Protein Oxidative Damage Research Articles

Multiple oxidative and advanced oxidative modifications of hemoglobin in gastric cancer patients measured by nanoflow LC-MS/MS

Overproduction of the reactive oxygen, nitrogen, and chlorine species by the immune systems during chronic infection and inflammation can cause structural and functional changes of cellular proteins. The high abundance of hemoglobin in blood makes hemoglobin adducts suitable biomarkers for assessing the damage of these reactive species in the body. In this study, a total of 23 types and sites of modification in human hemoglobin were simultaneously analyzed, including monooxygenation of histidine, tyrosine, methionine, and aspartate, conversion of histidine to aspartate and hydroxyaspartate, as well as chlorination and nitration of tyrosine residues. Hemoglobin was isolated from the blood of the study subjects, digested into peptides, and the extents of these modifications were quantified relative to their parent peptides using nanoflow liquid chromatography nanoelectrospray ionization tandem mass spectrometry under selected reaction monitoring (nanoLC − NSI-MS/MS-SRM). The extents of monooxygenation at β-His-77 and β-Tyr-130, chlorination at α-Tyr-24 and β-Tyr-130, and nitrosylation at α-Tyr-24 were elevated in gastric cancer patients. Conversely, conversion of histidine to aspartate at α-His-20, α-His-50, β-His-2, β-His-143, and monooxygenation at β-His-143 were decreased in gastric cancer patients. The areas under the receiver operating characteristics (AUC of ROC) curve of these ten types and sites of hemoglobin modifications were between 0.7644 and 0.9644. The ratio of conversion of histidine to hydroxyaspartate versus conversion of histidine to aspartate was significantly higher in gastric cancer patients at α-His-20, α-His-50, and β-His-143 (p < 0.05) with AUC of ROC ranging between 0.7689 and 0.9178. To our knowledge, this is the first report of simultaneous measurement of multiple types of oxidative and advanced oxidative hemoglobin modifications in gastric cancer patients. The results revealed elevated levels of oxidative stress-induced protein damage in gastric cancer patients and the potential of using these modifications of hemoglobin as biomarkers for evaluation of oxidative stress in one drop of blood.

α-Lipoic Acid Reduces Ceramide Synthesis and Neuroinflammation in the Hypothalamus of Insulin-Resistant Rats, While in the Cerebral Cortex Diminishes the β-Amyloid Accumulation.

BackgroundOxidative stress underlies metabolic diseases and cognitive impairment; thus, the use of antioxidants may improve brain function in insulin-resistant conditions. We are the first to evaluate the effects of α-lipoic acid (ALA) on redox homeostasis, sphingolipid metabolism, neuroinflammation, apoptosis, and β-amyloid accumulation in the cerebral cortex and hypothalamus of insulin-resistant rats.MethodsThe experiment was conducted on male cmdb/outbred Wistar rats fed a high-fat diet (HFD) for 10 weeks with intragastric administration of ALA (30 mg/kg body weight) for 4 weeks. Pro-oxidant and pro-inflammatory enzymes, oxidative stress, sphingolipid metabolism, neuroinflammation, apoptosis, and β-amyloid level were assessed in the hypothalamus and cerebral cortex using colorimetric, fluorimetric, ELISA, and HPLC methods. Statistical analysis was performed using three-way ANOVA followed by the Tukey HSD test.ResultsALA normalizes body weight, food intake, glycemia, insulinemia, and systemic insulin sensitivity in HFD-fed rats. ALA treatment reduces nicotinamide adenine dinucleotide phosphate (NADPH) and xanthine oxidase activity, increases ferric-reducing antioxidant power (FRAP) and thiol levels in the hypothalamus of insulin-resistant rats. In addition, it decreases myeloperoxidase, glucuronidase, and metalloproteinase-2 activity and pro-inflammatory cytokines (IL-1β, IL-6) levels, while in the cerebral cortex ALA reduces β-amyloid accumulation. In both brain structures, ALA diminishes ceramide synthesis and caspase-3 activity. ALA improves systemic oxidative status and reduces insulin-resistant rats’ serum cytokines, chemokines, and growth factors.ConclusionALA normalizes lipid and carbohydrate metabolism in insulin-resistant rats. At the brain level, ALA primarily affects hypothalamic metabolism. ALA improves redox homeostasis by decreasing the activity of pro-oxidant enzymes, enhancing total antioxidant potential, and reducing protein and lipid oxidative damage in the hypothalamus of HFD-fed rats. ALA also reduces hypothalamic inflammation and metalloproteinases activity, and cortical β-amyloid accumulation. In both brain structures, ALA diminishes ceramide synthesis and neuronal apoptosis. Although further study is needed, ALA may be a potential treatment for patients with cerebral complications of insulin resistance.

