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Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals

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A special issue of Antioxidants (ISSN 2076-3921). This special issue belongs to the section "Health Outcomes of Antioxidants and Oxidative Stress".

Deadline for manuscript submissions: closed (31 October 2022) | Viewed by 19060

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Prof. Dr. José Joaquín Cerón

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Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Regional Campus of International Excellence ‘Campus Mare Nostrum’, University of Murcia, 30100 Murcia, Spain
Interests: acute phase proteins; saliva; stress; biomarkers; oxidative status
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Dr. Camila Peres Rubio

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Interdisciplinary Laboratory of Clinical Analysis (Interlab-UMU), Veterinary School, Regional Campus of International Excellence ‘Campus Mare Nostrum’, University of Murcia, 30100 Murcia, Spain
Interests: analytical and clinical validation; biomarkers of health and welfare; non-invasive techniques; oxidative stress
Special Issues, Collections and Topics in MDPI journals

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Dear Colleagues,

Evaluation of oxidative stress using biomarkers is gaining attention with research into humans and different animal species. Changes in oxidative status are involved in the physiopathological mechanisms of many diseases and have a strong relationship with health and welfare. An accurate evaluation and quantification of these changes will allow one to determine the magnitude and possible influences in health and disease status.

The development of new assays and methods for the analysis of the different components of oxidative stress is of great importance to better evaluate this situation. These components involve assays for evaluation of different antioxidants, such as glutathione peroxidase, catalase, and thiol, and the antioxidant capacity, such as TEAC, FRAP and CUPRAC, or oxidants, such as assays for ROS estimation.

This Special Issue will deal with reports that involve the analytical validation of methods for the analysis of any component related to oxidative stress and the development of new or improved assays. This issue will cover works with humans and different animal species, as well as with different sample types, in order to establish a broad view of the possibilities that the applications of these assays and methods could have.

Prof. Dr. José Joaquín Cerón
Dr. Camila Peres Rubio
Guest Editors

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Keywords

Antioxidants
Assays
Health
Oxidants
Redox
Validation
Welfare

Published Papers (9 papers)

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13 pages, 2301 KiB

Open AccessArticle

Oxidative Modification Status of Human Serum Albumin Caused by Chronic Low-Dose Radiation Exposure in Mamuju, Sulawesi, Indonesia

by Masaru Yamaguchi, Yota Tatara, Eka Djatnika Nugraha, Yuki Tamakuma, Yoshiaki Sato, Tomisato Miura, Masahiro Hosoda, Shinji Yoshinaga, Mukh Syaifudin, Shinji Tokonami and Ikuo Kashiwakura

Antioxidants 2022, 11(12), 2384; https://doi.org/10.3390/antiox11122384 - 01 Dec 2022

Viewed by 1501

Abstract

The recently discovered high-level natural background radiation area (HBRA) of Mamuju in Indonesia provides a unique opportunity to study the biological effects of chronic low-dose radiation exposure on a human population. The mean total effective dose in the HBRA was approximately 69.6 mSv y⁻¹ (range: 47.1 to 115.2 mSv y⁻¹), based on a re-evaluation of the individual radiation exposure dose; therefore, proteomic analyses of serum components and oxidative modification profiling of residents living in the HBRA were reconducted using liquid chromatography-tandem mass spectrometry. The analysis of the oxidative modification sequences of human serum albumin revealed significant moderate correlations between the radiation dose and the modification of 12 sequences, especially the 111th methionine, 162nd tyrosine, 356th tyrosine, and 470th methionine residues. In addition, a dose-dependent variation in 15 proteins of the serum components was detected in the serum of residents exposed to chronic low-dose radiation. These findings suggest that the alterations in the expression of specific proteins and the oxidative modification responses of serum albumin found in exposed humans may be important indicators for considering the effects of chronic low-dose radiation exposure on living organisms, implying their potential utility as biomarkers of radiation dose estimation. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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9 pages, 1516 KiB

Open AccessArticle

Development of a New Colorimetric, Kinetic and Automated Ceruloplasmin Ferroxidase Activity Measurement Method

by Salim Neşelioğlu, Esra Fırat Oğuz and Özcan Erel

Antioxidants 2022, 11(11), 2187; https://doi.org/10.3390/antiox11112187 - 04 Nov 2022

