Protein Carbonyl Groups Research Articles

Adaptation of Ochrobactrum rhizosphaerae IMV B-7956 Bacteria to the Influence of Copper (II) Chloride

In technologically altered habitats, an increased content of organic compounds, nitrogen, phosphorus, sulfur compounds, antibiotic substances, etc. is found. Therefore, microorganisms that are systematically exposed to various stressors have developed adaptation mechanisms. The strain Ochrobactrum rhizosphaerae IMV B-7956, isolated from the infiltrate lakes of the Lviv solid waste landfill, is resistant to copper, chromium, manganese, and iron in concentrations exceeding the maximum permissible concentrations. The work aimed to study the response of O. rhizosphaerae IMV B-7956 cells to CuCl2 exposure by detecting changes in the content of lipid peroxidation products, products of the oxidative modification of proteins, activity of antioxidant defense system enzymes, and synthesis of extracellular polymers. Methods. To study the effect of CuCl2 on prooxidant indicators and the activity of enzymes of the antioxidant defense system, bacteria were pre-incubated in Tris-HCl buffer containing 2–10 mM CuCl2. After one hour of incubation, the bacterial cells were washed and cultured for 1, 12, 24, and 48 h in metal-free media. The copper content in the bacterial cells was determined by atomic absorption spectrometry. The content of lipid peroxidation indicators, carbonyl groups in proteins, total low-molecular-weight thiols, enzymatic activity, and the content of exopolysaccharides and extracellular proteins were determined photometrically. Results. Within an hour, O. rhizosphaerae IMV B-7956 bacteria accumulated 2.3–7.8 mg Cu/g of biomass. Under these conditions, an increased content of lipid peroxidation products was detected. During the first hour of growth in bacterial cells, enzymes with catalase and peroxidase activity were activated. During further cultivation, an increase in the activity of other antioxidant defense enzymes was detected. Carbonyl groups in proteins are probably formed due to an increase in the content of lipid peroxidation products, as they are formed later. Within 12–48 h of growth, the copper content in the bacterial cells decreases. This leads to the restoration of growth. Conclusions. The main damaging effect of CuCl2 on bacterial cells was detected during the first 24 h of growth. Activation of the enzymes of the antioxidant system and synthesis of exopolysaccharides are among the adaptations that ensure the survival of bacteria under these conditions.

Modulation of Redox and Inflammatory Signaling in Human Skin Cells Using Phytocannabinoids Applied after UVA Irradiation: In Vitro Studies.

UVA exposure disturbs the metabolism of skin cells, often inducing oxidative stress and inflammation. Therefore, there is a need for bioactive compounds that limit such consequences without causing undesirable side effects. The aim of this study was to analyse in vitro the effects of the phytocannabinoids cannabigerol (CBG) and cannabidiol (CBD), which differ in terms of biological effects. Furthermore, the combined use of both compounds (CBG+CBD) has been analysed in order to increase their effectiveness in human skin fibroblasts and keratinocytes protection against UVA-induced alternation. The results obtained indicate that the effects of CBG and CBD on the redox balance might indeed be enhanced when both phytocannabinoids are applied concurrently. Those effects include a reduction in NOX activity, ROS levels, and a modification of thioredoxin-dependent antioxidant systems. The reduction in the UVA-induced lipid peroxidation and protein modification has been confirmed through lower levels of 4-HNE-protein adducts and protein carbonyl groups as well as through the recovery of collagen expression. Modification of antioxidant signalling (Nrf2/HO-1) through the administration of CBG+CBD has been proven to be associated with reduced proinflammatory signalling (NFκB/TNFα). Differential metabolic responses of keratinocytes and fibroblasts to the effects of the UVA and phytocannabinoids have indicated possible beneficial protective and regenerative effects of the phytocannabinoids, suggesting their possible application for the purpose of limiting the harmful impact of the UVA on skin cells.