Plasma Total Antioxidant Capacity and Carbonylated Proteins Are Increased in Pregnant Women with Severe COVID-19.

Oxidative stress (OS) induced by SARS-CoV-2 infection may play an important role in COVID-19 complications. However, information on oxidative damage in pregnant women with COVID-19 is limited. Objective: We aimed to compare lipid and protein oxidative damage and total antioxidant capacity (TAC) between pregnant women with severe and non-severe COVID-19. Methods: We studied a consecutive prospective cohort of patients admitted to the obstetrics emergency department. All women positive for SARS-CoV-2 infection by reverse transcription-polymerase chain reaction (RT-qPCR) were included. Clinical data were collected and blood samples were obtained at hospital admission. Plasma OS markers, malondialdehyde (MDA), carbonylated proteins (CP), and TAC; angiogenic markers, fms-like tyrosine kinase-1 (sFlt-1) and placental growth factor (PlGF); and renin-angiotensin system (RAS) markers, angiotensin-converting enzyme 2 (ACE-2) and angiotensin-II (ANG-II) were measured. Correlation between OS, angiogenic, and RAS was evaluated. Results: In total, 57 pregnant women with COVID-19 were included, 17 (28.9%) of which had severe COVID-19; there were 3 (5.30%) maternal deaths. Pregnant women with severe COVID-19 had higher levels of carbonylated proteins (5782 pmol vs. 6651 pmol; p = 0.024) and total antioxidant capacity (40.1 pmol vs. 56.1 pmol; p = 0.001) than women with non-severe COVID-19. TAC was negatively correlated with ANG-II (p < 0.0001) and MDA levels (p < 0.0001) and positively with the sFlt-1/PlGF ratio (p = 0.027). Conclusions: In pregnant women, severe COVID-19 is associated with an increase in protein oxidative damage and total antioxidant capacity as a possible counterregulatory mechanism.

Ecotoxicological impacts of the Fundão dam failure in freshwater fish community: Metal bioaccumulation, biochemical, genetic and histopathological effects

This study investigates the ecotoxicological impacts of the Fundão dam rupture, one of the major environmental disaster that occurred in Brazil and in the world mining industry history, through multi-biomarkers responses and metals bioaccumulation in the fish community of different trophic levels. Specimens of the fishes (omnivorous/herbivorous and carnivorous) were collected along the Doce River channel and its affluent Guandú River, and in different lakes and coastal lagoons adjacent to the river channel, in the Espirito Santo State, Southeast of Brazil. Four sampling collections were carried out over two years (2018 to 2020, during dry and rainy seasons). For both trophic groups the biomarkers responses indicated physiological alterations related to metals exposure and showed strong seasonal variations. The principal component analysis and integrated biomarker response index showed that DNA damage and lipid peroxidation were more associated with dry season 2 (Sep/Oct 2019) and the oxidative damage in proteins, metallothioneins concentration and the activity of superoxide dismutase in the gills showed a greater association with rainy season 2 (Jan/Feb 2020). On the other hand, the enzymes of energy metabolism, catalase and histological damage in the liver and the gills, were more associated with the dry and rainy campaigns of the first year of monitoring. The multivariate approach also suggested a temporal intensification in the bioaccumulation of metals and biological effects in the lacustrine environments. Thus, these results demonstrate that the release of mineral residues from the rupture of the Fundão mine dam affects the health status of the fish from the Doce River basin, provoking metals bioaccumulation, hepatic and branchial damage in the fish besides inducing of enzyme activity related to metal contamination, even four years after the rupture.

Arousal from Torpor Increases Oxidative Damage in the Hibernating Thirteen-Lined Ground Squirrel (Ictidomys tridecemlineatus).

