A severidade da cárie na primeira infância aumenta atividade da SOD sensível ao KCN, reduz proteínas carboniladas e aumenta a biodisponibilidade do óxido nítrico na saliva / The severity of caries increases the activity of SOD sens KCN, reduces carbonylated proteins and increases the bioavailability of nitric oxide in saliva

O objetivo deste estudo foi avaliar se a progressão da severidade das lesões de cárie alteraria as atividades das isoformas da superóxido dismutase (SOD), sensível ou não ao cianeto de potássio (KCN), a carbonilação de proteínas e a biodisponibilidade do óxido nítrico (NO) salivar. Amostras de saliva, não estimulada, foram coletadas de crianças (1 a 3 anos) em uma creche municipal de Birigui, SP, Brasil. As crianças foram divididas em 4 grupos (n=30/grupo) de acordo com os critérios do ICCMSTM, sendo o grupo A (livres de cárie), grupo B (cárie inicial), grupo C (cárie moderada) e grupo D (cárie extensa). A atividade enzimática da SOD sensível ao KCN e da SOD insensível ao KCN (SOD sens KCN e SOD ins KCN, respectivamente) foram determinadas pelo método de oxidação do pirogalol. O método da carbonilação da proteína foi utilizado para determinar o dano oxidativo (DO), baseado no método alcalino de DNPH (2,4- Dinitrofenilhidrazina). A biodisponibilidade de NO salivar foi quantificada pelo método de Griess. Os dados foram analisados pelo teste de ANOVA e pós-teste de StudentNewman-Keuls (α = 0,05), e pela correlação de Pearson e Spearman. Os resultados demostraram que a progressão das lesões de cárie aumentou a atividade da SOD sens KCN, mas não alterou a atividade da SOD ins KCN, reduziu a carbonilação de proteína, e aumentou a biodisponibilidade do NO salivar. Em conclusão, os dados deste estudo mostraram que a progressão da cárie aumenta a atividade da SOD sens KCN que, possivelmente, reduz o dano oxidativo e favorece a biodisponibilidade de NO salivar. Embora exista um aumento da defesa antioxidante enzimática e da biodisponibilidade de NO, a progressão da cárie nestas condições se daria quando os fatores extrínsecos estiverem prevalecendo aos fatores de defesa antioxidante da saliva e ação antibacteriana do NO. Com isso, podemos reforçar a importância do acompanhamento odontológico preventivo da cárie desde a primeira infância para que sejam reduzidos os fatores de risco da cárie e prevaleça a defesa antioxidante salivar e ação antibacteriana do NO(AU)

The aim of this study was to assess whether caries progression alters the activity of superoxide dismutase (SOD) isoforms, protein carbonylation (a biomarker of oxidative damage) and the bioavailability of nitric oxide (NO) in children's saliva. Saliva samples, not stimulated, were collected from children (1 to 3 years old) in a municipal day care center in Birigui, SP, Brazil. The children were divided into 4 groups (n=30/group), according to the ICCMSTM criteria, being group A (caries free), group B (initial caries), group C (moderate caries) and group D (extensive caries). The enzymatic activity of potassium cyanide (KCN)- sensitive superoxide dismutase (SOD) and KCN-insensitive SOD (SOD sens KCN and SOD ins KCN, respectively) were determined by the pyrogallol oxidation method. The protein carbonylation method based on the alkaline method of DNPH (2,4- Dinitrophenylhydrazine) was used to determine oxidative damage (OD). Salivary nitric oxide (NO) bioavailability was quantified by the Griess method. Data were analyzed by ANOVA and Student-Newman-Keuls post-test (α = 0.05), and by Pearson and Spearman correlation. The results showed that with the progression of caries, there was an increase in SOD sens KCN activity, with no change in SOD ins KCN activity, a reduction in protein carbonylation, and an increase in salivary NO bioavailability. We can conclude that the greater the severity of caries, the greater the activity of SOD sens KCN, which leads to less oxidative damage and greater bioavailability of NO. However, it is possible that in the stages of greater caries severity, the antibacterial action of NO is superimposed by external cariogenic factors, favoring the process of demineralization and enamel and dentin damage. Thereby, we understand that the clinical dental care of children in early childhood is a key factor for controlling cariogenic factors and keeping in balance with the antioxidant defense and antibacterial action of NO in saliva(AU)

