Glutathione Supplementation Prevents Neonatal Parenteral Nutrition-Induced Short- and Long-Term Epigenetic and Transcriptional Disruptions of Hepatic H2O2 Metabolism in Guinea Pigs.

The parenteral nutrition (PN) received by premature newborns is contaminated with peroxides that induce global DNA hypermethylation via oxidative stress. Exposure to peroxides could be an important factor in the induction of chronic diseases such as those observed in adults who were born preterm. As endogenous H2O2 is a major regulator of glucose-lipid metabolism, our hypothesis was that early exposure to PN induces permanent epigenetic changes in H2O2 metabolism. Three-day-old guinea pigs were fed orally (ON), PN or glutathione-enriched PN (PN+GSSG). GSSG promotes endogenous peroxide detoxification. After 4 days, half the animals were sacrificed, and the other half were fed ON until 16 weeks of age. The liver was harvested. DNA methylation and mRNA levels were determined for the SOD2, GPx1, GCLC, GSase, Nrf2 and Keap1 genes. PN induced GPx1 hypermethylation and decreased GPx1, GCLC and GSase mRNA. These findings were not observed in PN+GSSG. PN+GSSG induced Nrf2 hypomethylation and increased Nrf2 and SOD2 mRNA. These observations were independent of age. In conclusion, in neonatal guinea pigs, PN induces epigenetic changes, affecting the expression of H2O2 metabolism genes. These changes persist for at least 15 weeks after PN. This disruption may signify a permanent reduction in the capacity to detoxify peroxides.

The potential effect of Moringa oleifera ethanolic leaf extract against oxidative stress, immune response disruption induced by abamectin exposure in Oreochromis niloticus.

Abamectin (ABM), a naturally fermented product of Streptomyces avermitilis, is applied to pest control in livestock and agriculture fields. The aim of the current study is to evaluate the protective effects of Moringa oleifera leaf ethanolic extract (MOE) on biochemical changes including oxidative stress indices, immune response marker, lipid profiles as well as mRNA expression of immune related genes, and abamectin (ABM, 5% EC) residue levels in Nile tilapia (Oreochromis niloticus) exposed to a sub-lethal concentration (0.5 µg/l) for 28 days. Disturbance in liver and kidney biomarkers was markedly increased in ABM-exposed fish compared to the control group. Malondialdehyde levels in the liver and brain tissues, as well as the activities of glutathione-s-transferase, superoxide dismutase, and glutathione peroxides, all increased significantly in ABM group. Additionally, ABM exposure increased the levels of interleukin 10 beta and growth factor gene expression. On the other hand, fish exposed to ABM had significantly lower serum alkaline phosphatase, creatinine, high-density lipoprotein, glutathione peroxides in brain, glutathione in liver and brain tissues, lysozyme activity, nitric oxide, immunoglobulin M, tumor necrosis factor, and interleukin 1 beta as compared to the control group. The recorded detrimental effects of ABM on tilapia have been overcome by the addition of MOE to the diet (1%) and ameliorating hepato-renal damage and enhancing antioxidant activity, innate immune responses, and upregulating the anti-inflammatory gene expression. Therefore, it could be concluded that MOE dietary supplementation at 1% could be used to counteract the oxidative stress, immune response disruption induced by abamectin exposure in Oreochromis niloticus, and reduce its accumulation in fish tissues.

Tomato seed oil-enriched tomato juice: Effect of oil addition type and heat treatment on lycopene bioaccessibility and oxidative stability.

In this study, tomato seed oil conventional emulsion (7 µm) and nanoemulsion (0.146 µm) with desirable stability were prepared, then the effect of tomato seed oil addition (bulk and emulsified forms) and thermal treatment on properties of tomato juice was evaluated. Tomato juice without oil and heat treatment exhibited the lowest bioaccessibility of lycopene (17.8 %). Incorporation of oil and applying heat treatment significantly increased the extent of lipid digestion and bioaccessibility of lycopene. In this regard, the nanoemulsion had the highest bioaccessibility (44.85 %) compared to conventional emulsion (33.90 %) and bulk oil (27.11 %), due to the smaller oil droplets. The oxidative stability of oil in heat-treated tomato juice samples decreased during 28 days of storage at 4 °C, whereas the nanoemulsion exhibited the highest peroxide value (4.43 meq O2/kg of oil) compared to conventional emulsion and bulk oil (3.91 and 3.49 meq O2/kg of oil, respectively) at the end of the period.

