Massive Reactive Oxygen Species Research Articles

Hexavalent chromium induces expression of mesenchymal and stem cell markers in renal epithelial cells.

Cr(VI) causes severe kidney damage. The patient's renal function could gradually recover by spontaneous kidney regeneration. The molecular effect of Cr(VI) on recovery of kidney cells, however, has not been clearly elucidated. Here we show that Cr(VI) induces expression of mesenchymal and stem cell markers, cell markers, such as paxillin, vimentin, α‐SMA, nanog, and CD133 of HK‐2 cells. Moreover, Cr(VI) activates epithelial‐to‐mesenchymal transition (EMT). By revealing that levels of dihydrodiol dehydrogenase were promptly reduced following Cr(VI) challenge, our data suggested that DDH could be involved in a Cr(VI)‐related oxidation to generate massive reactive oxygen species and H2O2, and to create intracellular hypoxia, which then increased levels of SUMO‐1 activating enzyme subunit 2, and sumoylation of eukaryotic elongation factor‐2, to mediate the subsequent molecular and cellular responses, e.g., expression of mesenchymal and stem cell markers. Pretreatment with vitamin C reduced Cr(VI)‐related cellular effects. However, no evident effect was observed when vitamin C was added following Cr(VI) challenge. © 2015 The Authors. Molecular Carcinogenesis published by Wiley Periodicals, Inc.

Yeast cytotoxic sensitivity to the antitumour agent β-lapachone depends mainly on oxidative stress and is largely independent of microtubule- or topoisomerase-mediated DNA damage

β-Lapachone (β-lap) is a promising antitumour drug currently undergoing clinical trials. Although it is known that β-lap generates reactive oxygen species (ROS), its actual mechanism of action is still controversial. Especially important is to determine whether concomitant DNA or microtubule damage is the key target of its antitumour properties and whether DNA damage is mediated by topoisomerases as previously suggested. Here, we have searched for determinants of β-lap cytotoxicity in the model organism Saccharomyces cerevisiae through a mechanism-driven approach whereby several pathways of the DNA and microtubule integrity responses, as well as the anti-oxidant response, were downregulated and the outcome of β-lap treatment examined. We also included in the analysis several β-lap derivatives expected to modify drug bioavailability and activity. We found that neither topoisomerase II nor microtubules contributed to yeast sensitivity to β-lap and its equitoxic derivative 3-bromo-β-lapachone. Instead, we found that oxidative and related environmental stresses were primarily responsible for toxicity. Accordingly, Yap1, the central transcription factor in the antioxidant response in yeast, together with several components involved in stress tolerance (i.e., Snf1 and Hog1) and chromatin remodelling (i.e., the SWR1 and RSC complexes), played major roles in protection against β-lapachone. Critically, we show that dioxygen enhanced toxicity and that ROS scavengers protected cells from it. Furthermore, we show that both quinones resulted in cell death in a manner which cytologically resembled apoptosis/necrosis. We thus conclude that β-lap is toxic to yeast through massive ROS production that either directly kills the cells or else triggers programmed cell death.

Synthetic mitochondria as therapeutics against systemic aging: a hypothesis.

We hypothesize herein that synthetic mitochondria, engineered, or reprogrammed to be more energetically efficient and to have mildly elevated levels of reactive oxygen species (ROS) production, would be an effective form of therapeutics against systemic aging. The free radical and mitochondria theories of aging hold that mitochondria-generated ROS underlies chronic organelle, cell and tissues damages that contribute to systemic aging. More recent findings, however, collectively suggest that while acute and massive ROS generation during events such as tissue injury is indeed detrimental, subacute stresses, and chronic elevation in ROS production may instead induce a state of mitochondrial hormesis (or "mitohormesis") that could extend lifespan. Mitohormesis appears to be a convergent mechanism for several known anti-aging signaling pathways. Importantly, mitohormetic signaling could also occur in a non-cell autonomous manner, with its induction in neurons affecting gut cells, for example. Technologies are outlined that could lead towards testing of the hypothesis, which include genetic and epigenetic engineering of the mitochondria, as well as intercellular transfer of mitochondria from transplanted helper cells to target tissues.

