Increase In Protein Carbonyl Content Research Articles

Fish injuries resulting from transient operating conditions in a Brazilian hydropower plant: morphological, physiological and biochemical evaluation in Pimelodus maculatus (Siluriformes: Pimelodidae)

Abstract This study investigated injuries sustained by Pimelodus maculatus caused by transient operating conditions in a Brazilian hydroelectric power plant. Fish (n = 43) that inhabited the tailrace and appeared on the water surface after changes in the operating conditions were evaluated (IF). Additionally, fish (n = 10) captured using longline in the vicinity of the tailrace were used for comparisons (NF). Blood samples were evaluated for cortisol, glucose, and lactate concentrations. Tissue samples (gill, liver, brain, and muscle) were analyzed for oxidative stress biomarkers (TBARS, protein carbonyl content, and catalase activity) and histology (gill, liver, and spleen). The most prominent findings in IF were barotraumas with intra-abdominal hemorrhage and swim bladder rupture. Cortisol and glucose concentrations were lower for IF when compared to NF. IF also suffered from oxidative stress, as indicated by increased TBARS in the liver and brain, as well as increased protein carbonyl content in the gills and liver. The main histological alterations induced by the adverse conditions were circulatory disorders, such as telangiectasia and gill aneurysm, hepatic congested veins, and hemorrhage in the liver and spleen. The abrupt drop of pressure resulting from changes in the operating conditions of the turbines caused barotraumas and oxidative stress, which were the leading causes of fish mortality

The anti-oxidative potential of ginger extract and its constituent on meat protein isolate under induced Fenton oxidation

Ginger extract has been reported to possess antioxidant properties. However, components isolated from ginger have been rarely reported to inhibit oxidation. Herein, the antioxidant properties of ginger and purified components derived from it (6-gingerol, zingerone, rutin, quercetin, and kaempferol) were confirmed by using HPLC and were further used to investigate its effect on lamb meat. Myofibrillar proteins isolated (MPI) from lamb meat were incubated with ginger and its constituents under induced Fenton oxidation (1.0 mmol/L FeCl3, 0.1 mmol/L Asc, and 20 mmol/L H2O2) for 1, 3,5, and 7 h. Incubating meat protein isolate in the absence of ginger extract or its components resulted in a substantial drop in sulfhydryl groups, an increase in protein carbonyl content, and a corresponding increase in TBARS content. However, ginger extract and its constituents demonstrated antioxidant properties, which might be attributed to their hydroxyl groups and suitable solubilizing side chains. Overall, ginger extract exhibited the highest antioxidant capabilities of all treated samples, suggesting that ginger extracts may be used as a natural antioxidant in meat and lipid/protein-containing processed products. Significance of the studyGinger extract is also frequently used as a herbal medicine due to its anti-inflammatory, anti-cancer, and antibacterial qualities. Nonvolatile pungent chemicals found in ginger, such as gingerol, shogaols, paradols, and zingerone, as well as kaempferol, rutin, and other phenolic compounds, have been confirmed in ginger extract and have been shown to have antioxidant action driven by free radical elimination. Despite these findings, ginger extract and its pure constituent components have seldom been shown to have the ability to slow protein and lipid oxidation in meat and meat-related products. The effect of ginger extracts on the oxidative stability of myofibriller protein isolate has never been investigated. Exploiting the phenolic content of ginger extract may result in a discovery that would have a huge influence on both the ginger and meat industries as well as other food processing sectors. The first aim of our study was to confirm the presence of six selected phenolic compounds (rutin, kaempferol, 6-gingerol, zingerone, naringenin, and quercetin) in ginger as reported by literature, and the second objective was to determine the efficacy of ginger extracts and its purified constituents on myofibrillar protein isolate treated under induced Fenton oxidation.

Premature Senescence and Telomere Shortening Induced by Oxidative Stress From Oxalate, Calcium Oxalate Monohydrate, and Urine From Patients With Calcium Oxalate Nephrolithiasis.