AbstractDuring hibernation, especially during arousal from torpor to interbout euthermia (IBE), blood flow changes drastically. In nonhibernating mammals, similar changes during ischemia/reperfusion lead to oxidative damage. We hypothesized that suppression of mitochondrial metabolism during hibernation protects against such damage. We compared markers of oxidative damage and total antioxidant capacity in eight tissues among summer, torpid, and IBE thirteen-lined ground squirrels. Overall, summer tissue had less lipid and protein oxidative damage than tissue from the hibernation season, but DNA damage (in four tissues) and total antioxidant capacity (in all eight tissues) were similar among all groups. During torpor, when mitochondrial metabolism is suppressed, lipid damage in heart, brown adipose tissue, and small intestine was lower than IBE by as much as fivefold. By contrast, oxidative damage to protein was at least twofold higher in liver and skeletal muscle in torpor compared with IBE. Our findings suggest that arousal from torpor creates oxidative damage similar to ischemia/reperfusion injury but that this damage is repaired during IBE. These differences cannot be explained by changes in antioxidant capacity, so they are likely due to differences is reactive oxygen species production among hibernation states that may relate to the well-characterized reversible suppression of mitochondrial metabolism during torpor.

Oxidative damage and mitochondrial functionality in hearts from KO UCP3 mice housed at thermoneutrality.

The antioxidant role of mitochondrial uncoupling protein 3 (UCP3) is controversial. This work aimed to investigate the effects of UCP3 on the heart of mice housed at thermoneutral temperature, an experimental condition that avoids the effects of thermoregulation on mitochondrial activity and redox homeostasis, preventing the alterations related to these processes from confusing the results caused by the lack of UCP3. WT and KO UCP3 mice were acclimatized at 30°C for 4weeks and hearts were used to evaluate metabolic capacity and redox state. Tissue and mitochondrial respiration, the activities of the mitochondrial complexes, and the protein expression of mitochondrial complexes markers furnished information on mitochondrial functionality. The levels of lipid and protein oxidative damage markers, the activity of antioxidant enzymes, the reactive oxygen species levels, and the susceptibility to in vitro Fe-ascorbate-induced oxidative stress furnished information on redox state. UCP3 ablation reduced tissue and mitochondrial respiratory capacities, not affecting the mitochondrial content. In KO UCP3 mice, the mitochondrial complexes activities were lower than in WT without changes in their content. These effects were accompanied by an increase in the level of oxidative stress markers, ROS content, and in vitro susceptibility to oxidative stress, notwithstanding that the activities of antioxidant enzymes were not affected by UCP3 ablation. Such modifications are also associated with enhanced activation/phosphorylation of EIF2α, a marker of integrated stress response and endoplasmic reticulum stress (GRP778 BIP). The lack of UCP3 makes the heart more prone to oxidative insult by reducing oxygen consumption and increasing ROS. Our results demonstrate that UCP3 helps the cell to preserve mitochondrial function by mitigating oxidative stress.

Inhibition of Peroxidation Potential and Protein Oxidative Damage by Metal Mangiferin Complexes

Background: Metal coordination complexes of polyphenolic compounds have been claimed to have better antioxidant and protection against protein oxidative damage effects than the isolated ligands. Whereas flavonoids have been extensively studied, xanthones such as mangiferin are lacking extensive research. Methods: Cu (II), Zn (II), and Se (IV) mangiferin complexes were synthesized with different stoichiometric ratios. Products were isolated by preparative chromatography and subjected to spectral analysis by FT-IR, HPLC-DAD, and HPLC-ESI-MS. The inhibition effects on peroxidation potential and protein oxidative damage were determined for all the metal–MF complexes. Results: Eight metal–MF complexes were isolated. Cu (II)–MF complexes did not improve MF antioxidant/protective effects; Zn (II) complexes (stoichiometric ratio 1:2) antioxidant/protective effects had no significant differences to MF; Zn (II)– and Se (IV)–MF complexes (stoichiometric ratio 1:3) showed the best inhibition effects on peroxidation potential (49.06% and 32.08%, respectively), and on the protection against protein oxidative damage (14.49% and 20.81%, respectively). Conclusions: The antioxidant/protective effects of Se (IV)– and Zn (II)–MF coordination complexes were significantly improved as compared to isolated MF, when the reaction between the metal salt and MF was performed with a stoichiometric ratio 1:3.