Effect of air pollution on carbonylated protein level in the stratum corneum and protective effect of pink pepper tree extracts and lipid mixtures on skin damage / 中华皮肤科杂志

Objective To evaluate the effect of air pollution on carbonylated protein level in the stratum corneum,and to assess the protective effect of pink pepper tree extracts and lipid mixtures on skin damage.Methods After the investigation of influencing factors in the preliminary experiment,fluorescence labelling assay was performed to detect the carbonylated protein level in the skin stratum corneum at different body sites of 34 healthy testees.Cigarette smoke was used to simulate pollutants,and the forearms of 15 healthy testees were exposed in the customized pollution simulation chamber with the flexor aspects facing upwards.After 0,1,2,4,5 hours of exposure,stratum corneum samples were collected by using D-squame tapes.In each of 14 selected healthy testees,3 adjacent areas on the flexor aspect of unilateral forearm were divided into 3 groups:pink pepper tree group treated with 1％ aqueous solution of pink pepper tree extracts,control group treated with deionized water,and blank group receiving no treatment.Then,the forearms of the 14 testees were exposed in the pollution simulation chamber for 5 hours,and stratum corneum samples were collected from the 3 areas before and after the exposure.Another 16 healthy testees were included,and 3 adjacent areas on the flexor aspect of their unilateral forearms were divided into 3 groups:lipid mixture group treated with 5％ lipid mixture emulsion,control group treated with lipid mixture-free placebo emulsion,and blank group receiving no treatment.Then,the forearms of the 16 testees were exposed in the pollution simulation chamber for 5 hours,and stratum corneum samples were collected from the 3 areas before and after the exposure.Moreover,twenty healthy testees were enrolled into the double-blind split-face clinical trial.That is,one half of the face was randomly chosen to be treated with 1％ emulsion of pink pepper tree extracts,and the other facial side was treated with placebo emulsion.Before and after 56-day treatment,stratum corneum samples were collected from the cheeks of testees by using D-squame tapes.Fluorescence labelling assay was conducted to detect the carbonylated protein level in the above stratum corneum samples.Results The analysis of 34 testees showed that carbonylated protein levels (average fluorescence intensity) significantly differed among different body sites (P ＜0.001),and the carbonylated protein levels were significantly higher in the cheeks (26.3 ± 7.1) and forehead (22.9 ± 7.9) than in the forearm (14.7 ± 4.9) and waist and back (12.6 ± 4.2) (P ＜ 0.001),and higher in the forearm than in the waist and back (P =0.046).In the short-term simulated accelerated exposure experiment,the carbonylated protein level increased along with the increase of the duration of exposure to pollution (R2 =0.995 9).After 5-hour exposure,the pink pepper tree group and lipid mixture group both showed significantly lower elevated levels of carbonylated protein in the stratum corneum (9.7 ± 5.2,5.8 ± 4.9) compared with the corresponding blank groups (19.0 ± 10.0,17.4 ± 8.8;P ＜ 0.005) and control groups (18.5 ± 7.3,15.9 ± 6.4;P ＜ 0.005).In the long-term human trial,the carbonylated protein levels significantly decreased in the facial side treated with 1％ emulsion of pink pepper tree extracts for 8 weeks compared with the placebo-treated facial side.Conclusion Air pollution aggravates skin damage induced by protein carbonylation in the stratum corneum,and pink pepper tree extracts and lipid mixtures can effectively reduce the carbonylated protein level.

Oxidation as an important factor of protein damage: Implications for Maillard reaction.

