Pro-oxidant Effects Research Articles

Report of the Scientific Committee of the Spanish Agency for Food Safety and Nutrition (AESAN) on the risk assessment of inadequate intake of antioxidant minerals (metals) in food supplements that can lead to prooxidant effects: copper, manganese, selenium and zinc

Report of the Scientific Committee of the Spanish Agency for Food Safety and Nutrition (AESAN) on the risk assessment of inadequate intake of antioxidant minerals (metals) in food supplements that can lead to prooxidant effects: copper, manganese, selenium and zinc

Stress responses in captive Crocodylus moreletii associated with metal exposure

Environmental pollution by metals has repercussions on wildlife health. It is known that some metals can have an influence on the neuroendocrine stress response, and at the same time, metals have pro-oxidant effects that can overwhelm the antioxidant system and cause oxidative stress. This study evaluates the association of metals with neuroendocrine stress activity and biomarkers of oxidative stress in 42 captive female Morelet's crocodiles (Crocodylus moreletii). We measured five metals of ecotoxicological importance (Hg, Cd, Pb, Cu and Zn), and three biomarkers of the oxidative stress response in the liver: glutathione (GSH) and glutathione disulfide (GSSG) as markers for antioxidant system and thiobarbituric acid reactive substances (TBARS) for oxidative damage. We also measured one biomarker of the neuroendocrine response to stress: corticosterone (B) in blood plasma. The mean ± SD concentrations of metals in the liver expressed in μg/g (dw) were: Cd: 0.004 ± 0.003, Hg: 0.014 ± 0.019, Cu: 0.017 ± 0.013, Zn: 0.043 ± 0.035, Pb: 0.16 ± 0.256. The mean ± SD of GSH was 0.42 ± 0.35 nmol/mg protein, the mean ± SD of GSSG was 0.24 ± 0.20 nmol/mg protein, the mean ± SD concentrations of TBARS were 0.36 ± 0.21 nmol/mg protein, and the mean ± SD of B was 393.57 ± 405.14 pg/mL. Hg presented a significant negative relationship with corticosterone. Cd had a negative relationship with both GSH and GSSG; meanwhile, Zn showed a negative relationship with TBARS levels, could be a protective element against hepatic oxidative damage. Finally, B had negative relationship with oxidative damage. The connection found between Hg and the neuroendocrine stress response, as well as the correlations of Cd and Zn with oxidative damage and antioxidant activity should be studied further, given their toxicological importance and implications for the conservation of C. moreletii and other crocodilians.

Sulfonamide-Derived Dithiocarbamate Gold(I) Complexes Induce the Apoptosis of Colon Cancer Cells by the Activation of Caspase 3 and Redox Imbalance.

Two new families of dithiocarbamate gold(I) complexes derived from benzenesulfonamide with phosphine or carbene as ancillary ligands have been synthesized and characterized. In the screening of their in vitro activity on human colon carcinoma cells (Caco-2), we found that the more lipophilic complexes—those with the phosphine PPh3—exhibited the highest anticancer activity whilst also displaying significant cancer cell selectivity. [Au(S2CNHSO2C6H5)(PPh3)] (1) and [Au(S2CNHSO2-p-Me-C6H4)(IMePropargyl)] (8) produce cell death, probably by intrinsic apoptosis (mitochondrial membrane potential modification) and caspase 3 activation, causing cell cycle arrest in the G1 phase with p53 activation. Besides this, both complexes might act as multi-target anticancer drugs, as they inhibit the activity of the enzymes thioredoxin reductase (TrxR) and carbonic anhydrase (CA IX) with the alteration of the redox balance, and show a pro-oxidant effect.