Protective Effect of SeMet on Liver Injury Induced by Ochratoxin A in Rabbits.

Ochratoxin A (OTA) is second only to aflatoxin in toxicity among mycotoxins. Recent studies have shown that selenomethionine (SeMet) has a protective effect on mycotoxin-induced toxicity. The purpose of this study was to investigate the protective effect and mechanism of SeMet on OTA-induced liver injury in rabbits. Sixty 35-day-old rabbits with similar body weight were randomly divided into five groups: control group, OTA group (0.2 mg/kg OTA), OTA + 0.2 mg/kg SeMet group, OTA + 0.4 mg/kg SeMet group and OTA + 0.6 mg/kg SeMet group. Rabbits were fed different doses of the SeMet diet for 21 d, and OTA was administered for one week from day 15 (the control group was provided the same dose of NaHCO3 solution). The results showed that 0.4 mg/kg SeMet could significantly improve the liver injury induced by OTA poisoning. SeMet supplementation can improve the changes in physiological blood indexes caused by OTA poisoning in rabbits and alleviate pathological damage to the rabbit liver. SeMet also increased the activities of SOD, GSH-Px and T-AOC and significantly decreased the contents of ROS, MDA, IL-1ß, IL-6 and TNF-α, effectively alleviating the oxidative stress and inflammatory response caused by OTA poisoning. In addition, OTA poisoning inhibits Nrf2 and HO-1 levels, ultimately leading to peroxide reaction, while SeMet activates the Nrf2 signaling pathway and enhances the expression of the HO-1 downstream Nrf2 gene. These results suggest that Se protects the liver from OTA-induced hepatotoxicity by regulating Nrf2/HO-1 expression.

Selenoprotein P - Selenium transport protein, enzyme and biomarker of selenium status.

The habitual intake of selenium (Se) varies strongly around the world, and many people are at risk of inadequate supply and health risks from Se deficiency. Within the human organism, efficient transport mechanisms ensure that organs with a high demand and relevance for reproduction and survival are preferentially supplied. To this end, selenoprotein P (SELENOP) is synthesized in the liver and mediates Se transport to essential tissues such as the endocrine glands and the brain, where the "SELENOP cycle" maintains a privileged Se status. Mouse models indicate that SELENOP is not essential for life, as supplemental Se supply was capable of preventing the development of severe symptoms. However, knockout mice died under limiting supply, arguing for an essential role of SELENOP in Se deficiency. Many clinical studies support this notion, pointing to close links between health risks and low SELENOP levels. Accordingly, circulating SELENOP concentrations serve as a functional biomarker of Se supply, at least until a saturated status is achieved and SELENOP levels reach a plateau. Upon toxic intake, a further increase in SELENOP is observed, i.e., SELENOP provides information about possible selenosis. The SELENOP transcripts predict an insertion of ten selenocysteine residues. However, the decoding is imperfect, and not all these positions are ultimately occupied by selenocysteine. In addition to the selenocysteine residues near the C-terminus, one selenocysteine resides central within an enzyme-like environment. SELENOP proved capable of catalyzing peroxide degradation in vitro and protecting e.g. LDL particles from oxidation. An enzymatic activity in the intact organism is unclear, but an increasing number of clinical studies provides evidence for a direct involvement of SELENOP-dependent Se transport as an important and modifiable risk factor of disease. This interaction is particularly strong for cardiovascular and critical disease including COVID-19, cancer at various sites and autoimmune thyroiditis. This review briefly highlights the links between the growing knowledge of Se in health and disease over the last 50 years and the specific advances that have been made in our understanding of the physiological and clinical contribution of SELENOP to the current picture.

