ABSTRACT
Glutathione (GSH) is indispensable for maintaining redox homeostasis in biological fluids and serves as a key component in cellular defense mechanisms. Accurate assessment of GSH relative to its oxidized counterpart, glutathione disulfide (GSSG), is critical for the early diagnosis and understanding of conditions related to oxidative stress. Despite existing methods for their quantification, the label-free and simultaneous measurement of GSH and GSSG in biological fluid presents significant challenges. Herein, we report the use of an alpha-hederin (Ah) nanopore for the direct measurement of the GSH:GSSG ratio in simulated biological fluid, containing fetal bovine serum (FBS). This system hinges on detecting characteristic relative ion blockades (ΔI/Io) as GSH and GSSG molecules pass through the Ah nanopore under an applied electric field. The distinct current blockage signals derived from the translocation of GSH and GSSG enabled us to determine the molar ratio of GSH and its oxidized form. Notably, the interactions between the hydroxyl groups of the sugar moiety lining the nanopore's inner surface and the sulfhydryl group of GSH significantly influence the translocation dynamics, resulting in a longer translocation time for GSH compared to GSSG. The Ah nanopore technology proposed in this study offers a promising approach for real-time, single molecule-level monitoring of glutathione redox status in biological fluids, eliminating the need for labeling or extensive sample preparation.
Subject(s)
Biosensing Techniques , Glutathione Disulfide , Glutathione , Nanopores , Oxidation-Reduction , Glutathione/chemistry , Glutathione/analysis , Glutathione/blood , Biosensing Techniques/methods , Glutathione Disulfide/analysis , Glutathione Disulfide/chemistry , Glutathione Disulfide/blood , Animals , Humans , Cattle , Oxidative StressABSTRACT
BACKGROUND: Periodontal diseases are the most frequently diagnosed problem in cats. It has been well-established that periodontal diseases could not only cause various oral health issues but could also contribute to systemic diseases. Oxidative stress is a possible link between systemic diseases and periodontitis. Our study aimed to illustrate the influence of periodontitis on oxidative stress development in cats. Furthermore, the changes in the bacterial flora of the gums were investigated. METHODS: Based on the clinical and laboratory examinations, fifty cats were divided into two groups normal (n = 25) and moderate to advanced periodontitis (n = 25). Serum total antioxidant capacity (TAC), total oxidant status (TOS), reduced (GSH) and oxidized glutathione (GSSG) were measured. In addition, samples were taken from the subgingival plaques of all cats for bacterial culture. RESULTS: Serum TOS, GSSG, GSSG to GSH ratio, and oxidative stress index (OSI), calculated as the ratio of TOS to TAC in cats with periodontal disease were significantly higher, and TAC was significantly lower (p < 0.05) compared with controls. The results of bacterial culture indicated that the number of isolated bacterial colonies is higher in patients than in the control group. Additionally, the analysis of these data showed a positive association between periodontal index and oxidative stress. CONCLUSIONS: Our results revealed that periodontitis in cats is related to a main oxidative stress. Furthermore, oxidant factors such as TOS and OSI, compared to antioxidant factors, may better indicate the presence of oxidative stress conditions in patients with periodontitis.
Subject(s)
Antioxidants , Cat Diseases , Glutathione , Oxidative Stress , Periodontitis , Animals , Cats , Cat Diseases/microbiology , Cat Diseases/blood , Cat Diseases/metabolism , Case-Control Studies , Periodontitis/veterinary , Periodontitis/microbiology , Female , Male , Antioxidants/metabolism , Glutathione/blood , Glutathione/metabolism , Glutathione Disulfide/blood , Glutathione Disulfide/metabolism , Oxidants/metabolism , Oxidants/bloodABSTRACT
OBJECTIVE: The present systematic review explored the involvement of enzymatic and nonenzymatic antioxidants in periodontitis, drawing from established literature. MATERIALS AND METHODS: The research approach encompassed an extensive electronic search from 2000 to 2023 across databases such as PubMed, Science Direct, and Wiley Online Library and cross-referencing using specific keywords. RESULTS: The initial literature exploration generated a total of 766 articles. After thoroughly examining the abstracts, 693 articles were excluded from consideration due to duplication and lack of relevance to the central research inquiry. Following that, 73 articles were left for in-depth evaluation. Following a qualitative assessment, 35 studies that satisfied the inclusion criteria were chosen, while 38 were removed for not meeting the necessary standards. Within this selection, a meta-analysis was conducted on 11 articles that provided consistent data for quantitative synthesis. Specifically, the analysis of glutathione (GSH) levels in serum samples revealed a standardized mean difference (SMD) of -5.552 µg/mL (CI 95%: -9.078 to -2.026; P-0.002). In contrast, the analysis of glutathione peroxidase (GPx) enzymes in gingival crevicular fluid (GCF) samples displayed an overall SMD of 2.918 ng/µL (CI 95%: 0.372-5.465; P-0.025), while salivary samples exhibited an overall SMD value of 0.709 U/l (95% CI: -1.907-3.325; P-0.596) which is of insignificant. CONCLUSION: The systematic review findings suggest a notable decrease in antioxidant enzymes across various systemic biological samples among patients with periodontitis, contrasting with the results from gingival tissue samples meta-analysis of GPx enzyme.