АНТИОКСИДАНТНІ ВЛАСТИВОСТІ ЕКСТРАКТІВ КАВОВИХ ЗЕРЕН

The antioxidant properties (AP) of 5 % aqueous extracts of roasted coffee and its spent grounds were investigated. The antioxidant activity (AA) of these extracts acting on rat liver hepatocytes in vitro was determined using two markers of oxidative stress (OS): the levels of thiobarbituric acid reactive substances and carbonyl groups in proteins. It was demonstrated that all examined extracts exhibited AA, characterized by a reduction in non-enzymatic OS markers. The formation of products of free radical damage to lipids and proteins was inhibited by both 5 % aqueous extracts of roasted coffee and 5 % aqueous extracts of used coffee grounds, which holds promise for the development of antioxidant agents based on them.

The effect of nettle extract on antioxidant defense system in piglets after weaning

Background. The effect of common nettle (Urtica dioica L.) extracts on the free radical processes and antioxidant system in piglets during the critical period of weaning from sows has been studied. Materials and Methods. Large white piglets were divided into 2 groups (control and experimental), 9 animals in each. Piglets of the experimental group from 14 days of age and before weaning received the standard diet and the nettle extract in the dose of 6 mg/kg of body weight for 22 days. The blood, as well as erythrocyte hemolysates and plasma of piglets obtained at 14, 36, and 42 days of age, were studied. Results. Our results have shown that weaning causes an oxidative stress in piglets. This state leads to an increase in the concentration of metabolites of free radical damage to protein molecules – carbonyl groups of proteins on the first day and primary products of lipid peroxidation on the seventh day after weaning. This activation of oxidative damage occurs in piglets against the background of a physiologically immature antioxidant system and is characterized by a decrease in the activity of the enzymatic chain – superoxide dismutase, glutathione peroxidase and catalase, as well as the concentration of its non-enzymatic antioxidant – reduced glutathione. Feeding piglets with nettle extract leads to activation of the antioxidant defense system in erythrocytes (higher activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and an increased reduced glutathione level compared to the control values) and a decrease in the concentration of oxidative damage products in the plasma (the content of lipid hydroperoxides, TBA-active products and carbonyl groups of proteins). Conclusion. The positive effect of nettle extract on the inhibition of free radical processes and activation of antioxidant systems indicates that this extract can be added to the standard diet of young animals to increase stress resistance and adaptability of their organism in critical periods of ontogenesis.

Comparison of the Regenerative Metabolic Efficiency of Lipid Extracts from Microalgae Nannochloropsis oceanica and Chlorococcum amblystomatis on Fibroblasts.

UVA radiation leads to oxidative stress and inflammation in skin cells. Therefore, the aim of this study was to compare the effect of lipid extracts from microalgae Nannochloropsis oceanica (N.o.) (marine) and Chlorococcum amblystomatis (C.a.) (freshwater) on the redox balance and PUFA metabolism in human skin fibroblasts modified by UVA. Lipid extracts from both types of microalgae introduced into the fibroblast medium after UVA irradiation significantly reduced the level of ROS and enhanced expression of Nrf2, which increased the activity/level of antioxidants (SOD1/2, CAT, GSH, Trx). The reduction in oxidative stress was accompanied by a decrease in the level of 4-HNE, its protein adducts and protein carbonyl groups. Microalgae also reduced the activity of COX1/2, FAAH and MAGL increased by UVA, and as a consequence, the level of lipid mediators (especially after N.o.) decreased, both from the group of endocannabinoids (AEA, 2-AG, PEA) and eicosanoids (PGE2, 15d-PGJ2, TXB2, 15-HETE), acting mainly through receptors related to G protein, the expression of which increases after UVA. This further contributed to the reduction in oxidative stress and pro-inflammatory signaling at NF-κB and TNFα levels. Therefore, it is suggested that lipid extracts from both N.o. and C.a. microalgae can be used to regenerate fibroblast metabolism disturbed by UVA radiation.