Protein oxidation, the process caused especially by reactive oxygen and nitrogen species, is thought to play a major role in various oxidative processes within cells and is implicated in the development of many human diseases. This review provides a brief overview of the protein oxidation with the emphasis on the types of oxidation (oxidation of protein backbone and amino acid residues side chains, site-specific metal-catalysed protein oxidation), oxidationdependent generation of protein hydroperoxides, carbonyl derivatives and protein–protein cross-linkages. Nonenzymatic glycoxidation (also known as Maillard reaction) as an important factor of protein damage, consequences of oxidative protein impairment and related diseases as well as means of monitoring and assessment of protein modifications are discussed.

The effect and mechanisms of 20-HETE on myocardial ischemia reperfusion injury / 重庆医学

Objective To investigate the effect of 20‐HETE on the isolated myocardial ischemia reperfusion injury and to ex‐plore its underlying mechanisms .Methods Experiments were performed in isolated rat hearts subjected to 35 min of ischemia fol‐lowed by 40 min of reperfusion in Langendorff preparations .HET0016 (1 μmol/L) and various concentrations (10 ,30 or 50 nmol/L) of 20‐HETE were infused 10 min before the onset of ischemia and throughout the reperfusion period .Cardiac hemodynamic changes and myocardial contractility were continuously recorded with the Powerlab /8P system .Myocardial infarct size was meas‐ured by TTC staining .The level of ROS and the protein carbonyl content were determined by DHE fuorescence and DNPH method , respectively .Results Perfusion with HET0016 significantly improved myocardial ischemia reperfusion injury reduction in cardiac contractility ,after inhibited the production of 20‐HETE significantly reduced the occurrence of myocardial infarction area (P<0 .05) ,but exogenous join 20‐HETE aggravated I/R‐induced myocardial injury (P<0 .05) .Myocardial ischemia reperfusion injury significantly increased production of ROS and oxidative stress ,both of which were significantly inhibited by HET 0016 and enhanced by 20‐HETE administration(P< 0 .05) .Conclusion 20‐HETE stimulates ROS production and enhance protein carbonylation , which aggravates myocardial ischemia reperfusion injury .

Blood carbonyl protein measurement as a specific oxidative stress biomarker after intestinal reperfusion in rats / Dosagem da proteína carbonilada sanguínea como biomarcador específico do estresse oxidativo após reperfusão intestinal em ratos

PURPOSE: An experimental study was performed to investigate the use of protein carbonyl group as a specific biological marker for oxidative stress in a rat model of intestinal ischaemia-reperfusion. METHODS: Twenty four male Wistar rats were randomly distributed into three groups with eight animals each: Group 1 - Control group; Group 2 - Sham; Group 3 - Intestinal ischaemia by clamping ileal branches of the superior mesenteric artery for one hour, followed by another hour of reperfusion. Blood samples were taken in order to analyze the protein carbonyl level by Slot blotting assay. RESULTS: In group 3 a significant increase of protein carbonyl level was observed if compared to the homogenous levels of groups 1 and 2. CONCLUSION: From the results it may be concluded that the protein carbonylation may be used as a specific marker for measuring oxidative stress in rat intestinal reperfusion model.

OBJETIVO: Realizou-se um estudo experimental com a finalidade de investigar o uso da proteína carbonilada como um marcador biológico específico do estresse oxidativo em um modelo de isquemia e reperfusão intestinal, em ratos. MÉTODOS: Vinte e quarto ratos da linhagem Wistar, machos foram distribuídos, aleatoriamente, em três grupos compostos por oito animais cada: Grupo 1 - Controle; Grupo 2 - Simulação e Grupo 3 - Submetido à isquemia, mediante clampeamento de ramos ileais da artéria mesentérica superior por uma hora, seguida de reperfusão, por igual período. Amostras sanguíneas obtidas foram utilizadas para analise dos níveis de proteína carbonilada, através do método Slot blotting. RESULTADOS: No grupo 3 houve uma elevação significante da concentração de proteína carbonilada sérica se comparada aos níveis sanguíneos homogêneos encontrados nos grupos 1 e 2. CONCLUSÃO: Fundamentado nos resultados é possível concluir que, a carbonilação protéica pode ser utilizada como um marcador específico para a mensuração do estresse oxidativo em modelos de reperfusão intestinal, em ratos.

