Ongoing Oxidative Stress Research Articles

Lung alveolar tissue destruction and protein citrullination in diesel exhaust-exposed mouse lungs.

Humanity faces an increasing impact of air pollution worldwide, including threats to human health. Air pollutants prompt and promote chronic inflammation, tumourigenesis, autoimmune and other destructive processes in the human body. Post-translational modification of proteins, for example citrullination, results from damaging attacks of pollutants, including smoking, air pollution and others, rendering host tissues immunogenic. Citrullinated proteins and citrullinating enzymes, deiminases, are more prevalent in patients with COPD and correlate with ongoing inflammation and oxidative stress. In this study, we installed an in-house-designed diesel exhaust delivery and cannabidiol vaporization system where mice were exposed to relevant, urban traffic-related levels of diesel exhaust for 14days and assessed integrity of alveolar tissue, gene expression shifts and changes in protein content in the lungs and other tissues of exposed mice. Systemic presence of modified proteins was also tested. The protective effect of phytocannabinoids was investigated as well. Data obtained in our study show subacute effects of diesel exhaust on mouse lung integrity and protein content. Emphysematous changes are documented in exposed mouse lungs. In parallel, increased levels of citrulline were detected in the alveolar lung tissue and peripheral blood of exposed mice. Pre-treatment with vaporized cannabidiol ameliorated some damaging effects. Results reported hereby provide new insights into subacute lung tissue changes that follow diesel exhaust exposure and suggest possible dietary and/or other therapeutic interventions for maintaining lung health and healthy ageing.

Low-Flow Domiciliary Oxygen as a Mechanism of Ongoing Oxidative Stress.

Oxidative stress occurs when imbalances exist between the production of oxygen free radicals and endogenous antioxidants that neutralize their harmful effects, causing irreversible tissue damage. Oxygen free radicals readily interact with DNA, proteins, and lipids, instigating conformational changes to cellular structures and leading to derangement and dysfunction. Oxidative stress is a key feature in the pathology of COPD. As disease progression occurs, supplemental oxygen is often warranted to ameliorate dyspnea. It has been established that supplemental oxygen at > 0.60 FIO2 is an instigator of oxidative stress. We sought to determine whether chronic exposure to low-flow domiciliary oxygen served as a mechanism of ongoing oxidative stress in this patient population. This study serves to inform best practices for low-flow domiciliary oxygen therapy. We utilized prospective data collection for this study. The exhaled-breath condensate (EBC) of 52 subjects with COPD (FEV1 < 70% of predicted normal for age and gender, height, weight, and smoking history) was evaluated for isofuran (IsoF), an oxidative stress biomarker synthesized in response to elevated tissue oxygen tension. The active control group (n = 26) was compared to the active treatment group receiving low-flow domiciliary oxygen for ≥ 6 h/d (n = 26). The active control group generated a mean ± SD EBC IsoF level of 35.81 ± 25.0 pg/mL compared to the active treatment group's level of 51.37 ± 42.2 pg/mL (P = .057). In subjects diagnosed with advanced COPD, chronic exposure to supplemental oxygen therapy at concentrations ≤ 36% did not appear to induce oxidative stress based on EBC IsoF levels. Our findings do not substantiate that chronic exposure to supplemental oxygen at concentrations < 36% instigates oxidative stress, and, therefore, changes to current domiciliary oxygen therapy prescription practices are not warranted.

Faktor Risiko Kerusakan Sel Beta Pankreas Melalui Deteksi Antibodi Gad (Glutamic Acid Decarboxylase) 65 pada Perempuan di Stikes Hang Tuah Surabaya

Pancreatic beta cell damage is irreversible due to ongoing oxidative stress on the formation of ROS underlying pathophysiology of the phase-onset diabetes mellitus. Prevention of damage to pancreatic beta cells in diabetes mellitus onset phase through the detection of GAD 65 as an autoantigen imonogenik diabetes mellitus in human pancreas reacts with GADA can be done early. The purpose of this study analyzed the age, physical activity, cholesterol, body mass index, and waist circumference as a risk factor in the positive expression of GAD 65 women's groups in serum STIKES Hang Tuah Surabaya. This study was cross sectional study. Samples are some women in STIKES Hang Tuah Surabaya as much 40 respondents who meet the inclusion and exclusion criteria were selected through probability sampling techniques. The result of the relationship with the independent variable dependent variable using Chi Square showed that age (p = 0.395), physical activity (p = 0.572), BMI (p = 0.388), and waist circumference (p = 0.411) so it is not a risk factor for damage pancreatic beta cells. While cholesterol (p = 0.035) so it is a risk factor for pancreatic beta cell destruction. The conclusion of this study was in women aged 19-37 years with risk factors that can be used as a prediction of pancreatic beta cell destruction via antibody detection GAD 65 is high cholesterol.Keywords: Pancreatic Beta Cell Destruction, GAD 65, Women

Persistent Oxidative Stress in Vestibular Schwannomas After Stereotactic Radiation Therapy.

