Oxidative stress in maternal milk and cord blood in gestational diabetes mellitus: a prospective study

ABSTRACT BACKGROUND: Reduced antioxidant defenses may reflect a poor protective response against oxidative stress and this may be implicated in progression of gestational diabetes mellitus (GDM). Oxidative stress induced by hyperglycemia plays a major role in micro and macrovascular complications, which imply endothelial dysfunction. OBJECTIVE: Our aim in this study was to investigate the association between GDM and oxidative stress markers measured in plasma, with regard to revealing changes to total antioxidant capacity (TAC) and total oxidant status (TOS) among mothers showing impairments in oral glucose tolerance tests (OGTTs). DESIGN AND SETTING: Prospective study at a university hospital in Turkey. METHODS: The study group consisted of 50 mothers with GDM, and 59 healthy mothers served as controls. Umbilical cord blood samples were taken from all mothers during delivery and breast milk samples on the fifth day after delivery. TAC, TOS, thiol and disulfide levels were measured. RESULTS: No statistically significant relationship between the blood and milk samples could be found. An analysis on correlations between TAC, TOS and certain parameters revealed that there were negative correlations between TOS and total thiol (r = -0.386; P < 0.001) and between TOS and disulfide (r = -0.388; P < 0.001) in milk in the control group. However, these findings were not observed in the study group. CONCLUSION: Our findings suggested that a compensatory mechanism of oxidative stress was expected to be present in gestational diabetes mellitus and that this might be ameliorated through good glycemic regulation and antioxidant supplementation.

Análise dos mecanismos pró-inflamatórios do ácido úrico solúvel: efeito sobre proteínas sensíveis à modulação redox e sobre a polarização de células THP-1 / Analysis of the pro-inflammatory mechanisms of soluble uric acid: effect on redox sensitive proteins and THP-1 cell polarization

Vários estudos epidemiológicos estabelecem correlação positiva entre os níveis de ácido úrico sérico e o aumento do risco para doenças cardiovasculares. Fatores dietéticos e socioeconômicos, além da presença de comorbidades estão diretamente associados aos níveis séricos de ácido úrico. Países desenvolvidos apresentam maior incidência e prevalência da gota e alguns grupos étnicos são particularmente susceptíveis à hiperuricemia. Cristais de ácido úrico são descritos por iniciar e perpetuar resposta inflamatória, e sinalizar um padrão de resposta molecular associado ao dano (DAMP), permitindo a diferenciação de macrófagos para perfis pró-inflamatórios. Por outro lado, os efeitos do ácido úrico em sua forma solúvel ainda carecem de estudos. Macrófagos derivados de precursores monocíticos apresentam diferenciação específica e respondem a um conjunto de fatores extrínsecos, resultando em perfis distintos, um fenômeno conhecido como polarização. Assim, os macrófagos podem ser classicamente ativados para uma resposta Th1 (T helper 1) e polarizados a um perfil pró- inflamatório (M1, resposta Th1) ou a um perfil alternativo e oposto, um perfil de resolução da inflamação (M2, resposta Th2, T helper 2). Nesse sentindo, buscamos analisar os efeitos do ácido úrico solúvel sobre vias de modulação da polarização fenotípica de macrófagos e modificação redox. Utilizamos a linhagem monocítica humana THP-1, a qual foi diferenciada em macrófagossímile por acetato miristato de forbol (PMA; 5 ng.mL-1) por 48 h, seguidas da incubação com ácido úrico em meio ausente de tióis e soro fetal bovino por 8h ou 24h (0-1000 µM). A expressão de fatores de transcrição e marcadores de polarização foi realizada através de citometria de fluxo, western-blotting e por microscopia de fluorescência com alto conteúdo de imagens (HCI). Em concentrações fisiológicas, verificamos que o ácido úrico solúvel regulou positivamente a frequência de células para receptor manose CD206, um marcador clássico de perfil alternativo/M2 e regulou negativamente a expressão óxido nítrico sintase induzível (iNOS), um marcador M1, sugerindo inicialmente uma modulação para o perfil de polarização M2. Além disso, as proteínas redoxsensíveis, heme oxigenase-1 (HO-1) e tiorredoxina (Trx) tiveram sua expressão reduzida e aumentada, respectivamente, pelo tratamento com ácido úrico. Os fatores de transcrição Nrf2 e STAT3 tiveram regulação negativa após a exposição ao ácido úrico solúvel. Os resultados apresentados nesta tese sugerem uma função do urato no priming de macrófagos através da alteração da polarização destas células

Several epidemiological studies have established a positive correlation between high serum uric acid levels and increased risk for cardiovascular diseases. Developed countries have a higher incidence and prevalence of gout and some ethnic groups are particularly susceptible to hyperuricemia. Although hyperuricemia is a prevalent condition, it has still controversy biological consequences. Uric acid crystals are described as capable of initiating and perpetuating inflammatory responses, by activating the damage-associated molecular response pattern (DAMP) cascade, allowing macrophage differentiation to inflammatory profiles. In spite of that, biological response to soluble uric acid are not completely understood. Monocyte-derived macrophages respond to a set of extrinsic factors that result in different profiles and can be polarized to a proinflammatory (M1) or anti-inflammatory (M2) profile. In this thesis, we analyzed the effects of soluble uric acid on redox-modulated pathways and the phenotypic polarization of macrophages. We used human monocytic THP-1 cell line, differentiated into macrophage by phorbol myristate acetate (PMA; 5 ng.mL-1) for 48 h. After differentiation, cells were incubated with soluble uric acid in medium without thiols and fetal bovine serum for 8 h and 24 h (0-1000 µM). The expression of transcription factors and polarization markers were assessed by flow cytometry, western-blotting and fluorescence microscopy with high content imaging (HCI). At physiological concentrations, soluble uric acid positively regulated the frequency of cells for mannose receptor CD206, a classic marker of the anti-inflammatory M2 profile and negatively regulated the inducible nitric oxide synthase (iNOS) expression, a proinflammatory M1 marker, suggesting that the soluble uric acid changes the polarization profile to M2 profile. In addition, the redox-sensitive proteins heme oxygenase-1 (HO-1) and thioredoxin (Trx) had their expression decreased and increased, respectively, after exposure to urate. STAT3 and Nrf2 transcription factors were downregulated upon soluble uric acid exposure. The results presented in this thesis suggest a role of uric acid in macrophage priming through the alteration of cell polarization

