Science topic

Oxidative Stress - Science topic

A disturbance in the prooxidant-antioxidant balance in favor of the former, leading to potential damage. Indicators of oxidative stress include damaged DNA bases, protein oxidation products, and lipid peroxidation products (Sies, Oxidative Stress, 1991, pxv-xvi).
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Hi all!
I would like to determine if there is leakage of mitochondrial DNA to the cytosol in cells (after oxidative stress) by qPCR by doing nDNA/mtDNA. According to the protocol in the reference, I have extracted cytosolic mtDNA and nDNA from cells.
But, the results of my experiment were not as expected.
Do I need to ensure that the concentration of DNA in each well is consistent? If the nDNA of each sample is diluted to the same concentration, such as 1 μg/μl, should my mtDNA change by the corresponding multiple with nDNA?
Thank you.
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This is your research project. You are responsible for knowing the literature in detail. You have an advisor, go talk with them.
You should read more about qPCR. Start with the MIQE standards to learn what is needed for reliable, publishable data.
If you have a specific question about qPCR, then come back and ask again.
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I plan to measure total antioxidant status (TAS) and total oxidant status (TOS) in tissue lysates using kits purchased from Rel Assay. I have measured the protein concentration in all my lysates and plan to use the same concentration for all samples. The kits say they are suitable for use with tissue lysates but don't provide a specific working concentration range.
Therefore I plan to run several dilutions to check my samples are within working range of the assay (comparing with the standards provided). However, I am unsure of what concentration range I should apply. I was thinking a top concentration of 200ug/ml, then 150ug/ml, 100ug/ml, 50ug/ml, 25ug/ml. I have four different tissues for each individual and therefore plan to run the same dilutions for all the tissues too.
I would appreciate if anyone could inform me if they think my dilution range is too low or high, or if I need to change my strategy.
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I was wondering what kind of way you applied for your study, David.
I am considering working with the same kit and would like to prepare lysate for suspended cells using RIPA.
Do you have any suggestion on this?
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I want to quantify oxidative stress or one of its indirect marker on frozen tissue (at -80°C for more than 6 months), I want to know if it is still possible with our samples since they are frozen for a long time. Can anyone help me by providing this with reference articles?
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Hi. In fact at such a long time, I think that the results will not be very reliable for you. Except maybe for hydrogen peroxide which is more stable than the others. On the other hand, malondialdehyde, which marks lipid peroxidation, may well pass
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The relevance of the Microbiota-Gut-Brain axis to Alzheimer’s and neurodegenerative diseases needs extensive analysis. The various articles indicate that there are various questions with relevance to microbiota-gut-brain axis that are relevant to the pathology, pathogenesis and treatment of neurodegnerative diseases.Several mechanistic studies are required to determine the underlying mechanisms for effective and safe probiotic treatment for AD and probiotic benefits remain to determined. The relevance of gut dysbiosis may induce inflammatory responses that may be the cause of the induction of the pathogenesis of AD and relevance of diet (unhealthy diets), probiotics and gut microbiota should be carefully assessed. The meta-analysis studies indicate that probiotics reduce inflammation and oxidative stress and enhances cognition in AD and MCI individuals. The effects of different types of probiotics on amyloid formation and deposition needs to be evaluated and probiotic mixture therapy may be unsafe. The safety of probiotic therapy for AD patients require investigation with relevance to neuron reprogramming and programmed cell death in AD. The risk of unsafe microbiota and probiotic use may lead to the inactivation of the anti-aging gene Sirtuin 1 and the generation of uncontrolled short chain fatty acid release that promote amyloid beta plaque formation.
The concerns with relevance to the induction of dyslipidemia and the role of safety of diet-microbiota-brain axis should be carefully assessed with relevance to the cholesterol-AD connections. The prebiotic, symbiotic and probiotic formulations should be carefully assessed for bacterial composition and living microorganisms such as gram negative and positive. The release of bacterial lipopolysaccharides (LPS) from gram negative bacteria needs to be controlled and the content of gram negative bacteria carefully assessed in these prebiotic, symbiotic and probiotic formulations. Unhealthy diets contain end products such as LPS and diets should be carefully assessed for LPS contents since LPS has been associated with the inactivation of Sirtuin 1. The gut microbiota based therapy is in progress and the relevance to the treatment of brain diseases such as AD is limited. The benefits, limitations and safety of gut microbiota and probiotics on Alzheimer’s disease needs to be placed under systematic review with relevance to dietary regulation and postbiotic supplementation that have the implications for amyloidosis and neurodegeneration. The role of probiotic therapies to create a health gut environment by balancing bacterial populations may require the activation of the anti-aging gene Sirtuin 1 to reverse the pathogenesis of Alzheimer’s disease. The literature indicates that yogurt is a prime source for probiotics and provide a healthy balance of live bacteria to provide health benefits to individuals in various countries of the world. However a recent article indicates that within 12 hours yoghurt can grow gram negative bacteria. The gram negative bacteria in yoghurt depending on daily or weekly intake can generate high levels of plasma LPS with relevance to prebiotic, synbiotic and probiotic quality products and ill health. Yoghurt products may need to be assessed for gram negative bacteria populations and LPS to determine the quality control of these products for international communities.
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RELEVANT REFERENCES:
A. Marzban A, Rahmanian V, Marzban A, Ramezani Siakhulak F. The Role of Probiotics in Improving Alzheimer's Disease. JNFS. 2022; 7 (2) :136-138.
B. de Rijke TJ, Doting MHE, van Hemert S, De Deyn PP, van Munster BC, Harmsen HJM, Sommer IEC. A Systematic Review on the Effects of Different Types of Probiotics in Animal Alzheimer's Disease Studies. Front Psychiatry. 2022 Apr 27;13:879491.
C. Guo L, Xu J, Du Y, Wu W, Nie W, Zhang D, Luo Y, Lu H, Lei M, Xiao S, Liu J. Effects of gut microbiota and probiotics on Alzheimer's disease. Transl Neurosci. 2021 Dec 27;12(1):573-580.
D. Ji HF, Shen L. Probiotics as potential therapeutic options for Alzheimer's disease. Appl Microbiol Biotechnol. 2021 Oct;105(20):7721-7730.
E. D’Argenio V, Sarnataro D (2021) Probiotics, prebiotics and their role in Alzheimer’s disease. Neural Regen Res 16(9):1768-1769.
F. Bonfili L, Cuccioloni M, Gong C, Cecarini V, Spina M, Zheng Y, Angeletti M, Eleuteri AM. Gut microbiota modulation in Alzheimer's disease: Focus on lipid metabolism. Clin Nutr. 2022 Mar;41(3):698-708.
G. Naomi, R.; Embong, H.; Othman, F.; Ghazi, H.F.; Maruthey, N.; Bahari, H. Probiotics for Alzheimer’s Disease: A Systematic Review. Nutrients 2022, 14, 20.
H. Arora K, Green M, Prakash S. The Microbiome and Alzheimer's Disease: Potential and Limitations of Prebiotic, Synbiotic, and Probiotic Formulations. Front Bioeng Biotechnol. 2020 Dec 14;8:537847. doi: 10.3389/fbioe.2020.537847.
