Nomenclature of the Ministry of Health of the Russian Federation (Order No. 804n): A09.05.238.001 "Determination of total antioxidant activity"

Biomaterial: Whole blood with heparin

Deadline (in the laboratory): 7 w.d. *

Description

Determination of antioxidant activity plays an important role in assessing the body's defense against oxidative stress. This allows you to: identify individuals with an increased risk of developing coronary artery disease, arterial hypertension, diabetes mellitus, cancer, retinopathy; identify premature aging, monitor the course of diseases, evaluate the effectiveness of therapy.

Also, the determination of antioxidant activity helps to identify the amount of antioxidants entering the human body, and whether there is a need for their additional introduction. Antioxidant activity is determined by the presence of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase, glutathioperoxidase) and non-enzymatic antioxidants (vitamins E, C, carotenoids, lipoic acid, ubiquinone).

Indications for appointment

• Assessment of the antioxidant status of the body and assessment of the risk of developing diseases associated with antioxidant deficiency (cancer, heart disease, rheumatoid arthritis, diabetes mellitus, retinopathy, early aging)
• Patients suffering from hypertension, atherosclerotic vascular disease, diabetes mellitus, coronary heart disease - as a monitoring of the course of the disease and assessing the effectiveness of the therapy received; determining the antioxidant defense of the body, and addressing the issue of the need for additional intake of antioxidant drugs.
• Elderly patients, with poor nutrition, smoking, alcohol abuse, stress - to assess the antioxidant defense of the body, and decide on the need for additional intake of antioxidant drugs.
• Patients on the background of chemotherapeutic treatment - to assess the antioxidant defense of the body, and decide on the need for additional intake of antioxidant drugs.
• Patients on a diet and food restriction - to assess the antioxidant defense of the body, and decide on the need for additional intake of antioxidant drugs.

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General section The state of the antioxidant system in Moscow residents with newly diagnosed thyropathies. Possibilities of using nutraceuticals for the correction of antioxidant and thyroid status

Traditionally, in the course of planning preventive programs, endemic goiter is considered as an isolated iodine-deficient microelementosis. At the same time, it is well known that in the genesis of this pathological condition, a violation of the optimal content and / or ratio of other macro- and microelements may be important (V.V. Kovalsky, 1974, De Groot L.Y. et al., 1996, M.V. Veldanova, 2000), among which selenium occupies an important place. The role of selenium in optimizing thyroid function has been identified relatively recently. It has been established that, on the one hand, selenium is a necessary component of monodeiodinase, an enzyme for the peripheral conversion of thyroxine to triiodteronine (G. Canettieri et al., 1999), on the other hand, it is a structural component of glutathione peroxidase, a key enzyme of the natural antioxidant defense system (J. Kvicala et al., 1995, R. Bercow, E. Fletcher, 1997, L.V. Anikina).