Effects of perfluorooctanoic acid in oxidative stress generation, DNA damage in cumulus cells, and its impact on in vitro maturation of porcine oocytes.

Perfluorooctanoic acid is a synthetic compound mostly used in a wide range of consumer products with several adverse effects on somatic cells and gametes. It has been linked to hepatotoxic and carcinogenic effects, alterations in the immune system, endocrine, and reproductive alterations. In vivo studies show an increase in reactive oxygen species and DNA damage. However, the mechanisms by which this compound affects fertility, remain contradictory. Therefore, the aim of the present study was to evaluate the effect of perfluorooctanoic acid on oocyte viability and maturation, as well as the viability, generation of oxidative stress, and genotoxic damage in the cumulus cells exposed during in vitro maturation. This compound had a negative effect on oocyte viability (lethal concentration, LC50 =269 μM) and maturation (inhibition maturation concentration IM50 =75 μM), while in cumulus cells the LC50 was 158 μM. The generation of reactive oxygen species evaluated in cumulus cells, protein carbonylation, and DNA damage, was significantly increased at 40 μM perfluorooctanoic acid. This study provides evidence that perfluorooctanoic acid causes reactive oxygen species generation, protein oxidation, and DNA damage in cumulus cells, compromising the maturation and viability of porcine oocyte, which may affect fertility.

LRP1 loss in airway epithelium exacerbates smoke-induced oxidative damage and airway remodeling

The LDL receptor-related protein 1 (LRP1) partakes in metabolic and signaling events regulated in a tissue-specific manner. The function of LRP1 in airways has not been studied. We aimed to study the function of LRP1 in smoke-induced disease. We found that bronchial epithelium of patients with chronic obstructive pulmonary disease and airway epithelium of mice exposed to smoke had increased LRP1 expression. We then knocked out LRP1 in human bronchial epithelial cells in vitro and in airway epithelial club cells in mice. In vitro, LRP1 knockdown decreased cell migration and increased transforming growth factor β activation. Tamoxifen-inducible airway-specific LRP1 knockout mice (club Lrp1−/−) induced after complete lung development had increased inflammation in the bronchoalveolar space and lung parenchyma at baseline. After 6 months of smoke exposure, club Lrp1−/− mice showed a combined restrictive and obstructive phenotype, with lower compliance, inspiratory capacity, and forced expiratory volume0.05/forced vital capacity than WT smoke-exposed mice. This was associated with increased values of Ashcroft fibrotic index. Proteomic analysis of room air exposed-club Lrp1−/− mice showed significantly decreased levels of proteins involved in cytoskeleton signaling and xenobiotic detoxification as well as decreased levels of glutathione. The proteome fingerprint created by smoke eclipsed many of the original differences, but club Lrp1−/− mice continued to have decreased lung glutathione levels and increased protein oxidative damage and airway cell proliferation. Therefore, LRP1 deficiency leads to greater lung inflammation and damage and exacerbates smoke-induced lung disease.

Impaired proteostasis in obese skeletal muscle relates to altered immunoproteasome activity.

Obesity-associated inflammation and/or oxidative stress can damage intramuscular proteins and jeopardize muscle integrity. The immunoproteasome (iProt) is vital to remove oxidatively modified proteins, but this function may be compromised with obesity. We sought to elucidate whether diet-induced obesity alters intramuscular iProt content and activity in mice to identify a possible mechanism for impaired muscle proteostasis in the obese state. Total proteasome content and activity and estimates of muscle oxidative damage, inflammation, muscle mass and strength were also assessed. Twenty-three male, 5-week-old C57BL/6J mice were fed a high-fat, high-sucrose (HFS; 45% kcal fat, 17% sucrose, n = 12) or low-fat, low-sucrose (LFS; 10% kcal fat, 0% sucrose, n = 11) diet for 12 weeks. Strength was assessed via a weightlifting test. Despite no change in pro-inflammatory cytokines (P > 0.05), oxidative protein damage was elevated within the gastrocnemius (P = 0.036) and tibialis anterior (P = 0.033) muscles of HFS-fed mice. Intramuscular protein damage coincided with reduced iProt and total proteasome activity (P < 0.05), and reductions in relative muscle mass (P < 0.001). Therefore, proteasome dysregulation occurs in obese muscle and may be a critical link in muscle oxidative stress. Novelty: Our results show for the first time that immunoproteasome and total proteasome function is significantly reduced within obese muscle. Visceral fat mass is a significant predictor of diminished proteasome activity in skeletal muscle. Proteasome function is inversely correlated with an intramuscular accumulation of oxidatively damaged proteins.