Subject(s)
Glutathione Peroxidase , Glutathione Reductase , Glutathione , Periodontitis , Humans , Periodontitis/diagnosis , Periodontitis/blood , Periodontitis/metabolism , Glutathione Peroxidase/metabolism , Glutathione/metabolism , Glutathione Reductase/metabolism , Glutathione Disulfide/metabolism , Glutathione Disulfide/blood , Gingival Crevicular Fluid/chemistry , Antioxidants/metabolism , Antioxidants/analysisABSTRACT
(1) Background: Autism, also known as autism-spectrum disorder, is a pervasive developmental disorder affecting social skills and psychological status in particular. The complex etiopathogenesis of autism limits efficient therapy, which leads to problems with the normal social integration of the individual and causes severe family distress. Injectable methylcobalamin was shown to improve the clinical status of patients via enhanced cell oxidative status and/or methylation capacity. Here we tested the efficiency of a syrup form of methylcobalamin in treating autism. (2) Methods: Methylcobalamin was administered daily at 500 µg dose to autistic children and young adults (n = 25) during a 200-day period. Clinical and psychological status was evaluated by parents and psychologists and plasma levels of reduced and oxidized glutathione, vitamin B12, homocysteine, and cysteine were determined before the treatment, and at day 100 and day 200 of the treatment. (3) Results: Good patient compliance was reported. Methylcobalamin treatment gradually improved the overall clinical and psychological status, with the highest impact in the social domain, followed by the cognitive, behavioral and communication characteristics. Changes in the clinical and psychological status were strongly associated with the changes in the level of reduced glutathione and reduced/oxidized glutathione ratio. (4) Conclusion: A high dose of methylcobalamin administered in syrup form ameliorates the clinical and psychological status of autistic individuals, probably due to the improved oxidative status.
Subject(s)
Autistic Disorder , Vitamin B 12 , Adolescent , Autistic Disorder/drug therapy , Child , Child, Preschool , Female , Glutathione Disulfide/blood , Humans , Male , Vitamin B 12/administration & dosage , Vitamin B 12/analogs & derivatives , Young AdultABSTRACT
The imbalance of high oxidative stress and low antioxidant capacities is thought to be a significant cause of the development and progression of hepatocellular carcinoma (HCC). However, the impact of oxidative stress, glutathione (GSH), and its related antioxidant enzymes on the recurrence of HCC has not been investigated. The purpose of this study was to compare the changes to oxidative stress and GSH-related antioxidant capacities before and after tumor resection in patients with HCC recurrence and non-recurrence. We also evaluated the prognostic significance of GSH and its related enzymes in HCC recurrence. This was a cross-sectional and follow-up study. Ninety-two HCC patients who were going to receive tumor resection were recruited. We followed patients' recurrence and survival status until the end of the study, and then assigned patients into the recurrent or the non-recurrent group. The tumor recurrence rate was 52.2% during the median follow-up period of 3.0 years. Patients had significantly lower plasma malondialdehyde level, but significantly or slightly higher levels of GSH, glutathione disulfide, trolox equivalent antioxidant capacity, glutathione peroxidase (GPx), and glutathione reductase (GR) activities after tumor resection compared to the respective levels before tumor resection in both recurrent and non-recurrent groups. GSH level in HCC tissue was significantly higher than that in adjacent normal tissue in both recurrent and non-recurrent patients. Decreased plasma GPx (HR = 0.995, p = 0.01) and GR (HR = 0.98, p = 0.04) activities before tumor resection, and the increased change of GPx (post-pre-resection) (HR = 1.004, p = 0.03) activity were significantly associated with the recurrence of HCC. These findings suggest there might be a possible application of GPx or GR as therapeutic targets for reducing HCC recurrence.
