2 . 2012

Methods of nanoparticles control in food and biological payloads Report 1. Use of microscopic and chromatography investigation methods

One of the promising applications of modern nanotechnology are food productions, which includes the improvement of food packaging, creation of new forms of nutrients that are characterized by improved assimilation and technological characteristics, quality control through the creation of compact and cheap test kits. All these applications of nanomaterials related to the risks of the possibility of receipt of potentially toxic nanoparticles in the diet. The task of regulation and hygienic standardization requires developing of the methods, their qualitative and quantitative analysis for such complex, multicomponent systems, which are the agricultural commodities and food products. The best hope in this plan are assigned to a group of approaches related to the microscopic visualization of artificial nanoparticles in the biological objects. While the typical size of nanoparticles (<100 nm) are below the theoretical maximum-resolution light optical methods, transmission electron microscopy often allows not only to identify nanoparticles on their size and shape, but also a qualitative and quantitative analysis their chemical composition with the use of additional analytical options. Another group of elaborate methods used in solving the problems of qualitative and quantitative analysis of nanoparticles are chromatographic methods, in particular, the exclusion, hydrodynamic, high-performance liquid chromatography, and the flow-field fractionation. Limitation of chromatographic approaches related with the need of complex sample preparation, as well as specific difficulties in nanoparticles detecting in chromatographic fractions. Transmission electron microscopy and high-performance liquid chromatography methods are officially recommended in Russia for the analysis of artificial nanoparticles in natural biological systems, including food products.
Biochemistry of nutrition

Body composition and resting metabolic rate

Indicators of body composition determined by bioelectrical impedance analysis (total body water, extracellular water mass, intracellular water mass, skeletal muscle mass, soft lean mass, protein, mineral mass, fat mass) were compared with the resting metabolic rate values (RMR). Studies were conducted in adults of different occupations, gender, age, weight and height. In total, the survey involved 180 people, residents of Nizhny Novgorod (70 men and 110 women) aged 17 to 63 years. It was shown that the normalization of the RMR with the parameters of the bioelectrical impedance analysis reduced the scatter of individual values. Comparison of the normalization results allowed us to estimate the interconnection between the RMR and the body composition. It was obtained statistical formulas for the relation of the RMR with the bioelectrical impedance. The greatest effect was found with the normalization of lean mass, as well as intracellular and extracellular water masses. The smallest effect was detected with the normalization of body fat. We obtained statistical formulas for the relation of the RMR with the bioelectrical impedance parameters. The highest correlations were found between the RMR and the lean mass, muscle mass and total water mass, the lowest with the fat mass. Role of the metabolically active tissue in energy metabolism, including the role of the intracellular and extracellular water is discussed. The results confirm the importance of the metabolically active tissues in the formation of the value of the RMR. Identified dependencies and high correlation between the RMR and the bioelectrical impedance parameters allows us to use these as a fast and convenient method for the approximate estimation of the RMR.

Influence of anatase titanium dioxide nanoparticles on protein expression profiles in rat liver microsomes

There was studied an influence of intragastric administration of titanium dioxide (anatase form) nanoparticles (NP) on protein expression profiles in rat’s liver microsomes by methods of proteomics. Animals received water suspension of NP in doses from 0,1 to 10 mg per kg body weight intragastirically daily during 28 days. Microsomes were isolated from liver by means of preparative ultracentrifugation. Proteins composition was studied by 2D-electrophoresis in acrylamide gel. Protein spots were identified by MALDI-TOF analysis. The results demonstrated appearance of 53 new protein spots and disappearance of 19 spots in animals subjected to NP irrespective of their dose. In addition 25 new spots appeared and 3 disappeared in higher doses of NP when compared to low dose group and control animals. Mass-spectrum analysis showed presence of few polypeptides registered in international database among proteins expressed under influence of NP. One of this dominant expressed proteins corresponded to enzyme glutathione transpherase Mu 2 isoform (M=41,55 kD, pI=8,0). The conclusion was made of well advances of proteomic analysis in artificial NP influences on biosynthetic processes estimation.

The Trp64Arg polymorphism of β3-Adrenoreceptor gene study in persons with overweight and obesity

The development of obesity is determined by lifestyle and genetic mechanisms. In particular, the polymorphisms in the adrenergic receptor genes (ADRB) have been extensively studied for association with obesity-related phenotypes. ADRB3 is an obvious candidate gene given its involvement in the regulation of lipolysis and thermogenesis. ADRB3 Trp64Arg polymorphism, a missense mutation in the first transmembrane domain of the R3-adrenergic receptor is associated with visceral obesity and insulin resistance in the Pima Indian, French, and Finnish populations. The recent meta-analysis that combined data of 6582 individuals from Japanese populations showed significant association the Arg64 allele with increased BMI. There are tested the polymorphisms in the β3-Adrenoreceptor (ADRB3) gene in associated with body mass index (BMI), fat mass and biochemical parameters.We have been examined 91 persons from Moscow region with BMI >25 kg/m2. The Trp64Arg polymorphism оf ADRB3 genes were genotyped with the use of an allelic discrimination assay. The TaqMan-based real-time PCR method was applied. There have been estimated of anthropometric and biochemical parameters. The frequencies of the Trp64Trp and Trp64Arg genotypes of ADRB3 gene were 82% and 12%, respectively, the frequencies of mutant allele was 6%. Trp64Arg genotypes of ADRB3 compared to Trp64Trp genotypes had significantly higher body fat percentage (respectively 48,6±0,96% and 43,8±1,72%, р<0,05), serum glucose (6,51±0,18 mmol/l and 5,67±0,09 mmol/l, р<0,01) and uric acid concentrations (0,46±0,02 mmol/l and 0,38±0,01 mmol/l, р<0,05).The test of the ADRB3 gene polymorphisms can be used for the personalization of diet in persons with obesity.