Intrinsically High Capacity of Animal Cells From a Symbiotic Cnidarian to Deal With Pro-Oxidative Conditions.

The cnidarian-dinoflagellate symbiosis is a mutualistic intracellular association based on the photosynthetic activity of the endosymbiont. This relationship involves significant constraints and requires co-evolution processes, such as an extensive capacity of the holobiont to counteract pro-oxidative conditions induced by hyperoxia generated during photosynthesis. In this study, we analyzed the capacity of Anemonia viridis cells to deal with pro-oxidative conditions by in vivo and in vitro approaches. Whole specimens and animal primary cell cultures were submitted to 200 and 500 μM of H2O2 during 7 days. Then, we monitored global health parameters (symbiotic state, viability, and cell growth) and stress biomarkers (global antioxidant capacity, oxidative protein damages, and protein ubiquitination). In animal primary cell cultures, the intracellular reactive oxygen species (ROS) levels were also evaluated under H2O2 treatments. At the whole organism scale, both H2O2 concentrations didn’t affect the survival and animal tissues exhibited a high resistance to H2O2 treatments. Moreover, no bleaching has been observed, even at high H2O2 concentration and after long exposure (7 days). Although, the community has suggested the role of ROS as the cause of bleaching, our results indicating the absence of bleaching under high H2O2 concentration may exculpate this specific ROS from being involved in the molecular processes inducing bleaching. However, counterintuitively, the symbiont compartment appeared sensitive to an H2O2 burst as it displayed oxidative protein damages, despite an enhancement of antioxidant capacity. The in vitro assays allowed highlighting an intrinsic high capacity of isolated animal cells to deal with pro-oxidative conditions, although we observed differences on tolerance between H2O2 treatments. The 200 μM H2O2 concentration appeared to correspond to the tolerance threshold of animal cells. Indeed, no disequilibrium on redox state was observed and only a cell growth decrease was measured. Contrarily, the 500 μM H2O2 concentration induced a stress state, characterized by a cell viability decrease from 1 day and a drastic cell growth arrest after 7 days leading to an uncomplete recovery after treatment. In conclusion, this study highlights the overall high capacity of cnidarian cells to cope with H2O2 and opens new perspective to investigate the molecular mechanisms involved in this peculiar resistance.

Free iron rather than heme iron mainly induces oxidation of lipids and proteins in meat cooking

In this study, the relationship between different forms of iron (free or binding) and oxidation of lipids, proteins in meat system were investigated. Pork tenderloin was heated in 80 °C water bath for 0, 30, 60, 120, 180, 240, 300 min. Compared with control group, Equal and Treble deferiprone group confirmed that free iron was the main oxidizing substance. Moreover, adding exogenous heme caused slight increase of meat oxidation (p < 0.05). At the same time, the antioxidant properties of deferiprone were also evaluated and it shows few antioxidant properties. This study also found that the oxidation of lipid by free iron was more serious than protein. These results suggested that controlling free iron and production of free iron from heme is a potential approach for reducing the oxidative damage of lipid and protein in meat cooking.