The use of vacuum ultraviolet irradiation to oxidize SO2 and NOx for simultaneous desulfurization and denitrification

A simple and efficient method for simultaneous desulfurization and denitrification via vacuum ultraviolet (VUV) irradiation and with no additional chemicals is presented. The simultaneous removal of 90% SO2 and 96% NOx (NO+NO2) was achieved from the simulated flue gas under the irradiation from a low-pressure mercury lamp with main wavelengths of 185 and 254nm, respectively. The composition, flow rate, and temperature of the simulated flue gas, as well as the VUV light intensity, were evaluated as the factors impacting on the efficiency of SO2 and NOx removal. The OH, HO2, O, and O3 produced from the photolysis of H2O and O2 were concluded as the major reactive oxygen species that oxidized SO2 and NOx. The additional OH and HO2 generated through the reactions of NO+HO2 and SO2+OH/HO2 improved treatment efficiency, while the oxidation products of NOx, e.g., NO2, HNO2, HNO3, and HNO4, consumed massive reactive oxygen species (such as O, OH, and HO2) and thereby reducing the removal efficiencies. The main reaction products were characterized as H2SO4 and HNO3 by ion chromatography, which could be used as chemical or fertilizer raw materials.

Mechanism of Cationic Phosphorus Dendrimer Toxicity against Murine Neural Cell Lines

The purpose of this manuscript is to study the toxic responses against murine embryonic hippocampal cells (mHippoE-18) and neuroblastoma cells (N2a) to treatment with cationic phosphorus dendrimers (CPD). Two low generations of CPD--generation 2 (G2) and generation 3 (G3)--were applied to cell cultures to monitor events leading to either apoptosis or necrosis. These processes were analyzed using several bioassays, which included the detection of reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm) alterations, morphology changes, apoptotic and dead cells, cytochrome c (Cyt c) release, caspase 3 activity, DNA fragmentation, as well as changes in cell cycle phases distribution. The results showed that CPD became highly cytotoxic at concentrations above 1 μM and at 0.7 μM in the case of G3 for mHippoE-18 cells. The toxicity was manifested by a pronounced decrease in cell viability, which is correlated with disturbances in cellular activities, such as massive ROS generation. The breakdown of cellular processes leads mainly to the necrotic cell death. Our findings are of high importance in the context of further biomedical studies on CPD.

Moderate extracellular acidification inhibits capsaicin-induced cell death through regulating calcium mobilization, NF-κB translocation and ROS production in synoviocytes

We previously show the expression of transient receptor potential vanilloid 1 (TRPV1) in primary synoviocytes from collagen-induced arthritis (CIA) rats. Capsaicin and lowered extracellular pH from 7.4 to 5.5 induce cell death through TRPV1-mediated Ca2+ entry and reactive oxygen species (ROS) production. However, under the pathological condition in rheumatoid arthritis, the synovial fluid is acidified to a moderate level (about pH 6.8). In the present study, we examined the effects of pH 6.8 on the TRPV1-mediated cell death. Our finding is different or even opposite from what was observed at pH 5.5. We found that the moderate extracellular acidification (from pH 7.4 to 6.8) inhibited the capsaicin-induced Ca2+ entry through attenuating the activity of TRPV1. In the mean time, it triggered a phospholipse C (PLC)-related Ca2+ release from intracellular stores. The nuclear translocation of NF-κB was found at pH 6.8, and this also depends on PLC activation. Moreover, the capsaicin-evoked massive ROS production and cell death were depressed at pH 6.8, both of which are dependent on the activation of PLC and NF-κB. Taken together, these results suggested that the moderate extracellular acidification inhibited the capsaicin-induced synoviocyte death through regulating Ca2+ mobilization, activating NF-κB nuclear translocation and depressing ROS production.