Associação entre Parâmetros Plasmático de Tiol e Níveis de Troponina em Pacientes com Síndrome Coronariana Aguda e Predição de Arritmia Ventricular Hospitalar / Association between Plasma Thiol Parameters and Troponin Levels in Patients with Acute Coronary Syndrome and Prediction of In-Hospital Ventricular Arrhythmia

Resumo Fundamento As arritmias ventriculares (AVs) são a principal causa de mortalidade e morbidade hospitalar em pacientes com síndrome coronariana aguda (SCA) e sua relação com o tiol é desconhecida. Objetivo Investigar a relação entre os níveis plasmáticos de tióis e os níveis de troponina em pacientes com SCA e estimar o desenvolvimento de AV intra-hospitalar durante a internação. Método O estudo incluiu 231 pacientes consecutivos com SCA com supradesnivelamento do segmento ST (SCA-SDST) e pacientes com SCA sem supradesnivelamento do segmento ST (SCA-SSDST). Após a aplicação dos critérios de exclusão, 191 pacientes foram incluídos na análise estatística. Os pacientes foram classificados em dois grupos: grupo SCA-SDST (n=94) e grupo SCA-SSDST (n=97). Os níveis plasmáticos de tiol, dissulfeto e troponina foram medidos e a razão de troponina para tiol nativo (RTTN) foi calculada. Considerou-se estatisticamente significativo um valor de p bilateral inferior a 0,05. Resultados Tiol nativo plasmático, tiol total, dissulfeto e suas razões foram semelhantes entre os grupos. A RTTN se mostrou significativamente maior no grupo SCA-SDST em comparação com o grupo SCA-SSDST. Houve correlação negativa significativa entre os níveis de troponina e tiol. Verificou-se que o tiol nativo é preditor independente do desenvolvimento de AV em pacientes com SCA-SDST e em todos os pacientes com SCA. Verificou-se que o RTTN é preditor independente do desenvolvimento de AV em pacientes com SCA-SSDST e em todos os pacientes com SCA. Conclusão Os níveis plasmáticos de tiol podem ser usados para identificar pacientes com alto risco de desenvolvimento de AV intra-hospitalar em pacientes com SCA. A correlação entre os níveis de troponina e tiol pode sugerir que os tióis possam ser marcadores importantes para o diagnóstico e prognóstico da SCA com a ajuda de estudos futuros.

Abstract Background Ventricular arrhythmias (VAs) are the main cause of in-hospital mortality and morbidity in acute coronary syndrome (ACS) patients and its relationship with thiol is not known. Objective To investigate the relationship between plasma thiol levels and troponin levels in patients with ACS and to estimate in-hospital VA development during hospital stay. Method The study included 231 consecutive ST-segment elevation ACS (STE-ACS) and non-ST-segment elevation ACS (NSTE-ACS) patients. After application of exclusion criteria, 191 patients were included in the statistical analysis. Patients were classified into two groups: STE-ACS group (n=94) and NSTE-ACS group (n=97). Plasma thiol, disulphide and troponin levels were measured and troponin-to-native thiol ratio (TNTR) was calculated. A two-sided p value of less than 0.05 was considered to be statistically significant. Results Plasma native thiol, total thiol, disulphide and their ratios were similar between the groups. TNTR was significantly higher in the STE-ACS group compared to the NSTE-ACS group. Troponin and thiol levels correlated negatively and significantly. Native thiol was found to be an independent predictor of VA development in STE-ACS patients and in all ACS patients. TNTR was found to be an independent predictor of VA development in NSTE-ACS patients and in all ACS patients. Conclusion Plasma thiol levels can be used to identify ACS patients at high risk for in-hospital VA development. Correlation between troponin and thiol levels may suggest that thiols may be an important marker for diagnosis and prognosis of ACS with the help of future studies.

Níveis de Tiol Sérico e Homeostase Tiol/Dissulfeto em Pacientes com Doença Valvar Mitral Reumatismal e em Sujeitos Saudáveis / Serum Thiol Levels and Thiol/Disulfide Homeostasis in Patients with Rheumatic Mitral Valve Disease and Healthy Subjects

