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How To Test For Oxidative Stress

Tests To Detect Effect Of Oxidative Stress On Sperm

Free Radical Testing, Oxidative stress

Unlike the somatic cells, the lipid bilayer in the plasma membrane of the human spermatozoa is rich in polyunsaturated fatty acids making them highly susceptible to damage caused by excess ROS .

The breakdown of polyunsaturated fatty acids to form lipid peroxides is known as lipid peroxidation . Lipid peroxides are unstable and decompose to form complex compounds such as 4-hydroxynonenal , acrolein and malondialdehyde which freely react and modify lipids, proteins and DNA as they are relatively more stable than free radicals. They act as cytotoxic second messengers of oxidative stress causing alterations in the sperm functions . Measurement of the end products of lipid peroxidation is a widely accepted marker of oxidative stress. Some of the common methods to measure lipid peroxidation and oxidative stress are described below.

15.3.1Measurement of 4-Hydroxynonenal Histidine Adduct

Highly reactive aldehyde 4-hydroxynonenal -protein adducts can be quantified using ELISA with a monoclonal antibody that rapidly detects the HNE-histidine adducts .

Hydroxynonenal modified BSA standards are prepared. Fatty acid free BSA is diluted in PBS . One mM of HNE is added to fatty acid free BSA to give 250 pmol/mg of ELISA with non-commercial antibody or 5000 pmol/mg of ELISA with commercial antibody. These stock solutions are incubated overnight at 4oC to allow binding of HNE to BSA and stored at -20oC. Clinical Interpretation

What Are Free Radicals And Antioxidants

A free radical can be defined as an oxygen-containing molecule that consists of unpaired electrons. For example superoxide radicals, hydroxyl radicals, and nitric oxide radicals. These free radicals are highly reactive with the other molecules.

Cells that use oxygen and enzymes for functioning are open to oxygen free radical reactions that can harm the cells. Antioxidants are defined as the molecules present in cells that avoid harmful reactions. They give an electron to free radicals without lessening themselves.

Neurological Disease And Oxidative Stress

Oxidative stress has been linked to several neurological diseases , amyotrophic lateral sclerosis , multiple sclerosis, depression, and memory loss) . In AD, several experimental and clinical researches showed that oxidative damage plays a pivotal role in neuron loss and progression to dementia . -amyloid, a toxic peptide often found present in AD patients’ brain, is produced by free radical action and it is known to be at least in part responsible for neurodegeneration observed during AD onset and progression .

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Cardiovascular Disease And Oxidative Stress

Cardiovascular diseases are clinical entities with a multifactorial etiology, generally associated with a very large amount of risk factors, the most broadly recognized of which are hypercholesterolaemia, hypertension, smoking habit, diabetes, unbalanced diet, stress, and sedentary life . During the last years, research data pointed out that oxidative stress should be considered either a primary or a secondary cause for many CVDs . Oxidative stress acts mainly as a trigger of atherosclerosis. It is well known that atheromatous plaque formation results from an early endothelial inflammation, which in turn leads to ROS generation by macrophages recruited in situ. Circulating LDL are then oxidized by reactive oxygen species, thus leading to foam cell formation and lipid accumulation. The result of these events is the formation of an atherosclerotic plaque. Both in vivo and ex vivo studies provided evidences supporting the role of oxidative stress in atherosclerosis, ischemia, hypertension, cardiomyopathy, cardiac hypertrophy, and congestive heart failure .

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Cancer And Oxidative Stress

[PDF] Laboratory tests for oxidative stress

Cancer onset in humans is a complex process, which requires both cellular and molecular alterations mediated by endogenous and/or exogenous triggers. It is already well known that oxidative DNA damage is one of those stimuli responsible for cancer development . Cancer can be driven and/or promoted by chromosomal abnormalities and oncogene activation determined by oxidative stress. Hydrolyzed DNA bases are common by-products of DNA oxidation and are considered one of the most relevant events in chemical carcinogenesis . The formation of such kind of adducts impairs normal cell growth by altering the physiological transcriptomic profile and causing gene mutations. Oxidative stress can also cause a variegated amount of modifications against DNA structure, for example, base and sugar lesions, DNA-protein cross-links, strand breaks, and base-free sites. For instance, tobacco smoking, environmental pollutants, and chronic inflammation are sources of oxidative DNA damage that could contribute to tumor onset . Oxidative stress resulting from lifestyle reasons can also play an important role in cancer development, as suggested by the strong correlation between dietary fat consumption and death rates from different types of cancer .