Effects of polyvitamin deficiency on the activity of xenobiotic-metabolizing enzymes in rat liver

The activity of xenobiotic-metabolizing enzymes was studied in the liver of male Wistar rats, which were fed for 4 weeks diets, containing 100 (control), 50 and 20% of vitamin adequate level. Moderate (50%) polyvitamin deficiency increased activity of EROD (by 13%), MROD (by 34%; p<0,05), 4-nitrophenol hydroxylase (by 16%), 6β-testosterone hydroxylase (by 17%), UDP-glucuronosyle transferase (by 26%, p<0,05) and quinone reductase (by 55%, p<0,05). Deep (20%) polyvitamin deficiency decreased in liver activity of MROD (to 78% of control level, p<0,05), 4-nitrophenol hydroxylase (to 74%, p<0,05), heme oxygenase-1 (to 83%, p<0,05) and quinone reductase (to 60%, p<0,05). At the same time a 22% increase in the UDP-glucuronosyle transferase activity compared to the control group was found; activities of EROD, PROD, 6β-testosterone hydroxylase and the total activity of glutathione S-transferase were unchanged. Deep polyvitamin deficiency had no significant effect on CYP1A1 mRNA and AhR mRNA level, whereas the expression of CYP1A2 mRNA and CYP3A1 mRNA were decreased to 62 and 79%, respectively, as compared with control.
Hygiene of nutrition

Characterization of silver nanoparticles migration from packagematerials destined for contact with foods

There was studied migration of artificial silver nanoparticles from polyethylene films destined for package of such foods as bread and poultry into model media reproducing physico-chemical properties and composition of said products. Modification of films was performed by 5- or 10-fold spraying of silver nanoparticles on the surface of package material. Model media were composed from water, alcohol and plant oil according to US FDA and Russian Federal Service for Surveillance of Consumer Rights Protection and Human Well-Being official recommendation. Nanoparticles were detected in model media by means of transmission electron and atomic force microscopy. Quantification of silver in nanoparticles migrating from films was performed by mass-spectrometry with inductively coupled plasma. The results obtained showed that silver migrated from films into test media in form of nanoparticles with mean diameter close to 10–20 nm. Migrated particles were partially aggregated to complexes with dimension about 50 nm with degree of aggregation depending on media composition. Quantification showed that amounts of silver nanoparticles migrating in foods did not exceed save level of this nanomaterial consumption even in aggravated conditions when almost all volume of product was consumed in form packaged in films modified with nanosilver.

Determination of the content dietary fiber in the various groups of foods of plant origin

The article presents data on the content of soluble, insoluble and total dietary fiber in foods Russian and CIS countries of origin (528 samples (47 types) of food). Conducted a comparative analysis of these data with data on the content ofdietary fiber in analogous products on the European market. Statistical analysis of the results was conducted on a PC programdesigned to package «Microsoft Excel office 2003», using the library of statistical functions. Comparative analysis revealed significant differences in content of various fractions of dietary fiber in plant food of domestic production compared to similar products of the production of the European Union, particularly Germany, more than 1/3 of the time. These differences can’t be explained by analytical errors, because, for the purposes of these studies used the method recommended by the Codex Alimentarius Commission to conduct similar studies. Consequently, the results are directly related to differences in the chemical composition of domestic food products, compared with foreign counterparts, which determines the further development and refinement of existing tables of chemical composition. Thus, our data on the content of soluble, insoluble and total dietary fiber can enhance the chemical composition of Russian food. In addition, studies show once again the need for the Russian Federation of their own, national tables of chemical composition of foods, consistent with modern scientific and technological level of development.

Hygienic assessment of food and beverages production in Krasnoyrsk Region

This article presents data on the prevalence of different types of sanitary violations, caused by failure to comply with sanitary regulations, on 573 enterprises that produce bakery products, confectionery, dairy, meat and fish products, drinks in the Krasnoyarsk Territory. It is shown that the greatest number of sanitary offenses are committed at manufactures of milk, fish and meat products. Finished goods at 21–37% of these enterprises do not meet the requirements of hygiene standards. The proportion of deposits of factors (such as type of food product, a form of ownership of the enterprise, the type of locality, where manufacture is) in the frequency of sanitary violations has been determined.