Subject(s)
Antioxidants/metabolism , Carcinoma, Hepatocellular/blood , Glutathione/blood , Liver Neoplasms/blood , Neoplasm Recurrence, Local/epidemiology , Oxidative Stress , Aged , Carcinoma, Hepatocellular/surgery , Cross-Sectional Studies , Female , Follow-Up Studies , Glutathione Disulfide/blood , Glutathione Peroxidase/blood , Glutathione Reductase/blood , Hepatectomy , Humans , Liver Neoplasms/surgery , Male , Malondialdehyde/blood , Middle Aged , Neoplasm Recurrence, Local/etiology , Oxygen Radical Absorbance Capacity , Postoperative Period , Predictive Value of Tests , PrognosisABSTRACT
Plant-rich diets alleviate oxidative stress and gut dysbiosis and are negatively linked to age-associated chronic disorders. This study examined the effects of consuming plant-based, antioxidant-rich smoothies and sesame seed snacks (PBASS) on antioxidant ability and gut microbial composition in older adults. Healthy and sub-healthy older adults (n = 42, 79.7 ± 8.6 years old) in two senior living facilities were given PBASS for 4 months. Blood and fecal samples were collected from these individuals at the baseline and after 2 and 4 months of PBASS consumption. After 2 months, serum levels of albumin and high-density lipoprotein-cholesterol and the ratio of reduced to oxidized glutathione (GSH/GSSG) had increased significantly and erythrocytic glutathione, GSH/GSSG and superoxide dismutase activity had decreased significantly compared with baseline levels (p < 0.05). After 4 months, red blood cells, hematocrit, serum blood urea nitrogen and erythrocyte glutathione peroxidase activity had decreased significantly, whereas plasma and erythrocyte protein-bound sulfhydryl groups had increased significantly. Furthermore, plasma glutathione and total antioxidant capacity were significantly greater after 2 months and increased further after 4 months of PBASS consumption. The results of next generation sequencing showed that PBASS consumption prompted significant decreases in observed bacterial species, their richness, and the abundance of Actinobacteria and Patescibacteria and increases in Bacteroidetes in feces. Our results suggest that texture-modified, plant-based snacks are useful nutrition support to benefit healthy ageing via the elevation of antioxidant ability and alteration of gut microbiota.
Subject(s)
Antioxidants/administration & dosage , Diet, Vegetarian/methods , Gastrointestinal Microbiome/physiology , Oxidative Stress/physiology , Snacks/physiology , Aged , Aged, 80 and over , Cholesterol, HDL/blood , Elder Nutritional Physiological Phenomena , Feces/microbiology , Female , Glutathione/blood , Glutathione Disulfide/blood , Homes for the Aged , Humans , Male , Seeds/chemistry , Serum Albumin/analysis , Sesamum/chemistry , Superoxide Dismutase/bloodABSTRACT
The reduced (GSH)-to-oxidized (GSSG) glutathione ratio represents a dynamic balance between oxidants and antioxidants. However, redox status in adolescents with obesity and anemia has not been investigated. This study investigated the association of erythrocyte GSH redox status (GSH, GSH:GSSG ratio, and glutathione peroxidase [GPx] activity) with anemia and adiposity in adolescents. This case-control study nested in a cross-sectional study enrolled 524 adolescents (268 boys; 256 girls). The prevalence of anemia in overweight and obesity (OWOB) was 5.2% in boys and 11.7% in girls. The GSH:GSSG ratio and GPx activity were significantly higher in girls than in boys (p < 0.001), in anemic than in non-anemic subjects (p < 0.001), and in OWOB than in normal-weight subjects (p < 0.001). Similarly, significantly higher GSH: GSSG level (p < 0.001) and GPx activity (p < 0.001) were found in subjects with 90th percentile waist circumference than in those with < 90th percentile. GPx and GSH:GSSG were positively associated with anemia after adjusting for age, sex, and body mass index (adjusted odds ratio, adjOR [95% confidence interval, CI] 2.18 [1.44-3.29]) or tertiles (adjOR [95% CI], T3 = 2.49 [1.03-6.01]). A similar association was noted for GSH and GPx. A compensatory increased redox defense mechanism exists in anemia and obesity among adolescents without metabolic disturbances.