Oxidative stress, a well-known cause of stress-induced premature senescence (SIPS), is increased in patients with calcium oxalate (CaOx) kidney stones (KS). Oxalate and calcium oxalate monohydrate (COM) induce oxidative stress in renal tubular cells, but to our knowledge, their effect on SIPS has not yet been examined. Here, we examined whether oxalate, COM, or urine from patients with CaOx KS could induce SIPS and telomere shortening in human kidney (HK)-2 cells, a proximal tubular renal cell line. Urine from age- and sex-matched individuals without stones was used as a control. In sublethal amounts, H2O2, oxalate, COM, and urine from those with KS evoked oxidative stress in HK-2 cells, indicated by increased protein carbonyl content and decreased total antioxidant capacity, but urine from those without stones did not. The proportion of senescent HK-2 cells, as indicated by SA-βgal staining, increased after treatment with H2O2, oxalate, COM, and urine from those with KS. Expression of p16 was higher in HK-2 cells treated with H2O2, oxalate, COM, and urine from those with KS than it was in cells treated with urine from those without stones and untreated controls. p16 was upregulated in the SA-βgal positive cells. Relative telomere length was shorter in HK-2 cells treated with H2O2, oxalate, COM, and urine from those with KS than that in cells treated with urine from those without stones and untreated controls. Transcript expression of shelterin components (TRF1, TRF2 and POT1) was decreased in HK-2 cells treated with H2O2, oxalate, COM, and urine from those with KS, in which case the expression was highest. Urine from those without KS did not significantly alter TRF1, TRF2, and POT1 mRNA expression in HK-2 cells relative to untreated controls. In conclusion, oxalate, COM, and urine from patients with CaOx KS induced SIPS and telomere shortening in renal tubular cells. SIPS induced by a lithogenic milieu may result from upregulation of p16 and downregulation of shelterin components, specifically POT1, and might contribute, at least in part, to the development of CaOx KS.

Tyrosinase deficiency increases protein carbonyl content in substantia nigra of mice administered retinol palmitate.

Tyrosinase is a key enzyme for the biosynthesis of melanin pigments in peripheral tissues such as skin and retina. Although tyrosinase activity is specifically detected in melanocytes, several studies have shown the expression and enzymatic activity of tyrosinase in the central nervous system, especially in the midbrain substantia nigra. In the present study, we investigated the antioxidative effects of tyrosinase on protein damage in the substantia nigra of mice. C57BL/10JMsHir (B10) and tyrosinase-deficient albino B10.C-Tyrc/Hir (B10-c) mice were intraperitoneally administered retinol palmitate to induce oxidative stress, and the protein carbonyl content, a hallmark of protein oxidative damage, was examined in the substantia nigra. Retinol palmitate administration was found to decrease catalase activity in the substantia nigra of both B10 and B10-c mice, suggesting the induction of oxidative stress due to imbalanced antioxidant systems. In this model, we found that tyrosinase deficiency markedly increases the protein carbonyl content in the substantia nigra. Thus, we concluded that tyrosinase activity prevents protein damage in the substantia nigra of mice that were challenged with oxidative stress. These findings provide novel insight into the physiological role of tyrosinase in the central nervous system.

Heat-induced endoplasmic reticulum stress in soleus and gastrocnemius muscles and differential response to UPR pathway in rats.

The present study aimed to investigate the differential response of oxidative (soleus) and glycolytic (gastrocnemius) muscles to heat-induced endoplasmic reticulum (ER) stress. It was hypothesized that due to compositional and functional differences, both muscles respond differently to acute heat stress. To address this, male Sprague Dawley rats (12/group) were subjected to thermoneutral (25°C) or heat stress (42°C) conditions for 1h. Soleus and gastrocnemius muscles were removed for analysis post-exposure. A significant increase in body temperature and free radical generation was observed in both the muscles following heat exposure. This further caused a significant increase in protein carbonyl content, AOPP, and lipid peroxidation in heat-stressed muscles. These changes were more pronounced in heat-stressed soleus compared to the gastrocnemius muscle. Accumulation of unfolded, denatured proteins results in ER stress, causing activation of unfolded protein response (UPR) pathway. The expressions of UPR transducers were significantly higher in soleus as compared to the gastrocnemius muscle. A significant elevation in resting intracellular calcium ion was also observed in heat-stressed soleus muscle. Overloading of cells with misfolded proteins in soleus muscle activated ER-induced apoptosis as indicated by significant upregulation of C/EBP homologous protein and Caspase12. The study provides a detailed mechanistic representation of the differential response of muscles toward UPR under heat stress. Data suggests that soleus majorly being an oxidative muscle is more prone to heat stress-induced insult indicated by enhanced apoptosis. This study may aid in devising mitigation strategies to improve muscle performance under heat stress.

