OXIDATIVE CHANGES OF LIPIDS, PROTEINS AND ANTIOXIDANTS IN YOGURT DURING THE SHELF LIFE

Background: Oxidation processes in milk and yogurt during the shelf life can result in an alteration of protein and lipid constituents. Therefore, the antioxidant properties of yogurt in standard conditions of preservation were evaluated. Results: Total phenols, free radical scavenger activity, degree of lipid peroxidation and protein oxidation were determined in plain and skim yogurts with or without fruit puree. After production, plain, skim, plain berries and skim berries yogurts were compared during the shelf life up to 9 weeks. All types of yogurts revealed a basal antioxidant activity that was higher when a fruit puree was present but gradually decreased during the shelf life. However, after five-eight weeks, antioxidant activity increased again. Both in plain and berries yogurts lipid peroxidation increased until the seventh week of shelf life and after decreased, while protein oxidation of all yogurts was similar either in the absence or presence of berries and increased during shelf life. Conclusion: During the shelf life, a different behavior between lipid and protein oxidation takes place and the presence of berries determines a protection only against lipid peroxidation

Mitochondrial Thioredoxin System as a Modulator of Cyclophilin D Redox State

The mitochondrial thioredoxin system (NADPH, thioredoxin reductase, thioredoxin) is a major redox regulator. Here we have investigated the redox correlation between this system and the mitochondrial enzyme cyclophilin D. The peptidyl prolyl cis-trans isomerase activity of cyclophilin D was stimulated by the thioredoxin system, while it was decreased by cyclosporin A and the thioredoxin reductase inhibitor auranofin. The redox state of cyclophilin D, thioredoxin 1 and 2 and peroxiredoxin 3 was measured in isolated rat heart mitochondria and in tumor cell lines (CEM-R and HeLa) by redox Western blot analysis upon inhibition of thioredoxin reductase with auranofin, arsenic trioxide, 1-chloro-2,4-dinitrobenzene or after treatment with hydrogen peroxide. A concomitant oxidation of thioredoxin, peroxiredoxin and cyclophilin D was observed, suggesting a redox communication between the thioredoxin system and cyclophilin. This correlation was further confirmed by i) co-immunoprecipitation assay of cyclophilin D with thioredoxin 2 and peroxiredoxin 3, ii) molecular modeling and iii) depleting thioredoxin reductase by siRNA. We conclude that the mitochondrial thioredoxin system controls the redox state of cyclophilin D which, in turn, may act as a regulator of several processes including ROS production and pro-apoptotic factors release

Cytotoxicity of a mitochondriotropic quercetin derivative: Mechanisms

AbstractThe mitochondriotropic compound 7-O-(4-triphenylphosphoniumbutyl)quercetin iodide (Q-7BTPI) in the μM concentration range caused necrotic death of cultured cells by acting as a prooxidant, with generation of superoxide anion in the mitochondria. Externally added membrane-permeating superoxide dismutase or catalase largely prevented death. Rescue by permeant catalase indicates that the toxicant is H2O2, or reactive species derived from it. Rescue by permeant dismutase suggests the possibility of a chain mechanism of H2O2 production, in which dismutation of superoxide constitutes a termination step. Oxidative stress was due to the presence of free phenolic hydroxyls and to accumulation in mitochondria, since the analogous mitochondriotropic per-O-methylated compound -3,3′,4′,5-tetra-O-methyl,7-O-(4-triphenylphosphoniumbutyl) quercetin iodide (QTM-7BTPI)—or Quercetin itself induced no or little superoxide production and cell death. Q-7BTPI did not cause a significant perturbation of the mitochondrial transmembrane potential or of respiration in cells. On the other hand its presence led to inhibition of glutathione peroxidase, an effect expected to accentuate oxidative stress by interfering with the elimination of H2O2. An exogenous permeable glutathione precursor determined a strong increase of cellular glutathione levels but did not rescue the cells. Death induction was selective for fast-growing C-26 tumoral cells and mouse embryonic fibroblasts (MEFs) while sparing slow-growing MEFs. This suggests a possible use of Q-7BTPI as a chemotherapeutic agent

Low in‑hospital mortality rate in patients with COVID‑19 receiving thromboprophylaxis: data from the multicentre observational START‑COVID Register

