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).
Questions related to Oxidative Stress
  • asked a question related to Oxidative Stress
Question
2 answers
Hello everyone! I am new to this platform, I am looking for this book: "Bioquimica del estres Oxidativo" (Diego Camps, 2010)
I would like to know if anyone has it in digital format to share it with me. Waiting for a reply. Thank you very much!
Relevant answer
  • asked a question related to Oxidative Stress
Question
1 answer
High to resist oxidative stress and still high until the exam time or must be consumed in this resistance therefor we found it in low levels
Relevant answer
Answer
In the context of oxidative stress, the levels of oxidative enzymes can vary depending on the phase and severity of the stress, as well as the adaptive response of the organism or cells involved.
Initially, upon the onset of oxidative stress, there is often an upregulation of antioxidant enzymes as part of the cell’s defensive response. This includes enzymes such as superoxide dismutase (SOD), catalase, and glutathione peroxidase. Their levels may rise in an attempt to neutralize the increased production of reactive oxygen species (ROS) and to restore redox homeostasis.
However, if the oxidative stress is sustained or intense, it can lead to a situation where the antioxidant defenses are overwhelmed. In chronic or severe cases, the levels of these enzymes may not be maintained, and their activity could eventually decrease due to the damage inflicted by persistent ROS, leading to the observed low levels of oxidative enzymes.
Furthermore, the expression of these enzymes can also be modulated by numerous factors, including genetic regulation, the presence of cofactors, and the availability of substrates. Therefore, assessing the levels of oxidative enzymes provides a snapshot of the dynamic balance between pro-oxidant and antioxidant forces at a given time.
  • asked a question related to Oxidative Stress
Question
1 answer
How does oxidative stress contribute to the initiation and progression of various arrhythmias?
Relevant answer
Answer
NO generated by oxidative stress could be a factor.
  • asked a question related to Oxidative Stress
Question
3 answers
I want to study the expression of some proteins related to oxidative stress with western blot. I wonder what microglia cell line is the best for it.
Relevant answer
Answer
If you want to correlate the data with humans, HMC-3 cells would be better to study oxidative stress and inflammation.
Please find the attached paper to know in detail.
  • asked a question related to Oxidative Stress
Question
4 answers
I am working on HUVEC cells in order to Establish the Oxidative Stress Injury Model Induced by H2O2. I want to treat the cells with 5mM H2O2 from 30%h2o2 stock solution for 16 hours. How to prepare 5 mM hydrogen peroxide from 30% H2O2 stock solution?
Relevant answer
Answer
Sofiane Benyamina hello Thank you for your attention.
so, If I want to make a 5mL hydrogen peroxide solution, according to your statement, should I dilute 2.5 microliter of 30% hydrogen peroxide with 4997.5 microliter of water?
  • asked a question related to Oxidative Stress
Question
3 answers
Hi, I want to evaluate oxidative stress in a neuroblastoma cell line, I know about SOD1, but I think it's not enough, so I was investigating and I found 4-hydroxynonenal, I have this in my lab 4-HNE B5605, but I don't find the concentration to use. Also I don't think it is the best marker for inmunofluorescence, It is more used in western blot . Do you know other types of marker that I can use for oxidative stress in a cell culture? I will appreciate your help.
Relevant answer
Answer
You could try using H2DCFDA. It is mainly used in fluorometry and flow cytometry but you can also find publications using it in immunofluorescence. I could provide you with a protocol but you would have to wait some weeks cause I am currently optimizing it. Use live cells, do a quick test using H2O2 as a positive control and I hope it works out for you.
I wish you best of luck,
George
  • asked a question related to Oxidative Stress
Question
7 answers
I am working on HUVEC cells in order to make the Oxidative Stress Injury Model Induced by H2O2.I treated the cells with 0.3% H2O2(30%) for 16 hours, then IC50 was determined by MTT test, but in the validation stage of the model, the oxidative stress indicators were not as expected. what stock of hydrogen peroxide I should be used to induce oxidative stress?
Relevant answer
Answer
the concentration of 0.3% (30%) H2O2 for 16 hours is relatively high and may potentially result in severe cellular damage or cytotoxicity, leading to unexpected oxidative stress indicators.