Hydrogen sulfide is a crucial element of the antioxidant defense system in Glycine max–Sinorhizobium fredii symbiotic root nodules

H2S is emerging as a signaling molecule involved in the regulation of many physiological processes in plants. Here, we investigated the potential antioxidant role of H2S in soybean (Glycine max)-rhizobia (Sinorhizobium fredii) symbiotic root nodules. An endogenous H2S production deficit rhizobia mutant ∆CSE was constructed to study the effect of decreased content of H2S in soybean nodules. Fluorescent probes and confocal microscope were used to observe the production and accumulation of H2S and reactive oxygen species. Transmission electronic microscopy was conducted to study the structural changes in ∆CSE soybean nodules. Finally, qRT-PCR, enzymatic activity, and oxidative damage parameters were measured. The results demonstrated that abundant H2S was generated in the nitrogen-fixing zone of soybean nodules. The deletion of the cystathionine γ-lyase (CSE) gene in S. fredii (∆CSE) caused a sharp decrease in H2S production in both free-living rhizobia and soybean nodules. We found that decrease in the H2S level in nodule cells inhibited nitrogenase activity. In addition, to elevated H2O2 and malondialdehyde accumulation, increased protein carbonyl content and decreased O2− scavenging ability was observed in ∆CSE root nodules. Transmission electron microscopy revealed that an H2S deficit caused the deformation of bacteroids and damage of peribacteroid membranes in nodule cells. Moreover, the expression of some rhizobial and soybean genes related to antioxidant defense was up-regulated in ∆CSE nodules. H2S is crucial for the nitrogen-fixation ability of soybean nodules by acting as an antioxidant element that protects nodule cells and bacteroids from oxidative damage.

Soybean β-conglycinin caused intestinal inflammation and oxidative damage in association with NF-κB, TOR and Nrf2 in juvenile grass carp (Ctenopharyngodon idella): varying among different intestinal segments

The current study aimed to investigate the effects and mechanisms of dietary soybean β-conglycinin in immune function and oxidative damage among different intestinal segments of juvenile grass carp (Ctenopharyngodon idella). 240 fish (13.77 ± 0.10 g) were fed control or 8% β‐conglycinin diet for 7 weeks. Dietary β-conglycinin caused inconsistent suppression effects on the innate immune by decreasing complement component, lysozyme, antimicrobial peptide and acid phosphatase among different intestinal segments. Meanwhile, dietary β-conglycinin caused inflammation in the mid and distal intestine by raising pro-inflammatory cytokines and declining anti-inflammatory cytokines mRNA levels, while more serious in the distal intestine than in the mid intestine. Furthermore, dietary β-conglycinin regulating inflammatory cytokines might be associated with transcription factors nuclear factor-κB P65 (NF-κB P65) nucleus translocation and target of rapamycin (TOR) phosphorylation in the distal intestine but only related to TOR phosphorylation in the mid intestine. Interestingly, in the proximal intestine, dietary β-conglycinin decreased both pro-inflammatory and anti-inflammatory cytokines mRNA level, and did not affect NF-κB P65 nucleus translocation and TOR phosphorylation. For oxidative damage, dietary β-conglycinin exposure elevated both malondialdehyde (MDA) and protein carbonyl (PC) contents in the distal intestine, which might be attributed to the suppression of the Mn-SOD, catalase (CAT) and glutathione peroxidase (GPx) activities. In the mid intestine, dietary β-conglycinin only increased PC content in association with the low activities of CAT, GPx and glutathione peroxidase (GR). Unexpectedly, in the proximal intestine, dietary β-conglycinin did not significantly change MDA and PC contents while decreased antioxidant enzyme activities. Furtherly, dietary β-conglycinin affect the antioxidant enzyme activity might be regulated by the varying pattern of nuclear factor-erythroid 2-related factor 2 (Nrf2) nucleus translocation among these three intestinal segments. In summary, dietary β-conglycinin caused intestinal inflammation and oxidative damage in association with NF-κB, TOR and Nrf2 signaling molecules, which were varying among the three intestinal segments of grass carp.