I. Peterson CT. Dysfunction of the Microbiota-Gut-Brain Axis in Neurodegenerative Disease: The Promise of Therapeutic Modulation With Prebiotics, Medicinal Herbs, Probiotics, and Synbiotics. J Evid Based Integr Med. 2020 Jan-Dec;25:2515690X20957225.
J. Kincaid HJ, Nagpal R, Yadav H. Diet-Microbiota-Brain Axis in Alzheimer's Disease. Ann Nutr Metab. 2021;77 Suppl 2:21-27. doi: 10.1159/000515700.
K. Alessio Vittorio Colombo Rebecca Katie Sadler Gemma Llovera Vikramjeet Singh Stefan Roth Steffanie Heindl Laura Sebastian Monasor Aswin Verhoeven Finn Peters Samira Parhizkar Frits Kamp Mercedes Gomez de Aguero Andrew J MacPherson Edith Winkler Jochen Herms Corinne Benakis Martin Dichgans Harald Steiner Martin Giera Christian Haass Sabina Tahirovic Arthur Liesz. (2021) Microbiota-derived short chain fatty acids modulate microglia and promote Aβ plaque deposition. eLife 10:e59826.
L. Anti-Aging Genes Improve Appetite Regulation and Reverse Cell Senescence and Apoptosis in Global Populations. Advances in Aging Research, 2016, 5, 9-26
M. Appetite Regulation and the Peripheral Sink Amyloid beta Clearance Pathway in Diabetes and Alzheimer’s Disease. Top 10 Commentaries in Alzheimer’s Disease (e-book). 2019;2:1-11. www.avidscience.com
N. Single Gene Inactivation with Implications to Diabetes and Multiple Organ Dysfunction Syndrome. J Clin Epigenet. Vol. 3 No. 3:24.
O. Sirtuin 1, a Diagnostic Protein Marker and its Relevance to Chronic Disease and Therapeutic Drug Interventions”. EC Pharmacology and Toxicology 6.4 (2018): 209-215.
P. Nutritional diets accelerate amyloid beta metabolism and prevent the induction of chronic diseases and Alzheimer’s disease. Photon ebooks. 2015.
Q. Wassenaar TM, Zimmermann K. Lipopolysaccharides in Food, Food Supplements, and Probiotics: Should We be Worried? Eur J Microbiol Immunol (Bp). 2018 Aug 21;8(3):63-69.
R. The Future of Genomic Medicine Involves the Maintenance of Sirtuin 1 in Global Populations. Int J Mol Biol . 2017. 2(1): 00013.
S. Bacterial Lipopolysaccharides and Neuron Toxicity in Neurodegenerative Diseases. Neurology Research and Surgery. 2018; 1(1): 1-3.
T. C.J. Hervert, N.H. Martin, K.J. Boor, M. Wiedmann. Survival and detection of coliforms, Enterobacteriaceae, and gram-negative bacteria in Greek yogurt, Journal of Dairy Science, Volume 100, Issue 2, 2017, Pages 950-960.
U. Fisberg M, Machado R. History of yogurt and current patterns of consumption. Nutr Rev. 2015 Aug;73 Suppl 1:4-7.
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Gerobiotics: probiotics targeting fundamental aging processes
Gerobiotics: probiotics targeting fundamental aging processes (nih.gov)
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I would like to measure H2O2 on tomato leaves. Just need a fast and quantitative complete protocol for the determination of H2O2.
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I'm setting up some assays to assess the effect of different compounds over a cell line. I started using the CM-H2DCFDA from Thermo, as it seems to be more sensitive, but it's turning very expensive if I try to scale it to an HTS. So, I'm looking for alternatives that could give me the same readout in a large-scale screening.
I appreciate your comments.
Many thanks,
Noni
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CM-H2DCFDA is a derivative of H2DCFDA, but with an additional thiol reactive chloromethyl group, which enhances the ability of the compound to bind to intracellular components, thereby prolonging the dye’s cellular retention.
So, this indicator exhibits much better retention in live cells than H2DCFDA.
You could refer to the link below for alternatives.
Best.
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I would like to know if there are any database or research study carried out mentioning about the proteins that doesn't have the cysteine aminoacid.
I knew few but would like to know more about them.
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,you can go through this article providing details of cysteine free protein family.
Potato Proteinase Inhibitor II Family,
These articles may be effective for your research
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There is a antioxidant defense system in cell.
Have any signalling pathway or other things can influence ROS level in a extracelluar way?
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Hi, the pathways of PAMPs and DAMPs mediated by pattern recognition receptors such as TLRs are missing. These can be directly oxidative stress in the intracellular pathway through the classical NFκB pathway or inflammasome-mediated inflammation, or they can induce inflammation through producing proinflammatory cytokines.Hypoxic response (mainly Hif-1α) and oxidative stress from mitochondria during ischemia-reperfusion should also be considered.
Thanks! 
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As a Biomarkers, I have protein concentration, protein carbonyl content, and GST activity in tadpoles. From this data result, I prepared plots and now I want to explain the relation between them. Does anyone know how they are related to each other?
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Thank you Azadeh Eshraghi for your answer.
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Seeking collaborators for the application of the Oxidative Stress Index (OSI) questionnaire in predicting the likelihood of disease onset and severity with increasing OSI.
Please contact Dr. Harold Zeliger at Zeliger Research, LLC.
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Hi Harold Zeliger can you explain more about what collaboration consists of?
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I would like to make an oxidative stress model for studying the antioxidant activity of a drug using C6 cells. Can I use the undifferentiated cells for the same.
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Dear colleague! It depends on the characteristics of the agent for oxidative stress and its concentration.
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How Campylobacter and Helicobacter species isolated from patients stored @-80 survive oxidative stress if they faced temperature variations?
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Dear Prof.Zaim,
Thank you so much for your detail explanation to my question.
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Hi everyone, and thanks in advance for reading!
I want to determine oxidative stress human serum/plasma using protein damage; I´ve seen there are a lot of kits and methods, I want to try an ELISA for this, but as I read more and more papers, the methods described are many times vague and unclear.
Please, If someone is doing this in his Lab or have experience on this issue, can you please reach me a protocol for sample preparation and protein carbonylation detection?
It will be very useful as I see there are a lot of steps and I will save a lot of time (and money) on starting it up making my own mess...
Thank you all!
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I. N. Popov thank you so much!!
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I need to carry out live cell imaging of the above gram-positive bacteria. However, when I transfer the cells from the liquid culture (cells are in exponential phase) to agar pad, the growth alts.
I do not have this issue with Gram-negatives. Any suggestion?
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Enrich them in BHI broth then you can subculture on agar media
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I am working of oxidative stress and found out one target protein form network pharmacology approach. We tried the molecular docking studies with some triazol derivatives. This gave us good result. Now we are planning to do wet work with this system but meanwhile i would like to so some simulation studies on the same. If any body is interested in collaboration pl mail me on
Dr. Manisha Modak
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Good Idea!!
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Dear all, I have been using blue light to induce oxidative stress in human cell line. I understand it's a phenomenon for the light irradiance to reduce after using of a period of time. However, other light panels that are not in use seemed to show a decrease as well. As for the cells, they were showing simialr response even though the irradiance has decreased. Could anyone please provide me with some advice on the possible reasons?