From Molecules to Behavior in Long-Term Inorganic Mercury Intoxication: Unraveling Proteomic Features in Cerebellar Neurodegeneration of Rats.

Mercury is a severe environmental pollutant with neurotoxic effects, especially when exposed for long periods. Although there are several evidences regarding mercury toxicity, little is known about inorganic mercury (IHg) species and cerebellum, one of the main targets of mercury associated with the neurological symptomatology of mercurial poisoning. Besides that, the global proteomic profile assessment is a valuable tool to screen possible biomarkers and elucidate molecular targets of mercury neurotoxicity; however, the literature is still scarce. Thus, this study aimed to investigate the effects of long-term exposure to IHg in adult rats' cerebellum and explore the modulation of the cerebellar proteome associated with biochemical and functional outcomes, providing evidence, in a translational perspective, of new mercury toxicity targets and possible biomarkers. Fifty-four adult rats were exposed to 0.375 mg/kg of HgCl2 or distilled water for 45 days using intragastric gavage. Then, the motor functions were evaluated by rotarod and inclined plane. The cerebellum was collected to quantify mercury levels, to assess the antioxidant activity against peroxyl radicals (ACAPs), the lipid peroxidation (LPO), the proteomic profile, the cell death nature by cytotoxicity and apoptosis, and the Purkinje cells density. The IHg exposure increased mercury levels in the cerebellum, reducing ACAP and increasing LPO. The proteomic approach revealed a total 419 proteins with different statuses of regulation, associated with different biological processes, such as synaptic signaling, energy metabolism and nervous system development, e.g., all these molecular changes are associated with increased cytotoxicity and apoptosis, with a neurodegenerative pattern on Purkinje cells layer and poor motor coordination and balance. In conclusion, all these findings feature a neurodegenerative process triggered by IHg in the cerebellum that culminated into motor functions deficits, which are associated with several molecular features and may be related to the clinical outcomes of people exposed to the toxicant.

Pharmacological ascorbate inhibits pancreatic cancer metastases via a peroxide-mediated mechanism.

Pharmacological ascorbate (P-AscH-, high-dose, intravenous vitamin C) is cytotoxic to tumor cells in doses achievable in humans. Phase I studies in pancreatic cancer (PDAC) utilizing P-AscH- have demonstrated increases in progression free survival, suggesting a reduction in metastatic disease burden. The purpose of this study was to determine the effects of P-AscH- on metastatic PDAC. Several in vitro and in vivo mechanisms involved in PDAC metastases were investigated following treatment with P-AscH-. Serum from PDAC patients in clinical trials with P-AscH- were tested for the presence and quantity of circulating tumor cell-derived nucleases. P-AscH- inhibited invasion, basement membrane degradation, decreased matrix metalloproteinase expression, as well as clonogenic survival and viability during exposure to fluid shear stress. In vivo, P-AscH- significantly decreased formation of ascites, tumor burden over time, circulating tumor cells, and hepatic metastases. Both in vitro and in vivo findings were reversed with the addition of catalase suggesting that the effect of P-AscH- on metastatic disease is mediated by hydrogen peroxide. Finally, P-AscH- decreased CTC-derived nucleases in subjects with stage IV PDAC in a phase I clinical trial. We conclude that P-AscH- attenuates the metastatic potential of PDAC and may prove to be effective for treating advanced disease.

Deciphering the in vivo redox behavior of human peroxiredoxins I and II by expressing in budding yeast.