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Enhance The Magnesium Intake

Magnesium is one of the most important components when it comes to keeping the levels of oxidative stress in control. It has its primary impacts on the oxidative stress which is exerted on the blood vessels. If you want to keep things in control and even get rid of oxidative stress, it is very important that you do keep the consumption of magnesium in check.

If you want to get the same through natural food sources, beans, pumpkin seeds, swiss chard and a few more fruits and veggies have been found to impose beneficial impacts on the body.

How To Test Oxidative Stress

  • 2021-11-20T21:59:11+00:00Added an answer on November 20, 2021 at 9:59 pmThere are a few tests that can be used to measure oxidative stress. Two of the most commonly used tests are the lipid peroxidation test and the nitric oxide test. Both tests are non-invasive and relatively inexpensive to perform. How you interpret the results from these tests depends on the individual test being run, but one of the most important factors is the baseline value of the test you are measuring. If a test is significantly elevated compared to the baseline, then the result is considered positive. The baseline value for a healthy individual can be determined by comparing it to a value for a person you know to be free from oxidative stress.
  • 2021-11-20T06:49:29+00:00Added an answer on November 20, 2021 at 6:49 amTo judge oxidative stress, you can order a simple blood test. This test will measure the total oxidative stress damage in the human body, so you can see how much potential damage you are doing to your body. This test is comparable to the urine test, but has advantages too. It is also much cheaper than the urine test. You can order this test here.
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    Kidney Diseases And Oxidative Stress

    Oxidative stress is involved in a plethora of diseases affecting renal apparatus such as glomerulo- and tubule-interstitial nephritis, renal failure, proteinuria, and uremia . The kidneys are negatively affected by oxidative stress mainly because of the fact that ROS production induces the recruitment of inflammatory cells and proinflammatory cytokine production, leading to an initial inflammatory stage. In this early phase, a predominant role is played by TNF-alpha and IL-1b, as proinflammatory mediators, as well as by NF-B as transcriptional factor required to sustain the inflammatory process. The latter stage is characterized by an increase in TGF-beta production, which orchestrates the extracellular matrix synthesis. So, when the oxidative stress stimuli act chronically on kidney tissues, the results will be an initial stage of inflammation and later the formation of abundant fibrotic tissue that impairs organ function potentially leading to renal failure. Certain drugs, such as cyclosporine, tacrolimus, gentamycin, and bleomycin, are known to be nephrotoxics mainly because of the fact that they increase free radical levels and oxidative stress via lipid peroxidation . Heavy and transition metals , acting as powerful oxidative stress inducers, are responsible for various forms of nephropathy, as well as for some types of cancers .

    Oxidative Stress In Clinical Samples Of Cancer Patients

    Dr. Marcus Cooke explains oxidative stress

    ROS is a contributing factor in the natural process of aging as well as in various pathological diseases including cancer. Oxidative stress has been reported in almost all types of cancers, promoting many aspects of tumor development and progression . During the process of carcinogenesis, an increase in ROS levels can occur due to elevated metabolic activity, oncogene activation, increased cellular receptor signaling, or mitochondrial dysfunction. Overproduction of ROS can also be induced exogenously by carcinogenic insults such as cigarette smoke, heavy metals, ionizing radiation, and asbestos . Alternatively, oxidative stress observed in cancer cells can arise from low levels or inactivation of antioxidant defense mechanisms as a result of mutations in tumor suppressor genes. For instance, mutant BRCA1 and p53 have been shown to attenuate the activation and function of nuclear factor -like 2 , a transcription factor that stimulates a stress response pathway by inducing expression of ROS-detoxifying enzymes .