Thank you very much!
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You should construct a device for measurements of light intensity. The simplest one is a biased silicon photodiode, for which you can measure current by a usual multimeter. You can use an AA or AAA battery as a source of bias. Of course, you should perform such measurement with a fixed distance between light source and photodiode and their orientation. Such a simple device will give you the numerical value of light intensity and the possibility to discuss difference or time evolution if you find any.
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complement factor receptor (C3aR and C5aR) activation under oxidative stress conditions on ARPE-19 cells.
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In a patient with hereditary desminopathy (mutation Thr341Pro DES in the heterozygous state) over the past three years, an increase in the blood uric acid level up to 440-480 µmol / l was established by 1.5 times (the norm is 428.4 µmol / l). With the progression of the disease, the level has risen and is above normal. It is known that uric acid is an antioxidant. Is it necessary to reduce the level of uric acid? The patient has no problems with the joints.
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The change in the level of uric acid and biochemical parameters in a patient with an identified case of desminopathy is presented in the article https://www.researchgate.net/publication/357311034_CHANGE_IN_REDOX_STATUS_AND_BIOCHEMICAL_PARAMETERS_IN_PATIENT_WITH_DESMINOPATHY_T341P_SEVERAL_YEARS_AFTER_DISEASE_SYMPTOMS_ONSET
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Dear all, I have been observing the activation of phosphorylated ERK1/2 in my healthy human retinal pigmented epithelium cells (ARPE-19) now and then during experiment. The healthy control group was treated similarly with the other treatment groups just that it was without oxidative stress induction. I have tried different harvesting methods (scrapping & trypsin) and also used a new vial of cell line but it didn't solve the problem. Could you please enlighten me about the possible reasons and how do I troubleshoot it?
Thank you very much!
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Thank you Prof Murakami for the suggestion :)
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So many researches have proved the effects of chronic inflammation or oxidative stress on human health, but no systematic discussion about the relationship between both. From what we can know now, the inflammatory process can induce oxidative stress, in return, the oxidative stress can also induce inflammation. Is it possible to decide who comes first? Or it is a new chicken-and-egg problem?
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Oxidative stress is mainly due to inflammatory mediators such as ROS, RNS free radicals produced by inflammatory cells such as macrophages and neutrophils activates NF-KB a key transcription factor involved in the process leads to tissue damage, cell aging, DNA damage, and cell death.
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Well, we read a lot about how in vitro manipulation of oocytes can induce stress and therefore reduce their potential or as people call it "quality". I've been looking into the studies mostly, they focus on the oxidative stress and reactive oxygen species relation with calcium regulation. Other studies investigated the heating as a stress factor. Well, how they established that this oocyte is healthy and this one is stressed? On what basis? What kind of methods they used? Are the polscope data useful?
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In vitro culture conditions never meets the body environment (in vivo conditions). Therefore, once the oocytes are retrieved for IVF purpose, they are always exposed to stress conditions (due to minor changes in physical, temperature and culture medium). Oocyte is more susceptible to the stress conditions due to larger surface area (100-120 microM in dia) and even minor stress conditions may initiate downstream signaling pathways to induce oocyte death under in vitro culture conditions. We have identified some morphological features that could be used to identify the stressed oocytes in vitro. The series of morphological changes that are identified in oocytes cutlured in vitro are:
1. The smoothness (clear) of oocyte cytoplam is reduced
2. Initiation of cytoplasmic granulation
3. Changes in Zona pelucida histoarchitecture
4. Polar body roughness (fragmentation)
5. Cytoplasmic dia shrinkage (more gap between zona and corona)
6. Membrane blabbings
7. Cytoplasmic fragmentation
and many more........
For more details, kindly see few of our published papers:
1. Apoptosis, 10: 863-875. IF=4.677
2. Fertility Sterility; 84; 1163-1172. IF= 7.329
3. Free Radical Research, 42:212-220 IF=4.148
4. Free Radical Research, 43, 287-294. IF=4.148
5. Eur J Pharmacol. 667; 419-424. IF =4.432
6. J Biomed Sci. 23;36,1-5. IF = 8.410
7. J Biomed Sci 25(1);36 (1-7). IF= 8.410
8. J Biomedical Science, 26;11 (1-6). IF=8.410
9. Stem Cell Reviews and Reports, 17(3): 777-784. IF=5.739
Good luck
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I am working on lung inflammatory disorder which is marked by oxidative stress. In order to measure mitochondrial ROS in bronchoalveolar lavage fluid (BALF) sample procured from mice, I am planning to utilize mitoSOX probe. I have a query if we can directly use mitoSOX on BALF cells and measure the fluorescent intensity through flow cytometry? OR
Do we need to use antibodies (along with mitoSOX) against specific markers of inflammatory cells involved in lung inflammatory disease?
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BALF from infectious mice has both Macs and Neutrophils. So, If you want to compare ROS in a particular cell type, it is recommended to add a marker antibody in the experiment.
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Hi, I'm trying to study the impact of reactive oxygen species on whole blood with respect to how it may induce proteolysis on an intracellular protein in PBMC's in a simulated extracorporeal circulatory model. Instead of exposure of PMBC's to hydrogen peroxide, I need to mimic the actual inflammatory environment such as cardiopulmonary bypass and therefore need whole blood. Trialed H2O2 diluted down to 0.05% without much success. Any suggestions would be much appreciated.
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It is not clear from your given details that which reactive oxygen species your are targeting for? For more focused suggestions, could you please make it bit more clear on your experimental targets? Anyway, with your given infromations, let me tell you that the hydrogen peroxide (H2O2) is a strong oxidizing agent and can generate OH- (hydroxyl ion) in a cell or solution. There are several reactive oxygen species and every one has its own pathway for damaging cellular histoarchtecture depending on the extent and nature of insult on the cellular/subcellular and macromolecules like protein and lipids. You can measure H2O2 level in blood serum/plasma using sensitive kits from R&D systems, USA. Total NO levels can also be measured accurately by using Nitrate/Nitrite kits in blood samples using kits from R&D systems, USA and other company make kits. Thus, you can analyze both H202 and total NO levels in blood samples.
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There are a few manuscripts implying redox regulation of the actin cytoskeleton. This makes me wonder if actin B can be used as a good loading control when investigating protein expression during oxidative stress. If not, what would be a more suitable loading control?
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Hello,
You can use GAPDH as loading control.
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Hi everyone;
is there any method, to differentiate this two products of oxidative stress in the same sample?
(apart from GC/MS)
thank you so much!
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How to go about it statistically?
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Most organic materials, in living systems or not, are susceptible to degradation at varying temperatures in the presence of atmospheric oxygen. This oxidation is at the origin of the deterioration of the mechanical properties of polymers, the rancidity of food or technical fatty substances, or various pathologies such as diabetes.
In biological systems, oxidative stress is the result of an imbalance between the production of free radicals or reactive oxygen species (ROS) and their destruction by antioxidant defense systems. Free radicals can cause significant damage to cell structure and metabolism by degrading proteins, lipids and nucleic acids. To cope with these attacks, organisms have developed antioxidant action systems: enzymatic and non-enzymatic defense systems.