Stereotactic radiation therapy is increasingly used to treat vestibular schwannomas (VSs) primarily and to treat tumor remnants following microsurgery. Little data are available regarding the effects of radiation on VS cells. Tyrosine nitrosylation is a marker of oxidative stress following radiation in malignant tumors. It is not known how long irradiated tissue remains under oxidative stress, and if such modifications occur in benign neoplasms such as VSs treated with significantly lower doses of radiation. We immunostained sections from previously radiated VSs with an antibody that recognizes nitrosylated tyrosine residues to assess for ongoing oxidative stress. Immunohistochemical analysis. Four VSs, which recurred after excision, were treated with stereotactic radiation therapy. Ultimately each tumor required salvage reresection for regrowth. Histologic sections of each tumor before and after radiation were immunolabeled with a monoclonal antibody specific to nitrotyrosine and compared. Two VSs that underwent reresection of a growing tumor remnant without previous radiation therapy served as additional controls. Irradiated tumors enlarged in volume by 3.16 to 8.62 mL following radiation. Preradiation sections demonstrated little to no nitrotyrosine immunostaining. Three of four of irradiated VSs demonstrated increased nitrotyrosine immunostaining in the postradiation sections compared with preradiation tumor sections. Nonirradiated VSs did not label with the antinitrotyrosine antibody. VSs exhibit oxidative stress up to 7 years after radiotherapy, yet these VSs continued to enlarge. Thus, VSs that grow following radiation appear to possess mechanisms for cell survival and proliferation despite radiation-induced oxidative stress.

Elevated circulating nitric oxide levels correlates with enhanced oxidative stress in patients with hyperemesis gravidarum

Since the biochemical and molecular mechanisms responsible for ongoing oxidative stress in hyperemesis gravidarum (HEG) patients have not yet been fully elucidated, the aim of this study was to evaluate the possible role of nitric oxide (NO), malondialdehyde (MDA) and other oxidative stress markers in the disease pathophysiology. Moreover, the relation between oxidative stress markers and Helicobacter pylori (H. pylori) infection was also investigated. Women with pregnancies complicated by HEG (n = 33) were compared with pregnant women without HEG (n = 30) and with healthy non-pregnant women (n = 31). Serum NO, MDA, total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI) and H. pylori infection status were determined for each subject. Serum NO levels and OSI index were found to be increased (p = .001 and .013, respectively) and TAS levels were decreased (p < .001) in HEG patients compared with both controls regardless of H. pylori infection status. Serum MDA and TOS levels were not different between the study groups. Helicobacter pylori infection rates were similar in each group. The reduced antioxidant activities, as well as the increased OSI and NO levels in HEG patients indicate possible oxidative stress conditions in HEG patients. Moreover, serum NO levels may be used as an adjunctive marker to distinguish HEG patients from other causes of emesis during pregnancy.Impact statementWhat is already known on this subject? Current evidence suggests that oxidative stress is a significant factor responsible for a number of complications during pregnancy.What do the results of this study add? Hyperemesis gravidarum is an oxidative stress condition, as reflected by increased nitric oxide (NO) and decreased total antioxidant status activity, regardless of H. Pylori infection.What are the implications for clinical practice and/or further research? Full disclosure of the association between circulating NO and hyperemesis gravidarum would shed light on underlying biological mechanisms and could help clinical management of similar pregnancy-associated morbidity states.

Association of heart rate variability and baroreceptor sensitivity with biochemical markers in breast cancer patients

Background and Aim: The global burden of breast cancer is on the rise, not sparing countries undergoing rapid urbanization. India is one among the top emerging countries to be affected by this disease. Breast cancer survivors are at increased risk of developing cardiovascular complications due to autonomic dysfunction and oxidative stress. Changes in autonomic balance can be assessed by measuring resting heart rate variability (HRV) and baroreceptor sensitivity (BRS). Serum levels of malondialdehyde and inflammatory markers give an estimate of ongoing oxidative stress. Methods: A study was conducted in two groups: (i) study group consisted of women with breast cancer who have undergone modified radical mastectomy and awaiting radiotherapy and (ii) control group consisted of normal healthy age-matched volunteers. Resting HRV and BRS were measured for participants of both the groups. Estimation of serum oxidative stress markers and inflammatory markers was also done. Results: Decrease in HRV and BRS accompanied by increase in oxidative stress markers and inflammatory markers in circulation was observed in cancer patients when compared to control group. Conclusion: Autonomic dysfunction, high oxidative stress, and decreased BRS were prominent in breast cancer patients, which could expose them to future cardiovascular events.