Resumo Fundamento A doença valvar mitral reumatismal (DVMR) é a apresentação mais comum das doenças cardíacas reumáticas (DCR). Os processos de inflamação e fibrose também têm papéis significativos em sua patogênese. Estudos recentes demonstram que os tióis e o tiol-dissulfeto são marcadores de stress oxidativo inéditos e promissores. Objetivos O objetivo deste estudo foi avaliar diferenças entre os níveis de tiol sérico e de tiol-dissulfeto em pacientes com DVMR e no grupo de controle. Métodos Noventa e dois pacientes com DVMR foram cadastrados no estudo. Cinquenta e quatro sujeitos saudáveis, e com correspondência de sexo e idade em relação ao grupo de estudo, também foram incluídos no estudo como um grupo de controle. Foram investigados os níveis de tiol nos pacientes com DVMR e o grupo de controle. Os p-valores menores que 0,05 foram considerados estatisticamente significativos. Resultados Os pacientes com DVMR apresentaram pressão sistólica da artéria pulmonar (PSAP) e níveis de diâmetro do átrio esquerdo (AE) mais altos que os do grupo de controle. Os níveis de tiol nativo (407±83 μmol/L vs. 297±65 μmol/L, p<0,001) e tiol total (442±82 μmol/L vs. 329±65 μmol/L, p<0,001) são mais altos no grupo de controle. Níveis de dissulfeto (16,7±4,9 μmol/L vs. 14,8±3,7 μmol/L, p=0,011) são mais altos no grupo de pacientes com DVMR. Foi identificada uma correlação positiva entre as razões dissulfeto/tiol nativo e dissulfeto/tiol total com PSAP, diâmetro de AE, e gravidade da EMi. A razão dissulfeto/tiol total é significativamente mais alta em pacientes com EMi grave que em pacientes com EMi leve a moderada. Conclusões Até onde se sabe, este é o único estudo que avaliou a homeostase tiol/dissulfeto como um preditor inédito, que está relacionado de forma mais próxima à DVMR e à gravidade da EMi.

Abstract Background Rheumatic mitral valve disease (RMVD) is the most common presentation of rheumatic heart disease (RHD). Inflammation and fibrosis processes also play significant roles in its pathogenesis. Recent studies showed that thiols and thiol-disulfide are promising novel oxidative stress markers. Objectives The present study aimed to evaluate differences in the serum thiol and thiol-disulfide levels in patients with RMVD and the control group. Methods Ninety-two patients with RMVD were enrolled in the study. Fifty-four healthy subjects, age, and gender-matched with the study group, were also included in the study as a control group. This study investigated thiol levels in patients with RMVD and the control group. P-values lower than 0.05 were considered statistically significant. Results The patients with RMVD presented higher systolic pulmonary artery pressure (SPAP) and left atrial (LA) diameter levels than the control group. Native thiol (407±83 μmol/L vs. 297±65 μmol/L, p<0.001) and total thiol (442±82 μmol/L vs. 329±65 μmol/L, p<0.001) levels were higher in the control group. Disulfide (16.7±4.9 μmol/L vs. 14.8±3.7 μmol/L, p=0.011) levels were higher in the group of patients with RMVD. A positive correlation was found between disulfide/native and disulfide/total thiols ratio with SPAP, LA diameter, and MS severity. Disulfide/total thiols ratio was significantly higher in patients with severe MS than with mild to moderate MS patients. Conclusions To the best of our knowledge, this is the only study of its kind that has evaluated thiol/disulfide homeostasis as a novel predictor, which was more closely related to RMVD and the severity of MS.

An evaluation of the relationship between vitamin D level and CTRP-9, tumor necrosis factor-alpha, thiol-disulfide hemostasis in women

SUMMARY OBJECTIVE: Many chronic diseases such as malignancy, cardiovascular diseases, endothelial dysfunction, and autoimmune diseases, which have been shown to be related to vitamin D in various studies; have similar relations with CTRP-9, TNFα, and thiol-disulfide hemostasis. We aimed to contribute to the literature by evaluating the relationship between CTRP-9, TNFα, and thiol-disulfide hemostasis and vitamin D levels, which we thought may have some effects on the pathogenesis of vitamin D deficiency. METHODS: In our study, 78 female volunteers older than 18 years were included. Volunteers were divided into three groups according to the reference values of vitamin D levels. Biochemical parameters, CTRP-9, TNFα, and thiol/disulfide hemostasis tests taken from all volunteers were studied. RESULTS: In this study, there was a significant difference in CTRP-9, TNFα, total thiol (TT), native thiol (NT), DIS (disulfide), TT/DIS, and NT/DIS levels in vitamin D groups (p<0.05). There was a significant negative correlation between vitamin D and TNFα and DIS, while a significant positive correlation was found with CTRP-9, TT, NT, TT/DIS, and NT/DIS (p<0.05). CONCLUSIONS: It was determined that vitamin D deficiency causes a significant decrease in CTRP-9 level and a significant increase in TNFα level, as well as an increase in thiol/disulfide hemostasis in favor of disulfide, which may be a risk factor for increased oxidative stress. We considered that these changes may play mediator roles for many chronic diseases and metabolic disorders that are increasing in frequency due to vitamin D deficiency.

The effect of low dose ionizing radiation exposure on dynamic thiol-disulfide homeostasis and ischemia modified albumin levels: an observational study / Efeito da exposição à radiação ionizante de baixa dose na homeostase dinâmica de tiol-dissulfeto e níveis de albumina modificada por isquemia: estudo observacional

Abstract Background: The primary objective of this study was to investigate the effect of low dose ionizing radiation exposure on thiol/disulfide homeostasis and ischemia modified albumin levels. The secondary objective is to compare thiol/disulfide homeostasis and ischemia modified albumin levels among the personnel exposed to low dose ionizing radiation in anesthesia application areas, in and out of the operation room. Methods: The study included a total of 90 volunteers aged between 18 and 65 years old, with 45 personnel working in a setting with potential for radiation exposure (Exposed Group) and 45 personnel in a setting without radiation exposure (Control Group). Their native thiol, total thiol, disulphide, albumine and IMA levels were measured. Exposed group included personnel who were exposed to radiation outside the operating room - Operation room (−) Group and inside the operating room - Operation room (+) Group. Results: Albumin, native and total thiol levels were significantly lower in the participants exposed to radiation in the anesthesia application area; no statistically significant difference was found in terms of disulfide and ischemia modified albumin levels. In the Operation room (−) Group exposed to radiation, native thiol and total thiol values were significantly lower compared to the Operation room (+) Group. Conclusion: Awareness of being in danger of oxidative stress should be established in personnel exposed to radiation in the anesthesia application area following low dose ionizing radiation exposure, and the necessary measures should be taken.