Role of growth differentiation factor-15 (GDF-15) in missed miscarriage and its relationship to oxidative stress markers

Aim. The study aimed to investigate the level of growth differentiation factor-15 (GDF-15) and its relationship to markers of oxidative stress (OS) as predictors of missed miscarriage (MM).
 Materials and methods. A prospective cohort study of 80 females was conducted. The main group included 40 patients with MM at 5–9 weeks of gestation; the control group included 40 females with elective termination at the same gestation period. OS markers, including malonic dialdehyde, carbonyl groups of proteins, and the superoxide dismutase antioxidant enzyme, as well as the GDF-15, were measured in serum, aspiration fluid, and chorionic tissues in women with normal pregnancy and MM at 5–9 weeks of gestation.
 Results. The results showed that the complex balance maintained between the production of reactive oxygen species and antioxidant activity in MM is shifted towards oxidative reactions (p0.05) and is associated with non-enzymatic lipid peroxidation, and OS can become a leading factor in the pathogenesis of MM. The GDF-15 levels were significantly higher in aspiration fluid and chorionic tissues in patients of the study groups; low concentrations of GDF-15 in MM patients in aspiration fluid and chorionic tissues highlight the role of this protein as an important regulator of proinflammatory molecules (p0.05). In addition, a decrease in the GDF-15 level negatively correlates with OS markers, such as malonic dialdehyde and the superoxide dismutase antioxidant enzyme, suggesting the role of this cytokine in peroxidation and OS development.
 Conclusion. The study of biomarkers of independent pathological pathways as predictors of miscarriage will allow clinicians to stratify and predict the risks of early reproductive loss more accurately.

Age-related hair denaturation related to protein carbonyls.

Hair ageing is accompanied by hair fibres becoming irregularly shaped causing them to stick out in irregular directions or have more curliness and being spread out. This is believed to be due to changes within the hair fibre structure which occur with ageing, and one of the causes of these changes could be an increase in the number of protein carbonyl groups present in the hair. The aim of this study is to investigate the internal denaturation of hair related to protein carbonyls in attempt to gain new insight into age-related changes that occur in hair. The degree of carbonylation of the hair structural protein as determined by fluorescent labelling and Western blotting analysis was used to investigate the primary structure of hair protein. The amount of helix, a common conformation in the secondary structure of proteins, in hair in groups of women with different ages was also analysed using infrared microscopy coupled with multivariate curve resolution (MCR). From the results of this, an image of the two-dimensional distribution of the α-helices was generated for the hair taken from each age group. Also, high-pressure differential scanning calorimetry (HPDSC) of the hair in water was performed on the hair taken from each age group to determine the peak temperature of endothermic effect and the enthalpy of denaturation. We found that the amino group content in hair proteins decreased and Type II keratin, one of the subunits of intermediate filament, was more carbonylated with age. The results of the MCR indicated eight separate components, including components of the secondary structure of proteins, such as α helices and β sheets. Two-dimensional images of the hair cross-sections revealed that the presence of α helices decreased with age. In addition, data from the HPDSC showed that the enthalpy associated with the denaturing temperature also significantly decreased with age. These results suggest that there is a negative correlation between age and structural integrity of the helix segment in intermediate filament. The results of this study also show that there is a positive correlation between age-related hair denaturation and protein carbonyls.

Carnosine supplementation and retinal oxidative parameters in a high-calorie diet rat model

BackgroundTo assess oxidative effects induced by a high-calorie diet on the retina of Wistar rats and test the antioxidative effects of carnosine supplementation.MethodsWistar rats were randomly divided into the following groups: standard diet (SD), high-calorie diet (HcD), standard diet + carnosine (SD + Car), and high-calorie diet + carnosine (HcD + Car). The body weight, adiposity index, plasma glucose, total lipids, high-density lipoprotein (HDL), low-density lipoprotein (LDL), uric acid, creatinine, and triglycerides of the animals were evaluated. The retinas were analyzed for markers of oxidative stress. Hydrogen peroxide production was assessed by 2',7'-dichlorodihydrofluorescein diacetate (DCF) oxidation. The total glutathione (tGSH), total antioxidant capacity (TAC), protein carbonyl, and sulfhydryl groups of the antioxidant system were analyzed.ResultsTAC levels increased in the retinas of the SD + Car group compared to the SD group (p < 0.05) and in the HcD + Car group compared to the HcD group (p < 0.05). The levels of GSH and the GSSH:GSSG ratio were increased in the HcD + Car group compared to the SD + Car group (p < 0.05). An increase in the retinal carbonyl content was observed in the HcD group compared to the SD group (p < 0.05) and in the HcD + Car group compared to the SD + Car group (p < 0.05). A high-calorie diet (HcD) was also associated with a decrease in retinal sulfhydryl-type levels compared to the SD group (p < 0.05).ConclusionThe results suggest that feeding a high-calorie diet to rats can promote an increase in carbonyl content and a reduction in sulfhydryl groups in their retinas. The administration of carnosine was not effective in attenuating these oxidative markers.Trial registrationAnimal Ethics Committee of Botucatu Medical School - Certificate number 1292/2019.