Peroxiredoxins (Prxs), scavenge cellular peroxides by forming recyclable disulfides but under high oxidative stress, hyperoxidation of their active-site Cys residue results in loss of their peroxidase activity. Saccharomyces cerevisiae deficient in human Prx (hPrx) orthologue TSA1 show growth defects under oxidative stress. They can be complemented with hPRXI but not by hPRXII, but it is not clear how the disulfide and hyperoxidation states of the hPrx vary in yeast under oxidative stress. To understand this, we used oxidative-stress sensitive tsa1tsa2Δ yeast strain to express hPRXI or hPRXII. We found that hPrxI in yeast exists as a mixture of disulfide-linked dimer and reduced monomer but becomes hyperoxidized upon elevated oxidative stress as analyzed under denaturing conditions (SDS-PAGE). In contrast, hPrxII was present predominantly as the disulfide in unstressed cells and readily converted to its hyperoxidized, peroxidase-inactive form even with mild oxidative stress. Interestingly, we found that plant extracts containing polyphenol antioxidants provided further protection against the growth defects of the tsa1tsa2Δ strain expressing hPrx and preserved the peroxidase-active forms of the Prxs. The extracts also helped to protect against hyperoxidation of hPrxs in HeLa cells. Based on these findings we can conclude that resistance to oxidative stress of yeast cells expressing individual hPrxs requires the hPrx to be maintained in a redox state that permits redox cycling and peroxidase activity. Peroxidase activity decreases as the hPrx becomes hyperoxidized and the limited protection by hPrxII compared with hPrxI can be explained by its greater sensitivity to hyperoxidation.

Modulation of selenium-dependent glutathione peroxidase activity enhances doxorubicin-induced apoptosis, tumour cell killing and hydroxyl radical production in human NCI/ADR-RES cancer cells despite high-level P-glycoprotein expression.

To define the role of glutathione peroxidase (GPx) in modulating the oxygen radical-related cytotoxicity of doxorubicin and H2O2 in cells that overexpress P-glycoprotein (Pgp), the GPx activity of NCI/ADR-RES cancer cells was altered by growth in 0.5% serum with (MR-30 subline) or without (MR-0 subline) selenium supplementation. GPx activity increased from 2.2 nmol/min/mg (MR-0) to 22.5 nmol/min/mg (MR-30) when cells were grown in 30-nM selenium, p < .01; the activities of other antioxidant enzymes were unchanged by selenium. By reverse transcriptase polymerase chain reaction, MR-30 and MR-0 cells expressed similar levels of the MDR1, GPx-1, BCL2 and TOP2A mRNA. The IC50 concentration for H2O2 in MR-0 cells was 10-fold lower than in the MR-30 subline, p < .01. Despite identical anthracycline accumulation and efflux in these two lines that expressed equivalent levels of Pgp, the doxorubicin IC50 decreased fivefold in MR-0 versus MR-30 cells, p < .01. Log-linear tumour cell killing by doxorubicin was observed only in selenium-deficient MR-0 cells. Doxorubicin exposure also produced substantially more apoptosis in MR-0 than MR-30 cells; this was not related to the presence of selenium per se. MR-0 cells generated ≈5-times more methane from dimethyl sulfoxide (a measure of reactive oxygen metabolism) than MR-30 cells in the presence of equimolar doxorubicin concentrations (p < .05). These studies suggest that GPx-mediated detoxification of peroxides can modulate the antitumor activity of doxorubicin in the presence of high levels of Pgp.

Effect of antioxidant supplements on lipid peroxidation levels in primary cortical neuron cultures.

Oxidative stress, specifically lipid peroxidation, is a major driving force in neurodegenerative processes. However, the exact role of lipid peroxidation remains elusive as reliable real-time detection and quantification of lipid peroxyl radicals proves to be challenging in vitro and in vivo. Motivated by this methodological limitation, we have optimized conditions for real-time imaging and quantification of lipid peroxyl radical generation in primary neuron cultures using the lipophilic fluorogenic antioxidant H4BPMHC (8-((6-hydroxy-2,5,7,8-tetramethylchroman-2-yl)-methyl)-1,5-di(3-chloropropyl)-pyrromethene fluoroborate), an α-tocopherol analog probe. By subjecting neurons to different antioxidant conditions in the presence and absence of lipid peroxidation inducing stressors (Haber-Weiss reagents), we maximized H4BPMHC sensitivity and confirmed its potential to temporally resolve subtle and marked differences in lipid peroxidation levels in real-time. Herein we report imaging and quantification of homeostatic and induced lipid peroxidation in primary neuron cultures, supporting the use of this probe for investigating healthy and diseased states. Overall these results provide the necessary foundation and impetus towards using H4BPMHC for elucidating and mapping lipid peroxyl radical contributions to ROS-associated pathological processes in neurons.

