2 . 2014

A review of current methods for nutrimetabolomic and proteomic research in biochemistry of nutrition

At present biochemistry of nutrition involves the use of OMICs to investigate food quality, safety, bioactivity and nutrition mechanisms. In this context, nutrimetabolomics is one of the latest directions of nutrition development and provides a better understanding of the influence of nutritional factors on the metabolic pathways of the organism. Proteomic methods play an important role in nutrimetabolomics and allow to detect, identify and quantify proteins under different conditions. Variety of technical and methodological advances, improvements in bioinformatics and possibility of tandem use of different methods helps to solve a number of basic and applied science’s problems. Currently huge amount of qualitative and quantitative data on the structure, functions and activities of proteins and their interactions is accumulated. Proteomics aims to establish and characterize a complete set of proteins of the organism. This review summarizes the basic applications of proteomics used in nutrimetabolomic researches. The advantages and disadvantages of the most common techniques of protein separation and sample ionization, types of mass analyzers, basic approaches to the identification of proteins and most widely used databases of known biological sequences are overviewed with a critical assessment of challenges and potential applications.
Biochemistry of nutrition

Phytoecdysteroids influence on the hormonal status and apoptosis in growing rats


The impact of the 15-day consumption of Serratula coronata extract containig phytoecdysteroids on some indicators of hormonal status and activity of apoptosis in various organs of growing male Wistar rats (initial body weight 127,8±2,5 г) has been studied. The extract from the leaves of Serratula сoronata was added to the water of animals of experimental groups 2 and 3 (n=8 in each group) daily at the dose of 5 and 15 mg phytoecdysteroids per kg of body weight respectively.

Animals of the control group 1 (n=8) received water alone throughout the experiment. Daily volume of drunk fluid was recorded. Аt the 15 th day of the experiment animals were taken out using the decapitation under the light ether anesthesia.

The content of corticosterone, prostaglandin E 2 and beta-endorphin in rat blood plasma were determined by ELISA test. Plasma level of noradrenaline was determined by HPLC. DNA damage and percentage of apoptotic cells (apoptotic index)were measured in isolated cells of the thymus, heart and brain by singlecell gel electrophoresis (the comet assay). Significantly lower concentration of norepinephrine was detected in plasma of experimental animals from groups 2 and 3 (10,3±1,1 and 7,2±0,8 ng/ml, respectively ) compared to the same index in the control group (20,4±3,4 ng/ml). Significant differences of other biochemical parameters for all groups of animals have not been identified. Statistical significant difference in the ratio of corticosterone/norepinephrine compared with control animals was detected for a group of rats consumed the highest dose of phytoecdysteroids. There was no statistically significant difference in DNA fragmentation and apoptosis index in animals consumed phytoecdysteroids in compare with the control group of animals. The absence of the activity of apoptosis in cells of the heart, brain and thymus of rats treated with phytoecdysteroid extract may indicate the safety of its use in the diet of the animals.

Soluble apoptosis markers in obese patients with food intolerance


For the soluble apoptosis markers study 151 patients with obesity (92 women and 59 men) aged between 18 and 63 years were examined. Diagnosis and degree of obesity was based on the body mass index (38,2±5,4 kg/m2). Generally food intolerance was identified in 36,4% of obese patients. Four groups of patients were formed: three groups of patients with obesity stage I (15 patients), II (18 patients) and III (22 patients), respectively, and with food intolerance, and a group of obese patients without food intolerance (control group, n=31).

Obese patients with food intolerance received standard version of hypocaloric diet with the exception of specific food allergens. Duration of observation was 39–43 days. Such soluble apoptosis markers as sFas-L, Caspase-9, Caspase-8 and sCD153 were significantly higher in stage III obesity patients compared obese patients without food allergy (0,120±0,030 vs 0,035±0,010; 13,2±3,2 vs 5,9±0,4; 1,4±0,18 vs 0,6±0,24; 0,123±0,010 vs 0,025±0,002 ng/ml respectively). Positive dynamic of sFas-L, Caspase-9 and Caspase-8 (decrease to 0,052±0,030; 7,7±2,2 and 0,4±0,18 ng/ml respectively) in patients with obesity stage III and intactness sCD153 during diet therapy course were revealed.

