Pro-oxidant Effects Research Articles

Simultaneously Determined Antioxidant and Pro-Oxidant Activity of Randomly Selected Plant Secondary Metabolites and Plant Extracts.

Oxidative stress is a well-known phenomenon arising from physiological and nonphysiological factors, defined by the balance between antioxidants and pro-oxidants. While the presence and uptake of antioxidants are crucial, the pro-oxidant effects have not received sufficient research attention. Several methods for assessing pro-oxidant activity, utilizing various mechanisms, have been published. In this paper, we introduce a methodology for the simultaneous determination of antioxidant and pro-oxidant activity on a single microplate in situ, assuming that the FRAP method can measure both antioxidant and pro-oxidant activity due to the generation of pro-oxidant Fe2+ ions in the Fenton reaction. Systematic research using this rapid screening method may help to distinguish between compounds with dominant antioxidant efficacy and those with dominant pro-oxidant effects. Our preliminary study has revealed a dominant pro-oxidant effect for compounds with a higher number of oxygen heteroatoms, especially sp2 hybridized compounds (such as those containing keto groups), such as flavonoids and plant extracts rich in these structural types. Conversely, catechins, carotenoids, and surprisingly, extracts from birch leaves and chestnut leaves have demonstrated dominant antioxidant activity over pro-oxidant. These initial findings have sparked significant interest in the systematic evaluation of a more extensive collection of compounds and plant extracts using the developed method.

Cu2+ -Anchored Carbon Nano-Photocatalysts for Visible Water Splitting to Boost Hydrogen Cuproptosis.

Both hydrogen (H2 ) and copper ions (Cu+ ) can be used as anti-cancer treatments. However, the continuous generation of H2 molecules and Cu+ in specific sites of tumors is challenging. Here we anchored Cu2+ on carbon photocatalyst (Cu@CDCN) to allow the continuous generation of H2 and hydrogen peroxide (H2 O2 ) in tumors using the two-electron process of visible water splitting. The photocatalytic process also generated redox-active Cu-carbon centers. Meanwhile, the Cu2+ residues reacted with H2 O2 (the obstacle to the photocatalytic process) to accelerate the two-electron process of water splitting and cuprous ion (Cu+ ) generation, in which the Cu2+ residue promoted a pro-oxidant effect with glutathione through metal-reducing actions. Both H2 and Cu+ induced mitochondrial dysfunction and intracellular redox homeostasis destruction, which enabled hydrogen therapy and cuproptosis to inhibit cancer cell growth and suppress tumor growth. Our research is the first attempt to integrate hydrogen therapy and cuproptosis using metal-enhanced visible solar water splitting in nanomedicine, which may provide a safe and effective cancer treatment.

Effects of Rosmarinus officinalis L. (Laminaceae) essential oil on adult and larvae of Drosophila melanogaster.

Rosmarinus officinalis (Lamiaceae family), also known as "alecrim," is a perennial herb, typical of the Mediterranean region and widely distributed in Brazilian territory. Despite having demonstrated several properties of human interest, insecticide/larvicidal effect of essential oil from R. officinalis on insects remains unclear. In this study, we tested the effects of R. officinalis essential oil on biomarkers of oxidative damage in Drosophila melanogaster. Exposure to R. officinalis essential oil increased adult mortality and decreased geotaxis behavior in adult fruit flies. In addition, essential oil increased of larval mortality and impaired the developmental success in D. melanogaster. R. officinalis essential oil showed a significant repellent effect, with duration time of about 6h. To understand the mechanism underlying the toxicity of essential oil both pro-oxidant effects and biomarkers of oxidative damage were evaluated in exposed flies. Exposure to essential oil caused a significant redox imbalance with impairment of both enzymatic and non-enzymatic antioxidant system and increased the lipid peroxidation levels. These results suggest that R. officinalis essential oil can be used as a bioinsecticide and/or larvicide as well as an alternative insect repellent.

Cerium Oxide Nanoparticles Protect Cortical Astrocytes from Oxygen-Glucose Deprivation through Activation of the Ca2+ Signaling System.