Diabetes-correcting and antioxidant effects of grape pomace extract rich in natural complex of polyphenols

Background. The positive health effects of polyphenols have led to an increased scientific interest in these natural compounds over the past decade. Many studies confirm the effectiveness of polyphenols as additional therapy in diabetes, especially due to the sugar-lowering effect of polyphenols. The aim of the research was to investigate the morphological and functional state of peripheral blood erythrocytes and the indices of oxidative stress in the liver of rats with experimental diabetes and after the administration of grape pomace extract rich in natural complex of polyphenols. Materials and Methods. We obtained grape pomace extract, which contains a variety of polyphenolic compounds. Rats of the following groups were used in the experiments: control animals, animals treated with grape pomace extract rich in natural complex of polyphenols for 14 days, animals with streptozotocin-induced diabetes mellitus, and animals with streptozotocin-induced diabetes mellitus treated with grape pomace extract rich in natural complex of polyphenols for 14 days. The number of erythrocytes and reticulocytes, the concentration of hemoglobin and glycated hemoglobin were determined in the peripheral blood of rats. The activities of catalase, superoxide dismutase and the content of thiobarbituric acid-reactive substance and carbonyl groups of proteins were determined in the liver tissues of rats. Results. The study has shown an increase in the number of erythrocytes and the level of hemoglobin, a decrease in the level of glycated hemoglobin and the number of reticulocytes in the peripheral blood of rats after administration of grape pomace extract rich in natural complex of polyphenols to rats with experimental diabetes. A decrease in the content of thiobarbituric acid-reactive substance and the content of carbonyl groups of proteins of neutral and basic character and an increase in the activity of catalase and superoxide dismutase in liver tissues were found under the same conditions. Conclusions. The results indicate that the extract of the natural complex of polyphenols is capable of correcting the morphological and functional state of erythrocytes, as well as improving the activity of antioxidant enzymes and the content of marker molecules of oxidative stress in hepatocytes of rats under experimental diabetes mellitus.

The Inflammatory and Oxidative Status of Newly Diagnosed Class III and Class IV Lupus Nephritis, with Six-Month Follow-Up.

Lupus nephritis (LN) is the most frequent and severe complication of systemic lupus erythematosus (SLE). A prospective cohort with a six-month follow-up was performed. Twelve SLE patients diagnosed with LN Class III, twelve NL Class IV patients, and twelve healthy control subjects (HC) were included. SLE data, renal function, oxidants, antioxidants, and inflammation were determined at baseline and six-month follow-up. During the six-month follow-up, the SLE Disease Activity Index (SLEDAI-2K) decreased in both LN Class III (20.08 ± 6.92 vs. 11.92 ± 5.87, p < 0.001) and LN Class IV (25.33 ± 6.01 vs. 13.83 ± 5.52, p < 0.001) patients. Furthermore, the values of the C4 component also increased during follow-up for LN Class III (25.36 ± 6.34 vs. 30.91 ± 9.22, p = 0.027) and LN Class IV (12.18 ± 3.90 vs. 20.33 ± 8.95, p = 0.008) groups. Regarding inflammation markers, both groups presented decreased C-reactive protein (CRP), but this was only significant for patients with LN class III (7.93 ± 1.77 vs. 4.72 ± 3.23, p = 0.006). Renal function remained stable in both groups, with no changes in eGFR. Patients with LN Class III and Class IV showed higher baseline levels for lipoperoxides (Class III p < 0.01, Class IV p < 0.1) and carbonyl groups in proteins (Class III p < 0.01, Class IV p < 0.1) compared to HC. Moreover, both groups presented lower baseline values of total antioxidant capacity (Class III p < 0.01, Class IV p < 0.1) and catalase (Class III p < 0.01, Class IV p < 0.1) compared to HCs. However, antioxidant and oxidant markers did not show significant differences between baseline values and at six months for either of the two study groups. In conclusion, patients show an imbalance in the oxidative state characterized by the increase in the oxidants LPO and protein carbonyl groups and the decrease in the activity of the antioxidant enzymes TAC and CAT compared to HC. However, the patients did not present an increase in disease activity and renal function improvement. The glomerular filtration rate did not change during the length of the study, and SLEDAI -2K, C3, and C4 improved. The early co-management between Rheumatologists and Nephrologists is essential to prevent the rapid progression of LN. It would be interesting to administer antioxidant supplements to patients with a recent diagnosis of LN and evaluate its effect in a follow-up study.