Significant differences for only Caspase-9 in patients with obesity stage II were obtained. The data obtained are considered as normalization of apoptosis due to nutritional correction of immunological disorders. Study of sFas-L, Caspase-9 and Caspase-8 allows to predict the course of disease, as immunological research for early detection of food allergy makes possible to implement the principles of personalized diet therapy.

Effect of wheat bran fiber on vitamin status of weaning rats with alimentary polyhypovitaminosis

Effect of wheat bran on the vitamin status of rats adequately provided with vitamins or insufficiently supplied with vitamins has been investigated. 32 male Wistar weaning rats (initial body mass – 49–67 g) were randomly divided into 4 groups and fed with complete semi-synthetic diet, containing 100 or 20% of vitamin mixture with or without addition of wheat bran (5% of diet mass) for 35 days. The animals of the control group received 100% of vitamin mixture without adding of wheat bran; 2 group – received those diet with wheat bran; 3 deficient group – 20% of vitamin mixture with full exclusion of vitamins Е, В1 and В2 ; 4 group – 20% of vitamin mixture and wheat bran. The inclusion of wheat bran in full semi-synthetic diet has been accompanied by significant decrease of α-tocopherol liver content on 17% (р=0,006), significant increase of vitamin B1 liver level on the 16% (р=0,027) and blood plasma vitamin D elevation on 19% (р=0,017), as well as a tendency (р=0,059) to increase the liver level of vitamin B2 . Indicators of vitamin A status as well as plasma vitamin E concentration, liver and blood plasma MDA levels were not changed in this group rats. The 5-fold reduction of the vitamin mixture quota and the exclusion of vitamins E, B1 and B2 resulted in a significant (p<0,05) 1,6–1,8 fold decreased in animal body weight and liver mass and the manifestation of the deep external signs of vitamin deficiency. Young animals were more sensitive than adult animals to a lack of vitamins in the diet. Vitamin A (retinol palmitate) liver content in rats from this group was 25,1-fold reduced, vitamin E (α-tocopherol) – 2,1-fold, vitamins B 1 and B 2 – by 57 and 38% compared with animals received a complete control diet (p<0,05). Blood plasma concentration of vitamins A, E, D was 19–34% decreased. Adding of bran in vitamin deficit diet led to increased consumption of vitamin B1 – on 40%, vitamins B 2 and E – 21%, both due to their natural content in the bran, and as a result of increased eatability of the feed by 16% relative to deficient group due to improved taste of the diet. Enrichment of vitamin scarce diet with wheat bran led to an increase in body weight by 56%, the efficiency of the diet by 67%. This circumstance didn’t allow to reveal the effect of dietary fiber on the vitamin status of rats with polyhypovitaminosis. The significant (p<0,05) increase of retinol plasma level by 34% and liver and blood plasma tocopherol content by 17% and 22% and reduction of MDA blood plasma level by 24% took place in animals from this group compared to a group of rats receiving vitamin deficit diet without any effect on liver MDA level, liver vitamin A, B1 and B2 content and heart coenzyme Q10 level. The results obtained suggest that wheat bran inclusion in the diet of rats adequately supplied with vitamins may lead to а deficiency of vitamin E.
Hygiene of nutrition

Regional features of obesity-associated gene polymorphism (rs9939609 FTO gene and gene Trp64Arg ADRB3) in Russian population