Influences of Light Intensity and ß-Carotene on Polycyclic Aromatic Hydrocarbons and Aldehydes in Vegetable Oil: A Case Study Using Palm Oil.

This study investigated the effects of three light intensities on four types of palm oils during consecutive storage for 12 months at 4 °C. The concentrations of 4-hydroxy-2- trans-hexenal (4-HHE), 4-hydroxy-2- trans-nonenal (4-HNE), polycyclic aromatic hydrocarbon (PAH)4, and PAH8 in the oils significantly increased with the increasing light intensity after storage. The red palm oil had the lowest rate of increase of 4-HNE, while 5° palm oil had the highest rate of increase of the PAH, OPAH, 4-HNE, and peroxide values during storage. For the same type of oil, OPAHs increased significantly under a light intensity of 6000 lx (lx) after storage. The increasing concentrations of 9FO, ATQ, and BaPO in the oils stored at 6000 lx showed a positive relation to their corresponding parent PAHs, indicating that PAH oxidation occurred at 6000 lx. The results suggest that light intensity and ß-carotene may control PAHs, OPAHs, and 4-hydroxy-trans- alkenals for vegetable oil storage, transportation, and retail.

Generation of hydroxyl radicals by Fe-polyphenol-activated CaO2 as a potential treatment for soil-borne diseases.

An Fe-polyphenol catalyst was recently developed using anhydrous iron (III) chloride and coffee grounds as raw materials. The present study aims to test the application of this Fe-polyphenol catalyst with two hydrogen peroxide (H2O2) sources in soil as a new method for controlling the soil-borne disease caused by Ralstonia solanacearum and to test the hypothesis that hydroxyl radicals are involved in the catalytic process. Tomato cv. Momotaro was used as the test species. The results showed that powdered CaO2 (16% W/W) is a more effective H2O2 source for controlling bacterial wilt disease than liquid H2O2 (35% W/W) when applied with an Fe-polyphenol catalyst. An electron paramagnetic resonance spin trapping method using a 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) assay and Fe-caffeic acid and Fe-chlorogenic acid complexes as models showed that these organometallic complexes react with the H2O2 released by CaO2, producing hydroxyl radicals in a manner that is consistent with the proposed catalytic process. The application of Fe-polyphenol with powdered CaO2 to soil could be a new environmentally friendly method for controlling soil-borne diseases.

Antioxidant Properties and Yield of Wood Ear Mushroom, Auricularia polytricha (Agaricomycetes), Cultivated on Rubberwood Sawdust.

Mushroom cultivation has become an important component of agriculture, providing food and contributing to the global economy. It uses vertical space and addresses issues of food quality, health improvement, and environmental sustainability. Auricularia mushrooms are popular ingredients in traditional Chinese cuisine. The objective of this study was to determine yield and evaluate radical scavenging capacity of A. polytricha cultivated on rubberwood sawdust on a large scale; we measured total phenolic content; DPPH, hydroxyl, superoxide anion, and peroxyl radical scavenging; and reducing power. Cultivation on rubberwood sawdust produces an average of 4 harvests per bag and a biological efficiency of 80-82%. The antioxidant capacity investigations revealed that the ethyl acetate fraction was the most potent radical scavenger in all assays except that for superoxide anions, whereas the aqueous fraction exhibited mild to moderate antioxidant capacity in scavenging the various radicals. Artificial cultivation of A. polytricha on rubberwood sawdust yields many sporophores with potent antioxidant capacity.

Endoperoxide-8-aminoquinoline hybrids as dual-stage antimalarial agents with enhanced metabolic stability.