To optimize your oxidative stress model and achieve the desired level of stress without excessive cytotoxicity, it is advisable to perform a dose-response experiment with a range of H2O2 concentrations. The following steps can guide you in determining the appropriate concentration of H2O2:
  1. Prepare a series of H2O2 dilutions: Start with a stock solution of high-concentration H2O2 (e.g., 30%) and prepare a series of dilutions using a suitable culture medium or buffer. Prepare a range of dilutions with decreasing concentrations (e.g., 0.1%, 0.05%, 0.01%, etc.). Ensure that the dilutions are freshly prepared to maintain the stability of H2O2.
  2. Treat cells with different H2O2 concentrations: Seed HUVEC cells in appropriate culture plates or dishes and allow them to reach an appropriate confluency. Treat the cells with different concentrations of H2O2, including the concentrations used previously (0.3%) and the newly prepared dilutions. Include a control group with no H2O2 treatment.
  3. Assess cellular viability and oxidative stress indicators: After the desired treatment duration, perform viability assays, such as the MTT assay, to assess cell viability and determine the IC50. Additionally, evaluate oxidative stress indicators such as reactive oxygen species (ROS) levels, antioxidant enzyme activity, lipid peroxidation, or DNA damage assays. Compare these indicators among different H2O2 concentrations to identify the optimal concentration for inducing oxidative stress without causing excessive cytotoxicity.
  4. Repeat and validate the results: It is crucial to repeat the experiments to ensure reproducibility and validate the findings. Consider performing multiple independent experiments to confirm the optimal H2O2 concentration and establish consistency in the oxidative stress model.
By systematically evaluating the effects of different H2O2 concentrations, you can identify the appropriate concentration that induces oxidative stress while maintaining cellular viability and obtaining the expected oxidative stress indicators. It is also important to consider the specific characteristics and sensitivity of HUVEC cells when selecting the H2O2 concentration.
  • asked a question related to Oxidative Stress
Question
2 answers
Mr. Guillermo Zalba did not participate in the research of the article "Prevalence of Hypertension and Obesity: Profile of Mitochondrial Function and Markers of Inflammation and Oxidative Stress". I ask you please to remove it from your information
Best regards
Alejandra Guillermina Miranda-Díaz, MD, PhD
Relevant answer
Answer
To correct Diabetes and other metabolic disorders syndrome Please go to the link and read the book
मधुमेह और अन्य उपापचय की असमान्य क्रिया को ठीक करने हेतु पढे:-
  • asked a question related to Oxidative Stress
Question
1 answer
As an antioxidant role of zinc, conflicting reports are pouring. It is recognized that Zn has prominent role in reducing oxidative stress by scavenging and decreasing production of ROS. On the other hand, there are reports that Zn in hypoxia can activate NADPH oxidase by acting as phosphorylation modulator and thereby initiate ROS accumulation. I seek kind proper explanation from the researchers in this regard.
Relevant answer
Answer
The actions of zinc are not straightforward owing to its numerous roles in biological systems. Zinc is known for its dual action, as either an antioxidant or a prooxidant, and the conditions under which each role is performed.
Zinc, an essential metal for life, plays important roles as an antioxidant to combat and suppress oxidative stress. However, please note that both zinc deficiency and zinc overload could contribute to oxidative stress.
Zinc homeostasis plays an important role in physiological functions. Disturbance of zinc homeostasis leads to various diseases. Hypoxic condition may induce abnormally high concentration of zinc accumulation. Intracellular zinc and ROS are intimately related. Zinc impairs mitochondrial function, leading to excessive ROS production, but it can also activate a variety of extra-mitochondrial ROS-generating signaling cascades. Zinc could act on mitochondria to increase superoxide production and induce NADPH oxidase activation.
Best regards,
Malcolm Nobre
  • asked a question related to Oxidative Stress
Question
2 answers
I'm trying to induce oxidative stress by using rotenone and SNP sodium nitroprusside in isolated mitochondria of rat brain. This is to study GSH assay from control (Mitochondria) group in comparison with { Mito + (SNP/Rotenone)}.