The effect of flavonoids on the reduction of cupric ions, the copper-driven Fenton reaction and copper-triggered haemolysis

Flavonoids are considered beneficial, but they may exhibit pro-oxidative effects likely due to metal reducing properties. For the first time, 24 structurally related flavonoids were compared for copper reduction, and modulation of the copper-triggered Fenton reaction and lysis of erythrocytes. The vast majority of flavonoids reduced cupric ions; their behaviour ranged from progressive gradual reduction through bell-shaped, neutral, to a blockade of spontaneous reduction. Similarly, different behaviours were observed with the Fenton reaction. Flavone was the only flavonoid that potentiated copper-triggered haemolysis (155 ± 81 % at twice the amount of Cu2+), while 18 flavonoids were at least partly protective in some concentrations. Only 5-hydroxyflavone did not reduce Cu2+ and behaved as an antioxidant in both assays (reduction of 60 ± 10 % and 88 ± 1%, respectively, at an equimolar ratio with Cu2+). In conclusion, relatively subtle structural differences resulted in very different anti/prooxidant behaviour depending on the model.

Pro-Oxidant and Cytotoxic Effects of Tucum-Do-Cerrado (Bactris setosa Mart.) Extracts in Colorectal Adenocarcinoma Caco-2 Cells

Colorectal cancer is one of the most common types of cancer. Bioactive natural compounds can act in cancer chemoprevention as tumor growth inhibitors. Tucum-do-cerrado (Bactris setosa Mart.) is a Brazilian fruit that contains several phenolic compounds. This study investigated the effect of tucum aqueous extract in Caco-2 cells in comparison to primary human intestinal organoids and fibroblasts. Cells were exposed to 0.5 and 1 mg/ml of tucum aqueous extract for 24 h. ROS production, mRNA levels for SOD1 and SOD2, CAT, GPX1, NFE2L2, HIF1A and NOS2 were evaluated in Caco-2 cells exposed to tucum extract. Cell viability of Caco-2 cells was decreased upon tucum extract exposure. Mitochondrial ROS levels increased in Caco-2 cells exposed to tucum extract. The mRNA levels of SOD1, SOD2, CAT, GPX, NFE2L2 and HIF1A were downregulated in Caco-2 cells exposed to tucum extract, while NOS2 mRNA levels remained unchanged. Protein levels of SOD2, CAT and NRF2 remained unchanged in Caco-2 cells treated with tucum extract, indicating that catalase and SOD2 cellular functions may be unaffected by the tucum extract at 24 h, of exposure. Aqueous extract of tucum-do-cerrado may induce cellular toxicity in a cancer cell-specific manner, possibly through increased mitochondrial ROS production and gene expression regulation.

Membrane Lipids in the Thyroid Comparing to Those in Non-Endocrine Tissues Are Less Sensitive to Pro-Oxidative Effects of Fenton Reaction Substrates.

Iron is an essential microelement for the proper functioning of many organs, among others it is required for thyroid hormone synthesis. However, its overload contributes to the increased formation of reactive oxygen species via Fenton chemistry (Fe2++H2O2→Fe3++˙OH + OH−), and it is potentially toxic. Individual organs/tissues are affected differently by excess iron. The excessive absorption of iron with subsequent deposition in various organs is associated with diseases such as hemochromatosis. Such an iron deposition also occurs in the thyroid gland where it can disturb thyroid hormone synthesis. In turn, melatonin is an effective antioxidant, which protects against oxidative damage. This study aims to check if lipid peroxidation resulting from oxidative damage to membrane lipids, is caused by Fenton reaction substrates, and if protective effects of melatonin differ between the thyroid and various non-endocrine porcine tissues (liver, kidney, brain cortex, spleen, and small intestine). To mimic the conditions of iron overload, Fe2+ was used in extremely high concentrations. Homogenates of individual tissues were incubated together with Fenton reaction substrates, i.e., FeSO4 (9.375, 18.75, 37.5, 75, 150, 300, 600, 1,200, 1,800, 2,100, 2,400, 3,000, 3,600, 4,200, and 4,800 µM)+H2O2 (5 mM), either without or with melatonin (5 mM). The concentration of malondialdehyde+4-hydroxyalkenals (MDA+4-HDA), as the LPO index, was evaluated by a spectrophotometrical method. Fenton reaction substrates increased concentrations of LPO products in all chosen tissues. However, in the thyroid, compared to non-endocrine tissues, the damaging effect was generally weaker, it was not observed for the two lowest concentrations of iron, and the LPO peak occurred with higher concentrations of iron. Melatonin reduced experimentally induced LPO in all examined tissues (without differences between them), and these protective effects did not depend on iron concentration. In conclusion, membrane lipids in the thyroid compared to those in non-endocrine tissues are less sensitive to pro-oxidative effects of Fenton reaction substrates, without differences regarding protective effects of melatonin.