Hybrid compounds may play a critical role in the context of the malaria eradication agenda, which will benefit from therapeutic tools active against the symptomatic erythrocytic stage of Plasmodium infection, and also capable of eliminating liver stage parasites. To address the need for efficient multistage antiplasmodial compounds, a small library of 1,2,4,5-tetraoxane-8- aminoquinoline hybrids, with the metabolically labile C-5 position of the 8-aminoquinoline moiety blocked with aryl groups, was synthesized and screened for antiplasmodial activity and metabolic stability. The hybrid compounds inhibited development of intra-erythrocytic forms of the multidrug-resistant Plasmodium falciparum W2 strain, with EC50 values in the nM range, and with low cytotoxicity against mammalian cells. The compounds also inhibited the development of P. berghei liver stage parasites, with the most potent compounds displaying EC50 values in the low µM range. SAR analysis revealed that unbranched linkers between the endoperoxide and 8-aminoquinoline pharmacophores are most beneficial for dual antiplasmodial activity. Importantly, hybrids were significantly more potent than a 1:1 mixture of 8-aminoquinoline-tetraoxane, highlighting the superiority of the hybrid approach over the combination therapy. Furthermore, aryl substituents at C-5 of the 8-aminoquinoline moiety improve the compounds' metabolic stability when compared with their primaquine (i.e. C-5 unsubstituted) counterparts. Overall, this study reveals that blocking the quinoline C-5 position does not result in loss of dual-stage antimalarial activity, and that tetraoxane-8- aminoquinoline hybrids are an attractive approach to achieve elimination of exo- and intraerythrocytic parasites, thus with the potential to be used in malaria eradication campaigns.

Rate of Lipid Peroxyl Radical Production during Cellular Homeostasis Unraveled via Fluorescence Imaging.

Reactive oxygen species (ROS) and their associated byproducts have been traditionally associated with a range of pathologies. It is now believed, however, that at basal levels these molecules also have a beneficial cellular function in the form of cell signaling and redox regulation. Critical to elucidating their physiological role is the opportunity to visualize and quantify the production of ROS with spatiotemporal accuracy. Armed with a newly developed, extremely sensitive fluorogenic α-tocopherol analogue (H4BPMHC), we report herein the observation of steady concentrations of lipid peroxyl radicals produced in live cell imaging conditions. Imaging studies with H4BPMHC indicate that the rate of production of lipid peroxyl radicals in HeLa cells under basal conditions is 33 nM/h within the cell. Our work further provides indisputable evidence on the antioxidant role of Vitamin E, as lipid peroxidation was suppressed in HeLa cells both under basal conditions and in the presence of Haber-Weiss chemistry, generated by the presence of cumyl hydroperoxide and Cu2+ in solution, when supplemented with the α-tocopherol surrogate, PMHC (2,2,5,7,8-pentamethyl-6-hydroxy-chromanol, an α-tocopherol analogue lacking the phytyl tail). H4BPMHC has the sensitivity needed to detect trace changes in oxidative status within the lipid membrane, underscoring the opportunity to illuminate the physiological relevance of lipid peroxyl radical production during cell homeostasis and disease.

Antioxidant and anti-osteoporotic effects of anthraquinones and related constituents from the aqueous dissolved Aloe exudates.

In this study, 25 known anthraquinones and related compounds were isolated from aqueous dissolved Aloe exudates. The antioxidant and anti-osteoporotic activities of the isolated compounds were then investigated. Compounds 8, 11, 20, and 23 showed good antioxidant activity based on peroxyl radical-scavenging and reducing capacity assays at a concentration of 10.0 µM. Additionally, compounds 7, 9, 15-16, 18, 21-22 and 24-25 showed potent peroxyl radical-scavenging activities with values ranging from 5.28 to 14.60 at 10.0 µM. Moreover, compounds 8, 11, 15, 20 and 22 exhibited significantly suppressed tartrate-resistant acid phosphatase (TRAP) activity in nuclear factor-κB ligand-activated osteoclastic RAW 264.7 cells, with values of 125.67, 118.54, 127.64, 125.82 and 124.98%, respectively. These results indicate that Aloe is an excellent source of antioxidant and anti-osteoporotic phytochemicals.

Improved detection of mineral oil toxicity using an extended mouse embryo assay.