In the enzymatic defense system, three types of antioxidant enzymes are used to destroy reactive oxygen species:
- Superoxidizedismutases (SOD) which catalyze the disproportionation of the superoxide anion to hydrogen peroxide (2📷+ 2📷+📷).
- Catalase which catalyzes the disproportionation of hydrogen peroxide into water allowing its elimination (📷O+📷).
- Gluthathione peroxidase (GPX) which also breaks down hydrogen peroxide using gluthation as a hydrogen donor (📷+ 2GSH📷O +GSSG).
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Hi All
s-glutathionylation is known to protect cysteines from irreversible oxidation and to keep the residue for reduction after oxidative stress is controlled.
I wonder, however, if the oxidative condition is persisting, even the glutathionylated cysteines can be further oxidized into an irreversible form particularly in the absence of enzymatic reactions.
Thanks for your answers in advance!
Sunjoo
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Tripeptide glutathione constitutes the first anti-radical defense barrier thanks to the nucleophilic thiol groups (SH) but in an oxidizing medium it can play the role of a highly reactive radical (GS-) which can cause irreversible cell damage.
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I am studying the influence of doses of plant extracts on rats, since the plant is toxic, the value of MDA (oxidative stress) increases with increasing doses. the problem is that for the maximum dose chosen the value of MDA decreased despite that at this dose an apoptosis occurred I have not found an explanation.
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You can also read the following paper, it may be helpful to you. With regards.
'Reactive Oxygen Species-Induced Lipid Peroxidation in Apoptosis, Autophagy, and Ferroptosis' https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/31737171/
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The tissue we are using is the cerebellum of black mice blc57, we are doing histopathology and oxidative stress assay, Should we use half the number of mice for each test or could we divide the cerebellum and do both tests for the whole number?
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Thank you Dr. shin and Dr. Manju for your answer.
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Serum TIBC and serum ferritin colorimetric assay protocol.
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Hi, arigo has a good Human Ferritin ELISA Kit for serum and plasma. Please refer to the detail at https://www.arigobio.com/Human-Ferritin-ELISA-Kit-ARG80501.html
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I have seen different results published online using CATALASE. I know that the outcome of the analysis depends on the type of tissue, and protocol used but I have seen gotten different results from my brain tissue sample using sample protocol.
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Please note that if you want check OXIDATIVE STRESS, you can not stick to the enzymes. You should measure lipid peroxidation, protein carbonyl, GHS:GSSG ratio. They indicate if oxidative stress occured or not.
However CAT results are somehow strange in in vivo conditions.
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oxidative stress, histopathology, leakage of enzymes, lipid profile, inflammation, apoptosis
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Many thanks for your responses
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Recently we treated retinal pigment epithelium (RPE) cells with hydrogen peroxide as oxidative stress, and the morphological changes do not look like apoptosis or necrosis. Does anyone give suggestions regarding oxidative stress-induced cell death? Many thanks.
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doi: 10.3892/ijmm.2012.1215. Epub 2012 Dec 18.. 2013 Feb;31(2):471-6.Int J Mol Med. The effects of exogenous H2O2 on cell death, reactive oxygen species and glutathione levels in calf pulmonary artery and human umbilical vein endothelial cells.
Can be useful
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I am working on oral cancer cell line and i would like to observe the in vitro antioxidant activity in treated cancer cell line to evaluate the enzymatic activity like SOD, CAT, GST. I have read many papers and they have tried inducing oxidative stress in the cancer cells to observe this activity. I would like to know why we need to induce it when the cancer cells can also produce free radicals.
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I think the purpose behind this is to make sure all the cells will produce extra ROS that can't be easily managed by the already existing cellular antioxidants, so the activities of the mentioned enzymes will show clearly
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I have tried with no success various UV-B stresses but I did not find an increase in beta galactosidase.
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Hi Anthony L, Yes: we often used the DNA-damaging drug etoposide to induce senescence. Soluble in DMSO. 10 uM was often ok, though the best concentration can vary with the cell type -- it gets toxic at higher concentrations (because extensive DNA damage is toxic). It was added to recently plated cells for 48 h, then removed, and the cells were left for 5 days to develop the senescent characteristics -- although this may also vary with cell type. So for a new cell type you might want to try lower and higher concentrations, and different timings. We published this method (used as a positive control) in Cairney CJ et al. 2017, PLoS Genetics. (See Supplementary File 2.)
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I am looking for the reagent which can scavenge superoxide in living cells. Most of ROS scavenger, like tempol, inhibit not only superoxide accumulation but also reactive nitrogen spices (RNS). But I am actually looking at the effect of RNS in oxidative stress but not superoxide. Dose anyone know the cell permeable superoxide scavenger?
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Hey Kazushi,
Tiron (1,2-dihydroxybenzene-3,5-disulfonate) is a vitamin E analog, cell-permeable
and acts as a global superoxide scavenger as an alternative for TEMPOL.
If you like you can have a look at our recent review:
"Functions of ROS in macrophages and antimicrobial immunity".
In this review, we give an introduction to ROS and their sources in macrophages, summarize the versatile roles of ROS in direct and indirect antimicrobial immune defense and provide an overview of commonly used ROS probes, ROS source inhibitors and ROS scavengers (also the difference between ROS scavengers and antioxidants, which are not synonymous, is explained).
Functions of ROS in Macrophages and Antimicrobial Immunity
February 2021, Antioxidants 10(2):313
All the best and stay healthy,
Marc
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I am searching for a lab to do In vitro study in neuronal cell lines like SH-SY5Y or N27 and the biochemical estimation by the following methods:
ELISA, Western blot
Oxidative stress markers (ROS, SOD, CAT and GPx)
Mitochondrial function (Mitochondrial Membrane Potential and Complex I activity)
Kindly suggest me a government lab.
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@Ehab Yehiea Jabber. Please tell me
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Adding a drug causes cells to undergo oxidative stress as indicated by DCFDA fluorescence and an increase of ABCG2. However, I get inconsistent results of SOD1 after 24 hours of exposure. Also, if you want to get the expression, do you collect only the remaining live cells or also collect the floating/apoptotic cells?
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Functioning of Superoxide Dismutase is increased, because generation of free radicals produce adverse effects. SOD catalyze the chemical reactions having single oxygen and slow down or prevent the generation of free radicals. So, automatically SOD activity increased...
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I have been trying to estimate some biochemical parameters in urinary bladder but homogenization is problem with me as indicated by very low amount of protein in homogenate.
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I'm a retired research chemist/physicist with Parkinson's disease and I've been working on upregulating Nrf2 with sulforaphane to fight oxidative stress.
The results I achieved on my PD and with a group of 8 people with PD have shown that sulforaphane strongly attenuated non-motor symptoms especially fatigue and lack of motivation, suggesting that the first target for Nrf2 seems to be mitochondria. I am not an expert in this subject and I would like to reach out to research groups and Parkinson's disease specialists to discuss how to take this idea further.
You can find the background here :
And a preprint I have written on RG here:
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I have been using 400 micrograms twice a day. Brand: Swanson.
