Imbalance of acquired immunity factors in newborns with cytomegaloviral infection depending on the severity of the disease.

OBJECTIVE: to assess the imbalance of the parameters of the immune status at the time of manifestation of CMVI in newborns in order to predict the development of a severe form of the disease. METHODS: 133 newborns with CMVI were examined. Depending on the severity of the condition, all children were divided into two groups: 1 - CMVI, severe form - 60 people (45.1%); 2 - CMVI, moderate form - 73 people (54.9%). An immunological examination was carried out, including the determination of T and B lymphocytes, as well as intercellular interaction between them. Using the "classification trees" method, we compared two differentiated approaches to the prediction of severe CMVI: based on a study of the relationships between T-lymphocytes (CD3+), T-helpers (CD4+), the content of cytotoxic lymphocytes (CD8+) and B lymphocytes (CD20+) and by studying the interaction of co-stimulation molecules CD28, CD3 + CD28 +, CD3-CD28 +, CD3 + CD28-. RESULTS: In this study, it was demonstrated that the "standard immunogram", limited to the definition of CD3+, CD4+, CD8+, CD20+ lymphocytes, shows less information in predicting the development of a severe form of the disease in this patient population than the combination of markers, including CD3 + CD28-, CD20 + CD40 +. FINDINGS: A model for predicting a severe form of cytomegalovirus infection in newborns based on the analysis of the content of CD 28 costimulation molecules has a higher information content, which makes it possible to start specific therapy on time.

[Nutriome as the direction of the "main blow": determination of physiological needs in macro- and micronutrients, minor biologically active substances].

One of the essential parts of fundamental research in Nutrition Science is the determination of the physiological requirements of humans for energy and food substances. Research that has been carried out in this area over the past 90 years, consistently develops and improves the norms of physiological requirements for energy and nutrients for various groups of the population of the Russian Federation. In the 50 years of the last century in this research field, determining the values of daily intake for macronutrients (proteins, lipids and carbohydrates), was in the first place. Then the Era of micronutrients (vitamins, minerals, trace elements) was started, and, finally, now there is the Era of minor food biologically active substances. More and more facts are accumulating about their leading role in regulating metabolism. They can be recognized as endogenous regulators, the primary vital components involved in the formation of human health. In recent years, the new definition of Nutriome is introduced into Nutrition Science. It is considered as a set of essential nutritional factors to maintain a dynamic equilibrium between human being and the environment, aimed to ensure viability, the preservation and reproduction of the species, keeping the adaptive capacity, the system of antioxidant defence, apoptosis, metabolism, and immune system function. The Nutriome is a formula for optimal nutrition, which is continually being improved and supplemented. Knowledge of this formula is the key to forming an optimal diet for a person, and, therefore, to save their health. It is evident that at the population level, the Nutriome has its characteristics, its structure for each age period of human life. The need to develop a formula for optimal nutrition and, consequently, updating nutrient-based dietary guidelines is induced by socio-economic and demographic changes in population, changes in anthropometric characteristics of children and adults, increasing prevalence of socially significant non-communicable diseases, developing studies of the significance of particular food substances and establishing the relationship between nutrition and health.

Effects of Quercetin on Expression of Genes of Carbohydrate and Lipid Metabolism Enzymes in the Liver of Rats Receiving High-Fructose Ration.

We studied the expression of genes encoding enzymes of carbohydrate and lipid metabolism ketohexokinase (Khk), glucokinase (Gck), pyruvate kinase (Pklr), acetyl-Co-carboxylase (Acaca), fatty acid synthase (Fasn), stearoyl-CoA desaturase (Scd), and their transcription regulators ChREBP (Mlxipl), SREBP-1c (Srebf1), and PPARα (Ppara) in rat liver. Control group rats received a semisynthetic ration over 20 weeks. Experimental group 1 received a semisynthetic ration and 20% fructose solution instead of drinking water. Experimental group 2 rats received a semisynthetic ration with quercetin (0.1% fodder weight) and 20% fructose solution. Consumption of 20% fructose solution (experimental group 1) led to an increase in Scd expression in comparison with the control and did not affect the expression of other genes. Addition of quercetin to the ration (experimental group 2) led to a decrease in the expression of Khk, Gck, Fasn, Scd, Mlxipl, and Ppara genes in comparison with experimental group 1. The results suggest that quercetin reduced the expression of genes of carbohydrate and lipid metabolism enzymes in the liver of rats receiving high-fructose ration.