Beta-eleostearic acid induce apoptosis in T24 human bladder cancer cells through reactive oxygen species (ROS)-mediated pathway

Beta-eleostearic acid (β-ESA, 9E11E13E-18:3), a linolenic acid isomer with a conjugated triene system, is a natural and biologically active compound. Herein, we investigated effects of β-eleostearic acid on T24 human bladder cancer cells. In this study, results showed that β-eleostearic acid had strong cytotoxicity to induce cell apoptosis, which was mediated by reactive oxygen species (ROS) in T24 cells. The cell viability assay results showed that incubation with β-eleostearic acid concentrations of 10–80μmol/L caused a dose- and time-dependent decrease of T24 cell viability, and the IC50 value was 21.2μmol/L at 24h and 13.1μmol/L at 48h. Annexin V/PI double staining was used to assess apoptosis with flow cytometry. Treatment with β-eleostearic acid caused massive ROS accumulation and GSH decrease, which lead to activation of caspase-3 and down-regulation of Bcl-2 indicating induction of apoptosis. Subsequently, N-acetyl-l-cysteine (NAC) and PEG–catalase effectively blocked the ROS elevated effect of β-eleostearic acid, which suggested that β-eleostearic acid-induced apoptosis involved ROS generated. Additionally, we found that treating T24 cells with β-eleostearic acid induced activation of PPARγ. A PPARγ-activated protein kinase inhibitor was able to partially abrogate the effects of β-eleostearic acid. These results suggested that β-eleostearic acid can induce T24 cells apoptosis via a ROS-mediated pathway which may be involved PPARγ activation.

Potential roles of PI3K/Akt and Nrf2–Keap1 pathways in regulating hormesis of Z-ligustilide in PC12 cells against oxygen and glucose deprivation

Many phytochemicals may ameliorate neurological disorders through a hormetic mechanism. The aim of this study was to characterize the hormetic role of Z-ligustilide in PC12 cells against oxygen glucose deprivation (OGD) induced cell death. We examined the interactions of Z-ligustilide with the pro-survival signals mediated by phosphatidylinositol 3-kinase (PI3K) and transcription factor nuclear factor-erythroid 2 p45-related factor 2 (Nrf2) pathways. We also investigated the effect of Z-ligustilide on the intracellular redox signaling system involving reactive oxygen species (ROS) and glutathione (GSH). Z-ligustilide not only triggered stress response by causing ROS formation and transient GSH depletion, but also activated survival-promoting signals via cross-talking of PI3K and Nrf2 pathways. A key finding was that Z-ligustilide preconditioning protected PC12 cells from OGD-induced injury either at a low concentration for a prolonged period of time or at a high concentration for a short period of time. Presumably, mild preconditioning stimulated moderate ROS production, but effectively activated hormetic signals and induced stress responsive genes. In contrast, higher concentrations of Z-ligustilide could be toxic over a prolonged period of time due to massive ROS production. These results suggest that the effect of Z-ligustilide may be regulated by a biphasic hormetic mechanism involving initial induction of oxidative stress and subsequent activation of stress response gene expression.

Neuronal Sirt3 Protects against Excitotoxic Injury in Mouse Cortical Neuron Culture

BackgroundSirtuins (Sirt), a family of nicotinamide adenine nucleotide (NAD) dependent deacetylases, are implicated in energy metabolism and life span. Among the known Sirt isoforms (Sirt1-7), Sirt3 was identified as a stress responsive deacetylase recently shown to play a role in protecting cells under stress conditions. Here, we demonstrated the presence of Sirt3 in neurons, and characterized the role of Sirt3 in neuron survival under NMDA-induced excitotoxicity.Methodology/Principal FindingsTo induce excitotoxic injury, we exposed primary cultured mouse cortical neurons to NMDA (30 µM). NMDA induced a rapid decrease of cytoplasmic NAD (but not mitochondrial NAD) in neurons through poly (ADP-ribose) polymerase-1 (PARP-1) activation. Mitochondrial Sirt3 was increased following PARP-1 mediated NAD depletion, which was reversed by either inhibition of PARP-1 or exogenous NAD. We found that massive reactive oxygen species (ROS) produced under this NAD depleted condition mediated the increase in mitochondrial Sirt3. By transfecting primary neurons with a Sirt3 overexpressing plasmid or Sirt3 siRNA, we showed that Sirt3 is required for neuroprotection against excitotoxicity.ConclusionsThis study demonstrated for the first time that mitochondrial Sirt3 acts as a prosurvival factor playing an essential role to protect neurons under excitotoxic injury.