The pathogenetic significance of lipid peroxidation in the occurrence and evolution of goiter transformation in iodine-deficient regions has been repeatedly discussed in the literature (N.Yu. Filina, 2003). This issue is of particular relevance in connection with the planning and implementation of mass iodine prophylaxis programs.
It is obvious that the intake of iodine in doses exceeding the traditional for the food chains of the area causes the activation of thyroid synthesis, which is the goal of preventive measures. However, in parallel, the formation of free radicals is activated due to the stimulation of redox processes directly regulated by thyroid hormones. With the weakness of enzymatic antioxidant systems against the background of a deficiency of selenium, zinc, copper and a number of other elements, this inevitably leads to the development of oxidative stress.
The purpose of this study was to study the features of the antioxidant status in Muscovites with newly diagnosed thyropathies, as well as to establish the possibilities of its correction using nutritional preparations.
Materials and methods. Determination of the antioxidant status was carried out in 38 patients who first turned to an endocrinologist about goiter transformation and who have not received therapeutic and prophylactic drugs that stimulate the natural antioxidant defense system over the past 6 months. There were 35 women (mean age 46 years) and 3 men (mean age 43 years) among the subjects. A comprehensive biochemical study using diagnostic reagents from Ranbox (Great Britain) included the determination of total antioxidant status (TAS), levels of glutathione peroxidase (GPO), superoxide dismutase (SOD), and lipid peroxidation (LPO) in blood serum. The thyroid status of the subjects was assessed by the results of a clinical examination, ultrasound examination of the thyroid gland, as well as by the content of antibodies to thyroglobulin and thyroid peroxidase, free thyroxine, free triiodothyronine and thyroid-stimulating hormone in the blood serum. Determination of antibodies and hormones of the "pituitary - thyroid gland" system was carried out by enzyme immunoassay using standard reagent kits "Immunotech RIO kit" (Czech Republic).
Results and its discussion. During the study of the thyroid status in the group of subjects, the following forms of thyroidopathy were diagnosed: diffuse enlargement of the thyroid gland - 5 patients, nodular goiter - 12 patients, mixed goiter - 8 patients, autoimmune thyroiditis - 12 patients, idiopathic hypothyroidism - 1 patient.
Certain changes in the indicators of the antioxidant status were detected in 36 subjects, which amounted to 94.7%. Among them, a decrease in TAS was observed in 76.8% of patients; decrease in the level of SOD - in 93.8%; GPO indicators as close as possible to the lower value of the range of normal fluctuations - in 50.0%; decrease in the level of GPO — in 12.5%; increase in LPO - in 15.6%.
The most significant disturbances in the system of natural antioxidant protection were found in patients with severe forms of goiter transformation (mixed goiter, autoimmune thyroiditis), however, given the insufficient representativeness of the sample, this result cannot be considered statistically significant.
Based on the obtained data, preparations of the VITALINE Corporation (USA), which have antioxidant activity, were added to the traditional treatment regimens for patients in the study group. All subjects with a decrease in TAS and/or an increase in lipid peroxidation received Pycnogenol, which is a mixture of bioflavonoids. In case of detection of reduced indicators of GPO and SOD in the blood serum, the preparations "Selenium" and "Zinc" were prescribed, respectively, in physiological doses for these elements.
Control studies of the antioxidant status were performed by the subjects 6 months after the start of therapy. As a result, normalization of TAS parameters was obtained in 85.6% of patients, normalization of lipid peroxidation — in 97.4%. In 50.4% of the subjects, the level of superoxide dismutase in the blood serum significantly increased compared to the initial level, in 30.2% it returned to normal. The level of glutathione peroxidase returned to normal compared to baseline in 100% of patients.
It is noteworthy that, against the background of the therapy, all subjects suffering from autoimmune thyroiditis had a significant decrease in the level of antibodies to thyroid peroxidase in the blood serum, and in 93.4% of patients this indicator decreased by 2-3 times compared with the baseline.
Thus, our studies revealed changes in the antioxidant status in the absolute majority of Muscovites suffering from thyroid pathology. this situation may be the result of pronounced technogenic pressure, which depletes the reserves of the natural antioxidant defense system. a clear trend towards a decrease in the levels of HCP in the blood serum of the subjects serves as an indirect confirmation of the deficiency of selenium in the food chains of Muscovites, caused by both natural and anthropogenic factors.
Obviously, in such a situation, enrichment of the diet with iodine without a simultaneous increase in the functional reserves of the antioxidant system of the population can lead to the development of oxidative stress and, as a result, to an increase in the incidence of the most severe forms of goiter transformation. Of particular concern are the prospects for the use of iodates, salts of iodic acid, which initially are strong oxidizing agents, for iodization of table salt. The risk of developing iodine-induced goiter pathomorphosis increases under conditions of technogenic stress, which is also accompanied by free radical aggression. The validity of the stated prognosis is confirmed by the long-term results of isolated iodine prophylaxis in many foci of endemic goiter (P.A.Rolon, 1986; E.Roti, L.E.Braverman, 2000, O.V. Terpugova, 2002).
Our studies allow us to recommend the use of antioxidant drugs, including physiological doses of selenium and zinc, which are coenzymes of the natural antioxidant defense system, to optimize programs for the prevention of iodine deficiency diseases, especially in ecologically unfavorable regions.
Biography:
Anikina L.V. The role of selenium in the pathogenesis and correction of endemic goiter: Abstract of the thesis. dis. … Dr. med. Sciences. - Chita, 1998. - 37 p.
Bercow R., Fletcher E. A guide to medicine. Diagnostics and therapy. T.1: Per. from English. — M.: Mir, 1997. — 667 p.
Veldanova M.V. The role of some strimogenic factors