Resumo Justificativa: O objetivo principal do estudo foi investigar o efeito de exposição à radiação ionizante de baixa dose nos níveis de homeostase tiol/dissulfeto e de albumina modificada por isquemia. O objetivo secundário foi comparar os níveis de homeostase tiol/dissulfeto e albumina modificada por isquemia entre indivíduos expostos à radiação ionizante de baixa dose nas áreas de procedimentos anestésicos, dentro e fora da sala de cirurgia. Método: O estudo incluiu um total de 90 voluntários com idades entre 18 e 65 anos, 45 profissionais que trabalhavam em ambiente de exposição potencial a radiação (Grupo Exposto) e 45 profissionais que trabalhavam em ambiente sem exposição à radiação (Grupo Controle). Foram medidos os níveis de tiol nativo, tiol total, dissulfeto, albumina e albumina modificada por isquemia. O Grupo Exposto era constituído por profissionais expostos a radiação fora da sala de cirurgia - Grupo sala de cirurgia (-) e na sala de cirurgia - Grupo sala de cirurgia (+). Resultados: Os níveis de albumina, tiol nativo e total foram significantemente mais baixos nos participantes expostos à radiação em área de realização de anestesia, e nenhuma diferença estatisticamente significante foi encontrada para os níveis de dissulfeto e albumina modificada por isquemia. No Grupo exposto sala de cirurgia (-), os valores de tiol nativo e tiol total foram significantemente mais baixos quando comparados ao Grupo sala de cirurgia (+). Conclusões: Os profissionais expostos à radiação em área de realização de anestesia devem ser conscientizados quanto ao perigo do estresse oxidativo após exposição à radiação ionizante de baixa dose e medidas cabíveis devem ser instituídas.

The effect of continuous ventilation on thiol-disulphide homeostasis and albumin-adjusted ischemia-modified albumin during cardiopulmonary bypass

Abstract Objective: To investigate the effect of continuous lung ventilation with low tidal volume on oxidation parameters, such as thiol/disulphide homeostasis and albumin-adjusted ischemia-modified albumin (AAIMA), during cardiopulmonary bypass (CBP) in coronary artery bypass grafting (CABG). Methods: Seventy-four patients who underwent elective CABG with CPB were included in the study. Blood samples were taken in the preoperative period, 10 minutes after CPB, and six and 24 hours postoperatively. Patients were assigned to the continuous ventilation group (Group 1, n=37) and the non-ventilated group (Group 2, n=37). The clinical characteristics, thiol/disulphide homeostasis, ischemia-modified albumin (IMA), and AAIMA levels of the patients were compared. Results: A significant difference was found between the groups regarding native thiol, total thiol, and IMA levels at the postoperative 24th hour (P=0.030, P=0.031, and P=0.004, respectively). There was no difference between the groups in terms of AAIMA. AAIMA levels returned to preoperative levels in Groups 1 and 2, at the 6th and 24th postoperative hours, respectively. Length of hospital stay was significantly shorter in Group 1 (P<0.001) than in Group 2. Conclusion: Continuous ventilation during CPB caused an increase in native and total thiol levels, an earlier return of AAIMA levels, and shorter hospital stay. Continuous ventilation may reduce the negative effects of CPB on myocardium (Table 2, Figure 1, and Reference 31).

Role of native thiol, total thiol and dynamic Disulphide in diagnosis of patient with prostate cancer and prostatitis

ABSTRACT Background: Our study investigates whether Native Thiol, Total Thiol and disulphide levels measured in serum of patients with prostate cancer and prostatitis and of healthy subjects, have any role in differential diagnosis. Materials and Methods: Patients followed up for histopathologically verified diagnosis of prostate cancer and prostatitis in 2016-2017 at the Medicalpark Gaziantep Hospital Urology Clinic were included in the study. Native Thiol (NT), Total Thiol (TT), Dynamic Disulphide (DD) levels in serum were measured by a novel automated method. Results: NT, TT, DD, NT / TT ratios, DD / TT ratio and DD / NT ratio were measured as 118.4 ± 36.8μmoL / L, 150.3 ± 45.3μmoL / L, 15.9 ± 7μmoL / L, 78.8 ± 7μmoL / L, 10.5 ± 3.5μmoL / L, 13.8 ± 5.8μmoL / L respectively in patients with prostate cancer; as 116.4 ± 40.5μmoL / L, 147.5 ± 50.1μmoL / L, 15.5 ± 8.7μmoL / L, 79.7 ± 9μmoL / L, 10.1 ± 4.5μmoL / L, 13.5 ± 7.2μmoL / L in patients with prostatitis and as 144.1 ± 21.2μmoL / L, 191 ± 32.3μmoL / L, 23.4 ± 10.1μmoL / L, 76.1 ± 98.3μmoL / L, 11.9 ± 4.1μmoL / L, 16.4 ± 6.9μmoL / L in healthy subjects. Significant difference was detected between groups of NT, TT and DD levels (p = 0.008, p = 0.001, p = 0.002). No significant difference was detected in terms of the NT / TT, DD / TT and DD / NT rates (p = 0.222, p = 0.222, p = 0.222). Conclusions: Serum NT, TT, DD levels in patients with prostatitis and prostate cancer were found significantly lower compared to the control group. This indicates that just as inflammation, prostate cancer also increases oxidative stress on tissues.