Propofol and N-Acetylcysteine attenuate oxidative stress induced by intestinal ischemia/reperfusion in rats: Protein carbonyl detection by immunoblotting / Propofol e N-Acetilcisteína atenuam o estresse oxidativo induzido pela isquemia/reperfusão intestinal em ratos: Detecção de proteína carbonilada por immunoblotting

PURPOSE: To evaluate the antioxidant effect of Propofol and N-Acetylcysteine (NAC) on intestinal ischemia/reperfusion (I/R) in rats by determining carbonyl protein level. METHODS: Forty Wistar rats were randomly assigned into the following groups: Control; Sham; I/R with Propofol; I/R with Propofol and NAC; I/R with Ketamine and Xylazine. The I/R groups underwent 60 minutes of ischemia and an equal period of reperfusion. Blood samples, collected by cardiac punction, were centrifuged for plasma obtainment. Protein carbonyl level in plasma samples was determined by immunoblotting. RESULTS: No significant difference in protein carbonyl level was found between Control and Sham groups (P>0.05). The highest reduction in protein carbonyl level (P<0.05) was obtained with the administration of Propofol and NAC (Group 4) in intestinal I/R procedure. CONCLUSION: The administration of Propofol and NAC showed the best antioxidant effect on oxidative stress in rats that underwent intestinal I/R procedure, suggesting a synergistic interaction.

OBJETIVO: Avaliar o efeito antioxidante do Propofol e N-Acetilcisteína (NAC) na isquemia/reperfusão (I/R) intestinal em ratos através da determinação do nível de proteína carbonilada. MÉTODOS: 40 ratos Wistar foram aleatoriamente distribuídos nos seguintes grupos: Controle; Sham; I/R com Propofol; I/R com Propofol e NAC; I/R com Ketamina e Xilazina. Os grupos I/R foram submetidos à isquemia durante 60 minutos e à reperfusão por igual período de tempo. Amostras de sangue, coletadas por punção cardíaca, foram centrifugadas para a obtenção de plasma. O nível de proteína carbonilada nas amostras de plasma foi determinado por imunoblotting. RESULTADOS: Nenhuma diferença significativa foi encontrada entre os grupos Controle e Sham (P>0.05). Uma redução marcante no nível de proteína carbonilada (P<0.05) foi obtida com a administração combinada de Propofol e NAC (Grupo 4) durante o procedimento de I/R intestinal, comparando-se com os demais grupos I/R testados. CONCLUSÃO: A administração de Propofol e NAC apresentou o melhor efeito antioxidante sobre o estresse oxidativo em ratos submetidos ao procedimento de I/R intestinal, sugerindo-se uma interação sinergística.

Relationships among Oxidative Stress Markers, Life Style Factors and Biochemical Findings / 대한진단검사의학회지

BACKGROUND: In many studies, oxidative stress markers have been employed to serve as a measure of a disease process or to reflect oxidative status. These oxidative stress markers must have some degree of predictive validity, but full substantiation of this relation is still lacking. This paper presents data on levels of three biomarkers, oxidized low-density lipoproteins (LDL), carbonyl, and 8-hydroxy-2'-deoxyguanosine (8-OHdG), and a number of life style factors associated with oxidative stress in healthy adults. METHODS: For 237 healthy adults aged 40-60 years, a number of life style factors, biochemical characteristics and oxidative status were evaluated. Markers of oxidative stress were measured by an ELISA method. RESULTS: Waist-hip ratio and use of vitamin supplement were associated with serum oxidized LDL (P<0.05). Body mass index and stress had a relationship (P<0.05) with protein carbonyl. Creactive protein was related to serum oxidized LDL (P<0.01). There was no correlation among three oxidative stress markers, oxidized LDL, carbonyl, and 8-OHdG. CONCLUSIONS: The oxidative stress markers used in this study could not be regarded as a general estimate of the healthy individual oxidative status. Further studies focusing on the development of biomarkers to reflect changes in the oxidative status under normal, non-pathological conditions in humans will be required.