Subject(s)
Anemia/blood , Antioxidants/metabolism , Glutathione/blood , Pediatric Obesity/blood , Adiposity/genetics , Adolescent , Adult , Anemia/genetics , Anemia/pathology , Body Mass Index , Child , Child, Preschool , Female , Glutathione/genetics , Glutathione Disulfide/blood , Glutathione Peroxidase/blood , Glutathione Reductase/blood , Homeostasis/genetics , Humans , Male , Oxidants/blood , Oxidation-Reduction , Oxidative Stress/genetics , Pediatric Obesity/genetics , Pediatric Obesity/pathology , Waist Circumference , Young AdultABSTRACT
Generating oxidative stress is a critical mechanism by which host cells defend against infection by pathogenic microorganisms. Radiation resistance is a critical problem in radiotherapy against cancer. Epstein-Barr virus (EBV) is a cancer-causing virus and its reactivation plays an important role in the development of EBV-related tumors. This study aimed to explore the inner relationship and regulatory mechanism among oxidative stress, EBV reactivation, and radioresistance and to identify new molecular subtyping models and treatment strategies to improve the therapeutic effects of radiotherapy. Methods: ROS, NADP+/NADPH, and GSSG/GSH were detected to evaluate the oxidative stress of cells. 8-OHdG is a reliable oxidative stress marker to evaluate the oxidative stress in patients. Its concentration in serum was detected using an ELISA method and in biopsies was detected using IHC. qPCR array was performed to evaluate the expression of essential oxidative stress genes. qPCR, Western blot, and IHC were used to measure the level of EBV reactivation in vitro and in vivo. A Rta-IgG ELISA kit and EBV DNA detection kit were used to analyze the reactivation of EBV in serum from NPC patients. NPC tumor tissue microarrays was used to investigate the prognostic role of oxidative stress and EBV reactivation. Radiation resistance was evaluated by a colony formation assay. Xenografts were treated with NAC, radiation, or a combination of NAC and radiation. EBV DNA load of tumor tissue was evaluated using an EBV DNA detection kit. Oxidative stress, EBV reactivation, and the apoptosis rate in tumor tissues were detected by using 8-OHdG, EAD, and TUNEL assays, respectively. Results: We found that EBV can induce high oxidative stress, which promotes its reactivation and thus leads to radioresistance. Basically, EBV caused NPC cells to undergo a process of 'Redox Resetting' to acquire a new redox status with higher levels of ROS accumulation and stronger antioxidant systems by increasing the expression of the ROS-producing enzyme, NOX2, and the cellular master antioxidant regulator, Nrf2. Also, EBV encoded driving protein LMP1 promotes EBV reactivation through production of ROS. Furthermore, high oxidative stress and EBV reactivation were positively associated with poor overall survival of patients following radiation therapy and were significant related to NPC patients' recurrence and clinical stage. By decreasing oxidative stress using an FDA approved antioxidant drug, NAC, sensitivity of tumors to radiation was increased. Additionally, 8-OHdG and EBV DNA could be dual prognostic markers for NPC patients. Conclusions: Oxidative stress mediates EBV reactivation and leads to radioresistance. Targeting oxidative stress can provide therapeutic benefits to cancer patients with radiation resistance. Clinically, we, for the first time, generated a molecular subtyping model for NPC relying on 8-OHdG and EBV DNA level. These dual markers could identify patients who are at a high risk of poor outcomes but who might benefit from the sequential therapy of reactive oxygen blockade followed by radiation therapy, which provides novel perspectives for the precise treatment of NPC.
Subject(s)
Chemoradiotherapy/methods , Epstein-Barr Virus Infections/therapy , Free Radical Scavengers/administration & dosage , Nasopharyngeal Carcinoma/therapy , Nasopharyngeal Neoplasms/therapy , Acetylcysteine/administration & dosage , Adolescent , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/blood , Biopsy , Cell Line, Tumor , DNA, Viral/blood , Epstein-Barr Virus Infections/blood , Epstein-Barr Virus Infections/mortality , Epstein-Barr Virus Infections/virology , Female , Follow-Up Studies , Glutathione Disulfide/blood , Glutathione Disulfide/metabolism , Herpesvirus 4, Human/genetics , Herpesvirus 4, Human/isolation & purification , Herpesvirus 4, Human/pathogenicity , Humans , Latent Infection/blood , Latent Infection/pathology , Latent Infection/therapy , Latent Infection/virology , Male , Middle Aged , Nasopharyngeal Carcinoma/blood , Nasopharyngeal Carcinoma/mortality , Nasopharyngeal Carcinoma/virology , Nasopharyngeal Neoplasms/blood , Nasopharyngeal Neoplasms/mortality , Nasopharyngeal Neoplasms/virology , Nasopharynx/pathology , Nasopharynx/virology , Oxidative Stress/drug effects , Patient Selection , Prognosis , Progression-Free Survival , Radiation Tolerance/drug effects , Reactive Oxygen Species , Viral Load , Viral Matrix Proteins/metabolism , Virus Activation/drug effects , Xenograft Model Antitumor Assays , Young AdultABSTRACT
In this pathbreaking study, we evaluated nitrosative stress in morbidly obese patients with and without metabolic syndrome. 62 women with class 3 obesity (BMI > 40 kg/m2) were divided into three subgroups: obese patients (OB), obese patients with hypertension (OB+HYP), and obese patients with metabolic syndrome (OB+MS). In comparison to the lean patients, OB had increased levels of serum myeloperoxidase (MPO), plasma nitric oxide (NO), S-nitrosothiols, and peroxynitrite (ONOO-), as well as nitrotyrosine, while oxidized glutathione (GSSG) rose only in OB+HYP group. Interestingly, ONOO- was significantly higher in OB+HYP and OB+MS as compared to OB group, while MPO only in OB+MS group. OB+MS had greater nitrotyrosine and S-nitrosothiol values than OB+HYP. Moreover, peroxynitrite could differentiate OB from OB+HYP and OB+MS (AUC 0.9292; p < 0.0001; 87.5% sensitivity, 90% specificity) as well as between OB and OB+MS group (AUC 0.9125; p < 0.0001; 81.25% sensitivity, 83.33%). In conclusion, we showed that MPO activity, NO formation, and nitrosative damage to proteins parallel the progression of metabolic disturbances of obesity. Evaluation of ONOO- concentrations may help predict the development of hypertension and metabolic syndrome in patients with morbid obesity; however, longer-term studies are required for larger numbers of patients.