[Effects of polyphenols on activity of glycosyl hydrolases in the cecum of rats fed obesity inducing diets].

The results of experimental studies indicate that the preventive and therapeutic effects of polyphenols in obesity are accompanied by a significant decrease in the severity of dysbiosis caused by the predominance of fats and simple carbohydrates in the diet, especially fructose, and the restoration of the functional state of the microbiota. The aim of the work was to study the effect of quercetin and resveratrol - polyphenols, widely represented in the daily human diet, on the activity of bacterial glycosidases in rats receiving diets high in fructose or fat and fructose. Material and methods. Using spectrophotometric analysis, the activity of ß-galactosidase (Gal), ß-glucosidase (Glu) and ß-glucuronidase (Gluс) was studied in the content of the cecum of Wistar rats receiving a semi-synthetic diet and a 20% solution of fructose instead of drinking water (hfr diet) or a semi-synthetic diet with a high (30%) fat content and a 20% solution of fructose instead of drinking water (hf/hfr diet). Results and discussion. Feeding rats with the hfr diet for 20 weeks led to the suppression of Gal activity by 35, Glu by 46 and Gluс by 31%. With the inclusion of quercetin in the hfr diet at a dose of 34 mg/kg b.w. enzyme activity was restored to the control values and exceeded the level of activity in rats fed hfr ration without quercetin by 60, 100 and 47%, respectively, for Gal, Glu, and Gluс. Feeding rats with the hf/hfr diet for 10 weeks did not have a significant impact on the activity of bacterial enzymes. The inclusion of resveratrol in the hf/hfr diet at a dose of 10 mg/kg b.w. resulted in a decrease in Glu activity by 58 and Gluс by 28%, and an increase in resveratrol dose to 100 mg/kg b.w. caused further suppression of Gal activity by 30, Glu by 76 and Gluc by 64% comparative to the activity in rats on the hf/hfr diet without resveratrol. Conclusion. The obtained data suggest that quercetin restores reduced by hfr diet activity of glycosyl hydrolases of the cecum microflora of rats, most likely due to an increase in the representation of the types of enzyme activity carriers. The suppressive effect of resveratrol on the activity of glycosyl hydrolases of the cecum microflora of rats fed a hf/hfr diet may be the result of its direct action on enzymes and is not associated with the effect on the composition of the intestinal microbiota.

[Effects of quercetin on protective capacity in rats fed a high-fructose diet].

The purpose of the study was to determine effects of quercetin on protective capacity parameters in the experiment on rats fed a high fructose diet. Rats of the control group received a semi-synthetic (s/s) diet and water; animals from the 1st experimental group - s/s diet and 20% fructose solution instead of drinking water; rats of the 2nd experimental group- s/s diet with quercetin (0.1% indiet) and 20% fructose solution instead of drinking water for 20 weeks. Parameters of antioxidant status [total antioxidant activity (AOA), the content of malondialdehyde (MDA) and lipids hydroperoxides, the level of reduced and oxidized glutathione, activity of superoxide dismutase, catalase, glutathione peroxidase, paraoxonase-1, hemeoxygenase-1, NAD(P)H-quinone oxidoreductase], the activity of xenobiotic-metabolizing enzymes [CYP1A1, CYP1A2, CYP2B1, CYP3A, UDP-glucuronosyltransferase (UDP-GT) and glutathione transferase] were studied in plasma and liver of rats. Consumption of the high-fructose diet led to changes in some parameters: diminution of AOA in blood plasma, decrease of AOA and MDA level, unsedimentable activity of lysosomal enzymes, increase of the UDP-GT activity in liver. The inclusion of quercetin in the diet did not affect the studied parameters, except for a more pronounced decrease of the unsedimentable activity of lysosomal enzymes in rat liver. The results of the study indicated that there was no significant effect of quercetin on the protective capacity of rats at the initial stage of obesity caused by high-fructose diet.

[The role of polymorphism of the interferon gene ɣ and interferonoproduction in the pathogenesis of infection caused by herpes 6 type virus in children of early age.]

