Marker Of Protein Oxidation Research Articles

Urinary Metabolomic Markers of Protein Glycation, Oxidation, and Nitration in Early-Stage Decline in Metabolic, Vascular, and Renal Health.

Glycation, oxidation, nitration, and crosslinking of proteins are implicated in the pathogenic mechanisms of type 2 diabetes, cardiovascular disease, and chronic kidney disease. Related modified amino acids formed by proteolysis are excreted in urine. We quantified urinary levels of these metabolites and branched-chain amino acids (BCAAs) in healthy subjects and assessed changes in early-stage decline in metabolic, vascular, and renal health and explored their diagnostic utility for a noninvasive health screen. We recruited 200 human subjects with early-stage health decline and healthy controls. Urinary amino acid metabolites were determined by stable isotopic dilution analysis liquid chromatography-tandem mass spectrometry. Machine learning was applied to optimise and validate algorithms to discriminate between study groups for potential diagnostic utility. Urinary analyte changes were as follows: impaired metabolic health—increased Nε-carboxymethyl-lysine, glucosepane, glutamic semialdehyde, and pyrraline; impaired vascular health—increased glucosepane; and impaired renal health—increased BCAAs and decreased Nε-(γ-glutamyl)lysine. Algorithms combining subject age, BMI, and BCAAs discriminated between healthy controls and impaired metabolic, vascular, and renal health study groups with accuracy of 84%, 72%, and 90%, respectively. In 2-step analysis, algorithms combining subject age, BMI, and urinary Nε-fructosyl-lysine and valine discriminated between healthy controls and impaired health (any type), accuracy of 78%, and then between types of health impairment with accuracy of 69%-78% (cf. random selection 33%). From likelihood ratios, this provided small, moderate, and conclusive evidence of early-stage cardiovascular, metabolic, and renal disease with diagnostic odds ratios of 6 – 7, 26 – 28, and 34 – 79, respectively. We conclude that measurement of urinary glycated, oxidized, crosslinked, and branched-chain amino acids provides the basis for a noninvasive health screen for early-stage health decline in metabolic, vascular, and renal health.

Sub-chronic exposure to Kalach 360 SL–induced damage in rats’ liver and hematological system

We investigated the effects of sub-chronic exposure to Kalach 360 SL (KL), glyphosate-based herbicide used in Tunisia, on liver and hematological system in different groups of female rats. Group 1 was used as a control, while animals of groups 2 and 3 received orally 0.07 mL and 0.175 mL of KL, respectively (126 and 315 mg of glyphosate/kg), for 60 days. As a result, the KL-exposed groups exhibited hypochromic microcytic anemia, systemic inflammation, cytolysis, decrease in hepatic enzyme activity, and cholestasis. Exposure to different doses of KL could induce erythrocyte destruction (hemolysis) in hematopoietic organs (bones). Moreover, lipid peroxidation contents and protein oxidation markers significantly increased in exposed groups, while enzymatic and non-enzymatic antioxidant activities decreased considerably, in both erythrocytes and liver tissues, compared with those in controls. Liver histological studies confirmed the presence of inflammatory reaction with pathology involving the damage or necrosis of hepatocytes, however, without fibrosis remodulation. Thus, KL sub-chronic exposure caused hepatonecrosis, systemic inflammation, and hemolysis.

Effect of the emulsion of Sacha Inchi (Plukenetia huayabambana) oil on oxidative stress and inflammation in rats induced to obesity

Abstract Sacha Inchi (Plukenetia huayllabambana), an oleoleguminosae with a high content ω-3, could help normalize alterations of obesity. We evaluated the effect of emulsion of Sacha Inchi (SI) oil on the lipid profile, oxidative stress and inflammation in serum and liver tissue of obesity-induced rats. Five groups induced to obesity by high-fat diet were analyzed. One group without treatment, two groups with the emulsion of SI oil (OSI1:0.25 ω-3/day, OSI2:0.5 ω-3/day), one group with atorvastatin and one group with atorvastatin plus the emulsion of SI oil. The treated groups lowered the values of total cholesterol, triglycerides, and LDLc, and increased HDLc. The lipid and protein oxidation marker, IL-6 and leptin decreased in serum, as well as TNF-α hepatic. In contrast, serum adiponectin, antioxidant capacity, and liver catalase activity increased in the OSI2 group. Therefore, the emulsion of SI oil normalizes the lipid profile and decreases oxidative stress and inflammation in an obesity model.