However, I'm not getting the promising results
could anyone suggest me why there is not decrease in GSH occurs significantly in (SNP or rotenone induced mitochondria comparision to control?
is there any matter of concentration?
or suggestions.
(doi : 10.14715/cmb/2014.60.2.6)
please fine protocol to perform GSH assay>
Relevant answer
Answer
I will answer the question in parts -
(1) How to decide the concentration need to induce oxidative stress in isolated mitochondria of rat brain by using either rotenone & sodium nitroprusside?
The term “how much concentration…induce oxidative stress” is inappropriate. Any amount of R or SNP (or combination R, SNP, R+SNP) will induce oxidative stress (OS) and that amount can vary between negligible to quintals (hypothetically). Negligible concentrations will be outpowered by antioxidants (AOX) (including GSH as you have asked) but there would be a time when the antioxidants would be overpowered by OS. Thus the amount of toxic chemicals is proportional to stress. So there is something called a toxicity test to understand what concentration YOU want to evaluate.
(2) I'm trying to induce oxidative stress by using rotenone and SNP sodium nitroprusside in isolated mitochondria of rat brain. This is to study GSH assay from control (Mitochondria) group in comparison with { Mito + (SNP/Rotenone)}.
Agreed that you are trying to induce OS by R and SNP, but you need to have something considerable so that you can judge the response of GSH alterations (check overflowing online articles to understand and conduct the test). First conduct isolated tests or R and SNP then combos.
(3) However, I'm not getting the promising results
What according to you is a promising result? Results are results. There is nothing promising about anything and that's the gambit of every research. If we knew the results, then what's the need for any research. Agreed? The catch here is first conduct the test, decide the conc of R and SNP, expose MC to chemicals and see outcomes on ETC dysfunction. Also, could it be that the R+SNP combo is nullifying each other?
(4) could anyone suggest me why there is not decrease in GSH occurs significantly in (SNP or rotenone induced mitochondria comparision to control?
This question forms discussion part of your manuscript. That's what you need to reason in your publication as to why it has happened or what are probable reasons. For that you need to see what are the other AOX responses? It is not mandatory that GSH must decrease if there is exposure to R and SNP, and if that be then it is definitely a very strong response. There can also be a tandem act with other AOX that you are evaluating. GSH is a bifunctional AOX, at times it is conjugated by (GST-GSSH system) or at times it can also act alone (Srikanth et al. 2013 attached)
  • asked a question related to Oxidative Stress
Question
10 answers
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.
Relevant answer
Answer
wow she is so rude. im just amazed on the responses. you could try to read this paper "Assessing mitochondrial DNA release into the cytosol and the subsequent activation of innate immune-related pathways in mammalian cells"
i hope it could help you.
this platform is to help each other as a researcher to overcome any difficulties. not to flaunt your degree.
  • asked a question related to Oxidative Stress
Question
5 answers
Dear community,
For my master's project, I want to design an experiment that will asses the efficiency of an antioxidant to stop/restore mitochondrial oxidative stress/redox balance in a PD mouse model, in both early and later stages of the disease. Redox balance will be assessed by using a roGFP lifetime measurements in vivo.
I was thinking to use MPTP model, due to it's effect on the mitochondria, simplicity and practicality. However I heard that this model can be difficult and dangerous to work with and might kill the mice faster than the experiment will end.
There is 6-ohda model, but I was reticent to use it because it has to be injected into the brain directly (which I believe might interfere with my fluorescence measurements) and the mechanisms behind it are not fully understood.
I would highly appreciate if anyone could share their experiences about working with these(or maybe other) models and give me an appreciation of their suitability in regard to my research question.
Thank you!
Relevant answer
Answer
* 6-OHDA and MPTP model are the most widely used animal models for screening of drugs against Parkinson's disease.
* 6-OHDA model requires stereotaxic surgery expertise, much more complex and mortality is high. Produces free radicals that acts as potent inhibitor of the mitochondrial respiratory chain complexes I and IV. Neuroinflammation and neuronal death may progress to other regions of the brain if surgery is not perfectly.