Effect of Chlorogenic Acid on the Physicochemical and Functional Properties of Coregonus Peled Myofibrillar Protein through Hydroxyl Radical Oxidation

The effects of chlorogenic acid (CA) (6, 30, and 150 μM/g protein) on the physicochemical and functional properties of Coregonus peled myofibrillar protein (MP) through oxidation using a hydroxyl radical oxidation system (0.01 mM FeCl3, 0.01 mM Asc, and 1 mM H2O2) were investigated. The result showed that CA inhibited the increase in protein carbonyl content but did not prevent losses in sulfhydryl and free amine contents caused by oxidation. The presence of CA also increased conformational changes in the secondary and tertiary structures of oxidized MP. Oxidized MP containing 6 μM/g CA had superior functional properties (solubility, emulsifying, foaming, and gel properties), while oxidized MP containing 150 μM/g CA aggregated, resulting in insolubility and a poor gel network.

Modification of endothelial nitric oxide synthase by 4-oxo-2(E)-nonenal(ONE) in preeclamptic placentas

Preeclampsia (PE) is a leading cause of pregnancy complications, affecting 3–7% of pregnant women worldwide. The pathophysiology of preeclampsia involves a redox imbalance, oxidative stress and a reduced nitric oxide (NO) bioavailability. The molecular and cellular mechanisms leading to the dysfunction of the placental endothelial NO synthase (eNOS) are not clarified. This study was designed to investigate whether aldehydes generated by lipid peroxidation products (LPP), may contribute to placental eNOS dysfunction in PE. The analysis of placentas from PE-affected patients and normal pregnancies, showed a significant increase in protein carbonyl content, indicative of oxidative stress-induced protein modification, as shown by the accumulation of acrolein, 4-hydroxynonenal (HNE), and 4-oxo-2(E)-nonenal (ONE) adducts in PE placentas. In contrast, the levels of these LPP-adducts were low in placentas from normal pregnancies. Immunofluorescence and confocal experiments pointed out a colocalization of eNOS with ONE-Lys adducts, whereas eNOS was not modified in normal placentas. LC-MS/MS analysis of recombinant eNOS preincubated with ONE, allowed to identify several ONE-modified Lys-containing peptides, confirming that eNOS may undergo post-translational modification by LPP. The preincubation of HTR-8/SVneo human trophoblasts (HTR8) with ONE, resulted in ONE-Lys modification of eNOS and a reduced generation of NO. ONE inhibited the migration of HTR8 trophoblasts in the wound closure model, and this was partly restored by the NO donor, NOC-18, which confirmed the important role of NO in the invasive potential of trophoblasts. In conclusion, placental eNOS is modified by ONE in PE placentas, which emphasizes the sensitivity of this protein to oxidative stress in the disturbed redox environment of preeclamptic pregnancies.

Maternal corticosterone in the mouse alters oxidative stress markers, antioxidant function and mitochondrial content in placentas of female fetuses.