The effect of short-term methionine restriction on glutathione synthetic capacity and antioxidant responses at the whole tissue and mitochondrial level in the rat liver.

Dietary methionine restriction (MR) where methionine is the sole source of sulfur amino acid increases lifespan in diverse species. Methionine restricted rodents experience a decrease in glutathione (GSH), a major antioxidant, in several tissues, which is paradoxical to longevity interventions because tissues with low GSH might experience more oxidative damage. Liver plays a key role in GSH synthesis and here we examined how MR influences GSH metabolism in the liver. We also hypothesised that low GSH might be subsidized by compensatory pathway(s) in the liver. To investigate GSH synthesis and antioxidant responses, Fischer-344 rats were given either a MR diet or a control diet for 8 weeks. Based on γ-glutamylcysteine synthetase activity, GSH synthetic capacity did not respond to low dietary methionine availability. Tissue level protein and lipid oxidation markers do not support elevated oxidative damage, despite low GSH availability. Whole tissue and mitochondrial level responses to MR differed. Specifically, the activity of glutathione reductase and thioredoxin reductase increase in whole liver tissue which might offset the effects of declined GSH availability whereas mitochondrial GSH levels were unperturbed by MR. Moreover, enhanced proton leak in liver mitochondria by MR (4 week) presumably diminishes ROS production. Taken together, we suggest that the effect of low GSH in liver tissue is subsidized, at least in part, by increased antioxidant activity and possibly by enhanced mitochondrial proton leak.

Aphid-Triggered Changes in Oxidative Damage Markers of Nucleic Acids, Proteins, and Lipids in Maize (Zea mays L.) Seedlings

Prior experiments illustrated reactive oxygen species (ROS) overproduction in maize plants infested with bird-cherry-oat (Rhopalosiphum padi L.) aphids. However, there is no available data unveiling the impact of aphids feeding on oxidative damages of crucial macromolecules in maize tissues. Therefore, the purpose of the current study was to evaluate the scale of oxidative damages of genomic DNA, total RNA and mRNA, proteins, and lipids in seedling leaves of two maize genotypes (Złota Karłowa and Waza cvs—susceptible and relatively resistant to the aphids, respectively). The content of oxidized guanosine residues (8-hydroxy-2′-deoxyguanosine; 8-OHdG) in genomic DNA, 8-hydroxyguanosine (8-OHG) in RNA molecules, protein carbonyl groups, total thiols (T-SH), protein-bound thiols (PB-SH), non-protein thiols (NP-SH), malondialdehyde (MDA) and electrolyte leakage (EL) levels in maze plants were determined. In addition, the electrical penetration graphs (EPG) technique was used to monitor and the aphid stylet positioning and feeding modes in the hosts. Maize seedlings were infested with 0 (control), 30 or 60 R. padi adult apterae per plant. Substantial increases in the levels of RNA, protein and lipid oxidation markers in response to aphid herbivory, but no significant oxidative damages of genomic DNA, were found. Alterations in the studied parameters were dependent on maize genotype, insect abundance and infestation time.

Effect of wooden breast condition on quality traits of emulsified chicken patties during frozen storage.

Wooden breast (WB) is a recent abnormality characterized by tougher consistency in raw broiler breast fillets. This myopathy has been associated with negative alterations in chicken meat quality. The aim of this study was to evaluate the influence of WB condition on quality parameters of emulsified chicken patties (ECP) during frozen storage. Three formulations of ECP were processed, namely, PN (100% normal breast), PW (100% WB) and PNW (50%:50% normal:WB). ECP were frozen stored during 90days and assessed by physicochemical and sensory analysis. Variations in redness were less pronounced in PW along the storage. TBARS were higher in PW and PNW until 30days of storage compared to PN samples. Incorporation of WB into ECP formulation resulted in reduced peroxide-value, p-anisidine index and carbonyl content at the end of storage. Despite the different alterations in lipid and protein oxidation markers along storage time, results did not influence sensory acceptability, since no effect of wooden breast condition and storage time in odor and color liking was found in our experiment. This study elucidates for the first time that the use of WB meat for the elaboration of ECP is a feasible strategy to minimize economic losses for the poultry industry.