La homeostasis de tiol/disulfuro es un indicador del estrés oxidativo en los niños críticamente enfermos con sepsis / Thiol-disulphide homeostasis is an oxidative stress indicator in critically ill children with sepsis

Objetivo. Evaluar un novedoso marcador del estrés oxidativo (la homeostasis de tiol /disulfuro) en la sepsis pediátrica y determinar sus efectos sobre el pronóstico de esta afección. Métodos. En el estudio, se incluyeron pacientes con diagnóstico de sepsis y controles sanos. Se midieron las concentraciones de tiol total, tiol nativo, disulfuro, disulfuro /tiol total, disulfuro /tiol nativo y tiol nativo/tiol total en los grupos con sepsis y de referencia. Se compararon los parámetros entre los supervivientes y los no supervivientes del grupo con sepsis. Se midieron las concentraciones de hemoglobina, leucocitos, trombocitos, lactato y proteína C-reactiva en los pacientes con sepsis al momento del diagnóstico. Se utilizaron el puntaje de riesgo de mortalidad pediátrico (Pediatric Risk of Mortality, PRISM) y el puntaje de disfunción orgánica (Pediatric Logistic Organ Dysfunction, PELOD) para estimar la gravedad de la enfermedad. Resultados. En el grupo con sepsis se incluyó a 38 pacientes y en el de referencia, a 40 niños sanos. Las concentraciones plasmáticas de tiol en los pacientes con sepsis fueron significativamente inferiores que las del grupo de referencia (p < 0,001). Conclusión. La homeostasis de tiol/disulfuro fue anormal en los niños con sepsis en la unidad de cuidados intensivos pediátricos.

The aim of this study is to evaluate a novel oxidative stress marker (thiol-disulphide homeostasis) in paediatric sepsis and to determine their effects on the prognosis of sepsis. Patients diagnosed with sepsis (n= 38) and healthy controls (n= 40) were incorporated in the study. Total thiol, native thiol, disulphide, disulphide/total thiol, disulphide/native thiol, and native thiol /total thiol levels were measured in the sepsis and control groups. Additionally, the parameters were compared between survivors and non-survivors in the sepsis group. The levels of hemoglobin, white blood cell, platelet, lactate, and C-reactive protein were measured in patients with sepsis at diagnosis. The paediatric risk of mortality and paediatric logistic organ dysfunction scores of the patients were used to estimate the disease severity. The plasma thiol levels of the patients with sepsis were significantly lower than the control group (p < 0.001). This study showed that thiol/disulphide homeostasis is abnormal in children with sepsis in Paediatric Intensive Care Unit.

Thiol/disulfide status regulates the activity of thiol-containing kinases related to energy homeostasis in rat kidney

ABSTRACT Considering that thiol-containing enzymes like kinases are critical for several metabolic pathways and energy homeostasis, we investigated the effects of cystine dimethyl ester and/or cysteamine administration on kinases crucial for energy metabolism in the kidney of Wistar rats. Animals were injected twice a day with 1.6 µmol/g body weight cystine dimethyl ester and/or 0.26 µmol/g body weight cysteamine from the 16th to the 20th postpartum day and euthanized after 12 hours. Pyruvate kinase, adenylate kinase, creatine kinase activities and thiol/disulfide ratio were determined. Cystine dimethyl ester administration reduced thiol/disulfide ratio and inhibited the kinases activities. Cysteamine administration increased the thiol/disulfide ratio and co-administration with cystine dimethyl ester prevented the inhibition of the enzymes. Regression between the thiol/disulfide ratio, and the kinases activities were significant. These results suggest that redox status may regulate energy metabolism in the rat kidney. If thiol-containing enzymes inhibition and oxidative stress occur in patients with cystinosis, it is possible that lysosomal cystine depletion may not be the only beneficial effect of cysteamine administration, but also its antioxidant and thiol-protector effect.

Chronic treatment with carvacrol improves passive avoidance memory in a rat model of Parkinson's disease / O tratamento com carvacrol melhora a memória de esquiva passiva em um modelo da doença de Parkinson em ratos

ABSTRACT The present study investigated the effects of carvacrol on motor and memory deficits as well as hyperalgesia in the 6-OHDA-lesioned rat model of Parkinson's disease. The animals were subjected to unilateral microinjection of 6-OHDA into the medial forebrain bundle and treated with carvacrol (25, 50 and 100 mg/kg, ip) for six weeks after surgery. The 6-OHDA-lesioned rats showed contralateral rotations towards the lesion side, which was accompanied by learning and memory deficits in a passive avoidance test and a decrease in tail withdrawal latency in a tail flick test at the end of week 6. The results also showed that treatment with carvacrol at a dose of 25 mg/kg ameliorated memory deficits, with no effect on rotations and hyperalgesia in lesioned rats. In conclusion, carvacrol improves memory impairments in rats with Parkinson's disease; therefore, it may serve as an adjunct therapy for the alleviation of memory deficits in Parkinson's disease patients.

RESUMO O presente estudo investigou os efeitos do carvacrol nos déficits motores e de memória, bem como na hiperalgesia, em um modelo da doença de Parkinson (DP) em ratos com lesões 6-OHDA. Os animais foram submetidos a microinjeção unilateral de 6-OHDA no feixe mediano do prosencéfalo e tratados com carvacrol (25, 50 e 100 mg / kg, ip) durante 6 semanas após a cirurgia. Os ratos com lesões 6-OHDA mostraram rotações contralaterais para o lado da lesão, que foram acompanhadas de déficits de aprendizagem e de memória em um teste de evitação passiva, e de uma diminuição da latência de retirada da cauda em um teste de cauda no final da semana 6. Os resultados também mostraram que o tratamento crônico com carvacrol a uma dose de 25 mg / kg aliviou os déficits de memória, sem efeito sobre rotações e hiperalgesia em ratos lesados. Em conclusão, o carvacrol melhora a deficiência de memória em ratos com DP e, portanto, pode servir como uma terapia complementar para aliviar os déficits de memória em pacientes com DP.