The purpose of the article is to reveal the immunological features of the infection caused by the herpesvirus type 6 virus, based on the level of the polymorphism IFNG geneγ (+874 A / T) and interferon production. Group I (n = 58) consisted of children with herpetic infection of type 6, severe form. Group II (n = 31) included children with herpetic infection of type 6, medium-heavy form. The control group consisted of 53 healthy newborns born. The allelic variants of the genes were determined by the PCR method followed by restriction analysis of the GosNII Genetics test systems (Moscow) Among the allelic variants of the interferon γ gene, patients with a severe form of herpetic infection of type 6 are significantly more likely than in children with a moderate-to-severe course of the disease, and the AA genotype of the polymorphism +874 of the IFNG gene is detected in the control group and significantly less than the control group, the allele T and the TT genotype. The level of interferon-γ had a significant decrease in children with severe infection in comparison with the control group. Exposure of the infection is associated with the A allele and the AA genotype of the polymorphic region +874 A / T of the IFNG gene.

[The influence of separate and combined supplementation with curcumin and quercetin on the protective capacity in rats].

The purpose of the study was to determine the effects of curcumin (CUR) and quercetin (QUER) on the expression of genes and activity of prototypical Nrf2/ARE- and AhR/ XRE-regulated enzymes. Investigation was carried out on male Wistar rats with initial body weight (230-235 g b.w.) that received for 14 days CUR (200 mg/kg b.w.) and QUER (200 mg/kg b.w.) separately or in combination within the standard semi-synthetic diet. The expression of genes and activity of Nrf2/ARE - regulated enzymes - heme oxygenase- 1(HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO1), AhR/XRE-regulated CYP1A1, CYP1A2 enzymes and the mRNA level of transcription factors Nrf2 and AhR were determined in rats liver. Also the expression of gene CYP3A1 and activity of CYP3A, UDP-glucuronosyltransferase, glutathione transferase were studied in rats liver. Along with this the total antioxidant activity (AOA), malondialdehyde and lipid hydroperoxides levels were determined in blood plasma and liver. The reduced and oxidized glutathione level, total and unsedimentable activity of lysosomal enzymes were investigated in rats' liver. QUER, especially in combination with CUR, increased the AOA of blood plasma and reduced the content of lipid hydroperoxides in it. CUR and QUER did not affect NQO1 activity, but the combined action caused an increase in the HO-1 activity without affecting the expression of the corresponding gene (Hmox1) and Nrf2 gene. CUR and, to a lesser extent QUER, had a strong inducing effect on CYP1A1, CYP1A2, CYP3A activity, but only the CYP1A1 activation was accompanied by the induction of CYP1A1 gene. The inducing effect of CUR and QUER on the activity of CYP450 enzymes greatly enhanced by their combined action. Membrane stabilizing action of CUR and QUER was also strongly expressed under its combined intake. Thus, we can conclude that CUR and QUER, especially in combination, contribute to the protective and adaptive capacity.

[The effect of rutin and hesperidin on the expression of Nrf2- and AhR-regulated genes and CYP3A1 gene in rats intoxicated with carbon tetrachloride].

The purpose of the study was to determine the effects of rutin (R) and hesperidin (Hes), the main representatives of two most studied subclasses of flavonoids - flavonols and flavanones, on the expression of prototypical Nrf2 and AhR-regulated genes and CYP3A1 gene in rats intoxicated with carbon tetrachloride (CCl4). Investigations were carried out on 5 groups of male Wistar rats with the initial body weight (b.w.) 180-200 g (n=40). Rats of the control group and the 1st experimental group received for 14 days the semisynthetic diet, rats of the 2nd experimental group - the same diet plus R (400 mg/kg b.w.), the animals of the 3rd experimental group received the diet with Hes in the same amount, of the 4th experimental group - diet with R (400 mg/kg b.w.) and Hes (400 mg/kg b.w.). Animals of the experimental groups 24 hours before the end of experiment were injected intraperitoneally CCl4 at a dose of 0.5 ml/kg b.w. in olive oil; rats of the control group were injected equal amount of olive oil. For gene expression assessment the mRNA content of NAD(P)H-quinone oxidoreductase (NQO1), heme oxygenase-1 (Hmox1), Nrf2 (Nrf2), AhR (AhR), CYP1A1, CYP1A2, CYP3A1 and ß-actin (Actb) in rat liver was determined by real-time RT-PCR. The results showed that in rats intoxicated with CCl4, enrichment of the diet with R, but not with Hes, led to a significant increase in the expression of genes Hmox1, NQO1 and CYP3A1. Combined intake of R and Hes with the diet led to additivity of their action on the expression of Hmox1 gene and to synergism in the effect on the expression of genes NQO1 and CYP3A1. A moderate increase in the levels of expression of AhR and CYP1A2 genes as compared to their expression in rats treated with CCl4 only, CCl4 and R or CCl4 and Hes has been noted. Thus, for the first time on the model of oxidative stress in rats the data have been obtained showing at the gene expression level a synergism of action of two flavonoids - R and Hes, widely present in the daily human diet.