Protective Effect of Semisynthetic and Natural Flavonoid on Aged Rat Microglia-enriched Cultures.

The ROS-mediated lysosomal dysfunction and coinciding deterioration of mitochondrial function are thought to be the prominent mechanisms responsible for aging. Microglia, the resident macrophages in the central nervous system, were postulated to belong to the major targets vulnerable to these detrimental processes, acting as principal drivers in brain aging. The present study investigated the potential protective effect of the semisynthetic flavonoid 3'-O-(3-chloropivaloyl) quercetin (CPQ) and quercetin (Q) on microglia-enriched mixed brain cultures (MBCs) established from aged Wistar rats. Both flavonoids tested suppressed the development of lipofuscin-related autofluorescence in aged cells. Further ensuing protective effects included reduction of protein oxidation markers in aged cells. Moreover, unlike Q, CPQ significantly suppressed sensitivity of aged cells to stimulation of superoxide burst. Other activation markers, cellular hypertrophy and isolectin B4 binding, were also downregulated by treatment with both CPQ and Q. In conclusion, results of our study suggest that both flavonoids tested may protect microglia with a quite comparable efficacy against aging-related accumulated alterations. The protective mechanism can include interference with the ROS-mediated vicious cycles involving lysosomal dysfunction. Nevertheless, the lipophilized quercetin, CPQ, a compound with proposed enhanced biological availability compared to parent molecule, can represent an agent potentially useful for new effective pharmaceutical intervention against brain aging, overcoming the limitations of clinical applicability of quercetin.

Propolis Extracts Inhibit UV-Induced Photodamage in Human Experimental In Vitro Skin Models.

The aim of this study was to assess the antioxidant, photoprotective, and antiaging effects of Greek propolis. Propolis was subjected to n-heptane or methanol extraction. Total phenolic/flavonoid content and antioxidant potential were determined in the extracts. Promising extracts were evaluated for their cytoprotective properties using human immortalized keratinocyte (HaCaT) or reconstituted human skin tissue following exposure to UVB. Assessment of cytotoxicity, DNA damage, oxidative status, and gene/protein expression levels of various matrix metalloproteinases (MMPs) were performed. The propolis methanolic fractions exhibited higher total phenolic and flavonoid contents and significant in vitro antioxidant activity. Incubation of HaCaT cells with certain methanolic extracts significantly decreased the formation of DNA strand breaks following exposure to UVB and attenuated UVB-induced decrease in cell viability. The extracts had no remarkable effect on the total antioxidant status, but significantly lowered total protein carbonyl content used as a marker for protein oxidation in HaCaT cells. MMP-1, -3, -7, and -9, monitored as endpoints of antiaging efficacy, were significantly reduced by propolis following UVB exposure in a model of reconstituted skin tissue. In conclusion, propolis protects against the oxidative and photodamaging effects of UVB and could be further explored as a promising agent for developing natural antiaging strategies.

Alterations of Antioxidant Enzymes and Biomarkers of Nitro-oxidative Stress in Tissues of Bladder Cancer.