Most of the works aimed at studying the cytoprotective properties of nanocerium are usually focused on the mechanisms of regulation of the redox status in cells while the complex effects of nanocerium on calcium homeostasis, the expression of pro-apoptotic and protective proteins are generally overlooked. There is a problem of a strong dependence of the effects of cerium oxide nanoparticles on their size, method of preparation and origin, which significantly limits their use in medicine. In this study, using the methods of molecular biology, immunocytochemistry, fluorescence microscopy and inhibitory analysis, the cytoprotective effect of cerium oxide nanoparticles obtained by laser ablation on cultured astrocytes of the cerebral cortex under oxygen-glucose deprivation (OGD) and reoxygenation (ischemia-like conditions) are shown. The concentration effects of cerium oxide nanoparticles on ROS production by astrocytes in an acute experiment and the effects of cell pre-incubation with nanocerium on ROS production under OGD conditions were studied. The dose dependence for nanocerium protection of cortical astrocytes from a global increase in calcium ions during oxygen-glucose deprivation and cell death were demonstrated. The concentration range of cerium oxide nanoparticles at which they have a pro-oxidant effect on cells has been identified. The effect of nanocerium concentrations on astrocyte preconditioning, accompanied by increased expression of protective proteins and limited ROS production induced by oxygen-glucose deprivation, has been investigated. In particular, a correlation was found between an increase in the concentration of cytosolic calcium under the action of nanocerium and the suppression of cell death. As a result, the positive and negative effects of nanocerium under oxygen-glucose deprivation and reoxygenation in astrocytes were revealed at the molecular level. Nanocerium was found to act as a "double-edged sword" and to have a strictly defined concentration therapeutic "window".

The Use of Soy and Egg Phosphatidylcholines Modified with Caffeic Acid Enhances the Oxidative Stability of High-Fat (70%) Fish Oil-in-Water Emulsions

This study investigated the effect of the combined use of sodium caseinate (CAS), commercial phosphatidylcholine (PC), and modified PCs on the physical and oxidative stability of 70% fish oil-in-water emulsions. Caffeic acid was covalently attached to both modified PCs (PCs originated from soy and eggs) in order to increase the antioxidant activity of PCs and investigate the advantage of bringing the antioxidant activity to the close proximity of the oil-water interface. Results showed that oxidative stability was improved when part of the PC was substituted with modified soy PC or egg PC. Emulsions containing a low concentration of modified PCs (10 wt.% of total PC) resulted in a prooxidative effect on the formation of hydroperoxides compared to emulsions with free caffeic acid. On the other hand, a decrease in the formation of volatile oxidation products was observed for emulsions containing higher levels of modified PCs (60 wt.% of total PC) compared to the emulsions with free caffeic acid added at its equivalent concentration. Increased concentrations of modified PCs provided better oxidative stability in high-fat emulsions, independent of the modified PC type. Moreover, when oxidation was initiated by producing singlet oxygen near a single oil droplet using a focused laser, fluorescence imaging showed that the oxidation did not propagate from one oil droplet to another oil droplet.

Effects of Dietary Supplementation of a Resin-Purified Aqueous-Isopropanol Olive Leaf Extract on Meat and Liver Antioxidant Parameters in Broilers.

Olive leaves are byproducts οf the agro-industrial sector and are rich in bioactive compounds with antioxidant properties. They could be supplemented in poultry diets powdered or less frequently as extracts to improve performance, health and product quality. The objective of this study was to investigate the possible beneficial effects of an aqueous isopropanol olive leaf extract-purified through filtration (250-25 µm) and a resin (XAD-4)-when supplemented in broiler chickens' diets, on meat quality parameters, focusing mainly on antioxidant parameters as there is limited published information. For this purpose, four-hundred-and-eighty-day-old broilers were randomly assigned to four dietary treatments: T1 (control: basal diet); T2 (1% olive leaf extract); T3 (2.5% olive leaf extract); T4 (positive control: 0.1% encapsulated oregano oil commercially used as feed additive). At the end of the experimental period (day 42), the birds were slaughtered, and samples from breast, thigh meat and liver were collected for antioxidant parameters evaluation. On day 1, after slaughter, in thigh meat, Malondialdehyde (MDA) was lower in T2 compared to T3, and total phenolic content (TPC) was higher in T2 compared to T3 and T4. Total antioxidant capacity (TAC) was increased in T2 and T4 breast meat compared to the control. In liver, T4 treatment resulted in higher TPC. The lack of dose-dependent effect for olive leaf extract may be attributed to the pro-oxidant effects of some bioactive compounds found in olive leaves, such as oleuropein, when supplemented at higher levels. In summary, it can be inferred that the inclusion of 1% olive leaf extract in the feed of broilers has the potential to mitigate oxidation in broiler meat and maybe enhance its quality.