Tests for total antioxidant status

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What is total antioxidant status?

In a healthy body, free radicals are formed a little, their negative impact is suppressed by the antioxidant defense of the body.

The study of inflammatory diseases has shown that inflammatory processes are often accompanied by a decrease in the level of antioxidants in the blood and the activation of free radicals that form reactive oxygen species (ROS). These include O 2 , OH, H 2 O 2 molecules containing oxygen ions and actively reacting with such cell components as proteins, lipids, nucleic acids. As a result of chemical (free radical) reactions, the cell membrane is destroyed, it is degraded, and the products formed as a result of the reaction penetrate into the blood.

Alien radicals are also formed in the body under the influence of ultraviolet and ionizing radiation, the ingress of toxic products into the body. Diets, malnutrition and deficiency of vitamins C, E, A, which are natural antioxidants, lead to a decrease in their level in cells, and an increase in CPP. Deficiency of antioxidants provokes the development of such pathologies as:

• diabetes;
• oncology, AIDS;
• cardiological diseases (myocardial infarction, atherosclerosis),
• diseases of the liver, kidneys.

Analysis for overall antioxidant status allows you to determine the rate of reaction processes by the number of free radicals in the bloodstream and the number of products of CPP reactions, and also shows the presence of antioxidants designed to block free radicals. Antioxidant enzymes include superoxide dismutase, definition which allows you to evaluate the antioxidant defense of the body. Superoxide dismutase (SOD) is produced in the mitochondria of human cells and is one of the antioxidant enzymes.

Why is it necessary to conduct a blood test for GGTP?

An increase or decrease in the level of certain enzymes in the bloodstream may indicate the appearance of certain pathologies in the body. One such enzyme is gamma glutamyl transpeptidase. This enzyme serves as a natural catalyst for chemical reactions in the body and is involved in metabolic processes. Gamma GTP blood test indicates the state of the gallbladder, liver. In addition, an increase in the level of this enzyme may indicate diseases such as:

• heart failure;
• systemic lupus erythematosus;
• hyperfunction of the thyroid gland;
• diabetes;
• pancreatitis;

For analysis, blood is taken from a vein.

The city medical center on the glider will conduct the most complex blood tests with high accuracy of indicators, which are guaranteed by modern laboratory equipment and professional experience of specialists.

Relatively recently, biochemists have identified a new criterion for assessing the state of the body - antioxidant status. What is hidden under this name? In fact, this is a set of quantitative indicators of how well the cells of the body can resist peroxidation.

What are antioxidants for?

There is a wide range of pathological conditions, the primary source of which are free radicals. Among the most famous are all the processes associated with aging and cancer. The presence of a large number of unpaired electrons triggers chain reactions that severely damage cell membranes. Thus, the cell is no longer able to cope with its duties normally, and failures begin in the work of first individual organs, and then entire systems. Substances that have antioxidant activity are able to suppress these reactions and prevent the development of formidable diseases.