Bladder cancer (BC) is one of the most common tumors found in the urinary bladder for both male and female in western countries. In vitro and in vivo studies suggest that high levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS) and oxidative stress play a crucial role in human cancer. Low concentration of ROS and RNS is indispensable for cell survival and proliferation. However, high concentration of ROS and RNS can exert a cytotoxic effect. Increased oxidative stress is a result of either increased ROS/RNS production or a decrease of antioxidant defense mechanisms. A literature search was carried out on PubMed, Medline, and Google Scholar for articles in English published up to May 2018 using the following keywords: oxidative stress, antioxidants, reactive oxygen species, lipid peroxidation, paraoxonase, urinary bladder cancer, and nitric oxide. Literature data demonstrate that BC is associated with oxidative stress and with an imbalance between oxidants and antioxidant enzymes. Markers of lipid peroxidation, protein and nucleic acid oxidation are significantly higher in tissues of patients with BC compared with control groups. A decrease of activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione, and paraoxonase) has also been demonstrated. The imbalance between oxidants and antioxidants could have a potential role in the etiology and progression of bladder cancer.

STUDY OF RELATIONSHIP BETWEEN THE PROTEIN OXIDATION MARKERS AND ADIPOKINES IN OBESE TYPE 2 DIABETIC PATIENTS

Objective: This study was done to explore the correlation if any, between obesity markers adiponectin, leptin, and protein oxidative stress (OS) status in obese with and without type 2 diabetic mellitus (T2DM) patients.
 Methods: In the present study, 30 healthy subjects, 30 obese non-diabetics, and 30 obese T2DM patients were enrolled. Protein OS parameters such as advanced oxidation of protein products (AOPPs) and protein carbonyl (PC) were estimated. Serum leptin, adiponectin, and insulin levels were measured by ELISA.
 Results: The AOPP, PC, leptin, leptin adiponectin ratio (LAR), insulin, and homeostasis model assessment of insulin resistance (HOMA-IR) levels were significantly higher in obese non-diabetic and obese T2DM as compared to healthy control (p<0.001). However, serum adiponectin levels were significantly lower in obese non-diabetic and obese T2DM as compared to control (p<0.001). HOMA-IR and LAR both the index of IR were increased in obese non-diabetic and obese T2DM. Positive correlations were observed for AOPP with body mass index, PC in obese non-diabetic and with fasting blood glucose, postprandial blood glucose, HOMA-IR, and PC in obese T2DM. A negative correlation was found between PC and adiponectin in obese non-diabetic and obese T2DM. A significant inverse correlation was obtained between leptin and adiponectin in obese T2DM only.
 Conclusion: The present study suggested that increased protein OS, hyperinsulinemia, and hyperleptinemia may lead to hypoadiponectinemia in obese with and without T2DM. Moreover, determination of protein oxidation markers can be useful for monitoring the dysregulation of adipokines and glucose metabolism in obesity and T2DM.

Immune Challenge During Reproduction Results in Tissue Specific and Intensity‐Dependent Responses for Mitochondrial Stress

Life‐history theory suggests that tradeoffs can occur between self‐maintance and an animal's capacity to allocate resources to reproduction. This effect is thought to be exacerbated when a female is faced with an immune challenge during a reproductive event. Ultimately, such tradeoffs are thought to curtail lifespan, but the bioenergetic underpinnings of this effect are not well understood. We determined the extent of any effects in mitochondrial function and oxidative damage in female mice when exposed to an immune challenge (keyhole limpet haemocyanin) during reproduction. Female ICR mice were paired with a male until just before giving birth and randomly assigned to one of three groups. Female mice in the control group (LC) were injected with saline 2 days after parturition, and then sacrificed 1 week after weaning. In the lactating immune challenge group (LI), females were treated similarly except they were injected with KLH. Finally, in the pregnant‐lactating immune challenge group (PLI), females were treated the same as LI group, except the male was left in the box with the female until 3 days post‐partum, so the female became pregnant again while lactating. The second litter was born within 3 days of weaning, immediately removed, and females were sacrificed 1 week later. Mitochondrial function and reactive oxygen species (ROS) emission were determined in the liver and hindlimb skeletal muscle. Antioxidant protein levels and oxidative damage markers were also analyzed in these tissues. A one‐way ANOVA was performed on all measures, with a Tukey's post hoc test to determine group differences. No significant differences were observed in liver or hindlimb skeletal muscle mitochondrial coupling as measured by the respiratory control ratio (RCR) for both complex I and complex II (p>0.05). Isolated mitochondrial ROS emission in the hindlimb skeletal muscle was significantly lower in LI compared to LC (p=0.041), but no differences were observed in the liver. Additionally, skeletal muscle PGC1‐α was significantly reduced in LI mice compared to LC (p=0.01). In skeletal muscle the antioxidant GPX was significantly increased in the PLI animals compared to LI animals (p=0.022). Lastly, liver SOD1 was significantly higher in LC compared to PLI (p=0.039). The opposite response was observed in HSP70, in which PLI had significantly increased expression compared to LC (p=0.018). In conclusion, females that experienced an immune challenge during reproduction did not appear to experience a decline in mitochondrial respiratory performance but a drop in PGC1‐a in skeletal muscle that could contribute to a reduction in future performance associated with a reduction in mitochondrial density.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Malondialdehyde and advanced oxidation protein products are not increased in psoriasis: a controlled study