I have had the drug checked at the University and they confirmed it is 87% pure sulforaphane. Not the 99% the lab says. But is good enough for me.
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for induction of oxidative stress
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I'm following the best answer.
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We are trying to design a clinical trial on type 2 diabetes patients. The main data that we want to assess include FBS, 2hpp, HbA1c, insulin, and HOMA-IR. Also, we will assess the lipid profile and stress oxidative indices (MDA and TAC). The problem is that we could not find any similar study to determine the sample size. In this situation is it possible to use the Cohen formula? If not what is the right way for determining the sample size?
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You can use G*Power calculation to determine your sample size without worrying if the sample size is under the attribution of statistical significance. G*Power is a common sample calculation particularly in setting up a clinical trial.
Please check out this publication for more details:
Use of G*Power Software | SpringerLink by:
Verma J.P., Verma P. (2020) Use of G*Power Software. In: Determining Sample Size and Power in Research Studies. Springer, Singapore. https://doi.org/10.1007/978-981-15-5204-5_5
Hope it would be helpful and all best luck!
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Hi all,
I have been using hydrogen peroxide, Potassium ferricyanide and cumene hydroperoxide for my studies involving proteins with a redox switch. The proteins I typically worked and working are transcription factors and polymerases with redox switches like HXXXCXXC motif and Fe-S clusters. I purify the proteins under anaerobic and/or reducing conditions, oxidise them to observe the changes for example structural and activity related. Ferricyanide is a coloured compound and thus I can not use it for some methods, whereas hydrogen peroxide leads to the formation of air bubbles in the cuvette. Given these issues, It would be of a great help if any of you could suggest me any other chemicals that can be tested for the same purpose.
Thanks in Advance
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Well the reasons for the bubbles with the peroxide is its decomposition by catalase, you could try adding less peroxide to the sample, or inactivate the catalase with something like aminotriazole (though inactivation of catalase is, in and of itself an oxidative stressor).
You could look at inactivating certain antioxidant enzymes such as glutathione reductase or peroxidase. There are available inhibitors for these enzymes.
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I am looking for a new method for measuring oxidative stress in animal tissues ... a previously unrecognized method .... Thank you for your cooperation
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i am working on Carum copticum extract and its seems it has itself induced oxidative stress at higher doses. 200mg/kg PO
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Sourbh Garg , could you explain please what is "the immunization of your compound?" In addition, it's not good at all to use bold fonts.
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Hello all,
What are some markers of oxidative stress in brain?
Thanks
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Yes, Various antioxidant enzymes such as Superoxide dismutase (SOD), Catalase (CAT), Glutathione (GSH) and oxidative damage biomarkers such as malondialdehyde, hydroperoxides, carbonyl groups etc. are considered for research work.
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Susceptibility of Covid-19 patients to drug-induced OS
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That's the worst scenario that could covid19 patient face, as a result doctors constantly recommend to covid19 patient to follow a diet rich of antioxidant such as tumeric, ginger, garlic ... etc
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I am planning to conduct a research on oxidative stress of the pregnant mothers. There I am planning to compare oxidative stress between mothers with PIH and mothers without PIH. I would like to know what would be the best research design to conduct this study. Is case control study design ok for this? Or is there a better way?
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Hello
a well-designed case-control
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Hallo experts,
How can I best measure damage after oxidative stress in liver tissue harvested 24h after exposure stop?
Im thinking about TBARS Elisa kit, but maybe anyone have advice regarding a better endpoint to measure? 4-HNE maybe, or Protein Carbonyls?
If someone have kits to recommend – I will be very happy. (ELISA is preferable).
All the best, and hope everyone is safe
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Hello, Hildegunn!
Spectrophotometric determination of tissues MDA, PCO, and GSH levels
MDA
This method depends on the formation of MDA as an end product of lipid peroxidation which reacts with thiobarbituric acid producing thiobarbituric acid reactive substance (TBARS), a pink chromogen, which can be measured spectrophotometrically at 532 nm, an MDA standard was used to construct a standard curve against which readings of the samples were plotted [22].
PCO
This method depends on the formation of a Schiff base from the reaction of dinitrophenylhydrazine with protein carbonyls to form protein hydrazones which was measured spectrophotometrically. Briefly, after precipitation of protein with an equal volume of 1% trichloroacetic acid, the pellet was resuspended in 10 mmol/L DNPH plus 2N HCl, or in 2N HCl as a control blank. Next, after the washing procedure with 1:1 ethanol-ethylacetate, the final palette was dissolved in 6 mol/L guanidine. The carbonyl group was determined from the absorbance at 370 nm. The carbonyl content was calculated in terms of nmol/mg protein [23,24].
GSH
The method is based on the reduction of 5,5 dithiobis (2-nitrobenzoic acid) (DTNB) with reduced glutathione (GSH) to produce a yellow compound. The reduced chromogen is directly proportional to GSH concentration and its absorbance can be measured at 405 nm by using a commercial kit was used (Biodiagnostic, Egypt) [25].
Enzymatic Assays
Tissues GST enzyme activity: it measures the conjugation of 1-chloro-2,4-dinitro benzene (CDNB) with reduced glutathione that produces a dinitrophenyl thioether which can be detected by spectrophotometer at 340 nm. One unit of GST activity is defined as the amount of enzyme producing 1 mmol of CDNB-GSH conjugate/min under the conditions of the assay according to the method described by Habig et al. [26].
Tissues GPx enzyme activity: it was measured as IU/gm wet tissue by the reaction between glutathione remaining after the action of GPx and 5, 5-dithiobis-(2-nitrobenzoic acid) to form a complex that absorbs maximally at 412 nm. Glutathione peroxidase activity of 1 U/mg protein was defined as 1 μg of GSH consumed/min/mg protein [27].
Determination of tissues CAT enzyme activity: it assayed by the method of Sinha which based on formation of chromic acetate from dichromate and glacial acetic acid in presence hydrogen peroxide, chromic acetate that produced was measures colorimetrically at 570 nm, one enzyme unit was defined as the amount of enzyme which catalyzed the oxidation of 1 μmole H2O2 per minute under assay conditions [28].
Tissues PON1 enzyme activity: PON1 activity towards paraoxon (O,O-diethyl-O-p-nitrophenyl phosphate) was determined by measuring the initial rate of substrate hydrolysis to p- nitrophenol, whose absorbance was monitored at 405 nm in the assay mixture, A PON1 activity of 1 U/mg protein was defined as 1 μmol p-nitrophenol formed per minute per mg protein.
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What media or salt solution (HBSS, EBSS, Tyrode) can be used for real-time intracelular h2o2 imaging? I am using a genetically-encoded biosensor based on roGFP and have to find the media that wouldn't have much buffering capacity as well as compounds acting like antioxidants, since it is important to see how the cells alone are coping with h2o2 adition.
Estimated duration of the experiment - 1-1.5 h.
Cells: ipsc-derived neurons. Standart Neurobasal+B27 media was designed to protect neurons from any kind of oxidative damage and contains loads of antioxidants, so h2o2 don't get the chance to get inside the cell.