Oxidative stress and Na,K-ATPase activity differential regulation in brainstem and forebrain of Wistar Audiogenic rats may lead to increased seizure susceptibility

The Wistar Audiogenic Rat (WAR) is a well-characterized seizure-prone, inbred rodent strain that, when acutely stimulated with high-intensity sounds, develops brainstem-dependent tonic-clonic seizures that can evolve to limbic-like, myoclonic (forebrain) seizures when the acoustic stimuli are presented chronically (audiogenic kindling). In order to investigate possible mechanisms underlying WAR susceptibility to seizures, we evaluated Na,K-ATPase activity, Ca-ATPase activity, Mg-ATPase activity, lipid membrane composition and oxidative stress markers in whole forebrain and whole brainstem samples of naïve WAR, as compared to samples from control Wistar rats. We also evaluated the expression levels of α1 and α3 isoforms of Na,K-ATPase in forebrain samples. We observed increased Na,K-ATPase activity in forebrain samples and increased oxidative stress markers (lipid peroxidation, glutathione peroxidase and superoxide dismutase) in brainstem samples of WAR. The Ca-ATPase activity, Mg-ATPase activity, lipid membrane composition and expression levels of α1 and α3 isoforms of Na,K-ATPase were unaltered. In view of previous data showing that the membrane potentials from naïve WAR’s neurons are less negative than that from neurons from Wistar rats, we suggest that Na,K-ATPase increased activity might be involved in a compensatory mechanism necessary to maintain WAR’s brains normal activity. Additionally, ongoing oxidative stress in the brainstem could bring Na,K-ATPase activity back to normal levels, which may explain why WAR’s present increased susceptibility to seizures triggered by high-intensity sound stimulation.

Differential exposure and acute health impacts of inhaled solid-fuel emissions from rudimentary and advanced cookstoves in female CD-1 mice

BackgroundThere is an urgent need to provide access to cleaner end user energy technologies for the nearly 40% of the world's population who currently depend on rudimentary cooking and heating systems. Advanced cookstoves (CS) are designed to cut emissions and solid-fuel consumption, thus reducing adverse human health and environmental impacts. Study premiseWe hypothesized that, compared to a traditional (Tier 0) three-stone (3-S) fire, acute inhalation of solid-fuel emissions from advanced natural-draft (ND; Tier 2) or forced-draft (FD; Tier 3) stoves would reduce exposure biomarkers and lessen pulmonary and innate immune system health effects in exposed mice. ResultsAcross two simulated cooking cycles (duration ~ 3h), emitted particulate mass concentrations were reduced 80% and 62% by FD and ND stoves, respectively, compared to the 3-S fire; with corresponding decreases in particles visible within murine alveolar macrophages. Emitted carbon monoxide was reduced ~ 90% and ~ 60%, respectively. Only 3-S-fire-exposed mice had increased carboxyhemoglobin levels. Emitted volatile organic compounds were FD ≪ 3-S-fire ≤ ND stove; increased expression of genes involved in xenobiotic metabolism (COX-2, NQO1, CYP1a1) was detected only in ND- and 3-S-fire-exposed mice. Diminished macrophage phagocytosis was observed in the ND group. Lung glutathione was significantly depleted across all CS groups, however the FD group had the most severe, ongoing oxidative stress. ConclusionsThese results are consistent with reports associating exposure to solid fuel stove emissions with modulation of the innate immune system and increased susceptibility to infection. Lower respiratory infections continue to be a leading cause of death in low-income economies. Notably, 3-S-fire-exposed mice were the only group to develop acute lung injury, possibly because they inhaled the highest concentrations of hazardous air toxicants (e.g., 1,3-butadiene, toluene, benzene, acrolein) in association with the greatest number of particles, and particles with the highest % organic carbon. However, no Tier 0–3 ranked CS group was without some untoward health effect indicating that access to still cleaner, ideally renewable, energy technologies for cooking and heating is warranted.

The mechanism of cone cell death in Retinitis Pigmentosa.