Insulin resistance enhances binge ethanol-induced liver injury through promoting oxidative stress and up-regulation CYP2E1

Alcoholic liver disease (ALD) has caused a serious burden on public and personal health in crowd with ethanol abuse. The effects of insulin resistance (IR) on ALD and the mechanisms underlying these responses are still not well understood. In this study, we investigated the changes of liver injury, inflammation, apoptosis, mitochondrial dysfunction and CYP2E1 changes in liver of mice exposed to ethanol with IR or not. We found IR increased the sensitivity of liver injury in mice exposed to ethanol, manifested as the increase serum activities of AST and ALT, the accumulation of triglycerides, the deterioration of liver pathology and increase of inflammatory factors. IR also exacerbated apoptosis and mitochondrial dysfunction in liver of mice exposed to ethanol. The increase of oxidative stress and the decrease of antioxidant defense ability might be responsible for the sensitizing effects of IR on ethanol-induced liver injury, supported by the increase of MDA levels and the decline of GSH/GSSG, the inactivation of antioxidant enzymes SOD, GR through the inhibition of Nrf-2 pathway. The activation of CYP2E1 might be also involved in the sensitizing effects of IR on ethanol induced liver injury in mice. These results demonstrated that IR exhibited a significant pro-oxidative and pro-apoptosis effects to aggravate alcoholic liver injury. Our study helped us to better understand the sensitive role of IR on ALD and suggested that alcohol intake may be more harmful for people with IR.

Complex interactions between nicotine and resveratrol in the Drosophila melanogaster wing spot test

Nicotine (NIC) and resveratrol (RES) are chemicals in tobacco and wine, respectively, that are widely consumed concurrently worldwide. NIC is an alkaloid known to be toxic, addictive and to produce oxidative stress, while RES is thought of as an antioxidant with putative health benefits. Oxidative stress can induce genotoxic damage, yet few studies have examined whether NIC is genotoxic in vivo. In vitro studies have shown that RES can ameliorate deleterious effects of NIC. However, RES has been reported to have both antioxidant and pro-oxidant effects, and an in vivo study reported that 0.011 mM RES was genotoxic. We used the Drosophila melanogaster wing spot test to determine whether NIC and RES, first individually and then in combination, were genotoxic and/or altered the cell division. We hypothesized that RES would modulate NIC’s effects. NIC was genotoxic in the standard (ST) cross in a concentration-independent manner, but not genotoxic in the high bioactivation (HB) cross. RES was not genotoxic in either the ST or HB cross at the concentrations tested. We discovered a complex interaction between NIC and RES. Depending on concentration, RES was protective of NIC’s genotoxic damage, RES had no interaction with NIC, or RES had an additive or synergistic effect, increasing NIC’s genotoxic damage. Most NIC, RES, and NIC/RES combinations tested altered the cell division in the ST and HB crosses. Because we used the ST and HB crosses, we demonstrated that genotoxicity and cell division alterations were modulated by the xenobiotic metabolism. These results provide evidence of NIC’s genotoxicity in vivo at specific concentrations. Moreover, NIC’s genotoxicity can be modulated by its interaction with RES in a complex manner, in which their interaction can lead to either increasing NIC’s damage or protecting against it.