Two-dimensional gel analysis to investigate the effect of hydroxyl radical oxidation on freshness indicator protein of Coregonus peled during 4 °C storage

The present study investigates the freshness indicator protein of Coregonus peled and the effects of oxidation on the quality of fish during 4 °C storage. Samples were stored at 4 °C after oxidation treatment and analyzed for changes in proteins, using two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS). A total of 382 differential proteins were identified, of which 20 proteins were quantified and studied including 12 proteins related to oxidation. The analysis of differential proteins was related to quality postmortem. Moreover, creatine kinase, malate dehydrogenase, and glyceraldehyde 3-phosphate dehydrogenase were identified as potential biomarkers for protein oxidative damage. Actin can be used to represent the softening of fish in postmortem storage. Pearson correlation analysis indicated that the changes in protein structure and biomarkers were influenced by oxidation. Bioinformatics suggested that differentially expressed proteins participated in the metabolic pathway via oxidation during 4 °C storage. The results of this study can contribute to a further understanding of the changes in fish quality during storage and the use of differential proteins as indicators of fish freshness.

Anti-α-Glucosidase and Antiglycation Activities of α-Mangostin and New Xanthenone Derivatives: Enzymatic Kinetics and Mechanistic Insights through In Vitro Studies.

Diabetes mellitus is characterized by chronic hyperglycemia that promotes ROS formation, causing severe oxidative stress. Furthermore, prolonged hyperglycemia leads to glycation reactions with formation of AGEs that contribute to a chronic inflammatory state. This research aims to evaluate the inhibitory activity of α-mangostin and four synthetic xanthenone derivatives against glycation and oxidative processes and on α-glucosidase, an intestinal hydrolase that catalyzes the cleavage of oligosaccharides into glucose molecules, promoting the postprandial glycemic peak. Antiglycation activity was evaluated using the BSA assay, while antioxidant capacity was detected with the ORAC assay. The inhibition of α-glucosidase activity was studied with multispectroscopic methods along with inhibitory kinetic analysis. α-Mangostin and synthetic compounds at 25 µM reduced the production of AGEs, whereas the α-glucosidase activity was inhibited only by the natural compound. α-Mangostin decreased enzymatic activity in a concentration-dependent manner in the micromolar range by a reversible mixed-type antagonism. Circular dichroism revealed a rearrangement of the secondary structure of α-glucosidase with an increase in the contents of α-helix and random coils and a decrease in β-sheet and β-turn components. The data highlighted the anti-α-glucosidase activity of α-mangostin together with its protective effects on protein glycation and oxidation damage.

Acutely applied melatonin prevents CCl4-induced testicular lesions in rats: the involvement of the oxidative capacity and arginine metabolism

Carbon tetrachloride (CCl4) represents an organic chemical that causes reactive oxygen species derived organ disturbances including male infertility. Melatonin (MLT) is a neurohormone with strong antioxidant capacity, involved in numerous physiological processes. In this study we evaluated the capability of MLT, administered in a single dose of 50 mg/kg, to preserve the testicular tissue function after an acute administration of CCl4 to rats. The disturbance in testicular tissue and the effects of MLT after CCl4 exposure were estimated using biochemical parameters that enabled us to determine the tissue (anti)oxidant status and the intensity of arginine/nitric oxide metabolism. Also, the serum levels of testosterone and the histopathological analysis of tissue gave us a better insight into the occurring changes. A significant diminution in tissue antioxidant defences, arginase activity and serum testosterone levels, followed by the increased production of nitric oxide and extensive lipid and protein oxidative damage, was observed in the CCl4-treated group. The application of MLT after the CCl4 caused changes, clearly visible at both biochemical and histological level, which could be interpreted mainly as a consequence of general antioxidant system stimulation and a radical scavenger. On the other hand, the application of MLT exerted a limited action on the nitric oxide signalling pathway.