Sustainable Exploitation of Waste Orange Peels: Enrichment of Commercial Seed Oils and the Effect on Their Oxidative Stability

The current project aimed at examining the effect of the enrichment of commercial seed oils with waste orange peel (WOP) extracts on their polyphenolic profiles and resistance against oxidation. Polyphenol-containing WOP extracts were produced using a novel combination of ethanol and triacetin, and they were incorporated into seed oils (sunflower, soybean, corn oil), at a level of 36.87 mg per kg of oil. The oils were then stored at 60 °C, for 58 days. By performing a Rancimat test, it was shown that enrichment of sunflower, soybean, and corn oils with WOP extracts did not provoke any prooxidant effects, but, to the contrary, exerted an antioxidant action, with protection factors varying from 1.01 to 1.61. Furthermore, in all cases examined, it was demonstrated that, during the storage period, the stabilizing effect of WOP extract against oxidation was comparable to that observed in oil samples containing 200 mg BHT per kg oil. This outcome was ascertained by measuring the onset of peroxide value, thiobarbituric-acid-reactive substances, and the TOTOX value. Furthermore, it was revealed that the Trolox-equivalent antiradical activity of the enriched oils exhibited a decline at the end of the examination period, a fact most probably attributed to the depletion of the antioxidants occurring in the oils. It was concluded that the method proposed might be a means of stabilizing commercial seed oils against oxidation, and of enhancing their nutritional value by enriching them with natural polyphenols.

The multifaceted role of quercetin derived from its mitochondrial mechanism.

Quercetin is a flavonoid with promising therapeutic applications; nonetheless, the phenotype exerted in some diseases is contradictory. For instance, anticancer properties may be explained by a cytotoxic mechanism, whereas antioxidant-related neuroprotection is a pro-survival process. According to the available literature, quercetin exerts a redox interaction with the electron transport chain (ETC) in the mitochondrion, affecting its membrane potential. It also affects ATP generation by oxidative phosphorylation, where ATP deprivation could partly explain its cytotoxic effect. Moreover, quercetin may support the generation of free radicals through redox reactions, causing a prooxidant effect. The nutrimental stress and prooxidant effect induced by quercetin might promote pro-survival properties such as antioxidant processes. Thus, in this review, we discuss the evidence supporting that quercetin redox interaction with the ETC could explain its beneficial and toxic properties.

P03-11: Prooxidative effect of non-thermal plasma generated from ambient air, nitrogen and oxygen on Schizosaccharomyces pombe

P03-11: Prooxidative effect of non-thermal plasma generated from ambient air, nitrogen and oxygen on Schizosaccharomyces pombe

Combined effects of valproate and naringin on kidney antioxidative markers and serum parameters of kidney function in C57BL6 mice.

Valproate is known to disturb the kidney function, and high doses or prolonged intake may cause serum ion imbalance, kidney tubular acidosis, proteinuria, hyperuricosuria, polyuria, polydipsia, and dehydration. The aim of this in vivo study was to see whether naringin would counter the adverse effects of high-dose valproate in C57Bl/6 mice and to which extent. As expected, valproate (150 mg/kg bw a day for 10 days) caused serum hyperkalaemia, more in male than female mice. Naringin reversed (25 mg/kg bw a day for 10 days) the hyperkalaemia and activated antioxidative defence mechanisms (mainly catalase and glutathione), again more efficiently in females. In males naringin combined with valproate was not as effective and even showed some prooxidative effects.

Exogenous Melatonin Protects against Oxidative Damage to Membrane Lipids Caused by Some Sodium/Iodide Symporter Inhibitors in the Thyroid.