Cited by 3 | Viewed by 1370

Abstract

Background: Ceruloplasmin plays an important role in the regulation of iron metabolism. Ceruloplasmin is an acute-phase protein known to have many metabolic effects. Its activity increases during infection, inflammation, and compensation of oxidation. In the current study, our aim is to develop a new method for the measurement of ferroxidase activity without requiring any chromogen. Methods: Venous blood samples were collected into serum separator tubes. Ferric iron ions formed by the enzyme ferroxidase were measured, both manually and fully automatically, at the 415 nm wavelength without using chromogen. These results were compared to conventional ferroxidase measurement methods and to the immunoturbidimetric ceruloplasmin measurement method. Results: The detection limit of the new assay was 14.8 U/L. The upper limit of the linearity was 1380 U/L. Precision values were calculated for high, medium, and low levels of ferroxidase activity in serum pool. The coefficient of variation was <5% for each level. Conclusion: In the present method, chromogens are not used. With its considerably low cost and short reaction time, this method is able to provide fast results, can be performed easily, and makes accurate measurements. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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8 pages, 1038 KiB

Open AccessArticle

Changes in Biomarkers of Redox Status in Saliva of Pigs after an Experimental Sepsis Induction

by María José López-Martínez, Damián Escribano, Alba Ortín-Bustillo, Lorena Franco-Martínez, Luis Guillermo González-Arostegui, José Joaquín Cerón and Camila Peres Rubio

Antioxidants 2022, 11(7), 1380; https://doi.org/10.3390/antiox11071380 - 16 Jul 2022

Cited by 5 | Viewed by 1437

Abstract

Saliva from pigs is gaining attention as an easy sample to obtain, being a source of biomarkers that can provide information on animal health and welfare. This study aimed to evaluate the changes that can occur in salivary biomarkers of the redox status of pigs with an experimentally induced sepsis. For that, the cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of saliva (FRAS), Trolox equivalent antioxidant capacity (TEAC), advanced oxidation protein products (AOPP), ferrous oxidation-xylenol orange (FOX), peroxide activity (POX-Act), and reactive oxygen-derived compounds (d-ROMs) were measured in the saliva of pigs with experimentally induced sepsis by endotoxin lipopolysaccharide (LPS), non-septic inflammation induced by turpentine, and in healthy individuals before and after 3 h, 6 h, 24 h, and 48 h. AOPP, POX-Act, and d-ROMs in the sepsis group were higher than in the control from 3 h to 24 h after the inoculation. CUPRAC, FRAS, and TEAC were higher in sepsis than the control group at 24 h. These changes were of higher magnitude than those that occurred in the turpentine group. In conclusion, our findings reveal that sepsis produces changes in salivary biomarkers of redox status, which opens the possibility of using them as potential biomarkers in this species. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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14 pages, 4680 KiB

Open AccessArticle

Influence of Diabetes-Induced Glycation and Oxidative Stress on the Human Rotator Cuff

by Tomoya Yoshikawa, Yutaka Mifune, Atsuyuki Inui, Hanako Nishimoto, Kohei Yamaura, Shintaro Mukohara, Issei Shinohara and Ryosuke Kuroda

Antioxidants 2022, 11(4), 743; https://doi.org/10.3390/antiox11040743 - 08 Apr 2022

Cited by 5 | Viewed by 1737

Abstract

Most shoulder rotator cuff tears (RCTs) are caused by non-traumatic age-related rotator cuff degeneration, of which hyperglycemia is a risk factor due to its glycation reaction and oxidative stress. We aimed to identify the influence of diabetes-induced glycation and oxidative stress in patients with non-traumatic shoulder RCTs. Twenty patients, aged over 50 years, with non-traumatic shoulder RCTs participated in this study. Patients with a history of diabetes mellitus or preoperative HbA1c ≥ 6.5% were assigned to the diabetic group (n = 10), and the rest to the non-diabetic group (n = 10). Cell proliferation; expression of genes related to oxidative stress, glycation reaction, inflammation, and collagen; intracellular reactive oxygen species (ROS) levels; and apoptosis rates were analyzed. The diabetic group had significantly lower cell proliferation than the non-diabetic group. In the diabetic group, the mRNA expression levels of NOX1, NOX4, IL6, RAGE, type III collagen, MMP2, TIMP1, and TIMP2 were significantly higher; type I collagen expression was significantly lower; and the rate of ROS-positive cells and apoptotic cells, as well as the expression of advanced glycation end-products (AGEs) and the receptor for AGEs (RAGE), was significantly higher. In conclusion, hyperglycemia caused by diabetes mellitus increased AGE and RAGE expression, and led to increased NOX expression, ROS production, and apoptosis in the human rotator cuff. This provides scope to find a preventive treatment for non-traumatic RCTs by inhibiting glycation and oxidative stress. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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11 pages, 979 KiB

Open AccessArticle

Measurement of Redox Biomarkers in the Whole Blood and Red Blood Cell Lysates of Dogs

by Luis G. González-Arostegui, Alberto Muñoz-Prieto, Asta Tvarijonaviciute, José Joaquín Cerón and Camila Peres Rubio

