitive dye 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) is an inexpensive and well-established tool for measuring intracellular ROS levels. However, it needs to be carefully controlled to be able to draw firm conclusions on the nature of ROS species produced and the cellular source of ROS generation such as the enzyme complex NADPH-oxidase 2 (NOX-2). In this protocol, a robust method to determine low intracellular ROS production using H2DCFDA was validated by several ROS-specific as well as NOX-2-specific inhibitors. Cells were treated with inhibitors or control substances prior to treatment with the ROS-inducer of interest. H2DCFDA was added only for the last 30 min of the treatment schedule. To terminate its conversion, we used a ROS-specific inhibitor until analysis by flow cytometry within the FITC-channel (Ex: 488 nm/Em: 519 nm). In summary, this protocol allows the detection of signaling-relevant intracellular ROS production in cell lines and primary immune cells (e.g., Mono Mac 6 cells and Bone marrow-derived dendritic cells, respectively). Using this method in combination with specific inhibitors, we were able to validate even exceptionally low amounts of ROS produced by NOX-2 and relevant for immune-regulatory signaling.

Cytotoxicity and Effects of a New Cacium Hydroxide Nanoparticle Material on Production of Reactive Oxygen Species by LPS-Stimulated Dental Pulp Cells.

The aim of this study was to synthesize and characterize calcium hydroxide (CH) nanoparticles [CH-NP] and compare the cytotoxicity of these materials with that of mineral trioxide aggregate (White MTA) in human dental pulp mesenchymal cells (hDPMCs) stimulated by lipopolysaccharide (LPS). The CH-NP were synthesized by the co-precipitation method, and the physical properties were investigated through X-ray diffraction, scanning electron microscopy (SEM) and energy dispersive x-ray spectrometry (EDS). LPS-stimulated hDPMCs were placed in contact with different dilutions of culture media previously exposed to CH-NP and white MTA for 24 h. The groups were tested for cell viability by MTT formazan and Alamar Blue assays, the production of nitric oxide (NO) by Griess method and the production of reactive oxygen species (ROS) by means of the fluorescent oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Control groups for viability test were maintained in DMEM (not LPS-stimulated). For NO and ROS production, negative control group was cells in DMEM, and positive control was cells stimulated by LPS. The results were statistically analyzed by two-way ANOVA, Tukey's test and Dunnett's test (ɑ=0.05). The results showed that the cell viability remained above 50% in all materials, independent of the dilution in MTT formazan and Alamar Blue tests. MTA showed a reduction in NO production at dilutions of 1:4 to 1:32 compared with the positive control group (P<0.05). The tested materials exhibited lower ROS production by DPMCs than that by cells in the positive control group (P<0.05), and similar ROS production to the negative control group (P>0.05). The outcomes of present in vitro study showed that MTA and [CH-NP] were not cytotoxic materials, with MTA closer to the results of control group (DMEM). MTA and [CH-NP] reduced ROS production at basal levels, with MTA inhibiting NO production at higher dilutions.

Lipoteichoic acid reduces antioxidant enzymes in H9c2 cells

Infective endocarditis (IE) is an illness where the heart is invaded by bacteria, like Streptococcal and Staphylococcal species that contain lipoteichoic acid (LTA) related to an essential role in this disease. This study is the first in evaluating antioxidant enzyme levels in embryonic cardiomyocyte cell line (H9c2) induced by LTA from Streptococcus sanguinis. LTA increased reactive oxygen species (ROS) and reduced the levels of the antioxidant enzymes glutathione peroxidase, superoxide dismutase (SOD)-1 and catalase (CAT) but did not affect glutathione content. At the highest LTA concentration (15 μg/ml), SOD-1 and CAT levels did not change, and this effect was related to the induction of mRNA levels of Nrf2 induced by LTA. These results suggest that low antioxidant enzyme levels and ROS production could be related to IE.

In vitro evaluation of berries of various Vitis genotypes for disease resistance to Botrytis cinerea

Berries of 41 Vitis genotypes were evaluated for resistance to B. cinerea. Evaluation revealed that four genotypes were highly resistant (HR), eight resistant (R), eighteen susceptible (S) and eleven highly susceptible (HS). We further evaluated HR genotype 'Dong fang zhi xing' and HS genotype 'Gold finger' by comparing the fungal growth, reactive oxygen species (ROS) responses, jasmonic acid (JA) levels, anti-oxidants, e.g., Peroxidase (POD), Superoxide dismutase (SOD) and Malondialhydrate (MDA) content changes after infection with B. cinerea. Our results confirmed that the elevated resistance of 'Dong fang zhi xing' was due to weak fungal development, low ROS production, timely elevation of anti-oxidative functions, and high JA levels. Moreover, HS 'Gold finger' infection was severe and sustained ROS production which may be due to its relatively unchanged anti-oxidative activities and low JA level. Our results could help grape breeders to select suitable germplasm for future research work.

RETRACTED ARTICLE: Chondro-protective effects of polydatin in osteoarthritisthrough its effect on restoring dysregulated autophagy via modulating MAPK, and PI3K/Aktsignaling pathways

Osteoarthritis (OA) is a degenerative disease of the cartilage that isprevalent in the middle-aged and elderly demography. Polydatin (PD), a naturalresveratrol glucoside, has shown significant anti-inflammatory and anti-arthriticpotential in previous studies. This study was designed to evaluate the therapeuticproperties of PD in vitro and in vivo, and elucidate their underlying mechanisms. Theexpression levels of all relevant factors were evaluated by qRT-PCR, westernblotting, and immunohistochemistry (IHC) where suitable. Reactive oxygen species(ROS) and apoptosis were analyzed using the suitable probes and flow cytometry. Thehistological evidence of cartilage was assessed in rat models, moreover, the severalserum cytokines levels and autophagy levels were evaluated. The result showed PDdisplayed significant chondro-protective effects, inferred in terms of reducedinflammation and cartilage degradation, apoptosis inhibition, and lower ROSproduction. The protective effects were attenuated by the autophagy inhibitor 3-MA,indicating a mediating role of autophagy in PD action. Mechanistically, PD exertedits effects by inhibiting the MAPK and PI3K/Akt signaling pathways which led to thedown-regulation of mTOR. In conclusion, PD protects against cartilage degenerationby activating the autophagy flux in the chondrocytes via the MAPK and PI3K/Aktsignaling pathways.