Retinitis Pigmentosa (RP) is a group of diseases in which one of a large number of mutations causes death of rod photoreceptors. After rods die, cone photoreceptors slowly degenerate in a characteristic pattern. The mechanism of rod cell death varies depending upon the gene that is mutated and the rate that rods degenerate is an important prognostic feature, because cones do not begin to degenerate until almost all rods have been eliminated. Rod cell death causes night blindness, but visual disability and blindness result from cone degeneration and therefore it is critical to determine the mechanisms by which it occurs. The death of rods reduces oxygen consumption resulting in high tissue levels of oxygen in the outer retina. The excess oxygen stimulates superoxide radical production by mismatches in the electron transport chain in mitochondria and by stimulation of NADPH oxidase activity in cytoplasm. The high levels of superoxide radicals overwhelm the antioxidant defense system and generate more reactive species including peroxynitrite which is extremely damaging and difficult to detoxify. This results in progressive oxidative damage in cones which contributes to cone cell death and loss of function because drugs or gene transfer that reduce oxidative stress promote cone survival and maintenance of function. Compared with aqueous humor samples from control patients, those from patients with RP show significant elevation of carbonyl content on proteins indicating oxidative damage and a reduction in the ratio of reduced to oxidized glutathione indicating depletion of a major component of the antioxidant defense system from ongoing oxidative stress. The first step in clinical trials will be to identify doses of therapeutic agents that reverse these biomarkers of disease to assist in design of much longer trials with functional and anatomic endpoints.

The Nutraceutic Silybin Counteracts Excess Lipid Accumulation and Ongoing Oxidative Stress in an In Vitro Model of Non-Alcoholic Fatty Liver Disease Progression.

Non-alcoholic fatty liver disease (NAFLD) is a major cause of liver-related morbidity and mortality. Oxidative stress and release of pro-inflammatory cytokines, such as tumor necrosis factor α (TNFα), are major consequences of hepatic lipid overload, which can contribute to progression of NAFLD to non-alcoholic steatohepatitis (NASH). Also, mitochondria are involved in the NAFLD pathogenesis for their role in hepatic lipid metabolism. Definitive treatments for NAFLD/NASH are lacking so far. Silybin, the extract of the milk thistle seeds, has previously shown beneficial effects in NAFLD. Sequential exposure of hepatocytes to high concentrations of fatty acids (FAs) and TNFα resulted in fat overload and oxidative stress, which mimic in vitro the progression of NAFLD from simple steatosis (SS) to steatohepatitis (SH). The exposure to 50 µM silybin for 24 h reduced fat accumulation in the model of NAFLD progression. The in vitro progression of NAFLD from SS to SH resulted in reduced hepatocyte viability, increased apoptosis and oxidative stress, reduction in lipid droplet size, and up-regulation of IκB kinase β-interacting protein and adipose triglyceride lipase expressions. The direct action of silybin on SS or SH cells and the underlying mechanisms were assessed. Beneficial action of silybin was sustained by changes in expression/activity of peroxisome proliferator-activated receptors and enzymes for FA oxidation. Moreover, silybin counteracted the FA-induced mitochondrial damage by acting on complementary pathways: (i) increased the mitochondrial size and improved the mitochondrial cristae organization; (ii) stimulated mitochondrial FA oxidation; (iii) reduced basal and maximal respiration and ATP production in SH cells; (iv) stimulated ATP production in SS cells; and (v) rescued the FA-induced apoptotic signals and oxidative stress in SH cells. We provide new insights about the direct protective effects of the nutraceutic silybin on hepatocytes mimicking in vitro NAFLD progression.

Effect of Dark Chocolate Extracts on Phorbol 12-Myristate 13-Acetate-Induced Oxidative Burst in Leukocytes Isolated by Normo-Weight and Overweight/Obese Subjects.

Oxidative and inflammatory stress represents a major risk factor for cardiovascular disease (CVD) in overweight and obese subjects. Between the different plant foods, chocolate has been shown to decrease CVD risk due to its antioxidant and anti-inflammatory properties. However, as we recently showed in epidemiological studies, meta-analyses, and human trials, dietary antioxidants resulted more effective in subjects characterized by an ongoing oxidative stress, than in healthy people. Aim of this work was to investigate the effect of different concentrations of chocolate phenolic extract (CPE) on in vitro free radical production, stimulated by phorbol 12-myristate 13-acetate (PMA), in leukocytes extracted from blood of normo-weight and overweight/obese subjects. Neutrophils from overweight/obese group had a significantly higher free radical production compared to the normo-weight group. In neutrophils, the lowest CPE concentration significantly reduced free radical production in overweight/obese group only, and higher CPE concentrations were effective in both groups. In monocytes, the CPE concentration that was significantly effective in reducing free radical production was lower in overweight/obese subjects than in normo-weight subjects. Chocolate polyphenol extracts inhibit oxidative burst in human neutrophils and monocytes with a higher efficiency in subjects characterized by an unphysiological oxidative/inflammatory stress, such as overweight and obese. Results of this study provide further evidence about a differential role of dietary antioxidant strictly related to the “stress” condition of the subjects.