* Whereas MPTP model is simple, 25mg/kg in C57BL/6 mice for 5 days intraperitoneal. MPTP is metabolized into the toxic cation 1-methyl-4-phenylpyridinium (MPP+) by the enzyme monoamine oxidase B (MAO-B) of glial cells, specifically astrocytes and accumulates within SNpc DA neurons, where it inhibits ATP production and stimulates superoxide radical formation.
So, you can study mitochondrial dynamics, dysfunction and redox changes according to your study design.
Animal mortality would be around 10-15%. Yes MPTP is quite dangerous to work with, however I have been using the same from the past 6 years, I have attached the paper that discusses in detail, precautions needed to work with MPTP. If you can these necessary precautions, there is nothing to worry about more then.
* I would recommend you the MPTP induced PD model for your work.
Hope it works.
Thanks
Samir Ranjan
Ref:
  • asked a question related to Oxidative Stress
Question
5 answers
Neurological disorders include a variety of conditions including Alzheimer’s, Motor Neuron and Parkinson’s disease, affecting longevity and quality of life and these have been associated with oxidative stress. Several of the chronic neurodegenerative pathologies of the central nervous system share some common features, such as oxidative stress, closely related to inflammation, synapse dysfunctions, protein misfolding, and defective autophagia. Sources of reactive oxygen species (ROS) that cause oxidative stress, relating oxidative damage with the pathogenesis of neurodegenerative disorders. Antioxidants can powerfully neutralise ROS and free radicals, decreasing oxidative damage. Antioxidant genes, like manganese superoxide dismutase (SOD-2) enzymes, can undergo epigenetic changes that reduce its expression, thus increasing oxidative stress in tissue or DNA can be altered by free radical damage. The epigenetic landscape of these genes can alter antioxidant function and can result in neurodegenerative disease. This imbalance of free radical production and antioxidant function increase the ROS that cause neuronal cell damage, often observed as an age-related event. Familial MND is associated with SOD-1 antioxidant gene polymorphism and function.
Increased antioxidant expression in mice is protective against ROS in neurons as is the exogenous supplementation of antioxidants. The associated manganese deficiency observed in Alzheimer’s suggests that this transition metal could be supplemented in deficient patients or SOD—2 therapy considered for age-related neurodegenerative disorders.
A new mimetic of SOD-2, Avasopasem manganese (GC4419 AVA), is described and suggested as putative treatment to reduce the oxidative stress that causes neurodegenerative diseases.
Relevant answer
Answer
Antioxidant enzyme mimetic is already being trialled for diseases involving ROS, with some success, and some antioxidant molecules have been adapted to cross the BBB, Harald.
  • asked a question related to Oxidative Stress
Question
3 answers
tissue infected with various parasitic disease
Relevant answer
Answer
You can measure oxidative stress by measuring the ROS (Reactive Oxygen Species) level, lipid peroxidation, reduced glutathione level, and other antioxidant levels.
  • asked a question related to Oxidative Stress
Question
3 answers
In order to save some energy, one decision our lab is examining would be to increase the temperature limit of the freezers at -70°C instead of -80°C. I guess 10°C difference in this very low temperatures may help to save substantial amount of energy supply. Therefore, we are wondering if samples can be stored at this temperature without causing any remaining molecular activities (oxidative stress and so on).
It is not super clear in the litterature why this threshold of -80°C has been selected, does it result from empirical measures or not?
If anyone has an idea on the reasons for the choice of -80°C, this could help us to take the best decision.
Thanks in advance
Relevant answer
Answer
The choice of -80°C as a freezer temperature for long-term storage of biological samples is a widely accepted standard that has been empirically established through many years of research and experience. It is believed to ensure that biological samples remain stable over time and that molecular activities, such as oxidative stress, are minimized.
While increasing the freezer temperature to -70°C may save energy, it is not recommended as it can potentially result in the degradation of biological samples over time. At -70°C, some cellular processes, such as oxidation and enzymatic activity, can still occur, although at a slower rate compared to warmer temperatures. However, it is important to keep in mind that the stability of biological samples depends on many factors, including the type of sample, the storage conditions, and the length of time in storage.