This study investigated oxidative stress in patients with psoriasis of low and medium disease activity. We measured advanced oxidation protein products (AOPP) and malondialdehyde (MDA) in plasma using UV-spectrophotometry and high performance liquid chromatography connected to a fluorescence detector in 84 patients and 84 matched healthy subjects. AOPP is a marker of protein oxidation due to inflammation, whereas MDA is a hydroxyl radical initiated lipid peroxidation product. Clinico-demographic variables including age, gender, disease severity, and fatigue were assessed in relation to AOPP and MDA. Disease severity was evaluated with the Psoriasis Area and Severity Index and the Dermatology Life Quality Index. Median (interquartile range, IQR) AOPP concentrations were 66µmol/l (IQR 54-102) in patients and 69µmol/l (IQR 55-87) in healthy subjects (P = 0.75). Median plasma MDA concentrations were significantly lower in patients than in healthy subjects (0.68µM, IQR 0.54-0.85 vs. 0.76µM, IQR 0.60-0.97; P = 0.03). Plasma levels of AOPP and MDA did not indicate oxidative stress in patients with mild psoriasis. Higher AOPP concentrations were associated with male gender, high body mass index, and high hemoglobin values. Elevated MDA concentrations were associated with advanced age and male gender. No associations with disease severity were detected. Although, the two selected biomarkers do not provide a complete measure of oxidative damage, our study demonstrates that a number of physiological and methodological factors influence the levels of MDA and AOPP. Such methodological issues are important to consider when interpreting results using these biomarkers in patients with psoriasis.

The utility of biomarkers in diagnosis of aspirin exacerbated respiratory disease

BackgroundAspirin-exacerbated respiratory disease (AERD) is a distinct eosinophilic phenotype of severe asthma with accompanying chronic rhinosinusitis, nasal polyposis, and hypersensitivity to aspirin. Urinary 3-bromotyrosine (uBrTyr) is a noninvasive marker of eosinophil-catalyzed protein oxidation. The lack of in vitro diagnostic test makes the diagnosis of AERD difficult. We aimed to determine uBrTyr levels in patients with AERD (n = 240) and aspirin-tolerant asthma (ATA) (n = 226) and to assess whether its addition to urinary leukotriene E4 (uLTE4) levels and blood eosinophilia can improve the prediction of AERD diagnosis.MethodsClinical data, spirometry and blood eosinophilis were evaluated. UBrTyr and uLTE4 levels were measured in urine by HPLC and ELISA, respectively.ResultsBoth groups of asthmatics (AERD, n = 240; ATA, n = 226) had significantly higher uBrTyr, uLTE4 levels, and blood eosinophils than healthy controls (HC) (n = 71) (p < 0.05). ULTE4 levels and blood eosinophils were significantly higher in AERD as compared to ATA (p = 0.004, p < 0.0001, respectively). whereas uBrTyr levels were not significantly different between both asthma phenotypes (p = 0.34). Asthmatics with high levels of uBrTyr (> 0.101 ng/mg Cr), uLTE4 levels (> 800 pg/mg Cr) and blood eosinophils (> 300 cells/ul) were 7 times more likely to have AERD.. However, uBrTyr did not increase the benefit for predicting AERD when uLTE4 and blood eosinophils were already taken into account (p = 0.57).ConclusionUBrTyr levels are elevated both in AERD and ATA as compared to HC, but they could not differentiate between these asthma phenotypes suggesting a similar eosinophilic activation. The addition of uBrTyr to elevated uLTE4 levels and blood eosinophils did not statistically enhance the prediction of AERD diagnosis.