Redox-triggered events in lipid biology: thiol modification and lipidome alteration in ALS / Eventos redox na biologia dos lipídios: modificação de tióis e alterações do lipidoma em ELA

Lipids encompass a wide range of hydrophobic molecules present in cells. The molecular characteristics of lipids determine their cellular localization and biological function. In general, lipids are regarded as essential components of membranes, as energy reservoir and modulators of signaling pathways linked to cellular metabolism and survival, among others. In mammals, a large part of the lipids are esterified to polyunsaturated fatty acids (PUFAs), especially docosahexaenoic (DHA) and arachidonic (ARA) acids, essential for several physiological processes, including normal brain development. However, PUFAs are very susceptible to oxidation by reactive oxygen species (ROS) generated endogenously. Once oxidized, lipids are able to modify thiol groups of peptides and proteins leading to modulation of signaling pathways and cellular redox balance. In the chapter 1, we investigated the mechanisms involved in modification of thiol groups of peptides and protein by autoxidation products derived from PUFAs. Here, we identified several glutathione (GSH) adducts covalently modified by hydroxy-endoperoxides derived from both DHA and ARA. Detailed inspection of MS/MS spectra of GSH-adducts revealed that GSH and hydroxy-endoperoxides are likely bonded through a sulfur-oxygen chemical bond in a reaction which involves a nucleophilic attack by the thiolate anion. Also, we suggest that the efficiency of modification of thiol by hydroxy-endoperoxides are also dependent of the thiol reactivity, as demonstrated by covalent modification of the most reactive cysteine residue (Cys111) of the antioxidant enzyme Cu,Zn-superoxide dismutase (SOD1). Chemical modifications of thiol groups by hydroxy-endoperoxides may modulate protein aggregation and cellular redox status, yieldingGSH adducts capable to modulate inflammation, as reported for the enzymatically generated counterparts. In the chapter 2, we investigated the role of lipids in amyotrophic lateral sclerosis (ALS), since inflammation and oxidative stress in motor neurons are hallmarks of this neurodegenerative disease. Using an untargeted lipidomics approach based on mass spectrometry coupled to liquid chromatography (UHPLC-MS/MS), we investigated the lipid metabolism in motor cortex and spinal cord tissues of a rodent model of ALS. Analysis of the motor cortex showed that the main lipid alterations were age-dependent and linked to metabolism of sphingolipids. In contrast, the major lipid alterations in the spinal cord were found in ALS symptomatic group, being the metabolism of ceramides, cholesteryl esters and cardiolipin the most affected. According to our findings and data reported in the literature, we proposed a mechanism based on neuroprotection that involves accumulation of cholesteryl esters esterified to PUFAs in astrocytes. Collectively, our findings suggest that lipids play a crucial role in modulation of cellular process linked to thiol metabolism and neurodegeneration

Os lipídeos abrangem uma ampla gama de moléculas hidrofóbicas presentes nas células. As características moleculares dos lipídios determinam sua localização celular e função biológica. Em geral, os lipídios são considerados componentes essenciais de membranas, reservatórios de energia e moduladores de vias de sinalização ligadas ao metabolismo celular, sobrevivência, entre outros. Em mamíferos, grande parte dos lipídios é esterificada em ácidos graxos poli-insaturados (PUFAs), especialmente os ácidos docosahexaenóico (DHA) e araquidônico (ARA), essenciais para vários processos fisiológicos, incluindo o desenvolvimento normal do cérebro. No entanto, os PUFAs são muito suscetíveis à oxidação por espécies reativas de oxigênio (ROS) geradas endogenamente. Uma vez oxidados, lipídios são capazes de modificar grupos tióis de peptídeos e proteínas, levando à modulação das vias de sinalização e alterando o balanço redox celular. No capítulo 1, foram investigados os mecanismos envolvidos na modificação de grupos tióis de peptídeos e proteínas por produtos de auto-oxidação de PUFAs. Com as análises realizadas foi possível identificar vários adutos de glutationa (GSH) covalentemente modificados por endoperóxidos cíclicos derivados de DHA e ARA. Uma análise detalhada dos espectros de MS/MS dos adutos de GSH revelou que GSH e endoperóxidos cíclicos são provavelmente ligados através de uma ligação química de enxofre-oxigênio, em uma reação que envolve um ataque nucleofílico do ânion tiolato. Além disso, sugerimos que a eficiência da modificação do tiol por endoperóxidos cíclicos também é dependente da reatividade do tiol, como demonstrado pela modificação covalente do resíduo de cisteína mais reativo (Cys111) da enzima antioxidante superóxido dismutase 1(SOD1). Modificações químicas de tióis por endoperóxidos cíclicos podem modular a agregação proteica e o status redox celular, produzindo adutos de GSH capazes de modular a inflamação, como relatado para os conjugados de GSH gerados enzimaticamente. No capítulo 2, nós investigamos o papel dos lipídios na esclerose lateral amiotrófica (ALS), uma vez que a inflamação e o estresse oxidativo nos neurônios motores contribuem para o desenvolvimento desta doença neurodegenerativa. Usando uma abordagem lipidômica não direcionada baseada em espectrometria de massa acoplada à cromatografia líquida (UHPLC-MS/MS), nós investigamos o metabolismo lipídico no córtex motor e na medula espinhal de um modelo de ratos com ALS. A análise do córtex motor mostrou que as principais alterações lipídicas foram dependentes da idade e ligadas ao metabolismo dos esfingolipídios. Em contraste, as principais alterações lipídicas na medula espinhal foram encontradas no grupo sintomático da ALS, sendo o metabolismo de ceramidas, ésteres de colesterol e cardiolipinas os mais afetados. De acordo com os resultados obtidos e dados relatados na literatura, propusemos um mecanismo baseado em neuroproteção que envolve o acúmulo de ésteres de colesterol esterificados em PUFAs em astrócitos. Coletivamente, nossos achados sugerem que os lipídios desempenham um papel crucial na modulação de processos celulares ligado à oxidação de tióis e à neurodegeneração