The experimental model of alimentary polyhypovitaminosis of different degree in rats

A model of the alimentary polyhypovitaminosis varying degrees basing on partitive simultaneous reduction of all vitamins in rats diet has been proposed. The model has a principal difference from other experimental models, based on complete exclusion of 1 or 2 vitamins from animal diet. The proposed model allows you to get as close to the actually observed combined deficiency of several vitamins among the population. 5-fold decrease of vitamin mixture resulted in the fact that animals received 20–23% of vitamins D, A, B2, 33% of vitamin B1 and 57% of vitamin E from the content of these vitamins in the diet of animals from control group because of some nature vitamins contained in such diet basic components as casein (vitamins D, A, B1, B2) and sunflower oil (vitamin E). After one month treatment a deep deficiency of all vitamins has developed in rats from this group. Liver level of vitamin A decreased 8,5-fold, vitamins E and B1 – approximately 2-fold, vitamins C and B2 by 22%. Urinary excretion of vitamin B1 and B2 declined 2 and 5,3 fold. Blood plasma concentration of vitamin A decreased 1,9 fold, and E – 1,4 fold, B2 – by 30%. Activities of blood plasma vitamin B6-dependent enzymes reduced 1,4-fold. 2-fold decrease in the amount of vitamin mixture ensured intake about 50–59% of vitamins D, A, B2, and B1 and about 71% of vitamin E of those contained in the diet of animals from control group. Vitamin status indexes of these animals occupied an intermediate position. They have developed a moderate deficit of these essential micronutrients. The proposed model may be useful for metabolic disorders identification, the study of vitamins and minerals assimilation, investigations of the influence of biologically active components of food on these processes, as well as the development of personalized approaches to the correction of vitamin insufficient accuracy.
Diet treatment

Clinical analysis effectiveness utilization nutritional factors of patients after operation on gastrointestinal tract

The analysis of use of a basic clinical nutrition with inclusion to its structure of nutritious mixes after operation on a gastroenteric path at patients is carried out. He allows to predict possible consequences stressful impacts of surgical intervention and an alimentary deprivation, and also to develop ways of their elimination and regulation in an organism. The prognostic interrelation between level of survival rate and indicators of clinical, biochemical and anhropometric researches is established. Clinical manifestations of the postresection diseases, being characterized permanent diverse functional changes, are connected not only with operational intervention and length of rehabilitation period, but also with nutritional disorder of the patient that leads to albuminous, lipid and mineral disorder, vitamin insufficiency and lengthening of rehabilitation period. The correlation analysis showed that decrease in level of a visceral pool of serum proteins in patients after operations on a gastrointestinal tract associated with the high risk of complications, increases of lethality and duration of rehabilitation period. Inclusion of nutritious mixes in a basic clinical nutrition allows to do leveling the deficiency of energy and plastic substances which is observed in the operated.
Nutrition of sportsmеn

Medical-biological approach to development of feeding ration for sportswomen – members of Russia weight lifting team

Physical status of 12 sportswomen – member of Russia weight lifting team during training period was analiysed. Anthropometric, bioimpedance methods with analyzer ABC-01 «Medass» in observation [body weight Index (index Ketle-2), waist circle/hip circle index] and body compositions were used (adipose, bone and muscle tissues masse). Data obtained when bioimpedance method was used indicated: normal body weight and relative adipose mass to body weight – in 92%, surplus – in 8% of cases; body weight Index and normal level of adipose mass – in 75%, surplus – in 25%; active cellular mass and bone mass and general body liquid were found to be normal. 14-day feeding ration for sportswomen were analysed and 3 times per day food intakes was found to be not optimal. Data obtained let to recommend set of daily food products to compose optimal ration for present sportswomen group when energetic value of 4500 kKal and high nutritional value would be provided.

Immune dysfunction in highly skilled athletes and nutritional rehabilitation

The present review examines the effects of strenuous and moderate exercise on various components of the immune system including humoral and cell immunity. Acute exercise have been found to affect lymphocyte subsets, immunoglobulin levels, the activity of mononuclear phagocytic system, polymorphonuclear leukocytes and cytokine levels, especially IL-1, IL-2, IL-6 and TNF. The extreme type of immune deficiency in sportsmen, in which there are not determine different isotypes of immunoglobulins, there are described as phenomenon of Ig disappearance. The immune suppression is associated with an increased risk of subclinical and clinical infection in athletes. The mechanisms immunodeficiency are multifactorial and include neuroendocrinologocal and metabolic disorders. The immunosuppression in sportsmen may be prevented by use of special diets. Although athletes need to eat a well-balanced basic diet, there are several nutritional factors that are difficult to obtain at a sufficient level from a normal diet since athletes require more nutrients than the recommended daily allowances. Thus, appropriate nutrition enrichment of special products and food supplements containing proteins, amino acids, carbohydrates, vitamins, minerals, antioxidants, probiotics is an essential prerequisite for effective improvement of athletic performance, recovery from fatigue after exercise and ward off immunodeficiency.

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