[Effect of rutin and hesperidin on the expression of Nrf2 gene and the activity of hemoxygenase-1 and NAD(P)H-quinone oxidoreductase at their separate and combined action].

Flavonoids rutin (R) and hesperidin (Hes) have a broad spectrum of the biological activity based on their antiradical properties and ability to increase the activity of antioxidant enzymes, including the activity of heme oxygenase-1 (HO-1) and NAD(P)H-quinone oxidoreductase (QR). It is supposed that the main regulator of the activity of HO-1 and QR is the transcription factor Nrf2. The purpose of the study was to determine the effects of R and Hes on the expression of Nrf2 gene and protein, on the activity and mRNA and protein expression of HO-1 and QR at their separate and combined action. Administration in diet of male Wistar rats (with initial body weight 180-200 g) R (400 mg/kg b.w.) and Hes (400 mg/ kg b.w.) during 14 days separately or in combination had no toxic or pro-oxidant effect, which were assessed by the level of liver MDA, hydroperoxides of lipids, reduced and oxidized glutathione. R and, to a lesser extent, Hes caused increased activity of HO-1 and QR. Their combined effect on the activity of HO-1 did not differ from the separate effect of each flavonoid. The combined action of R and Hes on the activity of QR was additive. According to Western blotting, changes in HO-1, Nrf2 and QR protein levels under the action of R and Hes separately or in combination were not statistically significant. The results of real time PCR demonstrated the presence of small, but statistically significant, changes in the level of expression of the genes Nrf2 (Nrf2), HO-1 (Hmox1) and QR (NQO1) both for separate and combined action of R and Hes. Thus, the obtained results showed that high-dose of R and Hes separately and in combination didn't significantly affect the gene and protein expression of transcription factor Nrf2 and that the increased activity of HO-1 and QR was not associated with the increased expression of Hmox1 and NQO1 genes.

[Toxicological evaluation of colloidal nano-sized silver stabilized polyvinylpyrrolidone. III. Enzymological, biochemical markers, state of antioxidant defense system].

Nanosized colloidal silver (NCS) with primary nanoparticles (NPs) size in the range of 10-80 nm in aqueous suspension was administered to rats with initial weight 80±10 gfor the first 30 day intragastrically and for lasting 62 days with the diet consumed in doses of 0.1; 1.0 and 10 mg/kg of body weight b.w) per day based on silver (Ag). The control animals received deionized water and carrier of NPs - aqueous solution of stabilizer polyvinylpyrrolidone. Activity (Vmax) was determined in liver of microsomal mixed function monooxygenase isoforms CYP 1A1, 1A2 and 2B1 against their specific substrates, the activity of liver conjugating enzymes (glutathione-S-transferase and UDP-glucuronosyltransferase) in the microsomal fraction and a cytosol, and the overall and non-sedimentable activities of lysosomal hydrolases. In blood plasma there were evaluated malonic dialdehyde, PUFA diene conjugates, in erythrocytes - the activity of antioxidant enzymes. A set of standard biochemical indicators of blood serum was also determined. The studies revealed changes in a number of molecular markers of toxic action. Among them - the increase in the activity of key enzymes I and II stages of detoxification of xenobiotics, indicating its functional overvoltage; reducing the activity of glutathione peroxidase (GP), the total arylsulfatase A and B, ß-galactosidase (in the absence of changes in their non-sedimentable activity), levels of uric acid, increased alkaline phosphatase activity. These changes occurred mainly at the dose Ag of 10 mg/kg b.w., except for the GP to which the threshold dose was 1 mg/kg b.w. No significant changes in the studied markers in a dose Ag 0,1 mg/kg b.w. were identified. Possible mechanisms of the toxic action of silver NPs are discussed.

[Effect of minor bioactive food substances ­ rutin and hesperidin in their separate and combined alimentary arrives on the immune system of rats and the activity of nuclear factor NF-kB liver cells].