The thyroid gland is the primary site of sodium/iodide symporter (NIS), an intrinsic plasma membrane protein responsible for the active uptake of iodine, which is indispensable for thyroid hormone synthesis. Since exposure of the thyroid to NIS inhibitors can potentially have harmful effects on the entire organism, it is important to investigate the potential protective effects of known antioxidants, such as melatonin and indole-3-propionic acid (IPA), against pro-oxidative action of classic NIS inhibitors. The study aimed to check if and to what extent melatonin and IPA interact with some confirmed NIS inhibitors regarding their effects on oxidative damage to membrane lipids in the thyroid. For comparison with the thyroid gland, in which NIS is typically present, the liver tissue-not possessing NIS-was applied in the present study. Thyroid and liver homogenates were incubated in the presence of tested NIS inhibitors (i.e., NaClO3, NH4SCN, KSeCN, KNO3, NaF, KClO4, and BPA) in different ranges of concentrations with/without melatonin (5 mM) or IPA (5 mM). The malondialdehyde+4-hydroxyalkenals (MDA + 4-HDA) concentration (LPO index) was measured spectrophotometrically. NaClO3 increased LPO in the thyroid and in the liver, but these pro-oxidative effects were not prevented by either melatonin or IPA. Instead, pro-oxidative effects of NH4SCN observed in both tissues were prevented by both indole substances. KSeCN and NaF increased LPO only in the thyroid, and these pro-oxidative effects were prevented by melatonin and IPA. KNO3, KClO4, and BPA did not increase LPO, which can be due to their low concentrations resulting from restricted solubility. In conclusion, as melatonin prevented oxidative damage to membrane lipids in the thyroid caused by some sodium/iodide symporter inhibitors, this indoleamine shoud be considered as a potential protective agent when produced appropriately in living organisms but also as an exogenous substance recommended to individuals overexposed to NIS inhibitors.

Polystyrene microplastic alters the redox state and arsenic metabolization in the freshwater bivalve Limnoperna fortunei.

Most organisms possess the capacity to metabolize arsenic (As) accumulating compounds to less toxic forms, thus minimizing the adverse effect induced by this metalloid. However, other contaminants may to interfere with As metabolism, contributing to the accumulation of more toxic compounds. Microplastics (MPs) are omnipresent in aquatic environment and may induce toxicological effects (alone or in combination with other contaminants) on living organisms. Therefore, the objective of the present study was to evaluate the effect of the exposure of the freshwater clam Limnoperna fortunei to a combination of MP (4 and 40μg/L of polystyrene microbeads, 1.05μm) and As (50μg/L) for 48h, evaluating the accumulation and metabolization of As and oxidative stress parameters, such as catalase (CAT), glutathione-S-transferase activities, total antioxidant competence, reduced glutathione (GSH), and lipid damage in the gills and digestive glands. Results revealed that low MP concentration disrupts the redox state of the digestive gland by a decrease in the antioxidant activity (CAT and total antioxidant capacity). GSH levels in the gills of animals exposed to MP (4μg/L) alone and the combination of MP+As increased, concomitant with an increase in the percentage of toxic compounds, indicating the effect of MP on As metabolism. Although, few studies evaluated the effect of coexposure to MP+As by considering metabolization of metalloid in freshwater bivalve, our results revealed that exposure to MP reduced the metabolization capacity of As, favoring the accumulation of more toxic compounds besides the MP alone, which showed a pro-oxidant effect in L. fortunei.

The Effect of Piperidine Nitroxides on the Properties of Metalloproteins in Human Red Blood Cells.

Nitroxides are stable, low molecular-weight radicals containing a nitroxide group that has an unpaired electron. The presence of a nitroxide group determines their redox properties. The effect of the piperidine nitroxides, Tempo, Tempol, and Tempamine, on metalloproteins (hemoglobin, superoxide dismutase, catalase) and lactate dehydrogenase in red blood cells was investigated in this research. In addition, the level of lipid peroxidation and the level of protein carbonyl groups were examined as indicators of the effect of oxidative stress. Nitroxides increased superoxide dismutase activity and oxidized hemoglobin to methemoglobin, and also slightly decreased the catalase activity of red blood cells treated with nitroxides. Tempol significantly decreased lactate dehydrogenase activity. All three nitroxides had no effect on membrane lipid peroxidation and protein oxidation. Our results confirm that nitroxides have both antioxidant and prooxidative effects in human red blood cells. The piperidine nitroxides do not initiate the oxidation of proteins and lipids in the membranes of human red blood cells.