Antioxidants 2022, 11(2), 424; https://doi.org/10.3390/antiox11020424 - 19 Feb 2022

Cited by 5 | Viewed by 2368

Abstract

The evaluation of the biomarkers of oxidative status is usually performed in serum, however, other samples, such as red blood cells (RBCs) lysates or whole blood (WB), can be used. The objective of this study was to evaluate if a comprehensive panel of redox biomarkers can be measured in the WB and RBCs of dogs, and their possible changes “in vitro” after the addition of different concentrations of ascorbic acid. The panel was integrated by biomarkers of the antioxidant status, such as cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of plasma (FRAP), Trolox equivalent antioxidant capacity (TEAC), thiol and paraoxonase type 1 (PON-1), and of the oxidant status, such as total oxidant status (TOS), peroxide-activity (POX-Act), reactive oxygen-derived compounds (d-ROMs), advanced oxidation protein products (AOPP) and thiobarbituric acid reactive substances (TBARS). All the assays were precise and accurate in WB and RBCs lysates. In addition, they showed changes after ascorbic acid addition that are in line with previously published results, being WB more sensitive to detect these changes in our experimental conditions. In conclusion, the panel of assays used in this study can be measured in the WB and RBCs of the dog. In particular, the higher sensitivity to detect changes in our experimental conditions and its easier sample preparation makes WB a promising sample for the evaluation of redox status in dogs, with also potential applications to other animal species and humans. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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16 pages, 2099 KiB

Open AccessArticle

Probing Cell Redox State and Glutathione-Modulating Factors Using a Monochlorobimane-Based Microplate Assay

by Rezeda A. Ishkaeva, Mohamed Zoughaib, Alexander V. Laikov, Plamena R. Angelova and Timur I. Abdullin

Antioxidants 2022, 11(2), 391; https://doi.org/10.3390/antiox11020391 - 15 Feb 2022

Cited by 9 | Viewed by 3429

Abstract

Thiol compounds including predominantly glutathione (GSH) are key components of redox homeostasis, which are involved in the protection and regulation of mammalian cells. The assessment of cell redox status by means of in situ analysis of GSH in living cells is often preferable over established assays in cell lysates due to fluctuations of the GSH pool. For this purpose, we propose a microplate assay with monochlorobimane (MCB) as an available fluorescent probe for GSH, although poorly detected in the microplate format. In addition to the new procedure for improved MCB-assisted GSH detection in plate-grown cells and its verification with GSH modulators, this study provides a useful methodology for the evaluation of cell redox status probed through relative GSH content and responsiveness to both supplemented thiols and variation in oxygen pressure. The roles of extracellular interactions of thiols and natural variability of cellular glutathione on the assay performance were emphasized and discussed. The results are of broad interest in cell biology research and should be particularly useful for the characterization of pathological cells with decreased GSH status and increased oxidative status as well as redox-modulating factors. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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11 pages, 3149 KiB

Open AccessArticle

Extensive Thiol Profiling for Assessment of Intracellular Redox Status in Cultured Cells by HPLC-MS/MS

by Jiandong Wu, Anna Chernatynskaya, Annalise Pfaff, Huari Kou, Nan Cen, Nuran Ercal and Honglan Shi

Antioxidants 2022, 11(1), 24; https://doi.org/10.3390/antiox11010024 - 23 Dec 2021

Cited by 2 | Viewed by 2357

Abstract

Oxidative stress may contribute to the pathology of many diseases, and endogenous thiols, especially glutathione (GSH) and its metabolites, play essential roles in the maintenance of normal redox status. Understanding how these metabolites change in response to oxidative insult can provide key insights into potential methods of prevention and treatment. Most existing methodologies focus only on the GSH/GSH disulfide (GSSG) redox couple, but GSH regulation is highly complex and depends on several pathways with multiple redox-active sulfur-containing species. In order to more fully characterize thiol redox status in response to oxidative insult, a high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) method was developed to simultaneously determine seven sulfur-containing metabolites, generating a panel that systematically examines several pathways involved in thiol metabolism and oxidative stress responses. The sensitivity (LOQ as low as 0.01 ng/mL), accuracy (88–126% spike recovery), and precision (≤12% RSD) were comparable or superior to those of existing methods. Additionally, the method was used to compare the baseline thiol profiles and oxidative stress responses of cell lines derived from different tissues. The results revealed a previously unreported response to oxidative stress in lens epithelial (B3) cells, which may be exploited as a new therapeutic target for oxidative-stress-related ocular diseases. Further application of this method may uncover new pathways involved in oxidative-stress-related diseases and endogenous defense mechanisms. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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14 pages, 707 KiB