English

Colorectal cancer (CRC) is the third leading cause of cancer-related death. Colorectal mucosa has high levels of oxidative stress (OS) landmarks. OS has deleterious effect on cell structures as lipids, proteins and DNA. Markers of protein oxidation are named as advanced oxidation protein products (AOPPs) which have been considered as novel disease biomarkers. OS may also induce DNA damage which leads to mutation of chromosomes. Paraoxonase 1 (PON1) and Arylesterase (ARE) metabolize different substrates and act as antioxidant enzymes. Disruption of epigenetic processes can lead to altered gene function and malignant transformation. Altered expression of genes that encode histone deacetylase (HDACs) are linked to tumor development. The aim of this study was to evaluate the PON1 and ARE activities in CRC patients and to assess the epigenetic fingerprint via estimation of HDAC. The study was carried out on 30 CRC patients. After excision of tumor, segments were divided into two groups: group A (Control group) included parts taken from safety margin; Group B: (CRC group) subdivided into 3 grades, grade I: well differentiated tumor, grade II: moderately differentiated tumor, and grade III: poor differentiated tumor. CRC groups showed elevation of OS landmarks (AOPPs and DNA damage), HDACs activity and significant decrease in the level of PON1 and activities of PON1 and ARE. The histopathological results were correlated with the laboratory results. It was concluded that the damage of AOPPs and DNA is novel biomarkers in diagnosis of CRC; PON1 is important protective antioxidant enzyme and HDAC activity estimation is a promising line in CRC diagnosis. &nbsp; Key words: Colorectal cancer, epigenetics, oxidative stress, advanced oxidation protein products, paraoxonase 1, arylesterase, histone deacetylase enzyme.

Markers of Oxidative Stress in the Exhaled Breath Condensate of Workers Handling Nanocomposites.

Researchers in nanocomposite processing may inhale a variety of chemical agents, including nanoparticles. This study investigated airway oxidative stress status in the exhaled breath condensate (EBC). Nineteen employees (42.4 ± 11.4 y/o), working in nanocomposites research for 18.0 ± 10.3 years were examined pre-shift and post-shift on a random workday, together with nineteen controls (45.5 ± 11.7 y/o). Panels of oxidative stress biomarkers derived from lipids, nucleic acids, and proteins were analyzed in the EBC. Aerosol exposures were monitored during three major nanoparticle generation operations: smelting and welding (workshop 1) and nanocomposite machining (workshop 2) using a suite of real-time and integrated instruments. Mass concentrations during these operations were 0.120, 1.840, and 0.804 mg/m3, respectively. Median particle number concentrations were 4.8 × 104, 1.3 × 105, and 5.4 × 105 particles/cm3, respectively. Nanoparticles accounted for 95, 40, and 61%, respectively, with prevailing Fe and Mn. All markers of nucleic acid and protein oxidation, malondialdehyde, and aldehydes C6–C13 were elevated, already in the pre-shift samples relative to controls in both workshops. Significant post-shift elevations were documented in lipid oxidation markers. Significant associations were found between working in nanocomposite synthesis and EBC biomarkers. More research is needed to understand the contribution of nanoparticles from nanocomposite processing in inducing oxidative stress, relative to other co-exposures generated during welding, smelting, and secondary oxidation processes, in these workshops.