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The Cell Meter™ Hydrogen Peroxide Assay Kit OxiVision™ Green hydrogen peroxide sensor to quantify hydrogen peroxide in live cells
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I am looking for a biological response of fish exposed to trinitrotoluene or its amino metabolites.
Maybe a more or less specific gene expression could help?
Oxidative stress is one good example but its not specific for nitroaromates.
Any suggestions?
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Yes, i searched for it in NCBI gene database to make sure. :) yes i noticed your name after i posted the answer. Great article by the way.
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My OD value had a huge variation between each trial, even blank one. The worst thing is blank two has higher OD than the corresponding sample no matter if it was diluted or not. The repeatability of the assay kit is not good. My protocol is as follows.
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Lara Ivanković why don't you try to analyze the samples with an additional method for verification of the results? Inhibition of 1,2,3-trihydroxybenzene is often used ( . Some comparisons with standard kits have also been published ( eg. https://faseb.onlinelibrary.wiley.com/doi/abs/10.1096/fasebj.21.6.a814)
Kits are often ungrateful as, in reality, each analytical method should be optimized for a specific purpose and quality parameters should be satisfied for given conditions (eg. not all kits work with all buffers, with the same sample preparation procedures, ...)
If you decide to double-check with the pyrogallol method I'd be happy to help.
Best,
Jan
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Hello,
I also want to do some biochemical analyses and need to take some blood of the rats. To take the blood I need to use anesthetics. But I want to evaluate TNF-a and oxidative stress in the brain. Which anesthetic does no influence on this? I'm working on the model of VPA to induce austism.
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Oi!
I think that it's actually essential to use an anesthetic because of the ethical concerns (you could be authorized to not use it if you can prove that is could interfere with the quality of your data). Normally, it's preferable to use an aesthetic cocktail to ovoid the single high dose monoanesthesia toxicity.
Here I found you a comparative study of different methods of animal euthanasia to collect brain samples for metabolic profiling.
Good luck,
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Hello dear scientists:
When I treated the Pc12 cell line with H2O2, after 2h the cells seemed to be dead.
I didn't observe crystals, so I didn't add DMSO and left it with mtt in the incubator.
Today I checked it (after 24h) all the wells contained formazans. Also, oD was ok too. Is it a reliable result?
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Rabeah Al-Temaimi after seeing formazan  I changed the media and added  5o microliter DMSO and reed plate at 570 nm. Thank you I will try your suggestion. Today I observed the same for my synergic test, at first cells, dead, but after 24h, Fromazans appeared.
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We are doing some biomarkers of oxidative stress research and need to construct calibration curves using BHA, BHT for comparison of antioxidative power in certain extracts, so I would like to know what quality (purity) the reagents (BHA and BHT) need to be.
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For routine antioxidant assay, your standards do not have to go as far as an HPLC analysis grade. What you can get from the likes of Sigma-Aldrich of about ≥98.5% purity should do. The analytical grade of over 99% is worthwhile if your assay is based on HPLC or equivalent instrumentation. I personally use the above-mentioned quality from Sigma source for antioxidant power assay.
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the relationships between oxidation and inflammation
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Inflammation and oxidative stress are closely related and tightly linked pathophysiological processes. One of them may appear before or after the other, but when one of them appears the other one is most likely to appear; and then both of them take part in the pathogenesis of many chronic diseases. Although identification and treatment of primary abnormality are of great clinical importance, treating only the primary abnormality may not always be successful, because once the process has been already started, both inflammation and oxidative stress act in concert to accentuate each other and to induce progressive damage. Thus, antioxidant therapy alone is unlikely to prevent diseases known to be induced by oxidative stress, like cardiovascular and diabetic complications, neurodegenerative diseases, cancer, or aging. However, great care should be taken in selection of antioxidant agents, selection of dosage of antioxidants not to produce harmful effects, and, most importantly, quantification of redox and inflammatory status to make appropriate interpretation of the findings.
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Hello Can anyone suggest a good way of inducing oxidative stress in mice to exhibit proteinurea phenotype in the mice or a good method to induce oxidative stress in the mice to study kidney disease progression due to oxidative stress?
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I'm looking for laboratories working with oxidative stress to test nitrocellulose redox permanganometry (NRP). NRP is a simple method for reductive capacity assessment based on the analysis of the MnO2 precipitate following redox reaction between KMnO4 and biological samples fixed onto the nitrocellulose. This method estimates how well the sample can give up its electrons to KMnO4, so it estimates the amount of chemical antioxidants. In other words this method can be used to measure total chemical antioxidant capacity of biological samples. Advantages of the method are: great precision and accuracy; the possibility to measure a lot of samples simultaneously; simplicity; cost (you only need a piece of nitrocellulose membrane and KMnO4). Moreover, the method can be used to obtain information on the spatial distribution of antioxidant capacity in tissue sections (both cryosections and FFPE can be used), providing unique information on the redox status of the tissue.
As the method is new, and we proposed it just recently, I'm looking for laboratories that are working with oxidative stress to test the method in their samples. We conducted thorough analyses and the method seems to be very robust. Nevertheless, I believe additional independent testing is always a good idea. Also, I believe comments of researchers closely working with oxidative stress would be beneficial for further development of this simple method.
The original paper can be found here:
A step by step protocol can be found here:
Best,
Jan
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In my opinion, there are no simple methods to evaluate antioxidizing capacities of potential antioxidants. General tests like DPPH, ABTS... only (roughly) measure the ability of the antioxidants to scavenge free radicals. An antioxidant is much more than that; it can act as a scavenger of free radicals, repair of the oxidized species (reduction by one electron transfer) or by cascade (cumulative) effect . Furthermore, the antioxidant capacity can also depend on the agent responsible for the oxidative stress conditions (selective antioxidants).I recommend the evaluation of the mutual behavior of both antioxidant and the target to be protected in every specific oxidizing environment.For details see e.g.:
  • Santos, P.M.P.; Telo, J.P.; Vieira, A.J.S.C. "Structure and Redox Properties of Radicals Derived from One-electron Oxidised Methylxanthines", Redox Report (2008), 13(3), 123; DOI: 10.1179/135100008X259231
  • Santos, P.M.P.; Antunes, A.M.M.; Noronha, J.; Fernandes, E.; Vieira, A.J.S.C. "Scavenging activity of aminoantipyrines against hydroxyl radical", Eur. J. Med. Chem. (2010), 45, 2258-2264; DOI: 10.1016/j.ejmech.2010.01.071
  • Santos, P.M.P.; SILVA, S.A.G., JUSTINO, G.C.; VIEIRA, A.J.S.C. "Demethylation of Theophylline (1,3-Dimethylxanthine) to 1-Methylxanthine: the First Step of an Antioxidising Cascade", Redox Report (2010), 15(3), 138; DOI: 10.1179/174329210X12650506623726
  • SANTOS, P.M.P.; VIEIRA, A.J.S.C.Antioxidising activity of cinnamic acid derivatives against oxidative stress induced by oxidising radicals”, J. Phys. Org. Chem. (2013), 26, 432; DOI: 10.1002/poc.3104
  • VIEIRA, A.J.S.C.; SANTOS, P.M.P. “A Tentative Classification of Antioxidants: Which Role They Play when Protecting Biological Targets from Oxidative Stress Induced Damage?”, J Med Chem Drug Des (2017), 1, 1-3; DOI: 10.16966/jmcdd.101
  • VIEIRA, A.J.S.C.; GASPAR, E.M.; SANTOS, P.M.P. “Mechanisms of potential antioxidant activity of caffeine”, Rad. Phys. Chem. (2020), 174, 108968; DOI 10.1016/j.radphyschem.2020.108968
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I have to measure oxidative stress and nitrite in tissue as well as plasma. For tissue we have the protocol in place but for plasma what is the procedure that should be followed for its preparation before proceeding for the assay?