Abstract COVID-19 infection causes respiratory pathology with severe interstitial pneumonia and extra-pulmonary complications; in particular, it may predispose to thromboembolic disease. The current guidelines recommend the use of thromboprophylaxis in patients with COVID-19, however, the optimal heparin dosage treatment is not well-established. We conducted a multicentre, Italian, retrospective, observational study on COVID-19 patients admitted to ordinary wards, to describe clinical characteristic of patients at admission, bleeding and thrombotic events occurring during hospital stay. The strategies used for thromboprophylaxis and its role on patient outcome were, also, described. 1091 patients hospitalized were included in the START-COVID-19 Register. During hospital stay, 769 (70.7%) patients were treated with antithrombotic drugs: low molecular weight heparin (the great majority enoxaparin), fondaparinux, or unfractioned heparin. These patients were more frequently affected by comorbidities, such as hypertension, atrial fibrillation, previous thromboembolism, neurological disease,and cancer with respect to patients who did not receive thromboprophylaxis. During hospital stay, 1.2% patients had a major bleeding event. All patients were treated with antithrombotic drugs; 5.4%, had venous thromboembolism [30.5% deep vein thrombosis (DVT), 66.1% pulmonary embolism (PE), and 3.4% patients had DVT + PE]. In our cohort the mortality rate was 18.3%. Heparin use was independently associated with survival in patients aged ≥ 59 years at multivariable analysis. We confirmed the high mortality rate of COVID-19 in hospitalized patients in ordinary wards. Treatment with antithrombotic drugs is significantly associated with a reduction of mortality rates especially in patients older than 59 years

Inhibition of thioredoxin reductase by lanthanum chloride

Lanthanum chloride (LaCl3) is a good inhibitor of both the cytosolic and mitochondrial thioredoxin reductase with IC50 values of 1.75 and 7.46 \u3bcM, respectively. On the contrary, the related enzyme glutathione reductase is not inhibited by lanthanum ions even at relatively high concentrations. In the presence of LaCl3, steady-state kinetics shows a non-competitive type of inhibition with 5,5\u2032-dithiobis-(2-nitrobenzoic acid) (DTNB) as a substrate suggesting no interaction of this ion with the thiol/selenol active site of thioredoxin reductase. Comparison of the electrostatic surface potential of thioredoxin reductase shows that the presence of a trivalent cation such as La3 + decreases the negatively charged area of the enzyme surface particularly in the region closed to the NADPH binding site. Human ovarian carcinoma cells (A2780 cells) incubated with lanthanum ions show a noticeable inhibition of thioredoxin reductase activity indicating the ability of this ion to reach the active site of the enzyme even in a cellular setting. In addition, A2780 cells treated with LaCl3 show an increase in reactive oxygen species production in part dependent on thioredoxin reductase inhibition

Comparative effects of selenate and selenite on selenium accumulation, morphophysiology, and glutathione synthesis in Ulva australis

The capacity of Ulva australis Areschoug to tolerate and accumulate selenium (Se) supplied in the form of selenate or selenite was investigated. The macroalga was provided for 3 and 7 days with concentrations of selenate (Na2SeO4) or selenite (Na2SeO3) ranging from 0 to 400 mu M. U. australis exhibited the highest ability to accumulate selenium when fed with 100 mu M selenate and 200 mu M selenite after 7 days, and accumulation values were respectively 25 and 36 ppm Se. At the same concentrations, stimulation of the synthesis of chlorophylls and carotenoids was observed. Elevated doses of selenate or selenite decreased Se accumulation inside algal cells, perhaps through repression of membrane transporters. This effect was more pronounced in thalli cultivated with selenate. There were no morphological and ultrastructural alterations in thalli exposed to Se. However, selenite induced the increase of the oxidized fraction of glutathione (GSSG), perhaps because of its capacity to bind the thiol group of reduced glutathione (GSH). In conclusion, this study highlights the capacity of U. australis to resist to very high concentrations of selenite and selenate, which are normally toxic to other organisms. Also, the lack of bioconcentration in U. australis indicates that this alga does not facilitate delivery of Se in the food chain and remains safe for consumption when it grows in water bodies contaminated with Se. Its potential for the removal of excess Se from water bodies appears limited

Myotonic dystrophy protein kinase (DMPK) prevents ROS-induced cell death by assembling a hexokinase II-Src complex on the mitochondrial surface

The biological functions of myotonic dystrophy protein kinase (DMPK), a serine/threonine kinase whose gene mutations cause myotonic dystrophy type 1 (DM1), remain poorly understood. Several DMPK isoforms exist, and the long ones (DMPK-A/B/C/D) are associated with the mitochondria, where they exert unknown activities. We have studied the isoform A of DMPK, which we have found to be prevalently associated to the outer mitochondrial membrane. The kinase activity of mitochondrial DMPK protects cells from oxidative stress and from the ensuing opening of the mitochondrial permeability transition pore (PTP), which would otherwise irreversibly commit cells to death. We observe that DMPK (i) increases the mitochondrial localization of hexokinase II (HK II), (ii) forms a multimeric complex with HK II and with the active form of the tyrosine kinase Src, binding its SH3 domain and (iii) it is tyrosine-phosphorylated by Src. Both interaction among these proteins and tyrosine phosphorylation of DMPK are increased under oxidative stress, and Src inhibition selectively enhances death in DMPK-expressing cells after HK II detachment from the mitochondria. Down-modulation of DMPK abolishes the appearance of muscle markers in in vitro myogenesis, which is rescued by oxidant scavenging. Our data indicate that, together with HK II and Src, mitochondrial DMPK is part of a multimolecular complex endowed with antioxidant and pro-survival properties that could be relevant during the function and differentiation of muscle fibers