A pH Dependant Switch in DHP Oxidation Mechanism

Dehaloperoxidase (DHP) is a multifunctional enzyme found in Amphitrite ornata, a sediment-dwelling marine worm. This enzyme possess the structure of a traditional hemoglobin enzyme and serves as the primary oxygen carrier in A. ornata; however, it also possesses peroxidase and peroxygenase capabilities. These secondary oxidative functions provide a remarkable ability for A. ornata to resist the effects of toxic metabolites secreted by other organisms that cohabit its benthic ecosystem. This study will analyze the novel catalytic switching between peroxygenase and peroxidase oxidation mechanisms employed by DHP in response to pH changes

Antioxidant and antihemolytic activities of methanol extract of Hyssopus angustifolius

This study was designed to evaluate antioxidant and antihemolytic activities of Hyssopus angustifolius flower, stem and leaf methanol extracts by employing various in vitro assays. The leaf extract showed the best activity in DPPH (63.2 ± 2.3 μg mL-1) and H2O2  (55.6 ± 2.6 μg mL-1) models compared to the other extracts. However, flower extract exhibited the highest Fe2+ chelating activity (131.4 ± 4.4 μg mL-1). The extracts exhibited good antioxidant activity in linoleic acid peroxidation and reducing power assays, but were not comparable to vitamin C. The stem (23.58 ± 0.7 μg mL-1) and leaf (26.21 ± 1 μg mL-1) extracts showed highest level of antihemolytic activity than the flower extract

A computational model for analysis of uncoupled NO synthase on nitric oxide and superoxide interaction in microcirculation

Nitric oxide (NO) produced by endothelial cells is a key component for blood-vessel dilation. Dilation is achieved through smooth muscle relaxation as a response to NO transport. Inhibition of this process occurs through the inactivation of NO by reactive oxygen species, especially superoxide (O2 -). NO and superoxide react quickly, forming peroxynitrite (ONOO-). Both superoxide and peroxynitrite apply oxidative stress on vascular tissue. Experimental studies investigating NO interactions are difficult since these reactions occur rapidly and over small distances. This study presents a computational model to describe the interactions of NO, superoxide, and peroxynitrite across an arteriole/venule pair. Based on principles of mass transport, and using knowledge of chemical concentrations and reaction rates, a mathematical model was developed to generate the concentration profiles for NO, O2 -, and ONOO-. We simulated excessive oxidative stress by uncoupled eNOS and determined its effect on NO concentration profiles throughout the region. Based on our understanding of the interactions involved, we predicted 1) increased oxidative stress in the venule decreases NO levels in regions of both the venule and neighboring arteriole, and 2) the amount of NO reduction will vary depending on the location of O2 - increase. The model demonstrates that different sources of O2 - have varied effects on NO concentration profiles, and excessive oxidative stress in the venule can impact NO levels in the venule as well as the arteriole. The results provide a more complete description of nitric oxide transfer, which is an important step toward understanding vascular complications in many pathological conditions

Oxidative stress, protein glycation and nutrition – interactions relevant to health and disease throughout the lifecycle

Protein glycation has been studied for over a century now and plays an important role in disease pathogenesis throughout the lifecycle. Strongly related to diabetic complications, glycation of Hb has become the gold standard method for diabetes diagnosis and monitoring. It is however attracting attention in normoglycaemia as well lately. Longitudinal studies increasingly suggest a positive relationship between glycation and the risk of chronic diseases in normoglycaemic individuals, but the mechanisms behind this association remain unclear. The interaction between glycation and oxidative stress may be particularly relevant in the normoglycaemic context, as suggested by recent epidemiological and in vitro evidence. In that context nutritional and lifestyle factors with an influence on redox status, such as smoking, fruit and vegetable and antioxidants consumption, may have the capacity to promote or inhibit glycation. However, experimental data from controlled trials are lacking the quality and rigour needed to reach firm conclusions. In the present review, we discuss the importance of glycation for health through the lifecycle and focus on the importance of oxidative stress as a driver for glycation. The importance of nutrition to modulate glycation is discussed, based on the evidence available and recommendations towards higher quality future research are made

7-Keto-Cholesterol and Cholestan-3beta, 5alpha, 6beta-Triol Induce Eryptosis through Distinct Pathways Leading to NADPH Oxidase and Nitric Oxide Synthase Activation