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Depends on what you want to measure. For example, TBARS (often used for lipid peroxidation assessment) is usually analyzed following n-butanol extraction of the coloured adduct. Quantification of protein thiols (or GSH) is usually done by reacting DTNB with thiols, so you have to precipitate the proteins prior to analysis and analyze the fraction of interest.... the same applies for plasma analysis. What exactly do you want to measure?
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I want to do study the oxidative stress induced by H2O2 in Keratinocytes cells. What is the volume and concentration of H202 added to the cells to study the H2O2 induced cell damage?
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This is one of my research in which I used hydrogen peroxide to induce oxidative stress in rats: I gave drinking water by special bottles containing 1% hydrogen peroxide for 15 days:
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Protein carbonyls are important compounds to measure progression of oxidative stress in tissues.
Available articles online do not have details about how to do final estimation/calculation of the carbonyl content.
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Have a look at the references of a review paper that summarize the recent work in this field of study, for example the spectrophotometric methods to optimize the amount of protein have been investigated in the following article
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Which of the following genes is more important for diagnosing and examining apoptosis?
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I think you can test the following gene: Nrf2, p38MAPK, NFkB and so on. I have listed relevant literature below for your reference:
Exacerbation of diabetes-induced testicular apoptosis by zinc deficiency is most likely associated with oxidative stress, p38 MAPK activation, and p53 activation in mice;
Nuclear Factor-κB Activation in Human Testicular Apoptosis;
Besides gene expression, I am recommend you to determined the content of ROS (reactive oxygen species) and some relatival enzyme activities, such as SOD.
Hope it helps you!
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There is an increasing interest in the role of reactive oxygen species (ROS) and oxidative stress in the causation of male infertility. Can any one give us the exact mechanisms
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ROS play a role in spermatic function and fertilisation. The literature describes both a physiological and a pathological role of ROS in fertility. A delicate balance between ROS necessary for physiological activity and antioxidants to protect from cellular oxidative injury is essential for fertility.
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So many reports mentioned the function of hemoglobin on ROS producction, but few reports could I found on "anti-ROS", does any researchers focus on this part. I think that is interesting to define the role of hemoglobin on redox reaction.
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In fact, the available references on this subject are as follows:
and a book titled:
Free Radicals and Antioxidant Protocols
Editors: Uppu, R.M., Murthy, S.N., Pryor, W.A., Parinandi, N.L. (Eds.)
I hope these references will help you in what you need....
with respect
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I want to induce oxidative stress by hydrogen peroxide in S. cerevisiae. The stress was measured by t-bar assay. But the cells are not taking stress.
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I treated the culture with H2O2 40mM for 1 hour and I could see the lipid peroxidation using T-bars. My culture were in YPGly (medium with glycerol), I don't know if the H2O2 concentration should be different in other media.
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Dear colleagues,
Since 2017, in Redox Research Center, we are working on the use of hydrogen gas as a protective agent for preserving food products. Up to now, we have found an extraordinary effect of hydrogen gas on protecting the property of foods (see our publications in google scholar).
Since 2007, many studies were performed for using hydrogen gas as a therapeutic agent in different illnesses. From these studies, many ones confirmed the antioxidant, anti-inflammatory and anti-apoptotic protective effects of hydrogen on cells and organs. Some studies reported the protective effect of hydrogen against irradiation lung damage (Terasaki et al., 2011), amelioration of hyperoxic lung injury (Kawamura et al., 2013), and reduction of the HBV DNA level in CHB patients (Xia et al., 2013)(refer to the links below). One of these studies reported that "patients receiving hydrogen treatment had improved tendencies in the liver function and HBV (hepatitis B virus) DNA level when compared with patients undergoing routine treatment" and they concluded that "Further study with long‐term treatment with hydrogen‐rich water is required to confirm the protective effect of hydrogen on the liver function and its suppressive effect on viral replication" and " hydrogen‐rich water may attenuate the oxidative stress and have the potential to improve the liver function and reduce the HBV DNA level in CHB patients." (Xia et al., 2013).
I invite the colleagues who work on the viral pathology, especially the COVID-19, to conduct assays on the possible application of hydrogen (in inhalation form, or intraperitoneal and oral administration of hydrogen‐rich water) as a potential and inexpensive treatment of COVID-19.
Please share this message with your colleagues to reach all the researchers and groups working on COVID-19 topics. Any potential use of this cheap treatment for COVID-19 treatment could help thousands of patients and all humanity to win the war against this fatal risk.
Best health for all
Duried Alwazeer
References:
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Dear Dr. Alwazeer,
Thank you for opening this discussion forum. Based on my reading, molecular hydrogen has been studied extensively for reducing oxidative stress and inflammation associated organ injury since the first publication in 2007 in Nature Medicine [1]. Please see a review article by Qiu et al.[2]. The Chinese medical team has recommended use of 66.6% H2 / 33.3% O2 for inhalation for treatment of COVID-19 patients in the 7th edition of new coronavirus pneumonia guideline [3]. Since molecular H2 is safe and can reduce free radicals and inflammatory cytokines [2], the inhalation treatment is expected to significantly improve the condition of cytokine storm.
This treatment is cheep and and effective, as you suggested, physicians in other countries surely can follow Chinese team's lead to apply H2 containing gas to treat in-patients.
[1] Ohsawa I, Ishikawa M, Takahashi K, et al. Hydrogen acts as a therapeutic
antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med. 2007;
13: 688-94.
[2] Qiu P, Liu Y2, Zhang J. Recent Advances in Studies of Molecular Hydrogen against Sepsis. Int J Biol Sci. 2019 May 11;15(6):1261-1275. doi: 10.7150/ijbs.30741. eCollection 2019.
[3] In Chinese
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I'm working on a project to evaluate chemotherapy-induced cognitive impairment with cisplatin administration. I'm planning to use a biomarker, evaluated serially over time. My questions are:
1. What is the single best biomarker for this scenario (i.e. best in terms of sensitivity, specificity, and exclusively pointing to neuronal inflammation and oxidative stress)? I want it to be very specific, displaying aberrant results only when the neurons are injured. Many biomarkers such as interleukin or MDA increases in the event of systemic inflammation, which is almost inevitably happen under this research scenario (i.e. chemo almost always induces systemic inflammation).
2. Should it be derived from CSF or plasma?
Thank you in advance for anyone who can provide me with answers.
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Hi And
What is the effects of chemotherapy?
According to the dose and time of exposure. It could be reversable mild inflammatory response leads to cloudy swelling, hydropic degeneration or fatty change with acute inflammatory cellular infiltrate. If it is a lethal dose the response will be irreversable cell death in the form of Apoptosis changes Or Necrosis.