Molecular mechanism of the uptake and toxicity of EGF-LipoAgNPs in EGFR-overexpressing cancer cells

The surface of silver nanoparticles (AgNPs) is characterized by high reactivity resulting in prooxidative and cytotoxic properties. These effects are observed both in normal and in cancer cells, which overexpress the Epidermal Growth Factor Receptor (EGFR). In our previous paper, we have demonstrated that, with the use of liposomes labeled with the Epidermal Growth Factor (EGF), it is possible to direct the toxic effect of AgNPs in EGFR-overexpressing cells. Unfortunately, the mechanism of uptake and toxicity induction by such liposomes is still unknown. Therefore, the aim of this study was to determine the impact of EGF-LipoAgNPs on certain genes related to endocytosis and toxicity induction by such liposomes in human lung (A549) and tongue (SCC-15) cancer cells. The siRNA knock-out gene method was used in this study to determine the engagement of EGFR in this process. The confocal microscopy study revealed that the number of liposomes in the cytoplasm of the A549EGFR- and SCC-15EGFR- cells was lowered by 51.99 × 103 RFU and 138.50 × 103 RFU, respectively, proving the crucial role of EGFR in the liposome uptake. Moreover, the expression of the SHH and ATM genes was significantly increased, whereas the expression of the NRF2 gene was decreased after the treatment with EGF-LipoAgNPs and native AgNPs. Furthermore, the expression of the CLTC, AP2M1, CAV1, and SH3GLB1 genes indicated that the tested liposomes are uptaken via the clathrin-dependent pathway with engagement of the AP-2 complex and endophilin in this process. Summarizing, the created targeted delivery system of AgNPs causes an increase in the prooxidative and toxic effect of such NPs and has an impact on endocytosis regulatory genes, especially those related to the clathrin-mediated endocytosis.

Effects of Laser Irradiation at 488, 514, 532, 552, 660, and 785 nm on the Aqueous Extracts of Plantago lanceolata L.: A Comparison on Chemical Content, Antioxidant Activity and Caco-2 Viability

In this study, six laser radiation (488 nm/40 mW, 514 nm/15 mW, 532 nm/20 mW, 552 nm/15 mW, 660 nm/75 mW, and at 785 nm/70 mW) were tested on the aqueous extracts of leaves of Plantago lanceolata L. to compare extraction efficacy and antioxidant and cell viability effects in vitro. Briefly, in comparison with the control extract, laser extracts at 488, 514, 532, and 552 nm revealed small acquisitions of total extractible compounds in samples (up to 6.52%; laser extracts at 488 and 532 nm also revealed minerals and micro-elements increases (up to 6.49%); the most prominent results were obtained upon Fe (up to 38%, 488 nm), Cr (up to 307%, 660 nm), and Zn (up to 465%, 532 nm). Laser extracts at 488, 514, 552, and 785 nm proved more intense antioxidant capacity than the control sample, while laser extract at 660 nm indicated clear pro-oxidant effects. Caco-2 cells study indicated stimulatory activity for the extracts at 488 nm, no effects at 532 nm, and the decrease of the cell viability in the case of extracts at 660 nm respectively. Further studies are necessary to understand the pro-oxidant effects observed in the case of extracts exposed to laser radiation at 660 nm.

Concentration Dependence of Anti- and Pro-Oxidant Activity of Polyphenols as Evaluated with a Light-Emitting Fe2+-Egta-H2O2 System.

Hydroxyl radical (•OH) scavenging and the regeneration of Fe2+ may inhibit or enhance peroxidative damage induced by a Fenton system, respectively. Plant polyphenols reveal the afore-mentioned activities, and their cumulative net effect may determine anti- or pro-oxidant actions. We investigated the influence of 17 phenolics on ultra-weak photon emission (UPE) from a modified Fenton system (92.6 µmol/L Fe2+, 185.2 µmol/L EGTA (ethylene glycol-bis(β-aminoethyl-ether)-N,N,N′,N,-tetraacetic acid) and 2.6 mmol/L H2O2 pH = 7.4). A total of 8 compounds inhibited (antioxidant effect), and 5 enhanced (pro-oxidant effect) UPE at all studied concentrations (5 to 50 µmol/L). A total of 4 compounds altered their activity from pro- to antioxidant (or vice versa) along with increasing concentrations. A total of 3 the most active of those (ferulic acid, chlorogenic acid and cyanidin 3-O-glucoside; mean UPE enhancement by 63%, 5% and 445% at 5 µmol/L; mean UPE inhibition by 28%, 94% and 24% at 50 µmol/L, respectively) contained catechol or methoxyphenol structures that are associated with effective •OH scavenging and Fe2+ regeneration. Most likely, these structures can determine the bidirectional, concentration-dependent activity of some phenolics under stable in vitro conditions. This is because the concentrations of the studied compounds are close to those occurring in human fluids, and this phenomenon should be considered in the case of dietary supplementation with isolated phenolics.