Maternal exposure to the stress hormone corticosterone is known to programme a range of sex specific disease outcomes in offspring. Sex differences in placental adaptations are thought to mediate these processes. Placental oxidative stress is implicated in a range of pregnancy disorders but the role of placental oxidative stress in sex specific disease outcomes following prenatal corticosterone exposure is unknown. This study demonstrates that maternal corticosterone reduced placental hydrogen peroxide and 8-hydroxy-2'-deoxyguanosine concentrations but increased protein carbonyl content and advanced glycation end product concentrations in placentas of female fetuses but not male fetuses. These results highlight that placentas of female fetuses respond differently to maternal corticosterone exposure, with oxidative stress a major finding in placentas of female fetuses. Maternal exposure to glucocorticoids during pregnancy increases offspring risk of developing a range of sex specific disease phenotypes. These sex specific disease outcomes are thought to be in part mediated by different placental adaptations in males and females. The placenta is a highly metabolic organ which is vulnerable to the effects of oxidative stress. In other tissues, males and females have been shown to respond differently to the pro-oxidant effects of glucocorticoids. This study therefore used a well characterized animal model of maternal corticosterone exposure to investigate sex specific alterations in reactive oxygen species production, antioxidant concentrations and mitochondrial properties that might contribute to sex differences in placental outcomes. C57BL/6 mice were implanted with osmotic minipumps containing corticosterone (33μgkg-1 h-1 ) at embryonic day (E) 12.5 and placentas collected at E14.5 for analysis. Corticosterone exposure reduced placental hydrogen peroxide (H2 O2 ) and 8-hydroxy-2'-deoxyguanosine concentrations but increased protein carbonyl content and advanced glycation end product concentrations in placentas of female fetuses but not male fetuses. This dysregulation of different markers of oxidative stress may be due to increased placental activity of thioredoxin reductase in female but not male fetuses. Corticosterone reduced placental mitochondrial content but increased protein expression of the autophagosome cargo protein p62. This study demonstrates that placentas of female fetuses respond differently to maternal corticosterone exposure and highlights an important role of reactive oxygen species, mitochondrial adaptations and antioxidant responses in glucocorticoid induced programmed disease.

Effect of hydroxyl-radical on the biochemical properties and structure of myofibrillar protein from Alaska pollock (Theragra chalcogramma)

The aim of the study was to investigate the effect of hydroxyl-radical on myofibrillar protein properties. Myofibrillar protein isolated from Alaska pollock muscles were incubated with oxidant agents containing different H2O2 concentrations of 0, 0.1, 1, 5, 10 and 20 mmoL/L for 1, 3 and 5 h respectively. Incubation at higher concentrations of H2O2 induced a significant decrease of protein total sulfhydryl group and an increase in protein carbonyl content in the myofibrillar fraction. Moderate H2O2 concentration (1 mmoL/L) enhanced water-holding capacity and textural properties of a gel, while lower (0.1 mmoL/L) and higher (5, 10 & 20 mmoL/L) H2O2 concentrations significantly reduced the water-holding capacity. At higher H2O2 concentration, oxidized gel revealed some resistance toward the change in gel properties. Additionally, the results from SDS-PAGE and microstructure confirmed the structural changes through aggregation and denaturation of myofibrillar components that induced changes on gelation of myofibrillar protein. The whiteness of the myofibrillar protein gel decreased significantly (p < 0.05) with increasing the concentration of H2O2. Oxidation of myofibrillar protein revealed the slight improvement of textural properties of the heat set gel at a moderate concentration of oxidant agent.

Geraniin Protects High-Fat Diet-Induced Oxidative Stress in Sprague Dawley Rats.

Geraniin, a hydrolysable polyphenol derived from Nephelium lappaceum L. fruit rind, has been shown to possess significant antioxidant activity in vitro and recently been recognized for its therapeutic potential in metabolic syndrome. This study investigated its antioxidative strength and protective effects on organs in high-fat diet (HFD)-induced rodents. Rats were fed HFD for 6 weeks to induce obesity, followed by 10 and 50 mg/kg of geraniin supplementation for 4 weeks to assess its protective potential. The control groups were maintained on standard rat chows and HFD for the same period. At the 10th week, oxidative status was assessed and the pancreas, liver, heart and aorta, kidney, and brain of the Sprague Dawley rats were harvested and subjected to pathological studies. HFD rats demonstrated changes in redox balance; increased protein carbonyl content, decreased levels of superoxide dismutase, glutathione peroxidase, and glutathione reductase with a reduction in the non-enzymatic antioxidant mechanisms and total antioxidant capacity, indicating a higher oxidative stress (OS) index. In addition, HFD rats demonstrated significant diet-induced changes particularly in the pancreas. Four-week oral geraniin supplementation, restored the OS observed in the HFD rats. It was able to restore OS biomarkers, serum antioxidants, and the glutathione redox balance (reduced glutathione/oxidized glutathione ratio) to levels comparable with that of the control group, particularly at dosage of 50 mg geraniin. Geraniin was not toxic to the HFD rats but exhibited protection against glucotoxicity and lipotoxicity particularly in the pancreas of the obese rodents. It is suggested that geraniin has the pharmaceutical potential to be developed as a supplement to primary drugs in the treatment of obesity and its pathophysiological sequels.