Effect of different thermal processing methods on sensory, nutritional, physicochemical and structural properties of Penaeus vannamei

The aim of this study was to investigate the effect of different thermal processing methods on the nutritional and physicochemical qualities of Penaeus vannamei. Three different thermal processing methods, namely, drying (DS, 120 °C/40 min), steaming (SS, 100 °C/2 min), and microwaving (MS, 600 W/2 min) were used to treat the shrimps. Low-field nuclear magnetic resonance data indicated that fixed water was the main component of Penaeus vannamei. The ratio of fatty acids in MS and DS samples was more in line with the FAO/WHO recommended health requirements; The myofibrillar protein carbonyl group increased, whereas sulfhydryl content decreased after thermal processing, indicating that the proteins were oxidized by thermal processing. The magnitude of oxidation is: MS > SS > DS. Different thermal processing methods can exert great influence on color texture and nutrition to Penaeus vannamei, which can provide a theoretical knowledge for consumers to choose the appropriate processing method.

The Effect of Piperidine Nitroxides on the Properties of Metalloproteins in Human Red Blood Cells.

Nitroxides are stable, low molecular-weight radicals containing a nitroxide group that has an unpaired electron. The presence of a nitroxide group determines their redox properties. The effect of the piperidine nitroxides, Tempo, Tempol, and Tempamine, on metalloproteins (hemoglobin, superoxide dismutase, catalase) and lactate dehydrogenase in red blood cells was investigated in this research. In addition, the level of lipid peroxidation and the level of protein carbonyl groups were examined as indicators of the effect of oxidative stress. Nitroxides increased superoxide dismutase activity and oxidized hemoglobin to methemoglobin, and also slightly decreased the catalase activity of red blood cells treated with nitroxides. Tempol significantly decreased lactate dehydrogenase activity. All three nitroxides had no effect on membrane lipid peroxidation and protein oxidation. Our results confirm that nitroxides have both antioxidant and prooxidative effects in human red blood cells. The piperidine nitroxides do not initiate the oxidation of proteins and lipids in the membranes of human red blood cells.

Sex-based responses of heat stress and subsequent recovery on the growth performance, metabolic changes, and redox status of broilers at market age.

This experiment investigated the sex responses of heat stress (HS) and subsequent recovery on growth performance, serum metabolic parameters, and redox status. Two hundred 38-day-old broilers were arranged in a completely randomized design with a 2 × 2 (temperatures and sexes) factorial arrangement in five replicates. Broilers were raised at 24 ± 1 °C or 32 ± 1 °C for 3 days and returned to 24 °C for 2 days. The study showed that HS decreased both average daily feed intake (ADFI), average daily gain (ADG), serum total glutathione peroxidase (GPx), and superoxide dismutase (T-SOD). However, it increased feed conversion ratio (FCR), rectal temperature (RT), respiratory rate (RR), serum glucose, blood urea nitrogen (BUN), low-density lipoprotein cholesterol, and the protein carbonyl group (PCG). Male broilers had higher ADFI, ADG, lactic acid (LA), high-density lipoprotein cholesterol (HDL-C), and PCG, but lower FCR, albumin, total antioxidant capacity, T-SOD, and GPx. Temperature and sex significantly interacted with ADFI, ADG, LA, and HDL-C. The effects of HS on RR, RT, glucose, albumin, BUN, PCG, T-SOD, and GPx recovered after 2 days. These results indicate that HS and subsequent recovery affect growth performance, which is accompanied by disturbances in serum nutrient metabolism and abnormalities in redox function and manifested by temporal and gender differences.