BACKGROUND/AIMS: We showed that patho-physiological concentrations of either 7-keto-cholesterol (7-KC), or cholestane-3beta, 5alpha, 6beta-triol (TRIOL) caused the eryptotic death of human red blood cells (RBC), strictly dependent on the early production of reactive oxygen species (ROS). The goal of the current study was to assess the contribution of the erythrocyte ROS-generating enzymes, NADPH oxidase (RBC-NOX), nitric oxide synthase (RBC-NOS) and xanthine oxido-reductase (XOR) to the oxysterol-dependent eryptosis and pertinent activation pathways. METHODS: Phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, reactive oxygen/nitrogen species (RONS) and nitric oxide formation from 2',7'-dichloro-dihydrofluorescein (DCF-DA) and 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM DA) -dependent fluorescence, respectively; Akt1, phospho-NOS3 Ser1177, and PKCζ from Western blot analysis. The activity of individual 7-KC (7 μM) and TRIOL (2, μM) on ROS-generating enzymes and relevant activation pathways was assayed in the presence of Diphenylene iodonium chloride (DPI), N-nitro-L-arginine methyl ester (L-NAME), allopurinol, NSC23766 and LY294002, inhibitors in this order of RBC-NOX, RBC-NOS, XOR and upstream regulatory proteins Rac GTPase and phosphoinositide3 Kinase (PI3K); hemoglobin oxidation from spectrophotometric analysis. RESULTS: RBC-NOX was the target of 7-KC, through a signaling including Rac GTPase and PKCζ, whereas TRIOL caused activation of RBC-NOS according to the pathway PI3K/Akt, with the concurrent activity of a Rac-GTPase. In concomitance with the TRIOL-induced .NO production, formation of methemoglobin with global loss of heme were observed, ascribable to nitrosative stress. XOR, activated after modification of the redox environment by either RBC-NOX or RBC-NOS activity, concurred to the overall oxidative/nitrosative stress by either oxysterols. When 7-KC and TRIOL were combined, they acted independently and their effect on ROS/RONS production and PS exposure appeared the result of the effects of the oxysterols on RBC-NOX and RBC-NOS. CONCLUSION: Eryptosis of human RBCs may be caused by either 7-KC or TRIOL by oxidative/nitrosative stress through distinct signaling cascades activating RBC-NOX and RBC-NOS, respectively, with the complementary activity of XOR; when combined, the oxysterols act independently and both concur to the final eryptotic effect

Mechanisms linking colorectal cancer to the consumption of (processed) red meat : a review

Colorectal cancer (CRC) is the third most commonly diagnosed cancer in the world. The vast majority of CRC cases have been linked to environmental causes rather than to heritable genetic changes. Over the last decades, epidemiological evidence linking the consumption of red and, more convincingly, of processed red meat to CRC has accumulated. In parallel, hypotheses on carcinogenic mechanisms underlying an association between CRC and the intake of red and processed red meat have been proposed and investigated in biological studies. The hypotheses that have received most attention until now include (1) the presence of polycyclic aromatic hydrocarbons and heterocyclic aromatic amines, two groups of compounds recognized as carcinogenic, (2) the enhancing effect of (nitrosyl)heme on the formation of carcinogenic N-nitroso compounds and lipid peroxidation. However, none of these hypotheses completely explains the link between red and processed red meat intake and the CRC risk. Consequently, scientists have proposed additional mechanisms or refined their hypotheses. This review first briefly summarizes the development of CRC followed by an in-depth overview and critical discussion of the different potential carcinogenic mechanisms underlying the increased CRC risk associated with the consumption of red and processed red meat

Secondary Hematoma Expansion and Perihemorrhagic Edema after Intracerebral Hemorrhage: From Bench Work to Practical Aspects.

Intracerebral hemorrhages (ICH) represent about 10-15% of all strokes per year in the United States alone. Key variables influencing the long-term outcome after ICH are hematoma size and growth. Although death may occur at the time of the hemorrhage, delayed neurologic deterioration frequently occurs with hematoma growth and neuronal injury of the surrounding tissue. Perihematoma edema has also been implicated as a contributing factor for delayed neurologic deterioration after ICH. Cerebral edema results from both blood-brain barrier disruption and local generation of osmotically active substances. Inflammatory cellular mediators, activation of the complement, by-products of coagulation and hemolysis such as thrombin and fibrin, and hemoglobin enter the brain and induce a local and systemic inflammatory reaction. These complex cascades lead to apoptosis or neuronal injury. By identifying the major modulators of cerebral edema after ICH, a therapeutic target to counter degenerative events may be forthcoming