The effect of rutin and hesperidin in their separate and combined admission to the immune system and the activity of nuclear factor NF-kB of rat liver cells has been investigated. Wistar male rats with an initial body weight of 224­225 g were divided into 4 groups of 6 rats in each. The rats of the 1st group (control) received a complete semi-synthetic diet, rats in group 2 ­ the same diet supplemented with rutine (400 mg/kg b.w.); the rats of group 3 ­ with the addition of hesperidin (400 mg/kg bw); group 4 ­ with the addition of rutin and hesperidin (400 mg/kg b.w. each) for 14 days. Animals received feed in free access mode in an amount of 25­30 g per rat per day, that corresponded to 15 g of dry formula. Animals received water also in free access. It has been found that rutin and hesperidin, included in the diet of rats both alone (groups 2 and 3) and together (group 4), have immunomodulatory impact which is a reduce of lymphocyte relative content [1st gr. ­ 70.55±1.58%, 2nd gr. ­ 63.62±2.85%, 3rd gr. ­ 62.03±3.16% (p1­3<0.05), 4th gr. ­ 65.75±1.08% (p1­4<0.05)] and an increase of percentage of neutrophil leukocytes [1st gr ­ 19.98±0.97%, 2nd gr. ­ 25.35±3.14%, 3rd gr. ­ 28.27±3.30% (p1­3<0.05), 4th gr. ­ 24.15±1.52% (p1­4<0.05)] and NK-cells in the peripheral blood [1st gr. ­ 3.29±0.45%, 2nd gr. ­ 6.91±0.70% (p1­2<0.05), 3rd gr. ­ 5.88±0.79% (p1­3<0.05), 4th gr. ­ 4.64±0.32% (p1­4<0.05)], that can be considered as a shift in the direction of innate immunity factors. In addition, the combined effect of high doses of rutin and hesperidin led to a change in erythrocyte parameters: an increase in the average volume of red blood cells [1st gr. ­ 56.00±1.06 fl, 2nd gr. ­ 56.67±0.42 fl, 3rd gr. ­ 58.50± 0.99 fl, 4th gr. ­ 59.50±0.99 fl (p1­4<0.05)], and the average content of hemoglobin [1st gr. ­ 18.97±0.45 pg, 2nd gr. ­ 19.10±0.19 pg, 3rd gr. ­ 19.73± 0.32 pg, 4th gr. ­ 20.08±0.33 pg (p1­4=0.07)], as well as increase in the level of TGF-ß1 in peripheral blood [1st gr. ­ 15.55±2.13 ng/ml, 2nd gr. ­ 14.81± 2.36 ng/ml, 3rd gr. ­ 17.02±2.53 ng/ml, 4th gr. ­ 22.14±2.29 ng/ml (p1­4<0.05)] and the expression of nuclear factor NF-kB in the liver cells [1st gr. ­ 16.10± 0.60 ng/ml; 2nd gr. ­ 15.14±2.28 ng/ml; 3rd gr. ­ 15.85±2.09 ng/ml; 4th gr. ­ 20.49±1.68 ng/ml (p1­4<0.05)].

[Effects of rutin on protective capacity in rats].

The purpose of the study was to determine effects of rutin dietary administration on the activity of xenobiotic-metabolizing enzymes and antioxidant status. The study has been carried out on 3 groups of male Wistar rats (n = 8 in each), with initial body weight 100-120 g. Animals of the control group (1st group) received standard semi-synthetic diet, the experimental groups--the same diet with rutin in the amount of 40 mg/kg b.w. (2nd group) or 400 mg/kg b.w. (3rd group). The duration of the experiment was 2 weeks. In rat liver the activity of quinone reductase (QR), heme oxygenase-1 (HO-1), paraoxonase-1 (PON-1), glutathione-S-transferase (GST), ethoxyresorufin-O-dealkylase (EROD) activity of CYP1A1, methoxyresorufin-O-dealkylase (MROD) activity of CYP1A2, tes- tosterone 6ß-hydroxylase (6ß-TG) activity of CYP3A, total antioxidant activity (AOA) and malondialdehyde (MDA) content have been investigated. The expression of genes CYP1A1, CYP1A2 and CYP3A has been measured by reverse transcription-polymerase chain reaction (RT-PCR). The stability of lysosome membranes was estimated by the change of unsedimentable activity of lysosomal enzymes--arylsulfatase, ß-galactosidase and ß-glucuronidase. Rutin administration led to dose-dependent increase in the activity of antioxidant enzymes. In rats of the 3rd group received high-rutin diet the activity of QR, HO-1, PON-1 and GST increased by 68, 29, 17 and 22%, respectively, compared to the control (1st group); MDA level and AOA have not changed. Activity of EROD and MROD in liver microsomes of rats treated with rutin at a dose of 40 mg/kg b.w. (2nd group) increased by 33 and 58%, respectively, with a moderate increase in mRNA level of CYP1A1 and CYP1A2. Increasing the dose of rutin up to 400 mg/kg b.w. (3rd group) resulted in the decrease of the degree of EROD and MROD activation by 18 and 15%, respectively, compared to the 2nd group. Rutin had no significant effect on the activity of 6ß-TG and on the expression of CYP3A1 gene. Rutin dietary administration led to dose-dependent reduction of the unsedimentable activity of lysosomal enzymes, indicating the strengthening of the stability of lysosomal membranes. Thus, the obtained results showed that in healthy, intact rats high doses of rutin in the diet moderately but statistically significantly activate enzyme systems responsible for the protective and adaptive capacity of the organism.