Subject(s)
Glutathione/metabolism , Hypertension/complications , Hypertension/pathology , Metabolic Syndrome/complications , Metabolic Syndrome/pathology , Nitrosative Stress , Obesity, Morbid/complications , Obesity, Morbid/metabolism , Adult , Area Under Curve , Female , Glutathione/blood , Glutathione Disulfide/blood , Humans , Hypertension/blood , Metabolic Syndrome/blood , Middle Aged , Nitric Oxide/blood , Obesity, Morbid/blood , Oxidation-Reduction , Peroxidase/blood , Peroxynitrous Acid/blood , ROC Curve , Sulfhydryl Compounds/blood , Tyrosine/analogs & derivatives , Tyrosine/bloodABSTRACT
Age, sex and diet are well-established risk factors for several diseases. In humans, each of these variables has been linked to differences in plasma redox potentials (Eh) of the glutathione/glutathione disulfide (GSH/GSSG) and cysteine/cystine (Cys/CySS) redox couples. Mice have been very useful for modeling human disease processes, but it is unknown if age, sex and diet affect redox couples in mice as they do in humans. The purpose of the present study was to examine the effects of these factors on plasma redox potentials in C57BL/6J mice. We found that age had no effect on either redox couple in either sex. Plasma Eh Cys/CySS and Eh GSH/GSSG were both more oxidized (more positive) in females than in males. A 24-hour fast negated the sex differences in both redox potentials by oxidizing both redox couples in male mice, while having no effect on Eh Cys/CySS and a smaller effect on Eh GSH/GSSG in female mice. A diet with excess sulfur amino acids reduced the plasma Eh Cys/CySS in females to a level comparable to that seen in male mice. Thus, sex-specific differences in plasma Eh Cys/CySS could be normalized by two different dietary interventions. Some of these findings are consistent with reported human studies, while others are not. Most strikingly, mice do not exhibit age-dependent oxidation of plasma redox potentials. Care must be taken when designing and interpreting mouse studies to investigate redox regulation in humans.
Subject(s)
Cysteine/blood , Cystine/blood , Glutathione Disulfide/blood , Glutathione/blood , Aging , Animals , Diet , Fasting/blood , Female , Humans , Male , Mice, Inbred C57BL , Oxidation-ReductionABSTRACT
Endogenous glutathione (GSH) and glutathione disulfide (GSSG) status is highly sensitive to oxidative conditions and have broad application as a surrogate indicator of redox status in vivo. Established methods for GSH and GSSG quantification in whole blood display limited utility in human plasma, where GSH and GSSG levels are ~3-4 orders of magnitude below those observed in whole blood. This study presents simplified sample processing and analytical LC-MS/MS approaches exhibiting the sensitivity and accuracy required to measure GSH and GSSG concentrations in human plasma samples, which after 5-fold dilution to suppress matrix interferences range from 200 to 500 nm (GSH) and 5-30 nm (GSSG). The utility of the methods reported herein is demonstrated by assay performance and validation parameters which indicate good sensitivity [lower limits of quantitation of 4.99 nm (GSH) and 3.65 nm (GSSG), and high assay precision (intra-assay CVs 3.6 and 1.9%, and inter-assay CVs of 7.0 and 2.8% for GSH and GSSG, respectively). These methods also exhibited exceptional recovery of analyte-spiked plasma samples (98.0 ± 7.64% for GSH and 98.5 ± 12.7% for GSSG). Good sample stability at -80°C was evident for GSH for up to 55 weeks and GSSG for up to 46 weeks, with average CVs <15 and <10%, respectively.
Subject(s)
Chromatography, Liquid/methods , Glutathione Disulfide/blood , Tandem Mass Spectrometry/methods , Glutathione/blood , Humans , Limit of Detection , Linear Models , Reproducibility of ResultsABSTRACT
AIM: The removal of cysteine during a dialysis procedure may affect glutathione (GSH) concentration, allowing haemodialysis (HD) patients to become more susceptible to oxidative damage. This study was performed to determine whether the change of GSH/glutathione disulfide (GSSG) redox state and GSH redox potential were linked with the change of cysteine or oxidative stress in patients receiving HD treatment. METHODS: Sixty-seven HD patients who had received regular HD treatment were recruited. Plasma GSH, GSSG, cysteine and malondialdehyde (MDA) were measured at both pre- and post-HD. RESULTS: Plasma cysteine, GSH and GSSG levels significantly decreased after the completion of HD, compared to the levels at pre-HD. Plasma MDA concentration, GSH/GSSG ratio and GSH redox potential remained constant during the dialysis session. Plasma GSH and GSSG were positively associated with plasma MDA at post-HD, while GSH redox potential was negatively associated with plasma MDA at post-HD. However, plasma GSH, GSSG, GSH/GSSG ratio and GSH redox potential were not associated with plasma cysteine at either pre- or post-HD. CONCLUSION: The GSH and GSSG levels were significantly utilized during a HD session, and their levels were significantly associated with increased oxidative stress. HD patients may require higher GSH demands to cope with increased oxidative stress during an HD session.