Mentha pulegium crude extracts induce thiol oxidation and potentiate hemolysis when associated to t-butyl hydroperoxide in human's erythrocytes

ABSTRACT Mentha pulegium (Lamiaceae) tea has been used as a traditional medicine; however, the modulatory effect of M. pulegium extracts on damage to human erythrocytes associated to t-butyl hydroperoxide (t-BHP) exposure remains to be investigated. Accordingly, we perform this study in order to test the hypothesis that aqueous and ethanolic extracts of M. pulegium could modulate the hemolysis associated to t-BHP exposure, non-protein thiol (NPSH) oxidation and lipid peroxidation (measured as thiobarbituric acid reactive substances - TBARS) in human erythrocytes. Samples were co-incubated with t-BHP (4 mmol/L) and/or aqueous or ethanolic extracts (10-1000 mg/mL) during 120 min to further analysis. We found that both extracts, when associated to t-BHP, potentiate NPSH oxidation and hemolysis. Moreover, both extracts significantly prevents against t-BHP-induced TBARS production. A significant correlation among hemolysis and NPSH levels was found. Taking together, our data points that the association of M. pulegium extracts with t-BHP culminates in toxic effect to exposed erythrocytes, besides its protective effect against t-BHP-induced TBARS production. So, we infer that the use of this extract may exert negative effect during painful crisis in sickle cell anemia. However, more studies are still necessary to better investigate/understand the mechanism(s) involved in the toxic effect resultant from this association.

Dynamic thiol/disulfide homeostasis in patients with age-related macular degeneration / Homeostase dinâmica de tiol/dissulfureto em pacientes com degeneração macular relacionada à idade

ABSTRACT Purpose: We evaluated dynamic thiol/disulfide homeostasis (TDH), malondialdehyde (MDA) levels, and catalase (CAT) activity in patients with age-related macular degeneration (AMD). All analyzes were conducted on plasma samples. Methods: Thirty-two patients with AMD and 38 age-matched healthy controls were included. Native thiol, total thiol, and disulfide levels and TDH status were determined using a novel, automated assay. MDA levels and CAT activity were determined. Percentages were compared using the chi-squared test. The Student's t-test and Mann-Whitney U-test were used to compare quantitative variables. Results: Native thiol levels were significantly lower (p=0.004) in patients with AMD (272.02 ± 52.41 µmol/l) than in healthy individuals (307.82 ± 47.18 µmol/l), whereas disulfide levels were significantly higher (p<0.001) in patients with AMD than in controls (21.64 ± 5.59 vs. 14.48 ± 5.37 µmol/L). Dynamic TDH was also significantly lower (p<0.001) in patients with AMD than in controls (13.41 ± 4.3 vs. 25.41 ± 14.52 µmol/l). No significant differences were evident in total thiol or MDA levels. Mean CAT activity was significantly higher (p=0.043) in patients with AMD compared with controls (0.035 vs. 0.018 k/ml). Conclusions: The antioxidant/oxidant balance demonstrated by dynamic TDH is shifted to the oxidative side in patients with AMD.

RESUMO Objetivo: Avaliar a homeostase dinâmica de tiol/dissulfureto e os níveis de malon dialdeído (MDA) e catalase (CAT) em pacientes com degeneração macular relacionada à idade (DMRI). Todas as análises foram realizadas em amostras de plasma. Métodos: Foram incluídos 32 pacientes com degeneração macular relacionada à idade e 38 controles saudáveis de idade similar. Os níveis de tiol, tiol total, dissulfureto e estado de homeostase de tiol/dissulfureto foram determinados utilizando um novo ensaio automatizado. Os níveis de atividade de MDA e CAT foram também determinados. As porcentagens foram comparadas pelo teste do qui-quadrado. O teste t de Student e o teste U de Mann Whitney foram utilizados para comparar variáveis quantitativas. Resultados: Os níveis de tiol nativo foram significativamente menores (p=0,004) nos pacientes com degeneração macular relacionada à idade (272,02 ± 52,41 µmol/l) do que nos indivíduos saudáveis (307,82 ± 47,18 µmol/l), enquan to os dissulfetos foram significativamente maiores em pacientes com degeneração macular relacionada à idade (21,64 ± 5,59 µmol/l versus 14,48 ± 5,37 µmol/l, respectivamente, p<0,001). A homeostase dinâmica de tiol/dissulfureto também foi significativamente menor nos pacientes com degeneração macular re la cionada à idade (13,41 ± 4,3 µmol/l) versus os controles (versus 25,41 ± 14,52 µmol/l, p<0,001). Não foram observadas diferenças significativas nos níveis de tiol total ou MDA. A atividade média de CAT foi significativamente mais elevada (p=0,043) em doentes com degeneração macular relacionada à idade (0,035 k/ml vs. 0,018 k/ml). Conclusões: O equilíbrio antioxidante/oxidante demonstrado pela homeostase dinâmica de tiol/dissulfeto é deslocado para o lado oxidativo em pacientes com de generação macular relacionada à idade.