Botanical Flavonoids on Coronary Heart Disease

Ischemic heart disease (IHD) is one of the leading causes of death in Western countries. Prevention rather than treatment of heart disease can significantly improve patients' quality of life and reduce health care costs. Flavonoids are widely distributed in vegetables, fruits and herbal medicines. Regularly consuming botanicals, especially those containing flavonoids, has been associated with a reduction in cardiovascualar disease; thus, it is important to investigate how flavonoids improve cardiac resistance to heart disease and their related mechanisms of action. It has been shown that cardiomyocyte injury and death can result from ischemia-reperfusion, which is pathognomonic of ischemic heart disease. Massive reactive oxygen species (ROS) release at the onset of reperfusion produces cell injury and death. "Programming" the heart to either generate less ROS or to increase strategic ROS removal could reduce reperfusion response. Additionally, profuse nitric oxide (NO) release at reperfusion could be protective in "preconditioning" models. Botanical flavonoids induce preconditioning of the heart, thereby protecting against ischemia-reperfusion injury. In this article, we will discuss two herbs containing potent flavonoids, Scutellaria baicalensis and grape seed proanthocyanidin, which can potentially offer cardiac protection against ischemic heart disease.

Free radical formation and mitochondrial damage in epilepsy – a reperfusion injury

Event Abstract Back to Event Free radical formation and mitochondrial damage in epilepsy – a reperfusion injury Richárd Kovács1*, Christine Huchzermeyer1, Ismini Papageorgiu1, Oliver Kann1 and Uwe Heinemann1 1 Charite-Universitatsmedizin Berlin, Germany Several lines of evidence indicate that reactive oxygen species (ROS) formation is strongly enhanced during epileptic activity leading to mitochondrial dysfunction, energy failure and neuronal cell death. Nevertheless, the sources and targets of ROS have not been identified yet. Here we studied kinetics and spatial pattern of ROS formation in an in vitro model of epilepsy by using electrophysiology, oxygen electrode recordings, microfluorimetry and confocal microscopy. Seizure-like events (SLEs) were associated with biphasic transients of NAD(P)H fluorescence, indicating initial oxidation (dip) followed by a lasting reduction (overshoot) of NAD(P)H. Tissue pO2 decreased by ~107 mmHg during SLEs, but it remained clearly hyperoxic. NAD(P)H dip and overshoot were significantly faster than the peak and recovery of pO2, respectively. Decreasing electron transport chain activity in the presence of high O2 and increased NAD(P)H/NAD(P)+ ratio was associated with an increase in ROS formation, as revealed by the oxidation of mitochondrialy targeted hydroethidine (MitoSox). Co-localisation of MitoSox with mitochondrial as well as with neuronal and astrocytic markers indicated ROS formation in neuronal mitochondria. Mitochondria showed considerable heterogeneity in shape, motility and MitoSox fluorescence. Mitochondria with high levels of ROS were less motile and occasionally underwent thread to grain transition. Remarkably, massive ROS formation was observed in mitochondria of capillary pericytes, leading to contraction of capillaries at late stages of the experimental status epilepticus. Thus we concluded that ROS formation occurs in neurons when electron transport chain activity decreases in the presence of high levels of reducing equivalents and O2. Such conditions may occur in vivo during seizure associated increases in blood flow, whereas ROS dependent capillary constriction might be responsible for disturbances of neurovascular coupling during lasting status epilepticus. Conference: IBRO International Workshop 2010, Pécs, Hungary, 21 Jan - 23 Jan, 2010. Presentation Type: Poster Presentation Topic: Disorders of the nervous system Citation: Kovács R, Huchzermeyer C, Papageorgiu I, Kann O and Heinemann U (2010). Free radical formation and mitochondrial damage in epilepsy – a reperfusion injury. Front. Neurosci. Conference Abstract: IBRO International Workshop 2010. doi: 10.3389/conf.fnins.2010.10.00039 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 20 Apr 2010; Published Online: 20 Apr 2010. * Correspondence: Richárd Kovács, Charite-Universitatsmedizin Berlin, Berlin, Germany, richard.kovacs@charite.de Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Richárd Kovács Christine Huchzermeyer Ismini Papageorgiu Oliver Kann Uwe Heinemann Google Richárd Kovács Christine Huchzermeyer Ismini Papageorgiu Oliver Kann Uwe Heinemann Google Scholar Richárd Kovács Christine Huchzermeyer Ismini Papageorgiu Oliver Kann Uwe Heinemann PubMed Richárd Kovács Christine Huchzermeyer Ismini Papageorgiu Oliver Kann Uwe Heinemann Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Candida albicans cell surface superoxide dismutases degrade host‐derived reactive oxygen species to escape innate immune surveillance