The relationship between the cholinergic mechanism of toxicity and oxidative stress in rats during subacute diazinon poisoning

Diazinon is an organophosphate pesticide (OP) that has significant potential for accidental and intentional poisoning of wildlife, domestic animals and humans.The aim of the study is to investigate the correlation between cholinesterase activity and oxidative stress parameters in liver and diaphragm by continuous monitoring as a function of time during prolonged use of diazinon.Wistar rats were treated orally with diazinon (55 mg/kg/day): 7, 14, 21 and 28 days. At the end of each period, blood, liver and diaphragm were collected to examine cholinesterase activity and enzymatic/non-enzymatic oxidative stress parameters: superoxide dismutase 1 (SOD1), catalase (CAT), thiobarbituric acid substances (TBARS), protein carbonyl groups.In all four time periods, there was a significant change in acetylcholinesterase (AChE) in erythrocytes and butyrylcholinesterase (BuChE) in blood plasma, CAT in liver and diaphragm and SOD1 in diaphragm. Parameters significantly altered during the cholinergic crisis included: cholinesterases and TBARS in liver and diaphragm and partially SOD1 in liver. Protein carbonyl groups in liver and diaphragm were significantly altered outside the cholinergic crisis. In the liver, there was a very strong negative correlation between BuChE and TBARS in all four time periods and BuChE and CAT on day 7. In the diaphragm, a very strong negative correlation was found between AChE and TBARS at days 7 and 14, and a very strong positive correlation between AChE and SOD1 at days 14, 21 and 28.A better understanding of the relationship between cholinergic overstimulation and oxidative stress may help to better assess health status in prolonged OPs intoxication.

Metformin and thymoquinone co-treatment enhance 5-fluorouracil cytotoxicity by suppressing the PI3K/mTOR/HIF1α pathway and increasing oxidative stress in colon cancer cells.

This study investigated the chemotherapeutic effects of 5-fluorouracil (5-FU), metformin (Met), and/or thymoquinone (TQ) single/dual/triple therapies in the HT29, SW480 and SW620 colon cancer (CRC) cell lines. Cell cycle/apoptosis were measured by flow cytometry. The gene and protein expression of apoptosis (PCNA/survivin/BAX/Cytochrome-C/Caspase-3) and cell cycle (CCND1/CCND3/p21/p27) molecules, the PI3K/mTOR/HIF1α oncogenic pathway, and glycolysis regulatory enzymes were measured by quantitative-PCR and Western blot. Markers of oxidative stress were also measured by colorimetric assays. Although all treatments induced anti-cancer effects related to cell cycle arrest and apoptosis, the triple therapy showed the highest pro-apoptotic actions that coincided with the lowest expression of CCND1/CCND3/PCNA/survivin and the maximal increases in p21/p27/BAX/Cytochrome-C/Caspase-3 in all cell lines. The triple therapy also revealed the best suppression of the PI3K/mTOR/HIF1α pathway by increasing its endogenous inhibitors (PTEN/AMPKα) in all cell lines. Moreover, the lowest expression of lactate dehydrogenase and pyruvate dehydrogenase kinase-1 with the highest expression of pyruvate dehydrogenase were seen with the triple therapy, which also showed the highest increases in oxidative stress markers (ROS/RNS/MDA/protein carbonyl groups) alongside the lowest antioxidant levels (GSH/CAT) in all cell lines. In conclusion, this is the first study to reveal enhanced anti-cancer effects for metformin/thymoquinone in CRC that were superior to all monotherapies and the other dual therapies. However, the triple therapy approach showed the best tumoricidal actions related to cell cycle arrest and apoptosis in all cell lines, possibly by enhancing oxidative glycolysis and augmenting oxidative stress through stronger modulation of the PI3K/mTOR/HIF1α oncogenic network.