Associations of Serum Carotenoids With Risk of Cardiovascular Mortality Among Individuals With Type 2 Diabetes: Results From NHANES.

Although carotenoids have been suggested to exhibit antioxidant properties, some experimental studies reported that β-carotene may show pro-oxidant effects under certain conditions. Current evidence regarding the cardiovascular effects of carotenoids among patients with type 2 diabetes (T2D) is scarce. This study aimed to prospectively examine the associations of individual serum carotenoid concentrations with cardiovascular mortality among adults with T2D. This analysis included 3,107 individuals with T2D from the Third National Health and Nutrition Examination Survey (NHANES III) and NHANES 2001-2006. Cardiovascular mortality was ascertained by linkage to National Death Index records through 31 December 2015. Cox proportional hazards regression models were used to estimate hazard ratios (HRs) and 95% CIs. During an average of 14 years of follow-up, 441 cardiovascular deaths occurred. After multivariate adjustment including lifestyles, dietary factors, glucose control, and other major carotenoids, higher serum β-carotene concentrations were significantly associated with an elevated risk of cardiovascular mortality in a dose-response manner. When extreme quartiles of β-carotene were compared, the multivariable-adjusted HR was 2.47 (95% CI 1.62, 3.76) for cardiovascular mortality (Ptrend = 0.002); and per one-unit increment in natural log-transformed serum β-carotene was associated with a 46% higher risk of cardiovascular mortality (P = 0.001). Other individual carotenoids (α-carotene, β-cryptoxanthin, lycopene, and lutein/zeaxanthin) were not significantly associated with the risk of cardiovascular mortality. Consistent results were observed when stratifying by age, sex, race, BMI, smoking status, diabetes duration, and glycated hemoglobin A1c levels. Higher concentrations of serum β-carotene, but not other individual carotenoids, were significantly associated with an increased risk of cardiovascular mortality among individuals with T2D. Our findings, if replicated, underscore the need to estimate the optimal serum β-carotene concentrations in individuals with T2D.

Glochidion wallichianum Leaf Extract as a Natural Antioxidant in Sausage Model System.

This study highlighted the role of an 80% ethanolic Mon-Pu (Glochidion wallichianum) leaf extract (MPE), a novel natural antioxidative ingredient, in controlling the oxidative stability and physicochemical properties of a cooked sausage model system (SMS). MPE had a total extractable phenolic content of 16 mg/100 g, with DPPH● scavenging activity, ABTS●+ scavenging activity, and ferric reducing antioxidant power of 2.3, 1.9, and 1.2 mmole Trolox equivalents (TE)/g, respectively. The effects of different concentrations of MPE (0.01–10%, w/w) formulated into SMS on lipid oxidation, protein oxidation, and discoloration were compared to synthetic butylated hydroxyl toluene (BHT; 0.003%, w/w) and a control (without antioxidant). The peroxide value (PV), thiobarbituric acid reactive substances (TBARS), and protein carbonyl contents of SMS tended to increase with increasing MPE concentration (p < 0.05), indicating that high MPE excipient has a pro-oxidative effect. The lowest lipid oxidation (PV and TBARS) and protein carbonyl contents were observed when 0.01% MPE was used to treat SMS (p < 0.05), which was comparable or even greater than BHT-treated SMS. High concentrations (1–10%) of MPE incorporation led to increases in the discoloration of SMS (p < 0.05) with a negligible change in pH of SMS. The water exudate was reduced when MPE was incorporated into SMS compared to control (p < 0.05). Furthermore, MPE at 0.01% significantly reduced lipid oxidation in cooked EMS during refrigerated storage. According to the findings, a low amount of MPE, particularly at 0.01%, in a formulation could potentially maintain the oxidative stability and physicochemical qualities of cooked SMS that are comparable to or better than synthetic BHT.