Evidence supporting oxidative stress in a moderately affected area of the brain in Alzheimer\u2019s disease

The pathogenesis of Alzheimer’s disease (AD) remains to be elucidated. Oxidative damage and excessive beta-amyloid oligomers are components of disease progression but it is unclear how these factors are temporally related. At post mortem, the superior temporal gyrus (STG) of AD cases contains plaques, but displays few tangles and only moderate neuronal loss. The STG at post mortem may represent a brain region that is in the early stages of AD or alternately a region resistant to AD pathogenesis. We evaluated expression profiles and activity of endogenous anti-oxidants, oxidative damage and caspase activity in the STG of apolipoprotein ε4-matched human AD cases and controls. Total superoxide dismutase (SOD) activity was increased, whereas total glutathione peroxidase (GPX), catalase (CAT) and peroxiredoxin (Prx) activities, were decreased in the AD-STG, suggesting that hydrogen peroxide accumulates in this brain region. Transcripts of the transcription factor NFE2L2 and inducible HMOX1, were also increased in the AD-STG, and this corresponded to increased Nuclear factor erythroid 2-related factor (NRF-2) and total heme-oxygenase (HO) activity. The protein oxidation marker 4-hydroxynonenal (4-HNE), remained unchanged in the AD-STG. Similarly, caspase activity was unaltered, suggesting that subtle redox imbalances in early to moderate stages of AD do not impact STG viability.

Impact of Long-Term RF-EMF on Oxidative Stress and Neuroinflammation in Aging Brains of C57BL/6 Mice.

The expansion of mobile phone use has raised questions regarding the possible biological effects of radiofrequency electromagnetic field (RF-EMF) exposure on oxidative stress and brain inflammation. Despite accumulative exposure of humans to radiofrequency electromagnetic fields (RF-EMFs) from mobile phones, their long-term effects on oxidative stress and neuroinflammation in the aging brain have not been studied. In the present study, middle-aged C57BL/6 mice (aged 14 months) were exposed to 1950 MHz electromagnetic fields for 8 months (specific absorption rate (SAR) 5 W/kg, 2 h/day, 5 d/week). Compared with those in the young group, levels of protein (3-nitro-tyrosine) and lipid (4-hydroxy-2-nonenal) oxidative damage markers were significantly increased in the brains of aged mice. In addition, levels of markers for DNA damage (8-hydroxy-2′-deoxyguanosine, p53, p21, γH2AX, and Bax), apoptosis (cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase 1 (PARP-1)), astrocyte (GFAP), and microglia (Iba-1) were significantly elevated in the brains of aged mice. However, long-term RF-EMF exposure did not change the levels of oxidative stress, DNA damage, apoptosis, astrocyte, or microglia markers in the aged mouse brains. Moreover, long-term RF-EMF exposure did not alter locomotor activity in aged mice. Therefore, these findings indicate that long-term exposure to RF-EMF did not influence age-induced oxidative stress or neuroinflammation in C57BL/6 mice.

Abstract LB-178: Oxidation markers in relation to prostate cancer and BPH and progression of prostate cancer

Abstract Prostate cancer (PCa) is the leading cancer affecting men of all races in the U.S. Benign prostatic hyperplasia (BPH) is prostate enlargement and is not malignant. BPH and PCa have similar symptoms and both have elevated PSA levels. A biomarker which can distinguish PCa from BPH will help the diagnosis of PCa and avoid unnecessary biopsies. Further, many PCa patients after prostatectomy can have recurrence. Currently, PSA is the only monitoring marker for recurrence; however, the underlying mechanism for recurrence is unknown. A mechanistic marker that can predict the progression and recurrence this will allow us to design prevention strategies. Many animal studies have linked oxidative stress with PCa progression and recurrence. The aim of this study is to compare the levels of several oxidation markers between PCa and BPH, and compare the levels of oxidation marker in different stages of PCa and determine their associations with PSA velocities after prostatectomy. We have measured multiple oxidation markers including lipid, protein, DNA, carbohydrate and global oxidation markers. We have collected blood samples among 15 BPH and 22 prostate cancer patients who have prostatectomy. We have found that levels of advanced glycation end products (AGEs), a marker of glucose oxidation, formed under nonenzymatic glycation of lipids and proteins, are higher in PCa patients than BPH. However, among PCa patients, AGEs levels are lower among those with Gleason score higher than 7 than those with Gleason score less than 7. Finally, we have found F2-isoprostanes, a well-established lipid oxidation marker, are associated with PSA velocities among PCa patients who had prostatectomy. Our results suggest that AGEs can help distinguish BPH and PCa but are associated with lower grade PCa. Measurement of F2-isoprostanes, a marker of lipid oxidation can predict PCa progression among men with prostatectomy. Citation Format: Tianying Wu, Bruce Bracken, Susan Kasper. Oxidation markers in relation to prostate cancer and BPH and progression of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-178.