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    How The Test Works

    The Test is easy to perform, using a dry buccal swab which you use to collect a sample of cells from the mucous membrane of the oral cavity. Once received by the Lifebrain Genetics Laboratory, the DNA is extracted and analysed to show which gene variants are present.

    Want to carry out the test in the comfort of your own home? Purchase the oxidative stress test online by following the instructions below!

    Discover the other tests of the Anti-Ageing line:

    Measuring Changes In Blood Pressure And Vascular Tone

    The free radical NO plays a key role in vasodilatation. Its action can be antagonized by O2, which removes NO by reacting fast with it to give ONOO . In the healthy arterial wall, there is a balance between NO and O2, but increased oxidative stress can diminish the bioactivity of NO . Sources of O2 in and around the vessel wall include the vascular endothelium , fibroblasts, lymphocytes, phagocytes and the enzyme xanthine oxidase . Upregulation of O2-generating NADH oxidases may play a key role in causing impaired vascular tone and hypertension .

    Many papers have claimed that high doses of infused or orally administrated antioxidants can scavenge ROS, restore NO bioavailability and ameliorate human endothelial dysfunction , although negative reports are now accumulating . The timing of antioxidant administration and the state of the vascular bed may be critical variables . Hence it is possible that examination of short-term vascular effects, which is easily achievable in human studies by measurements of forearm blood flow, can gather evidence about localized scavenging of RS in blood vessel walls by antioxidants.

    However, it must not be assumed that any beneficial effects of antioxidants on vascular function are necessarily due to free radical scavenging. For example, ascorbate may have a direct stimulating effect on eNOS activity by increasing endothelial tetrahydrobiopterin levels .

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    Effects Of Oxidative Stress On The Body

    Oxidation is a normal and necessary process that takes place in your body. Oxidative stress, on the other hand, occurs when theres an imbalance between free radical activity and antioxidant activity. When functioning properly, free radicals can help fight off pathogens. Pathogens lead to infections.

    When there are more free radicals present than can be kept in balance by antioxidants, the free radicals can start doing damage to fatty tissue, DNA, and proteins in your body. Proteins, lipids, and DNA make up a large part of your body, so that damage can lead to a vast number of diseases over time. These include:

    Exogenous Antioxidants And Human Health

    Why You NEED to Understand Oxidative Stress

    Human body put in place several strategies to counteract the effects of free radicals and oxidative stress, based on enzymatic and nonenzymatic antioxidant molecules, all of them being endogenous antioxidants. Beside these, there are several exogenous antioxidant molecules of animal or vegetal origin, mainly introduced by diet or by nutritional supplementation.

    Here, we will discuss the most relevant nutritional antioxidants and their protective effects for human health.

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    Effects Of Oxidative Stress

    As we did mention before, oxidative stress can have negative impacts on the body and especially if it is not tended to in the long run.

    Dont let it just get out of control and instead ensure to keep your health in check by keeping the balance between the free radicals and the antioxidants in the body.

    When it comes to the effects and the impacts of oxidative stress on the body, you should know that not only does it enhance the risks associated with the cancer and the chronic diseases but does have impacts on ones overall health as well.

    Inflammation is one of the most common impacts that is causes because of the prolonged impacts of the oxidative stress.

    It is very important to ensure that you do opt for effective treatment approaches to ensure the best of health without any kind of further negative impacts on the body.

    Measuring Rs In Vivo: Basic Principles

    Some fascinating techniques such as L-band electron spin resonance with nitroxyl probes and magnetic resonance imaging spin trapping are under development to measure RS directly in whole animals , but no probes are currently suitable for human use. Most RS persist for only a short time in vivo and cannot be measured directly. There are a few exceptions: examples include H2O2 , and perhaps, NO, in the sense that serum levels of NO2 have been claimed to measure vascular endothelial NO synthesis , despite the fact that NO2 is quickly oxidized to NO3in vivo. Essentially, there are two approaches to detecting transient RS:

    • attempting to trap these species and measure the levels of the trapped molecules and
    • measuring the levels of the damage done by RS, that is, the amount of oxidative damage.