The Impact of Whirlpool Hop Addition on the Wort Metal Ion Composition and on the Flavor Stability of American Style Pale Ales Using Citra® Hop Extract and Pellets

The characteristic hoppy flavor of pale ales is known to be unstable and deteriorates during the beer’s shelf life. Metal ions, in particular iron and copper, have a prooxidative effect and therefore accelerate beer flavor deterioration. Recent studies suggest that hop constituents may complex metal ions in beer thereby suppressing their negative effect on beer flavor stability. Using benchtop wort boiling experiments and quantitating metal ions in the cast wort, this study determined that different whirlpool additions of up to 3.0 g/L hop extract or up to 17.4 g/L pellets significantly reduced metal ion concentrations in wort, especially Fe, which was reduced by a factor of up to 6 compared to the control. Based on identified dose- and product-related effects, 2.5-hL brewing trials were performed using a selection of whirlpool hopping levels with either extract or pellets revealing significant differences in resinous and sweaty flavor. The flavor of the beer hopped with 5.8 g/L pellets was perceived as 8 times more resinous and 9 times sweatier than the control revealing clear differences between extract and pellets, especially at the lower hopping levels. The beer flavor stability was predicted using ESR spectroscopy and verified by performing forced aging experiments at 30 °C and monitoring beer flavor changes by sensory evaluation. Results indicate that both extract and pellet whirlpool hopping improve beer flavor stability by reducing the formation of age-related off-flavors and by supporting the retention of flavors related to freshness with only minor variations between the two hop products.

Redox Mechanisms in Cisplatin Resistance of Cancer Cells: The Twofold Role of Gamma-Glutamyltransferase 1 (GGT1).

Cisplatin (CDDP) is currently employed for the treatment of several solid tumors, but cellular heterogeneity and the onset of drug resistance dictate that suitable biomarkers of CDDP sensitivity are established. Studies on triple-negative breast cancer (TNBC) have recently confirmed the involvement of gamma-glutamyltransferase 1 (GGT1), whose enzyme activity expressed at the cell surface favors the cellular resupply of antioxidant glutathione (GSH) thus offering cancer cells protection against the prooxidant effects of CDDP. However, an additional well-established mechanism depends on GGT1-mediated matabolism of extracellular GSH. It was in fact shown that glycyl-cysteine – the dipeptide originated by GGT1-mediated GSH metabolism at the cell surface – can promptly form adducts with exogenous CDDP, thus hindering its access to the cell, interactions with DNA and overall cytotoxicity. Both mechanisms: mainainance of intracellular GSH levels plus extracellular CDDP detoxication are likely concurring to determine GGT1-dependent CDDP resistance.

Phytochemical Composition, Antioxidant and Antifungal Activity of Thymus capitatus, a Medicinal Plant Collected from Northern Morocco.

Thymus capitatus is a Mediterranean endemic plant commonly known as “Zaïtra” in northern Morocco. As T. capitatus is widely used in traditional medicine and food, this present work aims to investigate the chemical compositions and biological activities of the T. capitatus leaves essential oil (TcLEO), acetonic (TcLAE), and methanolic extract (TcLME). The spectrophotometric determination demonstrated that T. capitatus is a natural source rich in phenolic contents (TPC) and flavonoid contents (TFC) and that TcLME revealed the highest TPC and TFC than TcLAE and TcLEO. The LC-MS analysis of phenolic compounds showed that paraben acid was predominant in both TcLME and TcLAE, followed by cinnamic acid and p-hydroxybenzoic acid. GC-MS analysis of the TcLEO revealed the presence of a total of 10 compounds, which were predominated by carvacrol. The antioxidant activity by ORAC was observed to be significantly higher in TcLEO and TcLAE than in TcLME. All samples used to assess DNA degradation effectively prevented DNA oxidation and, at the same time, had a prooxidant effect. The genotoxicity test showed that the T. capitatus were devoid of any mutagenic activity. Concerning antifungal activity, all samples were able to inhibit the growth of all microorganisms tested at low concentrations. TcLAE showed higher activity than TcLME, and in general, dermatophytes were more susceptible, being Microsporum canis the most sensitive one. Overall, the results obtained from this study confirm the wide uses of T. capitatus. Furthermore, the finding results suggest that the T. capitatus essential oil and extracts can be highly useful for pharmaceutical industries.