Mammalian innate immune cells produce reactive oxygen species (ROS) in the oxidative burst reaction to destroy invading microbial pathogens. Using quantitative real-time ROS assays, we show here that both yeast and filamentous forms of the opportunistic human fungal pathogen Candida albicans trigger ROS production in primary innate immune cells such as macrophages and dendritic cells. Through a reverse genetic approach, we demonstrate that coculture of macrophages or myeloid dendritic cells with C. albicans cells lacking the superoxide dismutase (SOD) Sod5 leads to massive extracellular ROS accumulation in vitro. ROS accumulation was further increased in coculture with fungal cells devoid of both Sod4 and Sod5. Survival experiments show that C. albicans mutants lacking Sod5 and Sod4 exhibit a severe loss of viability in the presence of macrophages in vitro. The reduced viability of sod5Δ/Δ and sod4Δ/Δsod5Δ/Δ mutants relative to wild type is not evident with macrophages from gp91phox−/− mice defective in the oxidative burst activity, demonstrating a ROS-dependent killing activity of macrophages targeting fungal pathogens. These data show a physiological role for cell surface SODs in detoxifying ROS, and suggest a mechanism whereby C. albicans, and perhaps many other microbial pathogens, can evade host immune surveillance in vivo.

Propofol Attenuates the Decrease of Dynamic Compliance and Water Content in the Lung by Decreasing Oxidative Radicals Released from the Reperfused Liver

Remote pulmonary injuries after hepatic reperfusion are frequently caused by reactive oxygen species (ROS)-induced damage. The choice of anesthetics may affect the balance between oxidants and antioxidants, and propofol, a commonly used anesthetic, has an antioxidant effect. In this study, we developed a model to study pulmonary function with hepatic ischemia/reperfusion (I/R) manipulation, with the aim of defining remote pulmonary dysfunction after hepatic reperfusion and determining if propofol affects this dysfunction by altering ROS production from the liver or lungs. Adult male rats weighing 160-250 g were randomly divided into four groups according to the type of surgery (sham or I/R) and the anesthetic administered (pentobarbital or propofol). To induce I/R, the portal vein and hepatic artery to the left and medial lobes of the liver were clamped. All of the measurements were done after 5 h of reperfusion, after 45 min of ischemia. Pulmonary function after hepatic I/R was determined by dynamic compliance, resistance and wet-to-dry ratio, and by histopathology. Hepato-cellular injuries were confirmed by alanine aminotransferase, whereas ROS production was measured from the inferior vena cava, jugular vein, and carotid artery. Products of lipid peroxidation, thiobarbiturate acid reactive substances and malondialdehyde, were measured in lung and hepatic tissues. Remote lung injury after hepatic I/R was shown by a significant decrease of Cdyn, and increases in resistance and the wet-to-dry ratio. ROS production was significantly increased and was highest in samples from the inferior vena cava. Thiobarbiturate acid reactive substances and malondialdehyde in the liver and serum alanine aminotransferase were significantly increased only in the I/R+pentobarbital group. All of the changes were significantly attenuated in the I/R+ propofol group (P=0.05). With propofol infusion, there was decreased ROS production from the reperfused liver, with less hepato-cellular injury, followed by well-maintained pulmonary function. Remote pulmonary dysfunction and reperfusion injury in the liver were demonstrated in our rat model, as well as massive ROS production and lipid peroxidation. Propofol infusion attenuated remote pulmonary injury by lessening oxidative injury from the reperfused liver.