Recent studies have shown a significant association with obesity polymorphisms: rs9939609 gene due to fat mass and obesity FTO in European and some Asian and African American populations Trp64Arg ADRB3 gene in several European populations. Association of variants rs9939609 and Trp64Arg obesity was studied in 1244 the inhabitants of Moscow and Sverdlovsk regions. Genotyping was performed using allele-specific amplification, detection results in real time using TaqMan-probes complementary DNA polymorphic sites. The frequency of the mutant allele of the FTO gene in the population of Moscow and Sverdlovsk region was 45,1%, with the TT genotype was detected in 30,2% of cases, AT – 49,5%, AA – 20,3%. Women had the presence of the mutant allele more likely than men (48,4 vs. 42,5%). People with obesity were more genotypes AA (26,3%) and AT (52,8%) compared to the surveyed with a BMI of less than 30 kg/m 2 (respectively 18,1 and 50,7%). A significantly higher incidence of risk allele A was found in individuals with obesity (52,6 and 43,4%). The presence of the mutant allele of the gene ADRB3 among the population of Moscow and Sverdlovsk regions was noted in 7,4% of cases. While 15,5% of patients had a heterozygous genotype Trp64Arg ADRB3, that is consistent with international research. The frequency of the risk allele and genotype Arg64 Trp64Arg in women (9,3 and 18,5%) was significantly higher than men (6,2 and 12,2%).

The presence of the mutant allele and genotype Trp64Arg ADRB3 (respectively, 9,1 and 18,1%) were significantly more marked in the examined obese compared with those with a body mass index less than 30 kg/m 2 (7,4 and 14,9%), but these differences were not statistically significant. The results of these studies suggest that genetic variants of the FTO gene rs9939609 genotype and Trp64Arg ADRB3 contribute to the development of obesity among residents of Moscow and Sverdlovsk Region of Russia. The risk of obesity increases in the case of combined polymorphisms in both genes.

Toxicological assessment of nanostructured silica. The acute oral toxicity


Nanostructured amorphous silica (SiO2 ) is one of the priorities of nanomaterials, exposing human to the ever-increasing scale as a component of food additives, drugs and cosmetic products. According to numerous publications SiO2 nanoparticles (NPs) possess deleterious effects on animal and human cells in vitro and also exhibit inhalation toxicity. However, the biological effects in vivo of silica NPs taken orally are studied insufficiently. This article represents the first section of this study which aim is identification of silica preparation as nanomaterial and estimating of acute toxicity after oral administration in the form of aqueous suspension. Studies of size and shape of the particles in aqueous suspension of silica used in the study by electron and atomic force microscopy, spectroaqustic analysis and dynamic laser light scattering showed that the test substance is a nanomaterial. Estimation of acute toxicity of an aqueous suspension of nanostructured silica with a single intragastric gavage to male BALB/C mice allowed to conclude that the test material has LD 50 by the oral route greater than 10 000 mg/kg and consequently belongs to class IV (low danger agents). Pathological changes in jejunum and colon of mice in the acute experiment (at a dose of 10 000 mg/kg) haven’t been identified. Thus SiO2 NPs possess low toxicity when administered in the gastrointestinal tract.

The available data, however, do not exclude possible presence of adverse effects under their long-term administration by oral way.

Effect of phytosterols and alkylglycerols in the diet on morphometric indicators of liver structure in rats

In the experiment where rats were fed a diet with phytosterols and alkylglycerols for 1,5 months, changes were observed in morphometric parameters in the liver structure in rats. In animals, which were fed a diet with 20% replacement of the fat component (lard) on phytosterols (stanols derived from rapeseed and conifers), blood circulatory disorders of the liver were observed. There was dilatation of the lumens of the central veins and hepatic veins in the interlobular vascular bundles. On the periphery of the lobules, around the vascular bundles, abundant clusters of lymphocytes were revealed. In both groups of rats fed a diet containing various amounts of alkylglycerols obtained from Berrytenthis magister liver (7 and 50 mg per day) and lard as a fat component, in peripheral areas of hepatic lobules the reticuloendothelial cell count was increased as compared with the control group of animals fed a diet containing as fatty component a mixture of lard and sunflower oil (1:1). These cells contained polysaccharides in the cytoplasm and formed thin bands along the hepatic tubules. In addition, in all groups of rats receiving diets with lipid components (both stanols and alkylglycerols), the occurrence of reticuloendothelium proliferation foci in the middle and central zones of liver lobules were 1,8, 2,3 and 2,1 fold higher than in control group. As compared to control animals, the foci in the above groups contained 1,8, 1,7 and 1,6 fold more cells. Furthermore, the number of animals with reticuloendothelium proliferation foci in the groups receiving investigated lipid components was also increased by 2 fold, as compared to controls.