Beta-Carotene Affects the Effects of Heme Oxygenase-1 in Isolated, Ischemic/Reperfused Rat Hearts: Potential Role of the Iron.

Beta-carotene (BC) is a well-known antioxidant. However, increasing evidence shows that under severe oxidative conditions, BC can become pro-oxidant, an effect that may be enhanced in the presence of iron (II). In our earlier studies, we observed that despite increasing heme oxygenase-1 (HO-1) levels in the heart, the protective effects of BC have been lost when it was used at a high concentration. Since iron releases from heme as a consequence of HO-1 activity, we hypothesized that the application of an iron-chelator (IC) would reverse the lost cardiac protection associated with an elevated HO-1 level. Thus, in the present study, we investigated the effects of desferrioxiamine (DFO) in isolated, ischemic/reperfused rat hearts after long-term treatment with vehicle or high-dose (HD) BC. Vehicle or 150 mg/bw kg daily doses of BC were administered to the rats for 4 weeks, and then their hearts were removed and subjected to 30 min of global ischemia (ISA) followed by 120 min of reperfusion (REP). During the experiments, cardiac function was registered, and at the end of the REP period, infarct size (IS) and HO-1 expression were measured. The results show that DFO treatment alone during REP significantly ameliorated postischemic cardiac function and decreased IS, although HO-1 expression was not increased significantly. In hearts isolated from BC-treated rats, no cardioprotective effects, despite an elevated HO-1 level, were observed, while DFO administration after ISA resulted in a mild improvement in heart function and IS. Our results suggest that iron could have a role whether BC exerts antioxidant or pro-oxidant effects in ISA/REP-injured hearts.

The dimerization of methyl vanillate improves its effect against breast cancer cells via pro-oxidant effect

Phenolic phytochemicals are a group of organic compounds with potent antioxidant features but can also act as powerful pro-oxidants. These characteristics are effective in reducing metastatic potential in cancer cells, and this effect has been associated with reactive oxygen species (ROS). Methyl vanillate (MV) and its dimer, methyl divanillate (DMV), are potent antioxidants. In the present study, we investigated the effects of MV and DMV on breast cancer cell lines MCF-7 and MDA-MB-231 and compared the results using the non-tumor cell line HB4a. Our results indicated that the compounds performed a pro-oxidant action, increasing the generation of ROS. DMV decreased the viability cell, showing a higher apoptotic effect and inhibition of proliferation than MV on both cell lines, with significant differences between groups (p < 0.05). Some modulation of NOX4, NOX5, and DUOX were observed, but the results did not correlate with the intracellular production of ROS. The dimer showed more effectivity and pro-oxidant effect than MV, impacting cell line MCF-7 in higher extension than MDA-MB-231. In conclusion, and corroborating with reported works, the dimerization of natural phenolic compounds was associated with improved beneficial biological effects as a potential cytotoxic agent to tumor cells.

Biological effects of grape stem extracts on human cancer cell lines

Grapes are rich in polyphenolic compounds, which are known for their beneficial effects on human health. Grape stems are byproducts of the winemaking procedure. They exert antioxidant effects; however, they can also become pollutants when discarded without control into the environment due to their high organic content. The main aim of the present study was to examine the potential prooxidant/anticancer effects of grape stem extracts, promoting the rationale of the scientific exploitation of such compounds. Therefore, three grape stem extracts derived from the native Greek vine varieties, Mavrodaphne, Muscat and Rhoditis were examined for their ability to alter the redox status [i.e., endogenous levels of reduced glutathione and reactive oxygen species (ROS)] of HeLa and HepG2 human cancer cell lines. The results revealed that the extracts from Muscat and Rhoditis exerted prooxidant effects on the tested cell lines, whereas the extract of Mavrodaphne did not exert any such effects. The fact that the extracts functioned as prooxidants appears to be detrimental for the cancer cells, since they are more susceptible to high concentrations of ROS, contrary to healthy cells that possess more robust antioxidant defense mechanisms. On the whole, as demonstrated in the present study, it appears that the Muscat and Rhoditis extracts may prove to be promising agents against cancer cells in the specific <em>in vitro</em> model. However, further studies are required to confirm these findings, in <em>in vivo</em> settings.