Effects of two aerobic exercise training protocols on parameters of oxidative stress in the blood and liver of obese rats.

We evaluated the effects of moderate-intensity continuous training (MICT) and high-intensity interval training (HIIT) protocols on the alterations in oxidative stress parameters caused by a high-fat diet (HFD), in the blood and liver of rats. The HFD enhanced thiobarbituric acid reactive substances (TBA-RS) and protein carbonyl content, while reducing total sulfhydryl content and catalase (CAT) and glutathione peroxidase (GSH-Px) activities in the blood. Both trainingprotocols prevented an increase in TBA-RS and protein carbonyl content, and prevented a reduction in CAT. HIIT protocol enhanced SOD activity. In the liver, HFD didn't alter TBA-RS, total sulfhydryl content or SOD, but increased protein carbonyl content and CAT and decreased GSH-Px. The exerciseprotocols prevented the increase in protein carbonyl content and the MICT protocol prevented an alteration in CAT. In conclusion, HFD elicits oxidative stress in the blood and liver and both protocols prevented most of the alterations in the oxidative stress parameters.

Postconditioning with metformin attenuates apoptotic events in cardiomyoblasts associated with ischemic reperfusion injury.

Ischemic postconditioning is considered as a potent method to combat ischemic reperfusion injury than preconditioning and rapid initiation of reperfusion with potent antioxidant compounds can be one of the most effective treatments to reduce the infarct size and behavioral deficits as a result of ischemic insult. The present study aims to determine the postconditioning effects of metformin in experimental ischemia. H9C2 cells were subjected to experimental ischemia and reperfused with metformin. Viability, oxidative stress and apoptotic events were determined. Postconditioning with Metformin effectively maintained cell viability and membrane stability of H9C2 cardiomyoblast cells from ischemic injury at a concentration of 2.5 µg/mL. Decrease in cell death was established viz fluorescent staining and Annexin V-FITC flow cytometric analysis. In metformin treated ischemic cells the DNA fragmentation and comet length were significantly reduced (p=.0001) affirming antiapoptotic activity of metformin. There was an increase in protein carbonyl content and decrease in nitrate levels after ischemic injuries which were alleviated with metformin treatment. Wound closure was enhanced by metformin treatment. Metformin synchronized the apoptotic proteins such as FasL and antiapoptotic proteins such as Bcl2, Bcl-xL and p21 which can be attributed as the major mechanism of cardioprotection. The results envisage therapeutic potential of metformin at lower concentrations as an effective postischemic conditioning agent.

Cerebral Oxidative Stress and Microvasculature Defects in TNF-α Expressing Transgenic and Porphyromonas gingivalis-Infected ApoE-/- Mice.

The polymicrobial dysbiotic subgingival biofilm microbes associated with periodontal disease appear to contribute to developing pathologies in distal body sites, including the brain. This study examined oxidative stress, in the form of increased protein carbonylation and oxidative protein damage, in the tumor necrosis factor-α (TNF-α) transgenic mouse that models inflammatory TNF-α excess during bacterial infection; and in the apolipoprotein knockout (ApoE-/-) mouse brains, following Porphyromonas gingivalis gingival monoinfection. Following 2,4-dinitrophenylhydrazine derivatization, carbonyl groups were detected in frontal lobe brain tissue lysates by immunoblotting and immunohistochemical analysis of fixed tissue sections from the frontotemporal lobe and the hippocampus. Immunoblot analysis confirmed the presence of variable carbonyl content and oxidative protein damage in all lysates, with TNF-α transgenic blots exhibiting increased protein carbonyl content, with consistently prominent bands at 25 kDa (p = 0.0001), 43 kDa, and 68 kDa, over wild-type mice. Compared to sham-infected ApoE-/- mouse blots, P. gingivalis-infected brain tissue blots demonstrated the greatest detectable protein carbonyl content overall, with numerous prominent bands at 25 kDa (p = 0.001) and 43 kDa (p = 0.0001) and an exclusive band to this group between 30-43 kDa* (p = 0.0001). In addition, marked immunostaining was detected exclusively in the microvasculature in P. gingivalis-infected hippocampal tissue sections, compared to sham-infected, wild-type, and TNF-α transgenic mice. This study revealed that the hippocampal microvascular structure of P. gingivalis-infected ApoE-/- mice possesses elevated oxidative stress levels, resulting in the associated tight junction proteins being susceptible to increased oxidative/proteolytic degradation, leading to a loss of functional integrity.