Voluntary wheel running differentially affects disease outcomes in male and female mice with experimental autoimmune encephalomyelitis

Multiple sclerosis (MS) is an inflammatory neurodegenerative disease of the central nervous system. The primary symptoms of MS include the loss of sensory and motor function. Exercise has been shown to modulate disease parameters in experimental autoimmune encephalomyelitis (EAE), a mouse model of MS, by reducing immune cell infiltration and oxidative stress. However, these initial studies were carried out exclusively in female mice. The present study compared the effects of daily voluntary wheel running on several disease parameters in male and female mice with EAE.Male and female mice were given access to a running wheel for 1h a day for 30 consecutive days. Daily wheel running significantly improved clinical scores in males with EAE but had little effect on clinical signs in females with the disease. Direct comparison of inflammation, axonal injury, and oxidative stress in male and female mice with EAE revealed significant differences in the amount of T-cell infiltration, microglia reactivity, demyelination and axon integrity. Male mice with EAE given daily access to running wheels also had significantly less ongoing oxidative stress compared to all other groups. Taken together, our results indicate that the inflammatory response generated in EAE is distinct between the sexes and its modulation by daily exercise can have sex-specific effects on disease-related outcomes.

Changes in the Hippocampal Proteome Associated with Spatial Memory Impairment after Exposure to Low (20 cGy) Doses of 1 GeV/n 56Fe Radiation.

Exposure to low (∼20 cGy) doses of high-energy charged (HZE) particles, such as 1 GeV/n 56Fe, results in impaired hippocampal-dependent learning and memory (e.g., novel object recognition and spatial memory) in rodents. While these findings raise the possibility that astronauts on deep-space missions may develop cognitive deficits, not all rats develop HZE-induced cognitive impairments, even after exposure to high (200 cGy) HZE doses. The reasons for this differential sensitivity in some animals that develop HZE-induced cognitive failure remain speculative. We employed a robust quantitative mass spectrometry-based workflow, which links early-stage discovery to next-stage quantitative verification, to identify differentially active proteins/pathways in rats that developed spatial memory impairment at three months after exposure to 20 cGy of 1 GeV/n 56Fe (20/impaired), and in those rats that managed to maintain normal cognitive performance (20/functional). Quantitative data were obtained on 665-828 hippocampal proteins in the various cohorts of rats studied, of which 580 were expressed in all groups. A total of 107 proteins were upregulated in the irradiated rats irrespective of their spatial memory performance status, which included proteins involved in oxidative damage response, calcium transport and signaling. Thirty percent (37/107) of these "radiation biomarkers" formed a functional interactome of the proteasome and the COP9 signalosome. These data suggest that there is persistent oxidative stress, ongoing autophagy and altered synaptic plasticity in the irradiated hippocampus, irrespective of the spatial memory performance status, suggesting that the ultimate phenotype may be determined by how well the hippocampal neurons compensate to the ongoing oxidative stress and associated side effects. There were 67 proteins with expression that correlated with impaired spatial memory performance. Several of the "impaired biomarkers" have been implicated in poor spatial memory performance, neurodegeneration, neuronal loss or neuronal susceptibility to apoptosis, or neuronal synaptic or structural plasticity. Therefore, in addition to the baseline oxidative stress and altered adenosine metabolism observed in all irradiated rats, the 20/impaired rats expressed proteins that led to poor spatial memory performance, enhanced neuronal loss and apoptosis, changes in synaptic plasticity and dendritic remodeling. A total of 46 proteins, which were differentially upregulated in the sham-irradiated and 20/functional rat cohorts, can thus be considered as markers of good spatial memory, while another 95 proteins are associated with the maintenance of good spatial memory in the 20/functional rats. The loss or downregulation of these "good spatial memory" proteins would most likely exacerbate the situation in the 20/impaired rats, having a major impact on their neurocognitive status, given that many of those proteins play an important role in neuronal homeostasis and function. Our large-scale comprehensive proteomic analysis has provided some insight into the processes that are altered after exposure, and the collective data suggests that there are multiple problems with the functionality of the neurons and astrocytes in the irradiated hippocampi, which appear to be further exacerbated in the rats that have impaired spatial memory performance or partially compensated for in the rats with good spatial memory.