Subject(s)
Glutathione/blood , Oxidative Stress , Renal Dialysis/adverse effects , Renal Insufficiency, Chronic/therapy , Aged , Biomarkers/blood , Cross-Sectional Studies , Cysteine/blood , Female , Glutathione Disulfide/blood , Humans , Male , Malondialdehyde/blood , Middle Aged , Oxidation-Reduction , Renal Insufficiency, Chronic/blood , Renal Insufficiency, Chronic/diagnosis , Time Factors , Treatment OutcomeABSTRACT
Chronic obstructive pulmonary disease (COPD) is prevalent in patients infected with HIV. The purpose of this study was to test the hypothesis that systemic oxidation correlates with loss of lung function in subjects with COPD, and that HIV infection can contribute to creating such an environment. Subjects were recruited at the University of Louisville in the following groups: HIV-infected (nâ¯=â¯36), COPD (nâ¯=â¯32), HIV and COPD (nâ¯=â¯28), and uninfected controls with normal lung function (nâ¯=â¯34). HIV infection was assessed by viral load and CD4 cell counts. Pulmonary function was determined by spirometry, and plasma was collected for measurement of cysteine (Cys), cystine (CySS), glutathione (GSH) and GSH disulfide (GSSG) by HPLC followed by estimation of redox potentials (Eh) using the Nernst equation. Results showed that patients with COPD had more oxidized plasma Eh Cys/CySS than patients with normal lung function, but plasma Eh GSH/GSSG was unaltered. In addition, there was a correlation between the extent of plasma Eh Cys/CySS oxidation and loss of lung function, and this correlation remained even after correcting for age, sex, race and body mass index. HIV infection per se was not associated with increased oxidation of plasma Eh Cys/CySS, but plasma Eh Cys/CySS was more oxidized in patients with lower CD4-positve T cell counts. In patients with both HIV infection and COPD, there was a significant correlation between CD4 cell counts and lung function. Thus, systemic oxidation correlated with decreased lung function in subjects with COPD and decreased CD4 counts in subjects infected with HIV. Thus, factors contributing to plasma Eh Cys/CySS may represent novel mechanisms underlying the increased prevalence of COPD in people living with HIV.
Subject(s)
Cysteine/blood , Cystine/blood , Glutathione Disulfide/blood , Glutathione/blood , HIV Infections/blood , Pulmonary Disease, Chronic Obstructive/blood , Adult , Aged , CD4-Positive T-Lymphocytes , Disulfides/blood , Female , Humans , Lung/metabolism , Male , Middle Aged , Multivariate Analysis , Oxidation-Reduction , Oxidative Stress , Respiratory Function Tests , Spirometry , Young AdultABSTRACT
Obesity among children has emerged as a serious public health problem. The growing prevalence of childhood obesity has led to the appearance of serious complications, including a chronic systemic inflammation associated with oxidative stress. In the present study, we analysed the interaction between two genes related with iron metabolism - HFE and haptoglobin - and the plasmatic concentration of glutathione, as a way to evaluate the antioxidant response capacity in obesity. To achieve this, 118 obese children and 89 eutrophic children were recruited for the study. Results showed that although obese children present a significantly decreased tGSH levels, once we analysed separately children based on their haptoglobin phenotype, the decreased tGSH levels is significant only for the Hp 2 allele. Additionally, Hp 2.2 obese children carrying H63D polymorphism show significantly lower tGSH/GSSG values. Our results found an association of haptoglobin and HFE with oxidative stress in childhood obesity.