Applicability of 2,4-dinitrophenylhydrazine (DNPH) method of protein oxidative damage measurement in the seminal plasma of canine ( Canis lupus familiaris) and stallion ( Equus caballus).

Seminal plasma (SP) proteins are responsible for sperm functional quality. Developing a reliable method to determine the degree of oxidative damage of these proteins is important for establishing semen fertilizing ability. The main aim of the study was to verify the applicability of protein carbonyl derivatives measurement in the SP of canine and stallion, using a method with 2,4-dinitrophenylhydrazine (DNPH). The research material consisted of ejaculates obtained from eight English Springer Spaniels, and from seven half-blood stallions during the breeding and non-breeding season. The content of carbonyl groups in the SP was measured on the basis of the reactions with DNPH. The following reagent variants were used to dissolve protein precipitates: Variant 1 (V1) - 6M Guanidine solution and Variant 2 (V2) - 0.1M NaOH solution. It has been shown that to obtain reliable results for the measurement of protein carbonylated groups in the dog and horse SP, both 6M Guanidine and 0.1M NaOH may be used. A correlation was also found between the number of carbonyl groups and the total protein content in the canine (V1: r = -0.724; V2: r = -0.847) and stallion (V1: r = -0.336; V2: r = -0.334) SP. Additionally, the study showed a higher content (p≤0.05) of protein carbonyl groups in the stallion SP in the non-breeding season compared to the breeding season. The method based on the reaction with DNPH, due to its simplicity and cost effectiveness, appears to be suitable for large-scale application in the determination of the SP proteins oxidative damage in dog and horse semen.

In Vitro Antiglycation Potential of Erva-Baleeira (Varronia curassavica Jacq.).

Varronia curassavica Jacq. (Boraginaceae) is traditionally used in the treatment of inflammatory processes. The ethanolic extract of its leaves (EEVc) showed anti-inflammatory properties and low toxicity. Medicinal plants have aroused interest for their antiglycation activities. The formation and accumulation of advanced glycation end products (AGEs) are associated with several chronic diseases. The objective of this study was to evaluate the antiglycation potential of EEVc and two isolated compounds. The compounds brickellin and cordialin A were obtained by chromatographic methods and identified by spectrometric techniques. Analysis of fluorescent AGEs, biomarkers of amino acid residue oxidation, protein carbonyl groups and crosslink formation were performed in samples obtained from an in vitro model system of protein glycation with methylglyoxal. EEVc, brickellin and cordialin A significantly reduced the in vitro formation of AGEs, and reduced the damage caused by oxidative damage to the protein. According to the results, EEVc, brickellin and cordialin A are potential candidates against AGEs formation, which opens the way to expand the therapeutic arsenal for many pathologies resulting from glycoxidative stress.

Quercetin and Its Derivative Counteract Palmitate-Dependent Lipotoxicity by Inhibiting Oxidative Stress and Inflammation in Cardiomyocytes.

Cardiac lipotoxicity plays an important role in the pathogenesis of obesity-related cardiovascular disease. The flavonoid quercetin (QUE), a nutraceutical compound that is abundant in the "Mediterranean diet", has been shown to be a potential therapeutic agent in cardiac and metabolic diseases. Here, we investigated the beneficial role of QUE and its derivative Q2, which demonstrates improved bioavailability and chemical stability, in cardiac lipotoxicity. To this end, H9c2 cardiomyocytes were pre-treated with QUE or Q2 and then exposed to palmitate (PA) to recapitulate the cardiac lipotoxicity occurring in obesity. Our results showed that both QUE and Q2 significantly attenuated PA-dependent cell death, although QUE was effective at a lower concentration (50 nM) when compared with Q2 (250 nM). QUE decreased the release of lactate dehydrogenase (LDH), an important indicator of cytotoxicity, and the accumulation of intracellular lipid droplets triggered by PA. On the other hand, QUE protected cardiomyocytes from PA-induced oxidative stress by counteracting the formation of malondialdehyde (MDA) and protein carbonyl groups (which are indicators of lipid peroxidation and protein oxidation, respectively) and intracellular ROS generation, and by improving the enzymatic activities of catalase and superoxide dismutase (SOD). Pre-treatment with QUE also significantly attenuated the inflammatory response induced by PA by reducing the release of key proinflammatory cytokines (IL-1β and TNF-α). Similar to QUE, Q2 (250 nM) also significantly counteracted the PA-provoked increase in intracellular lipid droplets, LDH, and MDA, improving SOD activity and decreasing the release of IL-1β and TNF-α. These results suggest that QUE and Q2 could be considered potential therapeutics for the treatment of the cardiac lipotoxicity that occurs in obesity and metabolic diseases.