Impact of Cumin (Cuminum cyminum) Incorporation on the Generation of Heterocyclic Aromatic Amines in Meatballs

In the current study, the impacts of using cumin (0.5% and 1%, w/w) in beef meatballs on heterocyclic aromatic amines (HAAs) formation and some quality characteristics when cooked at 150 and 250 °C were investigated. It was found that using of cumin at different ratios in beef meatballs showed a significant (p < 0.01) effect on pH, thiobarbituric acid reactive (TBARS) value, and total HAA. The cooking process significantly (p < 0.01) affected the meatballs’ water content, pH, and TBARS values, while the cooking loss and total HAA content of meatballs were significantly (p < 0.01) affected by cooking temperature. It was found that the cumin usage rate in meatballs increased the pH value, while it was found to decrease the TBARS value. As expected, the cooking process was associated with a reduction in water, while it was found to result in an increase in the pH and TBARS values. On the other hand, both cooking temperature and cumin addition to meatballs led to an increase in their total HAA content. Diverse samples exhibited detectable levels of IQx, MeIQx, MeIQ, and PhIP compounds. The presence of MeIQx compound was found in all samples, except for the control group cooked at 150 °C. Additionally, the use of 0.5% and 1% cumin in meatballs cooked at temperature of 250 °C had an inhibitory effect on MeIQx compound. Our results revealed that the incorporation of cumin in the meatballs resulted in an increase in the total HAA content, likely due to its prooxidant effect. However, it was found that the use of cumin at certain rates could reduce the formation of MeIQx compound with an antioxidant effect. However, in this study found that even if 100 g of meatballs with the highest concentration of total HAA were consumed, the maximum HAA exposure limit value (0–15 µg/day) was not exceeded.

Enhanced Antioxidant Activity and Reduced Cytotoxicity of Silver Nanoparticles Stabilized by Different Humic Materials.

The current article describes the biological activity of new biomaterials combining the "green" properties of humic substances (HSs) and silver nanoparticles. The aim is to investigate the antioxidant activity (AOA) of HS matrices (macroligands) and AgNPs stabilized with humic macroligands (HS-AgNPs). The unique chemical feature of HSs makes them very promising ligands (matrices) for AgNP stabilization. HSs have previously been shown to exert many pharmacological effects mediated by their AOA. AgNPs stabilized with HS showed a pronounced ability to bind to reactive oxygen species (ROS) in the test with ABTS. Also, higher AOA was observed for HS-AgNPs as compared to the HS matrices. In vitro cytotoxicity studies have shown that the stabilization of AgNPs with the HS matrices reduces the cytotoxicity of AgNPs. As a result of in vitro experiments with the use of 2,7-dichlorodihydrofluorescein diacetate (DCFDA), it was found that all HS materials tested and the HS-AgNPs did not exhibit prooxidant effects. Moreover, more pronounced AOA was shown for HS-AgNP samples as compared to the original HS matrices. Two putative mechanisms of the pronounced AOA of the tested compositions are proposed: firstly, the pronounced ability of HSs to inactivate ROS and, secondly, the large surface area and surface-to-volume ratio of HS-AgNPs, which facilitate electron transfer and mitigate kinetic barriers to the reduction reaction. As a result, the antioxidant properties of the tested HS-AgNPs might be of particular interest for biomedical applications aimed at inhibiting the growth of bacteria and viruses and the healing of purulent wounds.