MO531: The Differential Effect of Third-Generation Intravenous Iron Preparations (Ferric Carboxymaltose, Ferric Derisomaltose) on Cardiovascular Parameters in Patients With Non-Dialysis Dependent CKD and Iron Deficiency Anaemia

Abstract BACKGROUND AND AIMS Intravenous (IV) iron is commonly used to alleviate iron deficiency anaemia in patients with non-dialysis chronic kidney disease (CKD). Treatment with IV iron in patients with iron deficiency and heart failure has demonstrated an improvement in functional status and cardiac biomarkers; however, concerns remain regarding the potential pro-oxidative effect of IV iron and possible cardiovascular injury. Newer agents [such as ferric carboxymaltose (FCM) and ferric derisomaltose (FDI)] are increasingly used due to their advantage in delivering large amounts of iron effectively and safely; however, they differ as to their effect on serum phosphate and fibroblast growth factor 23, factors that have been previously shown to affect cardiac function. Given the above potential concerns, it is important to explore the comparative effect of IV iron in this population in terms of cardiac injury and vascular function. METHOD The exploratory single-centre double-blinded randomized controlled trial ‘Iron and Phosphaturia—ExplorIRON-CKD’ aimed primarily to not only assess the differential effects of FCM and FDI on levels of fibroblast growth factor 23 and phosphate, but also explore any cardiovascular effect. Non-dialysis patients with CKD and iron deficiency with/without anaemia defined as serum ferritin &amp;lt;200 µg/L or transferrin saturation ≤20% and serum ferritin 200–299 µg/L were recruited and randomized in a 1:1 ratio. Participants received 1000 mg at baseline and 500–1000 mg at 1 month to achieve replenishment. Measurements of NT-proBNP were taken at baseline, 1 month, 2 months and 3 months, whilst measurements of Troponin T were taken at every visit (total of eight visits). Vascular function was assessed using pulse wave velocity measurements at baseline, 1 month, 2 months and 3 months, monitoring carotid to femoral pulse wave velocity (PWVcf) and aortic augmentation index (AIx). RESULTS Twenty-six patients were recruited; 14 were randomized to receive FDI and 12 to receive FCM. All patients received at least one iron dose (1000 mg), 10 patients received two FDI doses and 11 received two FCM doses. There was a trend to an increase in NT-pro BNP (10.3%) in the combined group, with no statistical difference between the FCM and FDI groups [median for the combined population: baseline: 875.5 (3033.0) ng/L versus Visit 8: 967.0 (2976.0) ng/L] (Figure 1). Troponin T values remained constant throughout the study period in the combined and individual groups (Table 1). A decrease in both PWVcf and AIx was noted in the participants receiving IV iron irrespective of the compound used. A greater decrease was seen in the FDI group in terms of PWVcf; however, there was no statistically significant difference for the percentage change from baseline to the end of the study between the groups (FCM: −4.3 (33.6)% versus FDI: −11.9 (23.4)%; P = .60) (Figure 2). CONCLUSION Treatment of non-dialysis CKD patients with FDI or FCM was not associated with any cardiovascular safety signals. Troponin T was unaffected and there was no worsening in vascular function. Pulse wave velocity improved, as noted by the decreasing trends for PWVcf and AIx. The decrease in PWVcf was greater with FDI than FCM, but the difference was not statistically significant. A small rise in NT-proBNP was noted in the combined group, but there was no statistically significant difference between the FDI and FCM groups. These results suggest that newer IV iron compounds are not associated with any detriment in terms of vascular function and potentially no resulting cardiac insult. The study sample is small and therefore the results should be interpreted with caution; nonetheless, the evidence provided from a comparative view directs to no negative sequela.