Subject(s)
Genetic Predisposition to Disease , Glutathione/blood , Haptoglobins/genetics , Hemochromatosis Protein/genetics , Obesity/blood , Obesity/genetics , Case-Control Studies , Child , Female , Glutathione Disulfide/blood , Humans , Male , PhenotypeABSTRACT
Analyses of reduced glutathione (GSH), oxidized glutathione (GSSG), and total glutathione (tGSH) in red blood cell samples from 30 children diagnosed with autism and 30 age, gender, and socioeconomic status matched controls were undertaken. The children's ages ranged from 2 to 9. Samples were obtained from subjects residing in Western Pennsylvania, an area of the United States greatly affected by high levels of mercury deposition and airborne PM 2.5 particulates. Liquid chromatography - mass spectrometry was utilized by following EPA Method 6800 for sample analyses. The children with autism had a significantly lower mean red blood cell (RBC) reduced to oxidized glutathione ratio (GSH/GSSG) compared to the control children (pâ¯=â¯0.025). In addition, compared to the controls, the children with autism had significantly higher RBC tGSH values (pâ¯=â¯0.0076) and GSH values (pâ¯=â¯0.022). These results suggest that exposure to toxic elements may prompt compensatory increases in production of GSH in children with autism in environments higher in toxins. The compensation did not fully correct the anti-oxidant properties of exposure to xenobiotics as demonstrated by the significantly lower GSH/GSSG in children with autism compared to controls. Out of a set of glutathione biomarkers, GSH/GSSG may best determine the degree of compensation for oxidative stress in children with autism.
Subject(s)
Autistic Disorder/blood , Environmental Pollutants/adverse effects , Erythrocytes/metabolism , Glutathione Disulfide/blood , Glutathione/blood , Oxidative Stress/drug effects , Xenobiotics/adverse effects , Autistic Disorder/chemically induced , Autistic Disorder/pathology , Biomarkers/blood , Case-Control Studies , Child , Child, Preschool , Erythrocytes/drug effects , Female , Humans , MaleABSTRACT
Variability in the levels of GSH and GSSG in plasma is suggested to derive from inadequate pre-processing methods. The aim of this study was to develop a protocol for comparable and reliable measurements of GSH/GSSG. Venous blood from 8 healthy individuals were collected and divided into 7 different pre-processing procedures. For three of the samples an extraction mixture was added after 0 (baseline), 4 and 8â¯min and for three of the samples the extraction mixture was added at different times during defrost. A worst case scenario where a sample was left in a cool box during 6â¯h was also included. The samples were analyzed with UHPLC-ESI-MSMS. A large difference in the levels of GSH and GSSG were identified and it was clearly associated with the sample handling procedures. A sample left untreated for 4â¯min will have significantly reduced amount of GSH. Stability tests showed that the level of GSH was reduced after 3â¯months in -80⯰C.
Subject(s)
Chromatography, High Pressure Liquid/methods , Glutathione Disulfide/blood , Glutathione Disulfide/chemistry , Glutathione/blood , Glutathione/chemistry , Drug Stability , Humans , Linear Models , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization , Tandem Mass Spectrometry/methodsABSTRACT
It is assumed that in the progression of obesity, complicated by fatty hepatosis in adolescents, a significant role is given to the reactions of oxidative stress with deficiency of antioxidant factors. However, an important factor that remains outside the field of view of researchers is the ethnicity of the patient, which is an important element in the development of a personalized approach in the treatment and prevention of diseases. In connection with this, the purpose of this study was to study the changes in the lipid peroxidation-antioxidant defense processes in adolescent Mongoloids with exogenously constitutional obesity complicated by fatty hepatosis. 18 adolescent boys with fatty hepatosis were examined on the background of exogenous-constitutional obesity of the first degree; 38 adolescent boys with exogenously constitutional obesity without changes in the liver and 37 practically healthy adolescents (control group). All subjects surveyed for ethnicity belonged to the Mongoloids. Spectrophotometric and fluorometric methods were used in the research. The results of the study indicated a high intensity of lipid peroxidation reactions in Mongoloid boys with obesity and fatty hepatosis relative to control values: an increase in blood plasma content of compounds with unsaturated double bonds (p<0.001), diene conjugates (p=0.0012), ketodienes and conjugated trienes (p<0.0001), in the absence of significant differences in the level of TBA-active products. Increased values of total antioxidant activity in blood plasma (p=0.0023) and erythrocyte superoxide dismutase activity (p=0.0072), decreased levels of fat-soluble vitamins in blood plasma [α-tocopherol (p<0.0001) and retinol (p=0.0011)] and oxidized form of glutathione in erythrocytes (p=0.0083) have been found. The pronounced insufficiency of fatsoluble vitamins - α-tocopherol and retinol in patients of the main group was also noted in comparison with patients without morphological changes in the liver. Thus, in teenage Mongoloids with exogenously constitutional obesity and fatty hepatosis, it is possible to recommend antioxidant drugs in addition to basic metabolic therapy.