PURPOSE: Successful in vitro fertilization (IVF) relies on sound laboratory methods and culture conditions which depend on sensitive quality control (QC) testing. This study aimed to improve the sensitivity of mouse embryo assays (MEA) for detection of mineral oil toxicity. METHODS: Five experiments were conducted to study modifications of the standard mouse embryo assay (MEA) in order to improve sensitivity using clinical grade mineral oil with known peroxide concentrations. Assessment of blastocyst development at either 96 h or in an extended MEA (eMEA) to 144 h was tested in each experiment. In experiment 1, ability to detect peroxides in oil was compared in the MEA, eMEA, and cell number at 96 h. In experiment 2, serial dilutions of peroxide in oil were used along with time-lapse imaging to compare sensitivity of the morphokinetic MEA to the eMEA. Culture conditions that may affect assay sensitivity were assessed in experiments 3-5, which examined the effect of group versus individual culture, oxygen concentration, and protein supplementation. RESULTS: Extended MEA and cell counts identified toxicity not detected by the routine endpoint of blastocyst rate at 96 h. The eMEA was fourfold more sensitive than the standard MEA, and this sensitivity was similar to the morphokinetic MEA. Group culture had a protective effect against toxicity, while oxygen concentration did not affect blastocyst development. Protein supplementation with HSA had a protective effect on blastocyst development in eMEA. CONCLUSIONS: The standard MEA used by manufacturers does not detect potentially lethal toxicity of peroxides in mineral oil. While group culture may mask toxicity, protein supplementation and oxygen concentration have minimal effect on assay sensitivity. The eMEA and time-lapse morphokinetic assessment are equally effective in detection of peroxide toxicity and thus provide manufacturers and end-users a simple process modification that can be readily adopted into an existing QC program.

[In vitro Modulating Activity of aqueous extracts from American Plants on Chlorpyrifos-induced toxicity on Murine Splenocytes]. / Actividad moduladora in vitro de extractos acuosos de plantas americanas sobre la toxicidad inducida por clorpirifos en esplenocitos murinos.

Background: Chlorpyrifos is an highly toxic pesticide, which can induce immunotoxicity with deleterious effects on health worldwide. On the other hand, American plants can provide derivatives with protective and immunostimulating activity. Thus, plant potential against chlorpyrifos should be assayed. Objective: To identify bioactive aqueous extracts from Lantana grisebachii (LG), Aspidosperma quebracho-blanco (AQ), Peumus boldus (PB), and Ilex paraguariensis (IP), against chlorpyrifos-induced toxicity on female Balb/c splenocytes. Material and method: Splenocytes were treated in vitro for 72 hours with 0-35 µg/mL of chlorpyrifos, 0-100 µg/mL of each extract (LG, AQ, PB, IP), and 0-5 µg/mL of concanavalin A. Then, cellular viability and death (resazurin-based and propidium iodide stainings), hydroperoxides, lipoperoxides (xylenol orange-based assay), ?-glutamyl transpeptidase activity (Szasz method) were measured and analyzed statistically. Results: Chlorpyrifos reduced splenocyte viability in a dose-dependent manner, which was counteracted by AQ and IP, which was less active in concanavalin A-responsive cells (p<0.05). Chlorpyrifos toxicity involved ?-glutamyltranspeptidase induction with a consequent peroxide reduction, whereas AQ and mainly IP antagonized these responses (p<0.05). Conclusions: The extracts of Ilex paraguariensis and Aspidosperma quebracho-blanco protected splenocytes in vitro against chlorpyrifos. This effect depended on cellular type, given that concanavalin A-responsive cells were more susceptible to this toxic.