Laser Ablated Citrate-Stabilized Silver Nanoparticles Display Size and Concentration Dependant Biological Effects

Silver nanoparticles (AgNPs) have been recognized for widespread biological applications due to their antimicrobial and anti-inflammatory effect, especially in dentistry and for wound healing. Many features determine their beneficial or toxic potential, such as their synthesis type, size, morphology, coating, and concentration. Most synthesis types rely on the use of synthetic chemicals, which contributes to their toxicity. We present an environmentally friendly method for “green” synthesis of AgNPs from the silver target by pulsed laser ablation in liquid (PLAL) using citrate as the stabilizing agent. Since AgNPs already have many dental applications, we examined their antibacterial effect against supragingival biofilm-forming bacteria and bacterial strains known to cause resistant dental infections. Their impact on human fibroblast cells’ cytotoxicity, proliferation (measured by XTT and Ki-67 immunofluorescence), pro/antioxidant balance, and lipid peroxidation (measured by PAB and LPP) was evaluated. AgNPs1 (21 nm) and AgNPs2 (15 nm) spherical nanoparticles with good overall stability were obtained. The highest tested dose of smaller nanoparticles (AgNPs2) displays not only an effective antibacterial effect against the tested oral bacteria strains but also a pro-oxidant and cytotoxic effect on fibroblast cells. Lower doses do not affect bacterial survival but increase the cell proliferation and metabolic activity and show an antioxidative effect, suggesting that different concentrations display a substantially opposite effect. Compared to larger AgNPs1, smaller AgNPs2 possess more potent biological effects, indicating that size plays a pivotal role in their activity. Such opposite outcomes could be important for their medical application, and high concentrations could be used for the inhibition of dental biofilm formation and resistant dental infections as well as proliferative conditions, while low doses could be beneficial in the treatment of atrophic and inflammatory disorders. Finally, we showed that silver-targeted PLAL, using citrate as a stabilizing agent, produces biologically potent nanoparticles that could have many applications depending on their size and concentration.

Effect of Co-administration of Vitamin A and Artesunate in Malaria-Infected Albino Mice

Malaria is a potentially fatal illness that results from infection by parasitic protozoa, which spread to humans via the bites of mosquitoes that have already been infected with the parasite. This research examines the impact of the simultaneous administration of Vitamin A and Artesunate on Plasmodium berghei-infected albino mice. A cohort of sixteen (16) albino mice, with a mean weight of 28g and indeterminate gender, were subjected to P. berghei inoculation. The mice were separated into four groups, each consisting of four animals. Artesunate was administered to Group A, distilled water was administered to Group B, Group C received antioxidant treatment in the form of vitamin A, and Group D was subjected to a combination treatment of Artesunate and Vitamin A. The study monitored the parasitemia level, hematocrit (PVC), percentage mortality rate, and change in body weight following the administration of various treatments, including an antimalarial drug (Artesunate), distilled water, Vitamin A, and a combination therapy of both Artesunate and Vitamin A. The co-administration of vitamin A and Artesunate may decrease the effectiveness of Artesunate in treating malaria infection. The research illustrates the intricate relationship between Artesunate and antioxidant (vitamin A) in mice infected with malaria. The efficacy of Artesunate in clearing the parasite was demonstrated by the reduction of Parasitaemia levels from +2(++) on the 5th day to +3 (+++) on the 7th day, followed by a decrease to +2(++) on the 9th day, and ultimately to +1(+) Parasitaemia on the 10th day. The co-administration of Vitamin A and Artesunate resulted in a reduction of the latter's efficacy in treating malaria, particularly in cases where the parasitemia level was at +3(+++) on the 5th, seventh, and 11th days and +2(++) on the ninth day. This procedure suggests that Vitamin A acted as an antagonist to the antimalarial effects of Artesunate.
 Furthermore, the mortality rate was 50%, and the PCV was reduced compared to Group A. Artesunate, a pro-oxidant that generates free oxygen radicals that act against the parasite. However, the presence of antioxidants like Vitamin A potentially counteracted the effects of pro-oxidants, indicating some form of pharmacological antagonism. The research findings indicate that the co-administration of vitamin A and Artesunate may reduce therapeutic effectiveness. Clinical interventions are recommended to be implemented to regulate the simultaneous administration of Artesunate and Vitamin A, as this may impact the therapeutic objective of malaria treatment.

The implications of exercise in Drosophila melanogaster: insights into Akt/p38 MAPK/Nrf2 pathway associated with Hsp70 regulation in redox balance maintenance.