Consequences of prenatal exposure to contaminants in elasmobranchs: Biochemical outcomes during the embryonic development of Pseudobatos horkelii

Coastal elasmobranchs are vulnerable to chemicals mostly due to their k-strategic life history characteristics and high trophic positions. Embryos might be particularly exposed through the maternal offloading of contaminants, possibly leading to disruptions during critical developmental phases. Yet, knowledge on biochemical outcomes of prenatal exposure in elasmobranchs is notably limited. Therefore, we aimed to investigate the effects of prenatal exposure to contaminants in embryos of the critically endangered Brazilian guitarfish, Pseudobatos horkelii. Polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), pharmaceuticals and personal care products, and metals were determined in embryos. Additionally, glutathione S-transferase activity (GST), glutathione (GSH), and metallothionein levels (MT) were analyzed. Finally, lipid peroxidation levels (LPO) and protein carbonyl groups (PCO) were assessed. Embryonic exposure depended on yolk consumption, which was conspicuous in earlier development. We observed a dilution effect of contaminants levels, potentially related to biotransformation of these compounds throughout the embryonic development. Nevertheless, GST was not correlated to contaminant concentrations. The multivariate relationship between antioxidant components (GSH and GST) and LPO and PCO was negative, suggesting the lack of efficient defense of these biomarkers in early development, leading to oxidative damage. In this context, our results indicate that prenatal exposure to contaminants might impact the redox status in embryos of P. horkelii, leading to oxidative damage. Furthermore, metal concentrations influenced MT levels, suggesting this as a potential detoxification pathway in this species.

Increased survival of honey bees consuming pollen and beebread is associated with elevated biomarkers of oxidative stress

IntroductionSignificant losses of honey bee colonies have been observed worldwide in recent decades. Inadequate nutrition is considered to be one of the factors that can reduce honey bee resistance to abiotic and biotic environmental stresses. Accordingly, we assessed the impact of food composition on worker bee survival.MethodsBees in cages were fed six different diets, and then their survival, levels of thiobarbituric acid reactive substances and protein carbonyl groups, catalase and lysozyme activities were evaluated.Results and DiscussionAfter 17 days of feeding, the lowest mortality was observed in the group of bees that received sucrose solution with the addition of willow pollen or artificial rapeseed beebread or artificial willow beebread (diets 4–6). The highest mortality was found in bees that consumed only sucrose solution (diet 1) or the sucrose solution supplemented with a mixture of amino acids (diet 2), which can be explained by the lack of vitamins and microelements in these diets. In the group of bees that received the sucrose solution with rapeseed pollen (diet 3), mortality was intermediate. To check whether the decrease in insect survival could be related to oxidative damage, we evaluated biomarkers of oxidative stress. Consumption of pollen (diets 3 and 5) and artificial beebread (diets 4 and 6) enhances protein carbonylation in worker bees. Feeding bees artificial beebread also resulted in increase in lipid peroxidation and catalase activity, which is probably due to the presence of hydrogen peroxide in the honey contained in beebread. Remarkably, the increase in biomarkers of oxidative stress was not accompanied by adverse but positive effects on insect survival. A lack of amino acids and proteins in the diet 1 did not cause oxidative stress, but led to an increase in lysozyme activity in hemolymph, a biomarker of immune system status. In conclusion, we believe that the increase in oxidative stress biomarkers we found do not indicate oxidative damage, but rather reflect the changes in redox balance due to consumption of certain dietary options.