Oxidative status in relation to metal pollution and calcium availability in pied flycatcher nestlings – A calcium manipulation experiment

Metal exposure can produce oxidative stress by disrupting the prooxidant/antioxidant balance. It has been suggested that calcium (Ca) may provide protection against metal toxicity in the organism. The objective of this study is to explore the effects of Ca availability and metal pollution on oxidative stress biomarkers in pied flycatcher (Ficedula hypoleuca) nestlings. For this purpose, we performed a Ca-supplementation experiment with birds inhabiting a Ca-poor and metal-polluted area in SW Finland. An array of oxidative stress biomarkers (GSH, GSH:GSSG ratio, GPx, GST, CAT, SOD, lipid peroxidation and protein carbonylation) was measured in red blood cells. The effects of antioxidant molecules and oxidative damage on nestling size, growth, fledging success and fledgling number were evaluated. We observed an up-regulation of GST activity and increased protein carbonyl content in the polluted zone, probably related to a combination of higher metal exposure and reduced food quantity and quality in this area. As expected, birds from the unpolluted zone showed higher GSH:GSSG ratio but, unexpectedly, also showed signs of higher lipid peroxidation (not statistically significant, p = 0.056), both responses likely being related with the lower Ca availability. Our study suggests that different measures of oxidative damage are affected by different factors: while damage to proteins was the target of metal exposure/food limitation, poor Ca availability may enhance damage to lipids in growing birds. The intercorrelations found between Ca in plasma, metal exposure and the different oxidative stress biomarkers show that the antioxidant defense is finely regulated to cope with increased oxidative challenges. Finally, our results suggest that the antioxidant status during early development, conditioned by environmental pollution and Ca availability, is one factor affecting nestling survival and fledgling number.

Oxidative stress indicated by elevated expression of Nrf2 and 8-OHdG promotes hepatocellular carcinoma progression.

Reactive oxygen species (ROS) is excessively generated in tumors creating an oxidative stress in tumor microenvironment. We investigated hepatic expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and 8-hydroxydeoxyguanosine (8-OHdG) in hepatocellular carcinoma (HCC) patients, and asked if ROS epigenetically upregulated Nrf2 and enhanced aggressiveness in HCC cells. Expression of Nrf2 (n=100) and 8-OHdG (n=53) was remarkably increased in HCC tissues compared with the noncancerous hepatic tissues. Elevated expression of 8-OHdG was associated with poor survival in HCC patients. H2O2, as ROS representative, provoked oxidative stress in HepG2 cells, indicated by increased protein carbonyl content and decreased total antioxidant capacity. Nrf2 expression and 8-OHdG formation were markedly increased in the H2O2-treated cells compared with the untreated control. Co-treatment with antioxidants, tocopheryl acetate (TA) and S-adenosylmethionine (SAM) effectively attenuated expression of Nrf2 and 8-OHdG in H2O2-treated cells. HepG2 cells treated with H2O2 had significantly higher migration and invasion capabilities than the untreated control cells, and this aggressiveness was significantly inhibited by TA and SAM. Bisulfite sequencing revealed that CpG dinucleotides in Nrf2 promoter were unmethylated in the H2O2-treated cells similar to the untreated control. In conclusion, robust histological evidence of increased antioxidative response and oxidative DNA damage in human HCC tissues was demonstrated. Elevated oxidative DNA lesion 8-OHdG was associated with shorter survival. Experimentally, ROS enhanced Nrf2 expression, 8-OHdG formation and tumor progression in HCC cells. These effects were inhibited by antioxidants. Therefore, oxidative stress-reducing regimens might be beneficial to diminish the ROS-induced HCC progression.