OXIDATIVE DAMAGE MARKERS IN ALCOHOLIC HEPATITIS

Alcohol hepatitis (AH) is a clinical syndrome in patients with excessive and chronic alcohol consumption. Ethanol metabolism produces free radicals: reactive oxygen species (ROS) and reactive nitrogen species (RNS). This contributes to cellular damage in AH. The malondialdehyde (MDA) and carbonylated proteins are markers of oxidative damage in lipids and proteins. Our objective is to evaluate the levels of serum MDA and carbonylated proteins in AH patients with. This transversal study included 147 individuals divided into two groups: the control group (n=100), which consists of individuals with alcohol consumption ≤ 10 g/day and AUDIT score ≤ 7, and the group with AH patients (n=47). We measured the serum levels of MDA (thiobarbituric acid method) and carbonylated proteins (DNPH reaction). The statistical analysis was performed by the Windows SPSS v22 software. Data were expressed as mean values ± SEM. Comparisons were carried out by analysis of variance (ANOVA). A p-value <0.05 was considered statistically significant. The control group (CT), with a mean of 30.47±0.52 years old, had a consumption of 2.32±0.21g of alcohol daily (gOH/day) and an AUDIT score of 2.24±0.10. The AH patients, with a mean of 41.68±1.3 years old, had a consumption of 354.25±39.54 gOH/day and an AUDIT score of 30±1.45. The AST, ALT, GGT, total and indirect bilirubin serum levels were higher in AH compared to CT (p<0.0001), with a ratio of AST/ALT ≥2. The albumin serum levels were lower (p<0.0001) in AH vs. CT. The carbonylated proteins serum levels were higher in patients with AH compared to CT (p<0.0001). No differences in MDA serum levels were found between both groups. We reported greater MDA serum levels (p=0.001) in alcoholic liver cirrhosis patients when compared to the control group; and an insignificant difference in carbonylated proteins serum level between both participant groups. The alcoholic hepatitis patients have an increase in carbonylated proteins oxidative damage while there is no lipidic damage. Our results suggest that carbonylated proteins may a damage oxidative marker in the AH Mexican population. This damage may increase the risk of malnutrition, susceptibility to infections and sepsis, deficient coagulation factors production, gastrointestinal bleeding, among other complications that increase mortality. It is necessary to counteract oxidative damage to improve and complement the actual treatment in alcoholic hepatitis. Competing interests. The authors declare they have no competing interests. This work was partially financed by CONACyT SALUD-2016-272579 and PAPIIT- UNAM TA200515

Characteristics of T lymphocyte content according to oxidative stress and antioxidant activity in patients with pulmonary tuberculosis and COVID-19 infection

In order to study the particularities of T lymphocyte content depending on the indicators of oxidative stress and antioxidant activity, in patients with pulmonary tuberculosis in association with COVID-19 coinfection, 42 patients were examined, of which: a) baseline - 21 patients with pulmonary tuberculosis and co-infection COVID-19, b) control group - 21 patients with pulmonary tuberculosis. At the patients included in the study, there were determined the leukocyte formula, the level of CD3, CD4, CD8 lymphocytes, the oxidative stress reaction as well as the antioxidant activity. As a result, it was determined that at patients with tuberculosis and COVID-19 coinfection: as the content of lymphocytes, and in particular, T lymphocytes decreases at patients with pulmonary tuberculosis and COVID-19 co-infection, the degree of oxidative damage of proteins and the predisposition of lipoproteins to oxidation increases significantly according to the data of the AOPP marker and the indicator of malonic dialdehyde of oxidative stress (malonic dialdehyde - DAM), which indicates the severity of oxidative stress reactions at these patients. The total antioxidant activity, determined by the ATS method at patients with pulmonary tuberculosis and COVID-19 co-infection, was significantly reduced, and according to the CUPRAC method it increased significantly as the content of lymphocytes and, in particular, lymphocytes decreased. The enzyme superoxide dismutase (SOD), catalase and the activity of the antioxidant protein ceruloplasmin at patients with pulmonary tuberculosis and COVID19 decreased significantly as the content of lymphocytes and, in particular, T lymphocytes decreased. Thus, the parameters of oxidative stress and antioxidant activity can be considered potential biomarkers markers of the evolution of pulmonary tuberculosis in association with COVID-19 infection.

Evaluation of Protein Carbonyl and Vitamin C in Seminal Plasma of Infertile Male: A Hospital-based Study in Bengali Population