Coenzyme Q10 or Creatine Counteract Pravastatin-Induced Liver Redox Changes in Hypercholesterolemic Mice.

Statins are the preferred therapy to treat hypercholesterolemia. Their main action consists of inhibiting the cholesterol biosynthesis pathway. Previous studies report mitochondrial oxidative stress and membrane permeability transition (MPT) of several experimental models submitted to diverse statins treatments. The aim of the present study was to investigate whether chronic treatment with the hydrophilic pravastatin induces hepatotoxicity in LDL receptor knockout mice (LDLr-/-), a model for human familial hypercholesterolemia. We evaluated respiration and reactive oxygen production rates, cyclosporine-A sensitive mitochondrial calcium release, antioxidant enzyme activities in liver mitochondria or homogenates obtained from LDLr-/- mice treated with pravastatin for 3 months. We observed that pravastatin induced higher H2O2 production rate (40%), decreased activity of aconitase (28%), a superoxide-sensitive Krebs cycle enzyme, and increased susceptibility to Ca2+-induced MPT (32%) in liver mitochondria. Among several antioxidant enzymes, only glucose-6-phosphate dehydrogenase (G6PD) activity was increased (44%) in the liver of treated mice. Reduced glutathione content and reduced to oxidized glutathione ratio were increased in livers of pravastatin treated mice (1.5- and 2-fold, respectively). The presence of oxidized lipid species were detected in pravastatin group but protein oxidation markers (carbonyl and SH- groups) were not altered. Diet supplementation with the antioxidants CoQ10 or creatine fully reversed all pravastatin effects (reduced H2O2 generation, susceptibility to MPT and normalized aconitase and G6PD activity). Taken together, these results suggest that 1- pravastatin induces liver mitochondrial redox imbalance that may explain the hepatic side effects reported in a small number of patients, and 2- the co-treatment with safe antioxidants neutralize these side effects.

Bevacizumab Diminishes Inflammation in an Acute Endotoxin-Induced Uveitis Model.

Introduction: Uveitis is an eye disease characterized by inflammation of the uvea and an early and exhaustive diagnosis is essential for its treatment. The aim of our study is to assess the potential toxicity and anti-inflammatory efficacy of Bevacizumab in an experimental uveitis model by subcutaneously injecting lipopolysaccharide into Lewis rats and to clarify its mechanism.Material and Methods: Blood–aqueous barrier integrity was assessed 24 h after endotoxin-induced uveitis (EIU) by analyzing two parameters: cell count and protein concentration in aqueous humors. Histopathology of all eye structures was also studied. Enzyme-linked immunosorbent analyses of the aqueous humor samples were performed in order to calculate the diverse chemokine and cytokine protein levels and oxidative stress-related markers were also evaluated.Results: The aqueous humor’s cellular content significantly increased in the group treated with only Bevacizumab, but it had no effect on retina histopathological grading. Nevertheless, the inflammation noted in ocular structures when administering Bevacizumab with endotoxin was mostly prevented since aqueous humor cell content considerably lowered, and concomitantly with a sharp drop in uveal, vitreous, and retina histopathological grading. The values of the multi-faceted cytokine IL-2 also significantly decreased (p < 0.05 vs. endotoxin group), and the protective IL-6 and IL-10 cytokines values rose with related anti-oxidant system recovery (p < 0.05 vs. endotoxin group). Concurrently, some related M1 macrophage chemokines substantially increased, e.g., GRO/KC, a chemokine that also displays any kind of protective role.Conclusion: All these results revealed that 24 h after being administered, Bevacizumab treatment in EIU significantly prevented inflammation in various eye structures and correct results in efficacy vs. toxicity balance were obtained.