Effect of polyunsaturated fatty acids ω-3 on the induction of activity and expression of CYP1A1 and CYP1A2 genes in the liver of rats under the influence of indole-3-carbinol.

Supplementation of the ration with eicosapentaenoic and docosahexaenoic ω-3 polyunsaturated fatty acids (PUFA) in doses of 0.3 and 1 g/kg body weight for 4 weeks had no effect on ethoxyresorufin O-dealkylase (EROD) activity and expression of the CYP1A1 gene in male Wistar rats, but caused a dose-dependent increase in methoxyresorufin O-dealkylase (MROD) activity of CYP1A2 (by 28 and 73%, respectively) without significant changes in CYP1A2 mRNA expression. ω-3 PUFA had no effect on the indole-3-carbinol-induced (20 mg/kg body weight over the last 7 days of the experiment) EROD activity and expression of CYP1A1 mRNA. The indole-3-carbinol-induced MROD activity was shown to increase by 6.2 times in rats not receiving ω-3 PUFA and only by 3.9 and 2.7 times in animals receiving ω-3 PUFA. The indole-3-carbinol-induced expression of CYP1A2 mRNA slightly increased in animals receiving ω-3 PUFA. Our results suggest that the effect of ω-3 PUFA on the induced and basal activity of CYP1A2 is not related to modulation of CYP1A2 gene expression.

[Effects of vitamins deficiency on the cytochrome P450 inducibility in rats].

The purpose of the study was to determine multi-vitamin deficiency effects on the inducibility of main isoforms of cytochrome P450 in the rat liver. The study was carried out on 4 groups of Wistar rats. Rats of the 1st and 3rd group received semi-synthetic diets containing adequate (100% of recommended vitamin level) level of vitamins, the 2nd and 4th--the semi-synthetic diet containing vitamins in the amount of 20% from adequate level. The duration of the experiment was 4 weeks. During the last week indole-3-carbinol (I-3-C) in dose of 20 mg/kg body weight was added to the diet of the 3rd and 4th group of rats. Vitamin E content in liver and blood serum declined by 59 and 34%, respectively in rats which were fed vitamin-deficient diet (2nd group); vitamin A level decreased by 5 times in the liver, but was not changed in blood serum. Multi-vitamin deficiency in the diet led to the increase in the liver ethoxyresorufin O-dealkylase (EROD) activity of CYP1A1, methoxyresorufin O-dealkylase (MROD) activity of CYP1A2 and testosteron 6beta-hydroxylase (6beta-TG) activity of CYP3A by 11, 80 and 53%, respectively, and gene expression of CYP1A1, CYP1A2, CYP3A and AhR by 8,5; 1,6; 2,4 and 3,6 fold. In rats fed diet with adequate levels of vitamins (3rd group) I-3-C increased activity of EROD and MROD by 4,4 and 5,5 fold, and the expression of CYP1A1, CYP1A2 and AhR genes by 148; 3 and 3,5 fold compared to the parameters of the 1st group (without I-3-C). Multi-vitamin deficiency increased I-3-C-related induction of EROD activity and expression of CYP1A1 and CYP1A2 genes, but decreased I-3-C-related induction of the MROD activity. Thus, 5-fold reducing of vitamin content in rat diet lead to significant changes in activity and inducibility of cytochrome P450 of CYP1A and 3A family, which play a key role in the detoxification and metabolism of drugs.