Subject(s)
Asian People , Fatty Liver/blood , Lipid Peroxidation , Obesity/blood , Adolescent , Antioxidants/metabolism , Erythrocytes/metabolism , Glutathione Disulfide/blood , Humans , Male , Vitamin A/blood , alpha-Tocopherol/bloodABSTRACT
OBJECTIVE: Evaluate the relationship between exogenous subclinical hyperthyroidism and oxidative stress through the analysis of the redox profile of patients with subclinical hyperthyroidism exogenous (SCH) grade I (TSH = 0.1 to 0.4 IU/mL) and grade II (TSH < 0.1 IU/mL). SUBJECTS AND METHODS: We analyzed 46 patients with SCH due to the use of TSH suppressive therapy with LT4 after total thyroidectomy along with 6 control euthyroid individuals (3M and 3W). Patients were divided into two groups, G1 with TSH ≥ 0.1-0.4 IU/mL (n = 25; and 7M 14W) and G2 with TSH < 0.1 IU/mL (n = 25; and 4M 21W). Venous blood samples were collected to measure the levels of markers for oxidative damage (TBARS, FOX and protein carbonylation), muscle and liver damage (CK, AST, ALT, GGT) and antioxidants (GSH, GSSG and catalase). RESULTS: Individuals in G2 showed a GSH/GSSG ratio ~ 30% greater than those in G1 (p = 0.004) and a catalase activity that was 4 times higher (p = 0.005). For lipid peroxidation, the levels measured in G2 were higher than both control and G1 (p = 0.05). No differences were observed for both protein carbonyl markers. G1 and G2 presented with greater indications of cell injury markers than the control group. CONCLUSION: TSH suppression therapy with LT4 that results in subclinical hyperthyroidism can cause a redox imbalance. The greater antioxidant capacity observed in the more suppressed group was not sufficient to avoid lipid peroxidation and cellular damage.
Subject(s)
Hyperthyroidism/drug therapy , Thyrotropin/antagonists & inhibitors , Thyroxine/pharmacology , Adult , Case-Control Studies , Catalase/blood , Cross-Sectional Studies , Female , Glutathione/blood , Glutathione Disulfide/blood , Humans , Hyperthyroidism/metabolism , Lipid Peroxidation/drug effects , Male , Middle Aged , Oxidation-Reduction/drug effects , Oxidative Stress/drug effects , Phenols/blood , Protein Carbonylation , Reference Values , Statistics, Nonparametric , Sulfoxides/blood , Thiobarbituric Acid Reactive Substances/analysisABSTRACT
Glutathione (γ-glutamyl-cysteinyl-glycine; also known as GSH) is an endogenous antioxidant that plays a crucial role in cell defense mechanisms against oxidative stress. It is thus not surprising that this molecule can serve as a biomarker for oxidative stress monitoring. As capillary blood is a highly accessible target for biomarking, it is a valuable bodily fluid for diagnosing human GSH levels. This study focused on the optimization of GSH measurements from micro volumes of capillary blood prior to using electrochemical detection. The optimization of experimental parameters, including the sample volume and its stability, was performed and evaluated. Moreover, we tested the optimized method as part of a short-term study. The study consisted of examining 10 subjects within 96 h of their consumption of high amounts of antioxidants, attained from a daily dose of 2 g/150 mL of green tea. The subjects' capillary blood (5 µL) was taken at 0 h, 48 h, and 96 h for subsequent analysis. The short-term supplementation of diet with green tea showed an increase of GSH pool by approximately 38% (between 0 and 48 h) within all subjects.
Subject(s)
Glutathione/blood , Tea/chemistry , Adult , Capillaries , Diet , Electrochemical Techniques , Female , Glutathione Disulfide/blood , Humans , MaleABSTRACT
Acetaminophen (APAP) is commonly used to relieve pain and fever in a clinical setting, but its excessive use can lead to serious hepatotoxicity. Our previous study demonstrated that polydatin (PD) can effectively attenuate d-galactose- and alcohol-induced hepatotoxicity, however, its effect on APAP-induced hepatotoxicity is still unknown. In this study, we explore the protective effect and potential mechanism of PD against APAP-induced hepatotoxicity in mice. The results indicate that PD effectively improves the survival of mice with APAP-induced hepatotoxicity, significantly alleviating histopathologic alterations in the liver, and decreasing the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). PD significantly and dose-dependently reduces oxidative stress by lowering the content of oxidized glutathione (GSSG), reactive oxygen species (ROS), nitric oxide (NO) and malonaldehyde (MDA), while enhancing the hepatic activities of glutathione (GSH), glutathione peroxidase (GSH-Px) and the GSH/GSSG ratio. Meanwhile, PD also substantially inhibits the levels and mRNA expressions of inducible nitric oxide synthase (iNOS) and NADPH oxidase 2 (NOX2). Additionally, PD markedly arrests apoptosis by assuaging TUNEL-positive hepatocytes and the apoptotic index, decreasing the levels and expression of cytochrome c (CytC), cleaved-caspase-9, apoptotic protease activating factor 1 (Apaf-1), cleaved-caspase-3, and Bax and increasing the level and expression of Bcl-2. Overall, PD pretreatment shows a potent protective effect against APAP-induced hepatotoxicity by relieving oxidative stress and inhibiting apoptosis.