378 - Overexpression of Glutaredoxin Averts F-Actin Loss in Spines of Primary Cortical Neurons Derived from Mouse Model of Alzheimer's Disease

Oxidative stress is known to be one of the major contributors towards the pathogenesis of Alzheimer’s disease (AD) potentially leading to amyloid plaques, neurofibrillary tangles and loss of dendritic spine morphology. As a marker for ongoing oxidative stress, we observed increased levels of reactive oxygen species (ROS) in primary cortical neurons from APPSwe/PS1ΔE9 (APP/PS1) transgenic mouse model of AD, compared to wild type littermates. Thiol oxidation and protein glutathionylation are important targets during oxidative stress, resulting in perturbation of cellular thiol homeostasis. Glutaredoxins and other thiol disuflide oxidoreductases are important enzymes involved in reversing protein thiol oxidation in the brain. We hypothesized that overexpressing Grx1 (glutaredoxin) may partially reverse the adverse effects of oxidative stress in AD neurons. We overexpressed human glutaredoxin using AAV6 virus and confirmed overexpression of glutaredoxin using biochemical and immunostaining methods. Actin, an important neuronal cytoskeletal filament, is important for maintenance of spine morphology and function. It is also a target of cellular oxidation undergoing gluathionylation, leading to its conversion from the functional fibrillar form (F-actin) to the globular (G-actin) form. Other studies from our laboratory have shown increased actin oxidation in early stages of AD-related pathology leading to synaptic dysfunction using a combination of cellular, in vivo and post-mortem human brain samples. We also observed loss of F-actin in APP/PS1 neurons compared to their wild-type littermates, as indicated by reduced phalloidin staining. Importantly, overexpression of Grx1 reversed this F-actin loss, and preserved the F-actin microarchitecture in spines. Our results suggest that thiol oxidation events, such as protein glutathionylation including that of actin, play an important role in the synaptic structural and functional deficits seen in AD.

Use of Metabolomics to Trend Recovery and Therapy After Injury in Critically Ill Trauma Patients.

Metabolomics is the broad and parallel study of metabolites within an organism and provides a contemporaneous snapshot of physiologic state. Use of metabolomics in the clinical setting may help achieve precision medicine for those who have experienced trauma, where diagnosis and treatment are tailored to the individual patient. To examine whether metabolomics can (1) distinguish healthy volunteers from trauma patients and (2) quantify changes in catabolic metabolites over time after injury. Prospective cohort study with enrollment from September 2014 to May 2015 at an urban, level 1 trauma center. Included in the study were 10 patients with severe blunt trauma admitted within 12 hours of injury with systolic blood pressure less than 90 mm Hg or base deficit greater than 6 mEq/L and 5 healthy volunteers. Plasma samples (n = 35) were obtained on days 1, 3, and 7, and they were analyzed using mass spectrometry. Principal component analyses, multiple linear regression, and paired t tests were used to select biomarkers of interest. A broad-based metabolite profile comparison between trauma patients and healthy volunteers was performed. Specific biomarkers of interest were oxidative catabolites. Trauma patients had a median age of 45 years and a median injury severity score of 43 (interquartile range, 34-50). Healthy fasting volunteers had a median age of 33 years. Compared with healthy volunteers, trauma patients showed oxidative stress on day 1: niacinamide concentrations were a mean (interquartile range) of 0.95 (0.30-1.45) relative units for trauma patients vs 1.06 (0.96-1.09) relative units for healthy volunteers (P = .02), biotin concentrations, 0.43 (0.27-0.58) relative units for trauma patients vs 1.21 (0.93-1.56) relative units for healthy volunteers (P = .049); and choline concentrations, 0.17 (0.09-0.22) relative units for trauma patients vs 0.21 (0.18-0.22) relative units for healthy volunteers (P = .004). Trauma patients showed lower nucleotide synthesis on day 1: adenylosuccinate concentrations were 0.08 (0.04-0.12) relative units for trauma patients vs 0.15 (0.14-0.17) relative units for healthy volunteers (P = .02) and cytidine concentrations were 1.44 (0.95-1.73) relative units for trauma patients vs 1.74 (1.62-1.98) relative units for healthy volunteers (P = .05). From trauma day 1 to day 7, trauma patients showed increasing muscle catabolism: serine levels increased from 42.03 (31.20-54.95) µM to 79.37 (50.29-106.37) µM (P = .002), leucine levels increased from 69.21 (48.36-99.89) µM to 114.16 (92.89-143.52) µM (P = .004), isoleucine levels increased from 20.43 (10.92-27.41) µM to 48.72 (36.28-64.84) µM (P < .001), and valine levels increased from 122.56 (95.63-140.61) µM to 190.52 (136.68-226.07) µM (P = .004). There was an incomplete reversal of oxidative stress. Metabolomics can function as a serial, comprehensive, and potentially personalized tool to characterize metabolism after injury. A targeted metabolomics approach was associated with ongoing oxidative stress, impaired nucleotide synthesis, and initial suppression of protein metabolism followed by increased nitrogen turnover. This technique may provide new therapeutic and nutrition targets in critically injured patients.