Natural Antioxidants

In a living organism, there are a number of substances that, in their normal state, are able to withstand the attacks of free radicals. A person has this:

- superoxide dismutase(SOD) is an enzyme that contains zinc, magnesium and copper. It reacts with oxygen radicals and neutralizes them. Plays a big role in protecting the heart muscle;

Glutathione derivatives that contain selenium, sulfur and vitamins A, E and C. Glutathione complexes stabilize cell membranes;

Ceruloplasmin is an extracellular enzyme that is active in blood plasma. It interacts with molecules that contain free radicals that are formed as a result of pathological conditions such as allergic reactions, myocardial infarction and some others.

For the normal functioning of these enzymes, the presence in the body of such co-enzymes as vitamins A, C, E, zinc, selenium and copper is mandatory.

Laboratory determination of antioxidant indicators

To determine the antioxidant status of the body, conduct a number of biochemical studies, which can be conditionally divided into direct and indirect. Direct determination methods include tests for:

- SOD;

lipid peroxidation;

Total antioxidant status or TAS;

Glutathione peroxidase;

The presence of free fatty acids;

Ceruloplasmin.

Indirect indicators include determining the level of vitamins in the blood - antioxidants, coenzyme Q10, malonaldehyde and some other biologically active compounds.

How the test is done

Determination of antioxidant status is carried out in native venous blood or in its serum using special reagents. The test takes an average of 5-7 days. Healthy people are advised to conduct it at least once every six months, and in the presence of visible violations or for the purpose of checking effectiveness of antioxidant therapy- every 3 months. The test results are deciphered exclusively by an immunologist, who can prescribe medications to correct the indicators.

Summary The state of lipid peroxidation (LPO) processes (plasma content of diene conjugates, TBA-active products) and antioxidant protection (total AOA, concentration of α-tocopherol, retinol in blood plasma and riboflavin in whole blood), determined by spectrophotometric and fluorometric methods, were assessed. in 75 practically healthy children living in Irkutsk. Children of 3 age groups were examined: 21 children of preschool age (3-6 years old, average age 4.7±1.0 years), primary school age (7-8 years old, average age 7.6±0.4 years) - 28 children and secondary school age (9-11 years old, average age 9.9±0.7 years) - 26 children. In children of primary school age, the content of primary lipid peroxidation products was significantly increased, in children of middle school age, the content of final TBA-active products was significantly increased in comparison with the indicators of preschool children. At the same time, children of primary and secondary school age showed a significantly increased level of total AOA and the content of fat-soluble vitamins and riboflavin compared with preschoolers. An assessment of the actual provision with vitamins showed a lack of α-tocopherol in half of the preschool children, 36% of primary school children and 38% of middle school children. Insufficiency of retinol and riboflavin was recorded in a small number of children of all ages. In this regard, the additional supply of vitamins to children of preschool and secondary school periods is extremely necessary.

Keywords: children, age periods, antioxidant protection, antioxidant vitamins, LPO

Question. nutrition. - 2013. - No. 4. - S. 27-33.

In recent years, there has been a high prevalence of somatic, neurological and mental disorders in children of preschool and school age, a sharp increase in stressful effects on the child, and a decrease in his adaptive capabilities. Among the conditions contributing to the formation of inadequate health of the child population, a special role is given to environmental problems against the background of a sharp deterioration in social and living conditions, primarily malnutrition with a lack of protein and vitamin and mineral components. In addition, as a result of massive antibiotic therapy, a significant part of children develop microbiont defects that disrupt the absorption of nutrients that are supplied in sufficient quantities with food. Studies conducted in the region showed a deterioration in the health of children of preschool and primary school age: an increase in the incidence (91.2%), a decrease in the number of people in the 1st health group (7.2%), morphofunctional deviations (33.2%), slow rate of development (33%), low level of neuropsychic development in 15.5% of practically healthy children, high psycho-emotional stress (30.6%). At the same time, there is an increase in school disadaptation and neuropsychosomatic disorders.

The most important component of the organism's adaptive responses is the "lipid peroxidation (LPO)-antioxidant protection (AOP)" system, which makes it possible to assess the resistance of biological systems to the effects of the external and internal environment.