[Effects of rutin on the activity of antioxidant enzymes and xenobiotic-metabolizing enzymes in liver of rats fed diets with different level of fat].

The study has been carried out on 6 groups of male Wistar rats, which received semi-synthetic diets within 28 days. Rats of 1st and 4th group received fat-free diet, 2nid.and 5th - diet containing standard amount of fat (10% by weight, 26% by caloric content; lard/sunflower oil - 1/1); 3rd and 6th group - a high-fat diet (30% by weight, 56% by caloric content). During the last 14 days of the experiment rats received rutin in the dose of 40 mg/kg b.w. AOA, MDA level and the activity of paraoxonase I have been evaluated in blood serum. In rat liver along with the parameters of the antioxidant status (MDA level, activity of paraoxonase 1, quinone reductase, heme oxygenase-1) the activity of xenobiotic-metabolizing enzymes (XME) (CYP1A1, CYP1A2, CYP3A1, CYP2B1, UDP-glucuronosyl transferase and glutathione transferase) and the activity of lysosomal enzymes (arylsulfatase A and B, ß-galactosidase and ß-glucuronidase) have been investigated. Elevation of the activity of antioxidant enzymes and XME in liver with the increase of diet fat content has been-noted. Rutin admihistration had no effect onparamete6rs of antioxidant status and decreased unsedimentable activity of lysosomal enzymes that did not depend on fat content in the diet. Rutin receiving increased the activity of all studied XME in rats fed standard diet, but practically did not effect on their activity in rats fed by fat-free and high-fat diets. Thus, rutin in pharmacological dose has no effect on the activity of antioxidant enzymes that doesn't depend on the level of fat in the diet, while the decrease or increase of diet fat content modulates (weakens) the influence of rutin on the XME activity.

[Toxicological assessment of nanostructured silica. II. Enzymatic, biochemical indices, state of antioxidative defence].

Nanostructured silica (SiO2) "Aerosil" with the size of the primary nanoparticles (NPs) of 5-30 nm, in the form of ultrasound treated water suspension was administered to rats of 80 ± 4 g initial body weight for the first 30 days by intragastric gavage and then for 62 days with diets consumed in doses of 0.1; 1.0; 10 and 100 mg/kg body weight per day. The control group received vehicle of nanomaterial (NM)--deionized water. There were measured in liver of ani- mals the content of total cytochromes P450 and b5 in the microsomal fraction of liver, activity (Vmax) of microsomal monooxygenases with the mixed func- tion of isoforms CYP1A1, 1A2 and 2B1 on their specific substrates, the activity of conjugating liver enzymes glutathione-S-transferase and UDP-glucuronosyl-transferase in microsomal fraction and cytosol, the total and non sedimentable activity of lysosomal hydrolases (ß-glucuronidase, ß-galactozydase, arylsulphatase A, B). The content of PUFA's diene conjugates and TBA-reactive substances in the blood plasma and the activity of antioxidative enzymes (glutathionperoxidase, superoxidedismutase, glutathionreductase, katalase) in erytrocytes were estimated. A set of standard biochemical parameters of blood serum was also examined (total protein, albumin, glucose, creatinine, urea, uric acid, activities of hepatic transaminases). The studies revealed changes of a number of molecu lar markers that could be interpreted as unfavorable. These include isoforms of CYP2B1 activity decrease at a dose HM 1-10 mg/kg of body weight, decrease in the serum content of total protein, albumin and glucose levels in a dose range of 0.1-10 mg/kg. These changes were absent at the maximum dose of NM, which did not allow to clearly establish the dose-response. The remaining studied fig ures resided in the normal range or experienced changes that could not be interpreted as toxic.

[Effect of indole-3-carbinol and rutin on rats' provision by vitamins' A and E with different fat content in its diet].