Abstract LB-368: Mitochondrial DNA and cancer in the normative aging study

Abstract Background: Mitochondrial DNA (mtDNA) is a potentially sensitive marker of damage by reactive oxygen species (ROS). Two biomarkers in mtDNA, copy number (mtDNAcn) and 8-OHdG, have therefore been proposed as potential markers of oxidative stress and therefore carcinogenesis. Prospective human studies subjects of mtDNAcn have found inconsistent results, but many suggest that the interval between mtDNAcn measurement and cancer diagnosis affects the association. Moreover, prior studies suggest that the interplay of mtDNAcn and 8-OHdG reflect ongoing oxidative stress related to carcinogenesis, suggesting their potential use in tandem as an early detection biomarker of cancer. Our objective was to examine longitudinal associations between mtDNAcn and 8-OHdG measured in whole blood in relation to both cancer incidence and mortality. Methods: We studied 526 Normative Aging Study participants who had blood drawn 1-4 times from 1999 through 2012 and who were cancer free at first blood draw (median 9.7 years between blood draw and cancer diagnosis/censoring). mtDNAcn and 8-OHdG were measured using quantitative real-time PCR. We used Cox proportional hazards regression models to examine associations between mtDNAcn and mitochondrial 8-OHdG measured at the first blood draw only as well as time-dependent associations across a total of 1043 blood draws and time to all-cancer diagnosis. We ran a second set of models stratified by time between blood draw and diagnosis/censoring, and a third set that explored possible interaction between mtDNAcn and 8-OHdG via inclusion of a product term. All models adjusted for age, BMI, white blood cell count, proportion neutrophils, race, education, smoking status, pack-years of smoking, and alcohol consumption. Results: We found no significant associations between pre-diagnostic mtDNAcn and time to cancer diagnosis or death. When measured at the first blood draw only, 8-OHdG was inversely associated with all-cancer (HR: 0.17; 95% CI: 0.03-0.87) and prostate cancer incidence (HR: 0.05; 95% CI: 0.00-0.97). In the time-dependent models, mitochondrial 8-OHdG was inversely associated with time to all-cancer diagnosis (HR: 0.31; 95% CI: 0.10-1.01) and positively associated with time to all-cancer death (HR: 1.83; 95% CI: 1.13-2.98), while mtDNAcn was positively and independently associated with time to all-cancer death (HR: 1.59; 95% CI: 1.17-2.15; pinteraction = 0.96). In the stratum of 4-6 years between blood draw and diagnosis, 8-OHdG was associated with cancer incidence (HR: 0.01; 95% CI: 0-1.00). Modeling 8-OHdG and mtDNAcn together produced no noteworthy changes in results. Similarly, the test for interaction was non-significant across all models. Conclusion: Pre-diagnostic mtDNAcn does not appear to be significantly associated with cancer, however mitochondrial 8-OHdG may be a significant predictor of cancer several years prior to diagnosis. Mitochondiral markers of oxidative stress may also be significant predictors of cancer mortality, possibly related to metastasis and/or treatment. Additional prospective studies in larger cohorts with greater racial/ethnic, gender, and socioeconomic diversity are needed to validate the utility of 8-OHdG as an early detection biomarker of cancer. Future studies of mitochondrial oxidative stress markers measurable in blood and cancer mortality are warranted. Citation Format: Brian T. Joyce, Tao Gao, Yinan Zheng, Lei Liu, Andrea Baccarelli, Joel Schwartz, Lifang Hou. Mitochondrial DNA and cancer in the normative aging study. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-368.

Ongoing Oxidative Stress Causes Subclinical Neuronal Dysfunction in the Recovery Phase of EAE.

Most multiple sclerosis (MS) patients develop over time a secondary progressive disease course, characterized histologically by axonal loss and atrophy. In early phases of the disease, focal inflammatory demyelination leads to functional impairment, but the mechanism of chronic progression in MS is still under debate. Reactive oxygen species generated by invading and resident central nervous system (CNS) macrophages have been implicated in mediating demyelination and axonal damage, but demyelination and neurodegeneration proceed even in the absence of obvious immune cell infiltration, during clinical recovery in chronic MS. Here, we employ intravital NAD(P)H fluorescence lifetime imaging to detect functional NADPH oxidases (NOX1–4, DUOX1, 2) and, thus, to identify the cellular source of oxidative stress in the CNS of mice affected by experimental autoimmune encephalomyelitis (EAE) in the remission phase of the disease. This directly affects neuronal function in vivo, as monitored by cellular calcium levels using intravital FRET–FLIM, providing a possible mechanism of disease progression in MS.