This study investigated the potential effects of exercise on the responses of energy metabolism, redox balance maintenance, and apoptosis regulation in Drosophila melanogaster to shed more light on the mechanisms underlying the increased performance that this emerging exercise model provides. Three groups were evaluated for seven days: the control (no exercise or locomotor limitations), movement-limited flies (MLF) (no exercise, with locomotor limitations), and EXE (with exercise, no locomotor limitations). The EXE flies demonstrated greater endurance-like tolerance in the swimming test, associated with increased citrate synthase activity, lactate dehydrogenase activity and lactate levels, and metabolic markers in exercise. Notably, the EXE protocol regulated the Akt/p38 MAPK/Nrf2 pathway, which was associated with decreased Hsp70 activation, culminating in glutathione turnover regulation. Moreover, reducing the locomotion environment in the MLF group decreased endurance-like tolerance and did not alter citrate synthase activity, lactate dehydrogenase activity, or lactate levels. The MLF treatment promoted a pro-oxidant effect, altering the Akt/p38 MAPK/Nrf2 pathway and increasing Hsp70 levels, leading to a poorly-regulated glutathione system. Lastly, we demonstrated that exercise could modulate major metabolic responses in Drosophila melanogaster aerobic and anaerobic metabolism, associated with apoptosis and cellular redox balance maintenance in an emergent exercise model.

Evidence of increased gluten-induced perturbations in the nucleophilic tone and detoxifying defences of intestinal epithelial cells impaired by gastric disfunction

It has been increasingly demonstrated over the past few years that some proteolytically resistant gluten peptides may directly affect intestinal cell structure and functions by modulating pro-inflammatory gene expression and oxidative stress. The relationship between oxidative cell damage and Celiac Disease (CD) is supported by several studies on human intestinal epithelial cell lines, such as the Caco-2 cell model, already shown to be particularly sensitive to the pro-oxidative and pro-apoptotic properties of gluten protein digests. Through providing valuable evidence concerning some of the pathophysiological mechanisms that may be at play in gluten-related disorders, most of these in vitro studies have been employing simplified digestion schemes and intestinal cell systems that do not fully resemble mature enterocytes in terms of their characteristic tight junctions, microvilli and membrane transporters. Herein the peptide profile and pro-oxidative effect of two different gastrointestinal gliadin digestions was thoroughly characterized and comprehensively compared: one following the complete INFOGEST workflow and a second one by-passing gastric processing, to assess the dependence of gliadin-triggered downstream cell effects on pepsin activity. In both matrices, gluten-derived immunogenic peptide sequences were identified by non-targeted LC-MS/MS. Altogether, this study provides first-hand data concerning the still unexplored peptide composition, gastric-dependence and immunogenicity of physiologically representative gliadin protein digests as well as foundational clues stressing the need for more complex and integrated in vitro cell systems when modelling and exploiting gluten-induced perturbations in the nucleophilic tone and inflammatory status of intestinal epithelial cells.

Piper nigrum extract suppresses tumor growth and enhances the antitumor immune response in murine models of breast cancer and melanoma

Although the antitumor effect of P. nigrum has been widely studied, research related to its possible immunomodulatory effects is relatively scarce. Here, the antitumor and immunomodulatory activity of an ethanolic extract of P. nigrum were evaluated in the murine models of 4T1 breast cancer and B16-F10 melanoma. In vitro evaluations showed that the P. nigrum extract has cytotoxic activity, induces apoptotic cell death, and has a pro-oxidant effect in both cell lines, but it regulates glucose uptake differently in both lines, decreasing it in 4T1 but not in B16-F10. P. nigrum extract significantly reduced tumor size in both models and decreased the occurrence of macrometastases in 4T1 model. Evaluation of immune subpopulations by flow cytometry revealed that the P. nigrum extract significantly increases the frequency of dendritic cells and activated CD8+ T cells and decreases the frequency of myeloid-derived suppressor like cells and Tregs in the tumor microenvironment of both models but with different dynamics. Our findings strongly suggest that the P. nigrum extract exerts immunomodulatory functions, slightly related to the modulation of cellular energy metabolism, which could ultimately contribute to the promising antitumor effect of P. nigrum.