Melatonin in Cardiac Ischemia/Reperfusion-Induced Mitochondrial Adaptive Changes

Due to its small size and amphiphilic nature, indoleamine melatonin is able to reach all cell compartments quickly. The highest Melatonin concentrations are to be found in the mitochondria, and cytochrome c (cyt c) represents a Melatonin target, since Melatonin is able to oxidise organic molecules in the presence of physiological amounts of hydrogen peroxide (H(2)O(2)). The mitochondrial cyt c-mediated oxidation of melatonin depends on oxoferrylheme in two one-electron steps. The protective effects of Melatonin against cardiac ischemia/reperfusion (I/R)-induced mitochondrial oxidative damage are well documented as an antioxidant agent which prevents and/or modulates the production of reactive oxygen species (ROS) responsible for the reperfusion injury. On the basis of our previous studies on ischemia/reperfusion (I/R) in human cardiomyocytes it has become clear that severe hypoxic and ischemic stimuli evoke a mitochondrial response in terms of the functional adaptation of cyt c oxidase (COX) to oxygen altered availability. Most, if not all, data are compatible with a mechanism of adaptation, in which nitric oxide (NO) generation is now considered as the major player. The NO-induced COX inhibition represents a key point in ischemia-induced, adaptive changes, since a balanced oxidative metabolism between lower O(2) concentrations and COX catalytic capacity down-regulation is thus obtained. This mechanism is a limiting factor in the cardiac ischemia-induced ROS mitochondrial generation. In the light of this concept and if we consider melatonin to be strongly active in cyt c up-regulation, it is obvious that the use of this molecule in the course of cardiac ischemia could be deleterious. On the contrary, the prompt restoration of the coronary blood flow is inexorably associated with the so-called "reperfusion syndrome" and/or with a condition of myocardial stunning. These phenomena are strictly linked to the aforementioned ischemia-induced cyt c adaptive changes, for the reason that a primary imbalance is induced between the persistent down-regulated enzymatic oxidative capacity and mitochondrial oxygen gradients are rapidly restored. This leads to a massive ROS neo-generation, which evokes well-known cellular disorders. In this scenario the melatonin treatment could play a pivotal role in terms of a cyt c up-regulating agent, crucial for counteracting most, if not all the biochemical and molecular phenomena, on which the clinical symptoms proper of myocardial reperfusion syndrome are based.

A simple but effective cancer vaccine consisting of an antigen and a cationic lipid.