If you are considering changing the freezer temperature, it is recommended to perform a risk assessment to determine the potential impact on your samples. You can consider running a pilot study to assess the stability of a representative subset of your samples stored at -70°C over a period of time, and then comparing the results to samples stored at -80°C. You may also want to consult with experts in the field, such as cryobiologists or biorepository managers, for advice and guidance.
  • asked a question related to Oxidative Stress
Question
2 answers
During examination of transmission electron microscopy images of skeletal muscle biopsies from patients suffering from different myopathies, sometimes I find somewhat unusual mitochondria with white spots. Initially, I assumed that is some sort of oxidative stress damage or even artifact, yet now I think I could be wrong; usually, in patients with frequent "white spotted" mitochondria, I find more pronounced pathological changes in mitochondria (such as paracrystalline inclusion or onion-like cristae) in other fibers, sooner or later.
That made me think, so I've tried to find similar images in other studies, but to no avail. I have no reasonable explanation as for now. Any ideas?
Relevant answer
Answer
Dear Rabeah Al-Temaimi, thank you for your response!
Images shown in the mentioned article are somewhat similar to ours, it's quite possible that our mitochondria may also lose their membrane potential. Early or late apoptotic events are also possible, as I find in muscles with severely disorganized myofibrils.
Cristae swelling is rather unlikely (yet still possible) - on image taken on higher magnification (attached below) I cannot see swollen cristae, those hyperintensities seem to be independent of visible cristae location. Specimens for current study were fixed in 2.5% glutaraldehyde right after biopsy - however, I am not present during tissue collection, it's possible that between biopsy and muscle fixation more time passed.
Alpers-Huttenlocher syndrome can be rather excluded - it has early onset and we have adult patients only - although, from what I see, muscle mitochondria looks similar indeed.
Thank you again for your response, it was very helpful! Now I have some foothold for further research.
  • asked a question related to Oxidative Stress
Question
5 answers
Are there publications on this?
Relevant answer
Answer
Tanja Kögel you are very welcome!
Just write me an email to my institute address and I will have a look through my archive: dbrugger@nutrivet.uzh.ch
Not at the office currently and I know I will forget if not remined in my inbox ;)
  • asked a question related to Oxidative Stress
Question
5 answers
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?
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
3 answers
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.
📷
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.
Relevant answer
Answer
Gerobiotics: probiotics targeting fundamental aging processes
Gerobiotics: probiotics targeting fundamental aging processes (nih.gov)
  • asked a question related to Oxidative Stress
Question
5 answers
I would like to measure H2O2 on tomato leaves. Just need a fast and quantitative complete protocol for the determination of H2O2.
Relevant answer
  • asked a question related to Oxidative Stress
Question
2 answers
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
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
1 answer
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.
Relevant answer
Answer
,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
  • asked a question related to Oxidative Stress
Question
1 answer
There is a antioxidant defense system in cell.
Have any signalling pathway or other things can influence ROS level in a extracelluar way?
Relevant answer
Answer
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! 
  • asked a question related to Oxidative Stress
Question
12 answers
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.
Relevant answer
If you can give us more details would be great!
  • asked a question related to Oxidative Stress
Question
4 answers
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?
Relevant answer
Answer
Thank you Azadeh Eshraghi for your answer.
  • asked a question related to Oxidative Stress
Question
6 answers
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.
Relevant answer
Answer
Dear colleague! It depends on the characteristics of the agent for oxidative stress and its concentration.
  • asked a question related to Oxidative Stress
Question
4 answers
How Campylobacter and Helicobacter species isolated from patients stored @-80 survive oxidative stress if they faced temperature variations?
Relevant answer
Answer
Dear Prof.Zaim,
Thank you so much for your detail explanation to my question.
  • asked a question related to Oxidative Stress
Question
3 answers
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!
Relevant answer
Answer
I. N. Popov thank you so much!!
  • asked a question related to Oxidative Stress
Question
2 answers
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
Relevant answer
Answer
Good Idea!!
  • asked a question related to Oxidative Stress
Question
6 answers
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!
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
4 answers
complement factor receptor (C3aR and C5aR) activation under oxidative stress conditions on ARPE-19 cells.
Relevant answer
Answer
  • asked a question related to Oxidative Stress
Question
4 answers
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.
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
2 answers
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!