    Depending on the method that is used to measure it, the plasma or serum total antioxidant capacityâ usually involves major contributions from urate, ascorbate and sometimes albumin SH groups , although different methods measure different things . Thus, for example, if plasma albumin levels fall, TAC will fall. If urate levels rise, TAC will rise. The multiple changes in blood chemistry that occur in sick people mean that TAC changes should be interpreted with caution. TAC is also influenced by diet, often because consumption of certain foods may produce changes in plasma ascorbate and/or urate levels .

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    What Is Oxidative Stress

    Free radicals and antioxidants play an important role in the healthy functioning of your body. Oxidative stress is defined as an imbalance between the production of free radicals and antioxidants in the body.

    The cells in our body develop free radicals when the metabolic processes are driven in a normal manner. Moreover, the cells are also responsible for producing antioxidants that balance these free radicals.

    Usually, our body manages to balance free radicals and antioxidants. Nevertheless, the occurrence of oxidative stress in the body is natural and affects the aging process.

    It can be both useful or toxic to the body.

    Poor Gut Health: Another Contributor To Oxidative Stress

    OXIDATIVE STRESS – why do we get sick!

    After trigger foods, the other major source of oxidative stress for our bodies is overgrowing bacteria in the large intestine.5

    We regularly hear about how important it is to take a probiotic but in many cases, the issue is NOT a lack of good bacteria, but an overgrowth of bacteria that either shouldnt be hanging out in your intestines, or shouldnt be thriving quite so much.

    This usually happens due to antibiotic use, which kills off good bacteria and creates an imbalance in the bacterial ecosystem within our bodies. It can also happen as a result of other medications, such as antacids and proton pump inhibitors, which decrease stomach acid as well as our ability to digest food. That undigested food then feeds the wrong bacteria.

    Same with stress itself! Stress decreases our ability to digest food, and thus increases the imbalance of bacteria as does gluten, pesticides, and many other factors.

    While we often assume that the garden of bacteria in our intestines are wiped out and barren, I find that the garden is more often overgrown with weeds. And that overgrowth of bacteria leads to leaky gut, as well as oxidative stress, which can then spread throughout your body.

    Thats why addressing oxidative stress is so key to healing other health conditions, including digestive troubles and impaired methylation.

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    Effects Of Rs On Other Probes

    A wide range of probes is available to study many cellular events, such as pH changes and ion movements and to identify various cellular organelles . For example, rises in Ca2+ frequently accompany oxidative stress . Several papers have indicated that ion probes can be affected by the generation of RS or can themselves contribute to RS formation, such as calcein . MitoTracker Red, a fluorescent probe used to identify mitochondria, can stimulate mitochondrial ROS generation , for example.

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    Adverse Effects Of Oxidative Stress On Human Health

    Oxidation is a natural process of our body. Oxidative stress occurs due to an imbalance between free radicals and antioxidants. Proper functioning between both these activities can lead to fighting off pathogens. Pathogens are harmful and can cause infections.

    When the free radicals are more than the antioxidants present in the body, it harms the DNA, fatty tissue, and proteins.

    The body is formed primarily from proteins, DNA, and lipids. Damage to these elements can cause several diseases later on in life, such as:

    Heart disease

    Chronic diseases

    Cardiovascular disease

    Oxidative stress can harm the formation of cellular structures including lipids, lipoproteins, deoxyribonucleic acid, proteins, and membranes. The harmful condition exists when the formation of free radicals and the ability of cells to fight them off is imbalanced.

    For example, an excess of peroxynitrite and hydroxyl radicals can result in lipid peroxidation that can damage the lipoproteins and cell membranes. The hydroxyl radical is extremely reactive and is caused due to the Fenton reaction and decomposition of peroxynitrite. This causes the production of conjugated diene and malondialdehyde compounds that are mutagenic and cytotoxic.

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