Natural antioxidants and essential nutritional factors are fat-soluble vitamins: α-tocopherol and retinol. α-Tocopherol is one of the most important fat-soluble antioxidants that exhibit membrane-protective and antimutagenic activity.

Interacting with natural antioxidants of other classes, it is the most important regulator of oxidative homeostasis of cells and the body. The antioxidant function of retinol is expressed in the protection of biological membranes from damage by reactive oxygen species, in particular superoxide radical, singlet oxygen, peroxide radicals. An important water-soluble antioxidant is riboflavin (vitamin B 2), which is involved in redox processes. Literature data show that the majority of the child population in all regions of the country is characterized by an insufficient supply of B vitamins, as well as vitamins C, E and A.

Insufficient activity of protective antioxidant factors and an uncontrolled increase in free radical components can play a decisive role in the development of a number of childhood diseases: respiratory tract infections, bronchial asthma, type 1 diabetes mellitus, necrotizing enterocolitis, arthritis, diseases of the gastrointestinal tract, disorders of the cardiovascular system, allergic pathologies, psychosomatic disorders.

In this regard, adequate provision of the body of children with food antioxidants, which are important factors in the formation of the protective status of the body, is one of the ways to prevent and treat diseases. Undoubtedly, to analyze the state of nonspecific protection of the child's body, it is necessary to take into account, including ontogenetic aspects, that is, the intensity of the processes of proliferation and differentiation in the child's body in a specific age period.

In this way, goal research was the study of the system "LPO-AOZ" in children of different ages.

Material and methods

The studies were carried out in 75 children of Irkutsk (a large industrial center) of 3 age groups: preschool age (3-6 years old, average age 4.7 ± 1.0 years) - 21 children (group 1), primary school age ( 7-8 years old, mean age 7.6±0.4 years) - 28 children (Group 2) and secondary school age (9-11 years old, mean age 9.9±0.7 years) - 26 children ( 3rd group).

Practically healthy children who had no history of chronic diseases and had not been ill for 3 months prior to the examination and blood sampling were selected for the examination. All children attended pre-school institutions or schools. The surveyed did not take vitamins at the time of blood sampling. Blood was taken in the morning on an empty stomach from the cubital vein.

The work followed the ethical principles of the Declaration of Helsinki of the World Medical Association (World Medical Association Declaration of Helsinki, 1964, 2000 ed.).

The method for determining the primary products of lipid peroxidation - diene conjugates in blood plasma - is based on the intensive absorption of conjugated diene structures of lipid hydroperoxides in the region of 232 nm. The content of TBA-active products in blood plasma was determined in the reaction with thiobarbituric acid by the fluorimetric method.

To assess the total antioxidant activity (AOA) of blood plasma, a model system was used representing a suspension of egg yolk lipoproteins, which makes it possible to assess the ability of blood plasma to inhibit the accumulation of TBA-active products in suspension. LPO was induced by adding FeSO 4 ×7H 2 O . The method for determining the concentrations of α-tocopherol and retinol in blood plasma involves the removal of substances that interfere with the determination by saponification of samples in the presence of large amounts of ascorbic acid and the extraction of unsaponifiable lipids with hexane, followed by fluorimetric determination of the content of α-tocopherol and retinol. While α-tocopherol has intense fluorescence with a maximum of excitation at λ=294 nm and emission at 330 nm; retinol - at 335 and 460 nm. Reference values ​​for α-tocopherol - 7-21 µmol/l, retinol - 0.70-1.71 µmol/l. The method for determining riboflavin is based on the principle of measuring the fluorescence of luminflavin to detect riboflavin in microquantities of blood, which makes it possible to determine the content of this vitamin in erythrocytes and whole blood with sufficient accuracy and specificity. Reference values ​​for riboflavin are 266-1330 nmol/l whole blood. The measurements were carried out on a Shimadzu RF-1501 spectrofluorimeter (Japan).