Developing a cancer vaccine with a potent adjuvant, which is safe for human use, remains to be an unmet need. In this study, we developed a simple, safe, yet efficient, peptide-based therapeutic cancer vaccine, DOTAP/E7 complex, which comprises only two molecules: a DOTAP cationic lipid and a peptide antigen derived from E7 oncoprotein of human papillomavirus (HPV) type 16. The anti-cancer activity of DOTAP/E7 against existing HPV positive TC-1 tumor was compared to that of our previous LPD/E7 formulation, which contains bacterial DNA CpG motifs. Tumor-bearing mice showed significant tumor inhibition following a single vaccination of either formulation at the optimal lipid dose, suggesting that DOTAP liposome alone can provide a potent adjuvant activity without plasmid DNA. E7 peptide formulated with DOTAP induced migration of activated dendritic cells (DC) to the draining lymph node (DLN) and efficiently generated functional antigen-specific CD8+ T lymphocyte responses. Accumulation of CD8+ tumor infiltrating T cells and apoptosis at tumor sites were observed after treatment with DOTAP/E7 complexes, which was also associated with a decreased amount of CD25(+)Foxp3(+) regulatory T cells in treated animals. Reactive oxygen species (ROS) induced by DOTAP cationic lipid in DLN revealed a plausible mechanism of the initial interaction between DC and DOTAP. An adequate amount of ROS generation was apparently required for the initiation of the vaccine mechanism; however, an overdose of DOTAP induced massive ROS production and apoptosis of DC in DLN, which led to diminished anti-cancer immunity. Overall, these results indicate that cationic lipid DOTAP alone serves as an efficient vaccine adjuvant for the induction of a therapeutic, antigen-specific anti-cancer activity.

A triterpenediol from Boswellia serrata induces apoptosis through both the intrinsic and extrinsic apoptotic pathways in human leukemia HL-60 cells

A triterpenediol (TPD) comprising of isomeric mixture of 3alpha, 24-dihydroxyurs-12-ene and 3alpha, 24-dihydroxyolean-12-ene from Boswellia serrata induces apoptosis in cancer cells. An attempt was made in this study to investigate the mechanism of cell death by TPD in human leukemia HL-60 cells. It inhibited cell proliferation with IC50 approximately 12 microg/ml and produced apoptosis as measured by various biological end points e.g. increased sub-G0 DNA fraction, DNA ladder formation, enhanced AnnexinV-FITC binding of the cells. Further, initial events involved massive reactive oxygen species (ROS) and nitric oxide (NO) formation, which were significantly inhibited by their respective inhibitors. Persistent high levels of NO and ROS caused Bcl-2 cleavage and translocation of Bax to mitochondria, which lead to loss of mitochondrial membrane potential (Deltapsim) and release of cytochrome c, AIF, Smac/DIABLO to the cytosol. These events were associated with decreased expression of survivin and ICAD with attendant activation of caspases leading to PARP cleavage. Furthermore, TPD up regulated the expression of cell death receptors DR4 and TNF-R1 level, leading to caspase-8 activation. These studies thus demonstrate that TPD produces oxidative stress in cancer cells that triggers self-demise by ROS and NO regulated activation of both the intrinsic and extrinsic signaling cascades.

New functions of XPC in the protection of human skin cells from oxidative damage.

Xeroderma pigmentosum (XP) C is involved in the recognition of a variety of bulky DNA-distorting lesions in nucleotide excision repair. Here, we show that XPC plays an unexpected and multifaceted role in cell protection from oxidative DNA damage. XP-C primary keratinocytes and fibroblasts are hypersensitive to the killing effects of DNA-oxidizing agents and this effect is reverted by expression of wild-type XPC. Upon oxidant exposure, XP-C primary keratinocytes and fibroblasts accumulate 8,5'-cyclopurine 2'-deoxynucleosides in their DNA, indicating that XPC is involved in their removal. In the absence of XPC, a decrease in the repair rate of 8-hydroxyguanine (8-OH-Gua) is also observed. We demonstrate that XPC-HR23B complex acts as cofactor in base excision repair of 8-OH-Gua, by stimulating the activity of its specific DNA glycosylase OGG1. In vitro experiments suggest that the mechanism involved is a combination of increased loading and turnover of OGG1 by XPC-HR23B complex. The accumulation of endogenous oxidative DNA damage might contribute to increased skin cancer risk and account for internal cancers reported for XP-C patients.