Influence of titanium dioxide and silica nanoparticles on accumulation and toxicity of lead in experiments with intragastric co-administration

The aim of this work was experimental verification of assumptions about the possibility of potentiation of accumulation and toxicity of lead (Pb) after its joint intragastric administration with nanoparticles (NPs) of titanium dioxide (TiO2 ) and silica (SiO2 ). Lead acetate was administered intragastrically to rats at a dose of 20 mg/kg body weight of lead over 21–23 days as a solution in water or in aqueous slurry of TiO2 or SiO2 NPs taken at 1 and 100 mg/kg body weight. The data was obtained that co-administration of Pb with NPs of SiO2 and TiO2 led to changes in a number of indicators that can be interpreted as a slight increase in the toxic effect of the tested substances. However, the size and direction of identified effects depended on the type and the dose of NPs of both kinds. In coadministration of Pb with NPs of TiO2 at both doses (rats with initial body mass 80±8 g) there was a decrease in hemoglobin concentration on 24% (p<0,05), number of lymphocytes on 13% (p<0,05), and platelets on 10% (p<0,05) in the blood, together with the activation of apoptosis in hepatocytes. Introduction of Pb with SiO2 NPs (rats with initial body mass 140±4 g) contrary resulted in increased concentration of hemoglobin on 24% (p<0,05) and significant decrease of urinary excretion of 5-aminolevulinic acid. Accumulation of Pb coadministered with TiO2 was not influenced in liver and decreased in spleen on 50% (p<0,05), testis on 79% (p<0,05) and brain on 38% (p<0,05). SiO2 had no influence on these indices. It is concluded, that the hypothesis about Pb toxicity facilitation due to its transport across the intestinal wall in the form adsorbed on the NPs, does not receive experimental verification, and the observed effects were most likely due to both the toxicity of the Pb, and toxicity (in the studied doses ) of NPs studied.

The structure of the nourishment of preschoolers during the weekend (short report)


To study the feeding of 190 children aged 3–7 years attending 4 preschool educational institutions in the city Mezhdurechensk, Kemerovo region, at home on weekends questionnaires were conducted by the parents. Studies have shown that multiple meals in 57% of children was 5 times a day, 43% of children – 4 times a day, including snacks; 97% of children have at breakfast main dish, drinks and sandwiches and only 3% of children have breakfast that did not meet standards for structure and was insufficient and inadequate to cover subsistence expenses of energy, because consisted of a sandwich and a drink. Only 7% of preschooler’s lunch met recommendations on its structure and included the first, main dish, salad and drink, lunch of 93% of children included only one hot dish (first course, or the second). Dinner in volume, compared with the recommended standards, was redundant on average 57% of preschoolers since consisted of sandwiches, salad, meat dish and side, drink. Children’s snacking during the day included the following suite of products: sandwiches (with sausage, cheese, butter), tea with sweets (candies, chocolate), fermented milk drink, fruits.

When examining the frequency of meals during the day on weekends, there is a tendency to use the same dishes (what children ate for lunch, they eat at dinner on the same day, and then for breakfast on Sunday). The study of the structure of meals in pre-school children at home during the days of the week (evening meal) found that in most cases (67,8%) their dinner was irrational, the structure was identical to the structure of the adults of the family (fried dishes: fried pies, fried potatoes, sodas). Irrationally organized meals for children at the weekend showed inadequate knowledge of parents on the healthy diet for their child at home.
History of medicine

Historical Journey to Infant Feeding


Feeding The article provides an overview of the available literature on problems of infant nutrition, and shows the historical development of the principles of infant feeding. It discusses in greater detail the use of goat milk as a basis for infant nutrition.

It notes the need for a comparative analysis of breast milk substitutes, and for clinical studies evaluating the value of goat milk in infant nutrition.

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Viktor A. Tutelyan
Full Member of the Russian Academy of Sciences, Doctor of Medical Sciences, Professor, Scientific Director of the Federal Research Centre of Nutrition, Biotechnology and Food Safety (Moscow, Russia)

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