Effect of indole-3-carbinol (I-3-C) and rutin (R) supplementation on vitamins A and E status of growing Wistar rats, receiving for 6 or 4 week semi-synthetic diets with different levels (1, 11 and 31%) of fat (lard and sunflower oil at a ratio of 1:1) has been studied. The content of vitamin E was 6, 9 and 15 IU, vitamin A - 400 IU in 100 g of ration. Against the various fat content during the last 7 or 14 days of the experiment rats received respectively I-3-C (20 mg per 1 kg of body weight per day) or R (0.4% of the feed weight). Rat tissues were analyzed for vitamins A (retinol and retinyol palmitate) and E (alpha-tocopherol) by HPLC. Reducing fat content in diet from 11 to 1% was associated with significant (p<0.05) decrease of hepatic retinyl palmitate and alpha-tocopherol (1,6-1,7 times) with constant plasma concentration of retinol and alpha-tocopherol. Raising fat content from 11 to 31% , in contrast, led to increased levels of hepatic retinyl palmitate and alpha-tocopherol respectively by 13% (p=0.248) and 89% (p=0.006) and plasma ROL of 26% (p=0,024), while the plasma concentration of alpha- tocopherol has not changed. I-3-C and R do not affect the availability of vitamin E in rats, regardless of the fat content in the diet. With excess fat content (31%) in the diet, supplementation of I-3-C and R lowered hepatic RP by 22-52% (p<0.05) compared to rats receiving a diet with adequate fat. Adding of I-3-C to the high-fat diets resulted to a significant reduction of vitamin A concentration in blood plasma by 12% (p=0.024) and in liver by 37% (p=0.002).

[Effects of omega-3 polyunsaturated fatty acids on antioxidant capacity in rats].

Dietary administration of omega-3 polyunsaturated fatty acids (EPA + DHA, 1,2:1) at dose 0.3 or 1 g/kg bw during 4 weeks led to minor (by 14% and 17%, p<0.05) decrease of serum antioxidant capacity and serum level of vitamin E (by 30% and 31%, p<0.05) and the activity of paraoxonase-1 (by 14% at 0.3 g/kg bw, p<0.05). The activity of antioxidant enzymes in liver increased in a dose-dependent manner. At higher dose of omega-3 polyunsaturated fatty acids a 45% increase in the activity of paraoxonase-1 (p<0.05), 21% -heme oxygenase-1 (p>0.05), 68% - quinone reductase (p<0.05), 19% - glutathione S-transferase (p<0.05) compared to the control group was found. The direct relationship between activities of enzymes and increase of MDA level in liver (by 47 and 107%, p

Indole-3-carbinol induction of CYP1A1, CYP1A2, and CYP3A1 activity and gene expression in rat liver under conditions of different fat content in the diet.

Wistar male rats were fed semisynthetic diet with different fat content for 6 weeks: fat-free, 10%, and 30% fat (sunflower oil and lard, 1:1). Increasing fat content was associated with an increase in ethoxyresofurin-O-dealkylase (EROD) activity of CYP1A1, methoxyresofurin-O-dealkylase (MROD) activity of CYP1A2, and 6ß-testosterone-hydroxylase (6ß-TH) activity of CYP3A1. EROD, MROD, and 6ß-TH activities in rats on high-fat diet exceeded those in rats on fat-free diet by 64, 58, and 140%, respectively. Addition of indole-3-carbinol to the diet led to an increase in CYP1A1, CYP1A2, and CYP3A1 activities and mRNA levels, the degree of this increase was lower with increasing fat content in the diet. In rats on fat-free, 10% fat, and 30% fat diets, indole-3-carbinol induced EROD activity by 4.7, 3.2, and 2.0 times, respectively; MROD activity by 5.9, 5.6, and 5.4 times; and 6ß-TH activity by 2.1, 1.9, and 1.3 times. A correlation was detected between activities of the studied cytochrome P-450 isoforms, their inducibility by indole-3-carbinol, and vitamin A and E levels in rat liver.

[Effects of dietary fat level on the xenobiotic metabolism enzymes activity and antioxidant enzymes in rats].

Male Wistar rats received fat-free diet or diets containing 5, 10 and 30% of fat (sunflower oil + lard, 1:1) for 4 weeks. The direct relationship between dietary fat level and ethoxyresorufin O-dealkylase activity of CYP1A1, methoxyresorufin O-dealkylase activity of CYP1A2, pentoxyresorufin O-dealkylase activity of CYP2B1 and testosterone 6beta-hydroxylase activity of CYP3A was found. Activities of key enzymes of phase II xenobiotic metabolism (total activity of glutathione transferase, activity of UDP-glucuronosyle transferase) and antioxidant enzymes (catalase, glutathione peroxidase, glutathione reductase, paraoxonase-1 and heme oxygenase-1) also increased with higher dietary fat level.