A lot of cellular and plasma chemical mediators will be released.
so it is difficult to find
the single best biomarker for this scenario (i.e. best in terms of sensitivity, specificity, and exclusively pointing to neuronal inflammation and oxidative stress)?
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I'm using h2o2 as a positive control for h2dcfda assay for the detection of ROS. There's five fold less (20%) Ros than untreated control (100%) Thp1 cells. I'm treating cells for 24 Hours. Can I use some other controls which are less toxic and less expensive.
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HI Sandeep, the radicals produced using H2O2 depend on the availability of Fe for the fenton reaction. If you want the cell's mitochondria to produce ROS you can add Menadion (Vitamin K). at a concentration of 50uM it will induce Apoptosis in the long run but also increase ROS. Your 24h time is probably a challenge if you want to add the positive control for that amount of time you might end up with a lot of dead cells. It is usually better to measure ROS production kinetics, that way you might be successful in detecting differences after a few h. see also https://www.jbc.org/content/278/15/12645.long
Best
Heiko
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a marker taking into consideration the specificity, sensitivity and cost of measuring it.
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Hi,
You can try to quantify FACs (fluorescent aromatic compounds in bile or urine) and associate those levels with lipid peroxidation (TBARs - thiobarbituric reactive species).
Cheers
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I am interested in researching intermittent fasting (IF) and caloric restriction (CR) in relation to hypothesized positive effects on oxidative stress in the nervous system, specifically in the aging, possibly pre-AD brain. I will be using a zebrafish model to study the topic. I am currently in the process of doing a general lit review on the topic, but will be thankful for any insight/difficulties with collecting similar research in the species. It'd be great to have a light shone on any articles I might miss (any species). Thank you for taking the time to read and answer!
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Hi.
It's been a few months. Any update for us here?
I thought I would suggest two themes:
1) Re AD, consider calibrating to emerging consensus (?) that there might be maybe 3 or even 5 distinct etiologies at play. For example, do amyloid plaques serve a protective role against certain pathogens, where that route is completely independent of, say, AD being a form of "brain diabetes", etc. This might prove relevant in the fish model.
2) Overall, consider autophagy as a primary hub of IF impact. Oxidative status could be considered, most fundamentally, downstream of mitochondrial health itself downstream of how much opportunity cells have had to engage in sufficient mitophagy. Growth vs repair/survival programming is astoundingly well conserved over 4 billion years. If there is any food around then reproductive fitness has everything to do with growing/reproducing fast enough for your line to consume as much of it as possible before its gone, before your competitors do. If a huge percentage of your proteins are misfolded and organelles dysfunctional in the process, then so be it: plenty of time to take care of that -- via autophagy -- when baseline normal starvation conditions return for super-long durations. This last part is what modern humans lack, and is the primary basis of all chronic disease. [Also, for what it is worth, my hunch is that amyloid plaques result largely from insufficient autophagy natural break down, most commonly associated with insulin resistance -- where increased insulin powerfully stimulates mTOR and prevents CPT/fat-burning, both seriously compromising autophagy.
3) In line with the above.... if IF / Zebrafish, I would strongly suggest developing a "chronic feeding" protocol in fish to reflect the human model.
Good luck.
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Dear colleagues
I am organizing a Special Issue entitled “Oxidative stress, classification and quantitation” in the Antioxidants (IF 4.52; ISSN 2076-3921, http://www.mdpi.com/journal/antioxidants).
The Special Issue aims to expand our understanding and perspective on all aspects of redox status, its evaluation, homeostasis related adaptation and oxidative damage and repair
. I am particularly interested in your view on the apparent controversy regarding the use of the term Oxidative Stress and the more recently defined terms "Distress" and "Eustress".
In addition, the use of these terms relates to the possibility of quantitating OS. The problem is that the OS, as evaluated by the use of different biomarkers do not correlate with each other, namely, OS cannot be quantitated by a universal criterion and it depends on the biomarker used to evaluate it. Three complementary specific questions are:
(i) Should the term "Oxidative Stress" be re-defined to relate to more specific conditions than the original definition (Acute situations? Diseases? What else ? )
(ii) What term should we use to describe (if at all) imbalance between pro-oxidative and antioxidative components of a system ?.
(iii) Why should we quantitate “Oxidative Stress” (Is there any reason to try “to quantitate OS”?).
If you wish your answer to be publish in the special issue, please let me know
Looking forward to your replays
Dov
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Subject: Promotional Materials: [Antioxidants: IF 4.520] Special Issue "Oxidative Stress, Classification and Quantitation"
Dear Researcher,
I am organizing a Special Issue entitled “Oxidative stress, classification and quantitation”
in the Antioxidants (ISSN 2076-3921, http://www.mdpi.com/journal/antioxidants).
The Special Issue aims to expand our understanding and perspective on all aspects of redox status, its evaluation, homeostasis related adaptation and oxidative damage and repair processes.
In view of your distinguished contribution to this area, I invite you to contribute a research paper and/or a review and /or a commentary to this special issue. I am particularly interested in your view on the apparent controversy regarding the use of the term Oxidative Stress and the more recently defined terms "Distress" and "Eustress". In addition, the use of these terms relates to the possibility of quantitating OS. The problem is that the OS, as evaluated by the use of different biomarkers do not correlate with each other, namely, OS cannot be quantitated by a universal criterion and it depends on the biomarker used to evaluate it. Three complementary specific questions are:
(i) Should the term "Oxidative Stress" be re-defined to relate to more specific conditions than the original definition (Acute situations? Diseases? What else ) ?
(ii) What term should we use to describe (if at all) imbalance between pro-oxidative and antioxidative components of a system ?.
And (iii) Why should we quantitate “Oxidative Stress”
(Is there any reason to try “to quantitate OS”?).
The deadline for manuscript submission is 30 April 2020 . You may send your manuscript now or up until the deadline. Submitted papers should not be under consideration for publication elsewhere.
Please let we know if you accept my invitation and send me a tentative title and, if possible, send a short abstract to the editorial office(antioxidants@mdpi.com or edith.fang@mdpi.com)
Antioxidants is committed to rapid publication. The time from submission to first decision *is 16.1 days*; from acceptance to publication *is 4.4 days*.
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Open access is supported by the authors and their institutes.
Please note that an article processing charge (APC) of 1200 CHF (Swiss Francs) applies to papers accepted after peer review.
Thank you for your consideration.
We look forward to hearing from you.
I wish you a wonderful 2020
Kind regards
Dov
Prof. Dov Lichtenberg, Tel Aviv University, Sackler School of Medicine Tel Aviv- Yafo 69978 Guest Editor of Special Issue Oxidative stress, classification and quantitation https://www.researchgate.net/profile/Dov_Lichtenberg
Dov Lichtenberg <physidov@tauex.tau.ac.il> ........................................................
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Does anyone know good references in order to understand oxidative stress better?
Thank you kindly for the help.
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I am sending you an article on oxidative stress I wrote but never published. In it you will find many references that may help you in your quest.
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We have now tried a number of commercially available antibodies to detect the receptor of AGEs (RAGE, sometimes