Ganglioside GM3 is involved in neuronal cell death

Gangliosides abundant in the nervous system have been implicated in a broad range of biological functions, including the regulation of cell proliferation and death. Glutamate-induced cell death, which is accompanied by an accumulation of reactive oxygen species (ROS), is a major contributor to pathological cell death within the nervous system. However, the mechanism underlying this neuronal cell death has not been fully elucidated. In this study, we report that ganglioside GM3 is involved in neuronal cell death. GM3 was up-regulated in the mouse hippocampal cell line HT22 death caused by glutamate. Increment in GM3 levels by both the exogenous addition of GM3 and the overexpression of the GM3 synthase gene induced neuronal cell death. Overexpression of GM3 synthase by microinjecting mRNA into zebrafish embryos resulted in neuronal cell death in the central nervous system (CNS). Conversely, RNA interference-mediated silencing of GM3 synthase rescued glutamate-induced neuronal death, as evidenced by the inhibition of massive ROS production and intracellular calcium ion influx. 12-lipoxygenase (12-lipoxygenase) (12-LOX) was recruited to glycosphingolipid-enriched microdomains (GEM) in a GM3-dependent manner during oxidative glutamate toxicity. Our findings suggest that GM3 acts as not only a mediator of oxidative HT22 death by glutamate but also a modulator of in vivo neuronal cell death.

Glutathione-Related Systems and Modulation of Extracellular Signal–Regulated Kinases Are Involved in the Resistance of AGS Adenocarcinoma Gastric Cells to Diallyl Disulfide–Induced Apoptosis

We have previously characterized the cytotoxic action of diallyl disulfide (DADS) on neuroblastoma cells, and we have shown the crucial role of an early and massive reactive oxygen species production in the induction of c-Jun NH(2)-terminal kinase-mediated apoptotic pathway. In the present work, we report that DADS is ineffective in inducing apoptosis in a human adenocarcinoma gastric cell line (AGS). In particular, we show that AGS cells are able to recover from the p53/p21-mediated cell cycle arrest in the G(2)-M phase upon DADS treatment without committing cells to death. This event is most likely due to a peculiar surviving pathway of these cells involving: (a) the formation of mixed disulfides between reduced glutathione (GSH) and protein thiols, (b) a higher and inducible glutathione peroxidase activity, and/or (c) an efficient modulation of the phospho-active levels of the extracellular signal-regulated kinases 1 and 2 (ERK 1/2). Moreover, by increasing glutathione peroxidase expression or GSH concentrations, cell cycle arrest is fully abolished; the apoptotic death is induced by either decreasing the availability of intracellular GSH or inhibiting the reactivation of ERK 1/2. Altogether, our data show that ERK 1/2 participates in the active proliferation of AGS cells and that an efficient reactive oxygen species buffering system makes these cells resistant to DADS-mediated detrimental effects.

Finally, proof of weapons of mass destruction.

Allelopathy (one species' use of chemicals to harm other species) may be a key ingredient in successful invasions of alien plants into established communities. Bais et al. show that in response to elicitation by common soil fungi, spotted knapweed (Centaurea maculosa) launches an ineffective defense against the fungi that results in extensive collateral damage to neighboring plants. Specifically, the flavonoid (-)-catechin, released from the roots of knapweed, produces a massive reactive oxygen species (ROS)-mediated oxidative burst, Ca2+ signaling, and rapid cell death in those unadapted native species whose habitats the plant regularly invades. The roots release both (+) and (-) enantiomers, but only the (-) enantiomer functions as a weapon of mass destruction; the (+) enantiomer inhibits the growth of numerous common soil-borne bacterial pathogens. Eliciting apoptotic response for a competitive advantage is an example of signal cross-talk between the genomes of interacting organisms and highlights how the internal signaling of one organism can be used by others to adjust their phenotypes in an adaptive manner. The study provides strong circumstantial evidence for an allelopathic interaction, but the genetic manipulation of (-)-catechin release would allow researchers to determine if these responses occur in nature. Precise genetic control over the release of secondary metabolites from plants would benefit ecological research.