Relevant answer
Answer
Thank you Prof Murakami for the suggestion :)
  • asked a question related to Oxidative Stress
Question
7 answers
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?
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
4 answers
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?
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
2 answers
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?
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
7 answers
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.
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
1 answer
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?
Relevant answer
Answer
Hello,
You can use GAPDH as loading control.
  • asked a question related to Oxidative Stress
Question
1 answer
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!
Relevant answer
  • asked a question related to Oxidative Stress
Question
6 answers
How to go about it statistically?
Relevant answer
Answer
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).
  • asked a question related to Oxidative Stress
Question
2 answers
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
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
4 answers
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.
Relevant answer
Answer
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/
  • asked a question related to Oxidative Stress
Question
3 answers
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?
Relevant answer
Answer
Thank you Dr. shin and Dr. Manju for your answer.
  • asked a question related to Oxidative Stress
Question
4 answers
Serum TIBC and serum ferritin colorimetric assay protocol.
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
5 answers
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.
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
4 answers
oxidative stress, histopathology, leakage of enzymes, lipid profile, inflammation, apoptosis
Relevant answer
Answer
Many thanks for your responses
  • asked a question related to Oxidative Stress
Question
7 answers
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.
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
4 answers
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.
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
8 answers
I have tried with no success various UV-B stresses but I did not find an increase in beta galactosidase.
Relevant answer
Answer
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.)
  • asked a question related to Oxidative Stress
Question
8 answers
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?
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
2 answers
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.
Relevant answer
Answer
@Ehab Yehiea Jabber. Please tell me
  • asked a question related to Oxidative Stress
Question
5 answers
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?
Relevant answer
Answer
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...
  • asked a question related to Oxidative Stress
Question
6 answers
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.
  • asked a question related to Oxidative Stress
Question
3 answers
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:
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
8 answers
for induction of oxidative stress
Relevant answer
Answer
I'm following the best answer.
  • asked a question related to Oxidative Stress
Question
3 answers
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?
Relevant answer
Answer
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!
  • asked a question related to Oxidative Stress
Question
4 answers
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
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
6 answers
I am looking for a new method for measuring oxidative stress in animal tissues ... a previously unrecognized method .... Thank you for your cooperation
Relevant answer
  • asked a question related to Oxidative Stress
Question
11 answers
i am working on Carum copticum extract and its seems it has itself induced oxidative stress at higher doses. 200mg/kg PO
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
6 answers
Hello all,
What are some markers of oxidative stress in brain?
Thanks
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
4 answers
the relationships between oxidation and inflammation
Relevant answer
Answer
Dear @ Laith,
There is definitely a relation of oxidative stress with inflammation. As you know very well, Increase of free radicals results the oxidative stress. And, oxidative stress ultimately cause the inflammation. i.e., Decrease of antioxidants ---) Increase of Free radicals ----) Increase of Oxidative Stress ----) Inflammation.
  • asked a question related to Oxidative Stress
Question
8 answers
Susceptibility of Covid-19 patients to drug-induced OS
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
4 answers
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?
Relevant answer
Answer
Hello
a well-designed case-control
  • asked a question related to Oxidative Stress
Question
3 answers
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
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
2 answers
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.
Relevant answer
Answer
The Cell Meter™ Hydrogen Peroxide Assay Kit OxiVision™ Green hydrogen peroxide sensor to quantify hydrogen peroxide in live cells
  • asked a question related to Oxidative Stress
Question
12 answers
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.
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
3 answers
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.
Relevant answer
Answer
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,
  • asked a question related to Oxidative Stress
Question
3 answers
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?
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
4 answers
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.
Relevant answer
Answer
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.
  • asked a question related to Oxidative Stress
Question
3 answers
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?
Relevant answer
Answer
  • asked a question related to Oxidative Stress
Question
4 answers
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
Relevant answer
Answer
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
  • asked a question related to Oxidative Stress
Question
6 answers
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?
Relevant answer
Answer
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?
  • asked a question related to Oxidative Stress
Question
4 answers
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?
Relevant answer
Answer
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:
  • asked a question related to Oxidative Stress
Question
9 answers
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.
Relevant answer
Answer
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