Open AccessArticle

Correlations between Salivary Immuno-Biochemical Markers and HbA1c in Type 2 Diabetes Subjects before and after Dental Extraction

by George-Alexandru Maftei, Maria-Alexandra Martu, Marius-Cristian Martu, Dora Popescu, Petra Surlin, Diana Tatarciuc, Cristina Popa and Liliana-Georgeta Foia

Antioxidants 2021, 10(11), 1741; https://doi.org/10.3390/antiox10111741 - 30 Oct 2021

Cited by 20 | Viewed by 2131

Abstract

Dental extraction can trigger certain sequences of complex processes that involve both hard (alveolar bone) and soft tissue (periodontal ligament, gingiva) remodeling. Type 2 diabetes is a serious risk factor for many oral pathologies, both in terms of progression and severity, but also regarding subsequent rehabilitation possibilities. The aim of this study was to establish whether certain molecules: osteoprotegerin (OPG), kappa B nuclear factor receptor activator ligand (RANKL), hepatocyte growth factor (HGF), tumor necrosis factor-α (TNF-α), interleukin 18 (IL-18), matrix metalloproteinase 9 (MMP-9) and oxidative stress markers—total oxidant status (TOS), total antioxidant capacity (TAC)—evaluated in saliva are modified post-extraction in type 2 diabetes mellitus subjects and whether there is a correlation with HbA1c levels. The aforementioned markers plus HbA1c were investigated in a group of systemically healthy subjects (n = 45) and in a type 2 diabetes mellitus group (n = 41) before and three months after a tooth extraction. Diabetes patients’ recorded increased levels of OPG, RANKL, TNF-α, MMP-9, IL-18 and TOS compared to controls both pre- and post-extraction. In both study groups, the average OPG, HGF and TAC level recorded an upward trend three months post-extraction. TNF-α registered a statistically significant decrease only in the diabetes group after dental extraction, together with a decrement of mean HbA1c levels in the diabetes group. By plotting the ROC (receiver operating characteristic) curve, at baseline RANKL, TNF-α, IL-18, MMP-9, TOS and OPG were good predictors of HbA1c levels. Post-extraction, there was a significant correlation between HbA1c and oxidative status biomarkers, however the linear regression model indicated the influence of all studied salivary markers in HbA1c determinism, in a considerable proportion. In conclusion, our study demonstrated that several oxidative status markers and proinflammatory biomarkers are modified in the saliva of diabetic patients and they correlate to HbA1c levels, thus being potential indicators of the post-extraction healing status in the oral cavity. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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10 pages, 1628 KiB

Open AccessSystematic Review

Paraoxonase-1 Concentrations in Obstructive Sleep Apnoea: A Systematic Review and Meta-Analysis

by Maria Carmina Pau, Angelo Zinellu, Elisabetta Zinellu, Gianfranco Pintus, Ciriaco Carru, Alessandro G. Fois, Arduino A. Mangoni and Pietro Pirina

Antioxidants 2022, 11(4), 766; https://doi.org/10.3390/antiox11040766 - 12 Apr 2022

Cited by 2 | Viewed by 1509

Abstract

Obstructive sleep apnoea (OSA) is characterized by overproduction of reactive oxygen species and oxidative stress. The antioxidant enzyme paraoxonase-1 (PON-1) may be useful for monitoring the antioxidant defence systems and the effect of treatments in OSA patients. We investigated, by means of systematic review and meta-analysis, the serum concentrations of PON-1 in OSA patients and non-OSA controls. A literature search was conducted in PubMed, Web of Science, Scopus and Google Scholar databases, from the outset to November 2021, utilizing the terms: “paraoxonase” or “PON” or “paraoxonase-1” or “PON-1” and “obstructive sleep apnoea syndrome” or “OSAS” or “OSA”. Eleven studies in 429 OSA patients and 258 non-OSA controls were involved in the meta-analysis. The pooled serum PON-1 concentrations were significantly lower in OSA (standardized mean difference (SMD) = −0.70, 95% CI −1.13 to −0.28; p = 0.001). Despite the extreme between-study heterogeneity, the SMD values were not substantially affected by the sequential omission of individual studies. There was no publication bias. Our systematic review and meta-analysis supports the presence of an impaired antioxidant defence system in OSA, possibly the consequence of intermittent hypoxia. Further studies are required to determine the clinical use of PON-1 measurements for risk stratification and monitoring in OSA patients. Full article

(This article belongs to the Special Issue Development and Validation of Methods for Analysis of Oxidative Stress in Humans and Animals)

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