Glycemic variability predicts inflammation in adolescents with type 1 diabetes.

Adolescents with type 1 diabetes (T1D) have increased risk of cardiovascular disease as well as elevations in biomarkers of systemic inflammation, plasma protein oxidation and vascular endothelial injury. It is unclear whether hyperglycemia itself, or variations in blood glucose are predictors of these abnormalities. This study was designed to determine the relationship of inflammatory (C-reactive protein, CRP), oxidative (total anti-oxidative capacity, TAOC) and endothelial injury (soluble intracellular adhesion molecule 1, sICAM1) markers to glycemic control measures from 3 days of continuous glucose monitoring (CGM) and to hemoglobin A1c (HbA1c), and HbA1c×duration area under the curve (A1cDur). Seventeen adolescents (8 F/9M; age, 13.1±1.6 years (mean±SD); duration, 4.8±3.8 years, BMI, 20.3±3.1 kg/m2; A1c, 8.3±1.2%) were studied. Log CRP but was not related to age, duration, body mass index (BMI), HbA1c, or A1cDUR. TAOC increased as logA1cDUR increased (n=13, r=0.61, p=0.028). CRP and sICAM were not related to CGM average glucose but log CRP increased as 3 day glucose standard deviation increased (r=0.66, p=0.006). TAOC increased as glucose standard deviation increased (r=0.63, p=0.028). Increased glucose variability is associated with increased inflammation in adolescents withT1D. Increased TAOC with increasing variability may be an effort to compensate for the ongoing oxidative stress.

Increased oxidative stress and oxidative DNA damage in non-remission schizophrenia patients

Increasing evidence shows that oxidative stress plays a role in the pathophysiology of schizophrenia. But there is not any study which examines the effects of oxidative stress on DNA in schizophrenia patients. Therefore we aimed to assess the oxidative stress levels and oxidative DNA damage in schizophrenia patients with and without symptomatic remission. A total of 64 schizophrenia patients (38 with symptomatic remission and 26 without symptomatic remission) and 80 healthy volunteers were included in the study. 8-hydroxydeoxyguanosine (8-OHdG), total oxidant status (TOS) and total antioxidant status (TAS) were measured in plasma.TOS, oxidative stress index (OSI) and 8-OHdG levels were significantly higher in non-remission schizophrenic (Non-R-Sch) patients than in the controls. TOS and OSI levels were significantly higher in remission schizophrenic (R-Sch) patients than in the controls. TAS level were significantly lower and TOS and OSI levels were significantly higher in R-Sch patients than in Non-R-Sch patients. Despite the ongoing oxidative stress in patients with both R-Sch and Non-R-Sch, oxidative DNA damage was higher in only Non-R-Sch patients compared to controls. It is suggested that oxidative stress can cause the disease via DNA damage, and oxidative stress plays a role in schizophrenia through oxidative DNA damage.

Bronchial epithelial cells: The key effector cells in the pathogenesis of chronic obstructive pulmonary disease?

The primary function of the bronchial epithelium is to act as a defensive barrier aiding the maintenance of normal airway function. Bronchial epithelial cells (BEC) form the interface between the external environment and the internal milieu, making it a major target of inhaled insults. However, BEC can also serve as effectors to initiate and orchestrate immune and inflammatory responses by releasing chemokines and cytokines, which recruit and activate inflammatory cells. They also produce excess reactive oxygen species as a result of an oxidant/antioxidant imbalance that contributes to chronic pulmonary inflammation and lung tissue damage. Accumulated mucus from hyperplastic BEC obstructs the lumen of small airways, whereas impaired cell repair, squamous metaplasia and increased extracellular matrix deposition underlying the epithelium is associated with airway remodelling particularly fibrosis and thickening of the airway wall. These alterations in small airway structure lead to airflow limitation, which is critical in the clinical diagnosis of chronic obstructive pulmonary disease (COPD). In this review, we discuss the abnormal function of BEC within a disturbed immune homeostatic environment consisting of ongoing inflammation, oxidative stress and small airway obstruction. We provide an overview of recent insights into the function of the bronchial epithelium in the pathogenesis of COPD and how this may provide novel therapeutic approaches for a number of chronic lung diseases.