Antecedentes: Clorpirifos es un pesticida altamente tóxico, que puede producir inmunotoxicidad con efectos deletéreos sobre la salud a nivel mundial. Por otro lado, las plantas americanas pueden tener derivados con actividad protectora e inmunoestimulante. Por lo tanto, debe evaluarse el potencial de estas plantas frente a clorpirifos. Objetivo: Identificar extractos acuosos bioactivos de Lantana grisebachii (LG), Aspidosperma quebracho-blanco (AQ), Peumus boldus (PB), e Ilex paraguariensis (IP), contra la toxicidad de clorpirifos sobre esplenocitos de hembras Balb/c. Resultados: Esplenocitos fueron tratados in vitro por 72 horas con 0-35 µg/mL de clorpirifos, 0-100 µg/mL de cada extracto (LG, AQ, PB, IP) y 0-5 µg/mL de concanavalina A. Luego, se midió y analizó estadísticamente viabilidad y muerte celular (tinciones de resazurina y yoduro de propidio), hidroperóxidos, lipoperóxidos (ensayos basados en naranja de xilenol), actividad de la ?-glutamiltranspeptidasa (método de Szasz). Conclusiones: Los extractos de Ilex paraguariensis y Aspidosperma quebracho-blanco protegieron in vitro a los esplenocitos frente a clorpirifos. Este efecto dependió del tipo celular, dado que las células inducibles por concanavalina A fueron más susceptibles a este tóxico.

Antioxidant capacity of foods for scavenging reactive oxidants and inhibition of plasma lipid oxidation induced by multiple oxidants.

Unregulated oxidation of biological molecules induced by multiple oxidants has been implicated in the pathogenesis of various diseases. Consequently, the effects of antioxidants contained in foods, beverages and supplements on the maintenance of health and prevention of diseases have attracted much attention of the public as well as scientists. However, recent human studies have shown inconsistent results and failed to demonstrate the beneficial effects of antioxidants. The mechanisms and dynamics of antioxidant action and assessment of antioxidant capacity have been the subject of extensive studies and arguments. In the present article, the antioxidant capacity has been reviewed focusing on two main issues: the capacity of antioxidants to scavenge multiple reactive oxidants and to inhibit plasma lipid oxidation induced by different biological oxidants. It is emphasized that the capacity of antioxidants to scavenge reactive oxidants does not always correlate linearly with the capacity to inhibit lipid oxidation and that it is necessary to specify the oxidant to assess the efficacy of antioxidants, since multiple oxidants contribute to oxidative damage in vivo and the effects of antioxidants depend on the nature of oxidants. A convenient and rapid method using a microplate reader is discussed for assessing the antioxidant capacity against plasma lipid oxidation induced by multiple oxidants including peroxyl radicals, peroxynitrite, hypochlorite, 15-lipoxygenase, and singlet oxygen.

Impact of glutathione supplementation of parenteral nutrition on hepatic methionine adenosyltransferase activity.

BACKGROUND: The oxidation of the methionine adenosyltransferase (MAT) by the combined impact of peroxides contaminating parenteral nutrition (PN) and oxidized redox potential of glutathione is suspected to explain its inhibition observed in animals. A modification of MAT activity is suspected to be at origin of the PN-associated liver disease as observed in newborns. We hypothesized that the correction of redox potential of glutathione by adding glutathione in PN protects the MAT activity. AIM: To investigate whether the addition of glutathione to PN can reverse the inhibition of MAT observed in animal on PN. METHODS: Three days old guinea pigs received through a jugular vein catheter 2 series of solutions. First with methionine supplement, (1) Sham (no infusion); (2) PN: amino acids, dextrose, lipids and vitamins; (3) PN-GSSG: PN+10µM GSSG. Second without methionine, (4) D: dextrose; (5) D+180µM ascorbylperoxide; (6) D+350µM H2O2. Four days later, liver was sampled for determination of redox potential of glutathione and MAT activity in the presence or absence of 1mM DTT. Data were compared by ANOVA, p<0.05. RESULTS: MAT activity was 45±4% lower in animal infused with PN and 23±7% with peroxides generated in PN. The inhibition by peroxides was associated with oxidized redox potential and was reversible by DTT. Correction of redox potential (PN+GSSG) or DTT was without effect on the inhibition of MAT by PN. The slope of the linear relation between MAT activity and redox potential was two fold lower in animal infused with PN than in others groups. CONCLUSION: The present study suggests that prevention of peroxide generation in PN and/or correction of the redox potential by adding glutathione in PN are not sufficient, at least in newborn guinea pigs, to restore normal MAT activity.