The change in serum Thiol/Disulphide homeostasis after transrectal ultrasound guided prostate biopsy

ABSTRACT Objectives The aim of this prospective clinical study was to investigate variations in a novel oxidative stress marker (thiol/disulphide homeostasis) in men who underwent transrectal ultrasound guided prostate biopsy (TRUSB). Materials and Methods A total of 22 men undergoing TRUSB of the prostate were enrolled in the study. Patients with abnormal digital rectal examination and/or total prostate specific antigen (PSA) over 4ng/mL underwent TRUSB with 12 cores. Serum samples were obtained before and just after the procedure to evaluate the possible changes in thiol/disulphide homeostasis. Mean age, total PSA and free PSA, prostate volume and histopathological data were also recorded. Results Mean age of the study population was 65.05±8.89 years. Significant decreases in native and total thiol levels were documented after the biopsy procedure. However, serum disulphide levels and disulphide/native thiol, disulphide/total thiol and native/total thiol ratios did not significantly change after TRUSB. No correlation was observed between oxidative parameters and total PSA and free PSA levels, prostate volume and histopathology of the prostate. However, mean patient age was significantly correlated with mean native and total thiol levels. Conclusion Significant decreases in serum native and total thiol levels related to the prostate biopsy procedure suggest that TRUSB causes acute oxidative stress in the human body. Since our trial is the first in the current literature to investigate these oxidative stress markers in urology practice, additional studies are warranted.

Herbal extracts differentially inhibit oxidative effects caused by the biotransformation of nifurtimox, nitrofurantoin and acetaminophen on rat liver microsomes / Extractos herbales inhiben diferencialmente los efectos oxidativos causados por la biotransformación de nifurtimox, nitrofurantoína y acetaminofeno en microsomas hepáticos de rata]

Inflammation is a cellular defensive mechanism associated to oxidative stress. The administration of nitrofurantoin, nifurtimox and acetaminophen generates oxidative stress by their biotransformation through CYP450 system. The main adverse effect described for the first two drugs is gastrointestinal inflammation and that of the last, hepatitis. Therefore, standardised dry extracts from Rosmarinus officinalis, Buddleja globosa Hope, Cynara scolymus L., Echinacea purpurea and Hedera helix were tested to evaluate their capacity to decrease drug-induced oxidative stress. For that, rat liver microsomes were incubated with drugs in the presence of NADPH (specific CYP450 system cofactor) to test oxidative damage on microsomal lipids, thiols, and GST activity. All drugs tested induced oxidation of microsomal lipids and thiols, and inhibition of GST activity. Herbal extracts prevented these phenomena in different extension. These results show that antioxidant phytodrugs previously evaluated could alleviate drugs adverse effects associated to oxidative stress.

Inflamación es un mecanismo de defensa el cual está asociado a estrés oxidativo. La administración de nitrofurantoína, nifurtimox y paracetamol genera estrés oxidativo al metabolizarse a través del sistema CYP450. El principal efecto adverso de los dos primeros fármacos es inflamación gastrointestinal y del tercero, hepatitis. Por lo tanto, utilizamos diversos extractos herbales para disminuir el estrés oxidativo inducido por estos fármacos. Para esto se incubaron microsomas hepáticos de rata con dichos fármacos en presencia de NADPH (cofactor específico del sistema CYP450) y se evaluó el daño oxidativo generado sobre los lípidos, los tioles y la actividad GST microsómica. Todos los fármacos indujeron oxidación de los lípidos y los tioles microsómicos e inhibieron la actividad GST. Los extractos herbales previnieron estos fenómenos oxidativos en diferente extensión. Estos resultados indican que fitofármacos antioxidantes previamente evaluados, podrían aliviar los efectos adversos asociados a estrés oxidativo de los fármacos.

Evaluation of salivary oxidative parameters in overweight and obese young adults

ABSTRACT Background Obesity is characterized by a deposition of abnormal or excessive fat in adipose tissue, and is linked with a risk of damage to several metabolic and pathological processes associated with oxidative stress. To date, salivary oxidative biomarkers have been minimally explored in obese individuals. Thus, the aim of this study was to assess the concentrations of salivary oxidative biomarkers (ferric-reducing antioxidant power, uric acid, sulfhydryl groups) and lipid peroxidation in obese and overweight young subjects. Materials and methods Levels of lipid peroxidation, ferric-reducing antioxidant power, uric acid, and SH groups were determined in the saliva and serum of 149 young adults, including 54 normal weight, 27 overweight, and 68 obese individuals. Anthropometric measurements were also evaluated. Results Salivary levels of ferric-reducing antioxidant power, sulfhydryl groups, and lipid peroxidation, as well as serum levels of ferric-reducing antioxidant power, uric acid, and lipid peroxidation were higher in obese patients when compared with individuals with normal weight. There were correlations between salivary and serum ferric-reducing antioxidant power and salivary and serum uric acid in the obese and normal-weight groups. Conclusions Our results indicate that the increase in salivary levels of ferric-reducing antioxidant power, sulfhydryl groups, and lipid peroxidation, and serum levels of ferric-reducing antioxidant power, uric acid, and lipid peroxidation could be related to the regulation of various processes in the adipose tissue. These findings may hold promise in identifying new oxidative markers to assist in diagnosing and monitoring overweight and obese patients.

In vitro effect of N-acetyl-L-cysteine on glutathione and sulfhydryl levels in X-linked adrenoleukodystrophy patients

Introduction: Recent evidence shows that oxidative stress seems to be related with the pathophysiology of X-linked adrenoleukodystrophy (X-ALD), a neurodegenerative disorder. Methods: In the present study, the in vitro effect of N-acetyl-L-cysteine (NAC) on glutathione (GSH) and sulfhydryl levels in X-ALD patients was evaluated. Results: A significant reduction of GSH and sulfhydryl content was observed in X-ALD patients compared to the control group. Furthermore, 5 mM of NAC, in vitro, led to an increase in GSH content and sulfhydryl groups in these patients. Conclusion: These data probably indicate that an adjuvant therapy with the antioxidant NAC could improve the oxidative imbalance in X-ALD patients(AU)