Introduction: Male infertility has been coupled with the imbalance between production of Reactive Oxygen Species (ROS) and antioxidant (e.g.,: vitamin C) level. Elevated concentrations of ROS in the semen can lead to oxidative protein damage as they counter with the amino acids' side chains in the protein, leading to the production of carbonyl groups. Aim: To assess if there is any difference of seminal plasma Protein Carbonyl (PC) and vitamin C level in male infertile and fertile subjects in the midst of their correlation with other relevant seminal parameters. Materials and Methods: This was a hospital-based case- control study of a Bengali population. Semen samples of 124 males (Group A; 68 infertile males, Group B; 56 fertile males) were tested. Seminal fluid analysis was done with Makler counting chamber. PC and vitamin C were measured by Levin’s and Roe’s photometric methods respectively. To evaluate the differences in the mean ranks of these parameters Mann- Whitney U test was used. Results: Both in group A and group B sperm count was positively correlated with motility and vitamin C but negatively correlated with PC at significant level p&lt;0.05. Statistically significant differences of mean ranks of these parameters (sperm count: 52.82 and 74.26, motility: 52.10 Vs 75.13, PC: 76.57 and 45.41, vitamin C:55.99 and 70.40, Mann Whitney U:1245, 1197, 947 and 1461, respectively) between the two groups were found. Hence, indicate that in infertile subjects the balance between PC and vitamin C is disturbed. Conclusion: Assessment of oxidative status may serve the clinician in additional management of idiopathic male infertility.

Effects of orchiectomy and testosterone replacement therapy on redox balance and salivary gland function in Wistar rats

The objective of this study was to investigate the effects of orchiectomy (ORX) and testosterone replacement therapy (TRT) on redox balance and function of salivary glands. Forty-five young adult male Wistar rats (3 months old) were either castrated bilaterally or underwent fictitious surgery (SHAM) and were subsequently distributed into 3 groups: SHAM, ORX, and TRT (castrated rats that received an intramuscular injection of testosterone cypionate 10 mg/kg/weekly). All treatments started 4 weeks after castration (4 months old) and lasted 4 weeks (5 months old). At the end of treatment, pilocarpine-induced salivary secretion was collected to analyze salivary flow rate and biochemistry composition, and the parotid (PG) and submandibular (SMG) glands were sampled for redox balance markers and histomorphometric analyses. ORX increased salivary flow rate, calcium, phosphate, and chloride, and decreased total protein and amylase, while not changing the salivary buffer capacity, pH, sodium, and potassium compared to SHAM. TRT restored all salivary parameters to SHAM values. ORX increased oxidative lipid and protein damage, total antioxidant capacity, and uric acid in both salivary glands compared to SHAM. Superoxide dismutase, catalase, and glutathione peroxidase activities were greater only in the SMG of the ORX group in relation to SHAM. ORX decreased duct and acini area, while increasing connective tissue in the PG. On the other hand, ORX reduced duct area and increased acini area in the SMG compared to SHAM. TRT restored the redox balance and histomorphometric parameters to close to SHAM values in both salivary glands. Orchiectomy-induced salivary gland dysfunction was characterized by an increase in the salivary flow rate and changes in the secretion of total protein, amylase, and electrolytes, which are key factors, considered important for maintaining oral health status. To sum up, orchiectomy impaired the redox balance of the salivary glands. Our results also showed that TRT reversed the oxidative damage, morphological alterations, and salivary gland dysfunction induced by orchiectomy. Therefore, these results suggest an important action of testosterone on the redox balance and secretory ability of salivary glands.

Causation of Oxidative Stress and Defense Response of a Yeast Cell Model after Treatment with Orthodontic Alloys Consisting of Metal Ions.

Misaligned teeth have a tremendous impact on oral and dental health, and the most efficient method of correcting the problem is orthodontic treatment with orthodontic appliances. The study was conducted to investigate the metal composition of selected orthodontic alloys, the release of metal ions, and the oxidative consequences that the metal ions may cause in the cell. Different sets of archwires, stainless steel brackets, and molar bands were incubated in artificial saliva for 90 days. The composition of each orthodontic material and quantification of the concentration of metal ions released were evaluated. Metal ion mixtures were prepared to determine the occurrence of oxidative stress, antioxidant enzyme defense system, and oxidative damage to proteins. The beta titanium alloy released the fewest metal ions and did not cause oxidative stress or protein damage. The metal ions from stainless steel and the cobalt-chromium alloy can cause oxidative stress and protein damage only at high concentrations. All metal ions from orthodontic alloys alter the activity of antioxidant enzymes in some way. The determined amounts of metal ions released from orthodontic appliances in a simulated oral environment are still below the maximum tolerated dose, and the concentrations of released metal ions are not capable of inducing oxidative stress, although some changes in antioxidant enzyme activity were observed at these concentrations.