Protein carbonyl content: a novel biomarker for aging in HIV/AIDS patients

BackgroundThe major complications of “treated” Human Immunodeficiency Virus (HIV) infection are cardiovascular disease, malignancy, renal disease, liver disease, bone disease, and perhaps neurological complications, which are phenomena of the normal aging process occurring at an earlier age in the HIV-infected population. The present study is aimed to explore protein carbonyl content as a biomarker for detecting oxidative DNA damage induced ART toxicity and/or accelerated aging in HIV/AIDS patients. ObjectiveTo investigate the potential of carbonyl content as a biomarker for detecting oxidative Deoxyribonucleic acid (DNA) damage induced Antiretroviral Theraphy (ART) toxicity and/or accelerated aging in HIV/AIDS patients. MethodsIn this case–control study a total 600 subjects were included. All subjects were randomly selected and grouped as HIV-negative (control group) (n=300), HIV-infected ART naive (n=100), HIV-infected on first line ART (n=100), and HIV-infected on second line ART (n=100). Seronegative control subjects were age- and sex-matched with the ART naive patients and the two other groups. Carbonyl protein was determined by the method described in Levine et al. DNA damage marker 8-OH-dG was determined using 8-hydroxy-2-deoxy Guanosine StressXpress ELA Kit by StressMarq Biosciences. ResultsProtein carbonyl content levels and oxidative DNA damage were significantly higher (p<0.05) in HIV-infected patients on second line ART and HIV-infected patients on first line ART than ART naive patients and controls. In a linear regression analysis, increased protein carbonyl content was positively associated with increased DNA damage (OR: 0.356; 95% CI: 0.287–0.426) p<0.05. ConclusionsCarbonyl content may has a role as a biomarker for detecting oxidative DNA damage induced ART toxicity and/or accelerated aging in HIV/AIDS patients. Larger studies are warranted to elucidate the role of carbonyl content as a biomarker for premature aging in HIV/AIDS patients.

Evidence of oxidative stress in brain and liver of young rats submitted to experimental galactosemia.

Galactosemia is a disorder of galactose metabolism, leading to the accumulation of this carbohydrate. Galactosemic patients present brain and liver damage. For evaluated oxidative stress, 30-day-old males Wistar rats were divided into two groups: galactose group, that received a single injection of this carbohydrate (5μmol/g), and control group, that received saline 0.9% in the same conditions. One, twelve or twenty-four hours after the administration, animals were euthanized and cerebral cortex, cerebellum, and liver were isolated. After one hour, it was found a significant increase in TBA-RS levels, nitrate and nitrite and protein carbonyl contents in cerebral cortex, as well as protein carbonyl content in the cerebellum and in hepatic level of TBA-RS, and a significant decrease in nitrate and nitrite contents in cerebellum. TBA-RS levels were also found increased in all studied tissues, as well as nitrate and nitrite contents in cerebral cortex and cerebellum, that also present increased protein carbonyl content and impairments in the activity of antioxidant enzymes of rats euthanized at twelve hours. Finally, animals euthanized after twenty-four hours present an increase of TBA-RS levels in studied tissues, as well as the protein carbonyl content in cerebellum and liver. These animals also present an increased nitrate and nitrite content and impairment of antioxidant enzymes activities. Taken together, our data suggest that acute galactose administration impairs redox homeostasis in brain and liver of rats.

(−)-Epicatechin in vitro ameliorates erythrocyte protein carbonyl content in hypertensive patients: comparison with L-ascorbic acid

Context: Oxidative stress plays a key role in the patho-physiology of hypertension. (−)-Epicatechin has many important biological properties. Objective: The present study was undertaken to evaluate effect of (−)-epicatechin on protein carbonyl content in gender-based hypertensive patients and normal subjects. Methods: The study was carried out on 83 normal (male: 42; female: 41) and 62 hypertensive subjects (male: 32; female: 30). In vitro effect on (−)-epicatechin and L-ascorbic acid was estimated on protein carbonyl content. Results: Result showed a significant (p < 0.001) increase in protein carbonyl content in hypertensive patients but no gender-based difference was observed. (−)-epicatechin shows significant (p < 0.001) dose-dependent effect as compared to L-ascorbic acid, which is manifested as decrease in protein carbonyl content. Conclusion: We hypothesizes that a higher intake of (−)-epicatechin may provide protection against hypertension in males and females.