[The study of energy balance in individuals with the Trp64Arg polymorphism of the ß3-adrenoreceptor gene].

The study involved 104 people living in the Moscow region, including 18 men and 86 women aged 18 to 67 years. Genotyping of rs4994 ADRB3 polymorphisms was performed using allele-specific amplification, with result detection in real time and using TaqMan-probes complementary to polymorphic DNA regions. The frequency of the mutant allele in individuals was 8.0%, while the Trp64Trp genotype was detected in 84.0% of cases, Trp64Ar - in 16.0%, AA - in 19.0%. Compared with men with genotype Trp64Trp, the men with the Trp64Ar polymorphisms (rs4994) of ADRB3 gene had significantly lower energy expenditure at rest value, calculated per kg of body muscle mass that was associated with higher fat mass, levels of blood serum leptin and LDL cholesterol. The data obtained suggested that leptin could be a possible intermediary contributing to the association between the rs4994 polymorphism of ADRB3 gene and energy disbalance.

[Vitamin status of citizens from Moscow Region].

Evaluation of vitamin status in healthy individuals (68 men and 70 women) aged from 18 to 60 years (median - 37 years), residents of Moscow and the Moscow region has been performed by means of determination of vitamin C, A, E, B2, B12 and folic acid level in blood serum. The nutrition was investigated by questionnaire method on frequency of food consumption. Both diet of men and women had excessive fat content (41.7 and 42.7% of total calories), saturated fatty acids (14.1 and 13.6%), added sugars (11.1 and 11.0%), sodium, and had lack of dietary fiber (2.5-fold reduced level comparing with RDA). Daily intake of vitamin B1 was 1.37±0.04 mg for men and 1.06±0.07 mg for women, vitamin B2 - respectively 1.72±0.06 and 1.62±0.07 mg, niacin - 18.5±0.72 and 14.8±0.88 mg and did not reach the optimal level. All persons were sufficiently supplied with vitamins A, C, E and B12: mean and median of blood serum level of retinol, tocopherols, ascorbic acid and cobalamins were in the range of optimum values. The lack of vitamins A and B12 has not been found in any person. The frequency of vitamin C and E insufficiency was insignificant and amounted to 2 and 8% respectively. The lack of vitamin B2, and ß-carotene was most pronounced and took place in about a half of individuals. Only 34% of healthy people of working age were sufficiently supplied with all vitamins. A combined lack of two vitamins was detected in 26%, of three vitamins - in 8%. Women were better supplied with riboflavin and ß-carotene. The blood serum level of ß-carotene and vitamin E was significantly higher in individuals older than 30 years compared with persons of younger age. Individuals with overweight or obesity were worse supplied with ß-carotene and folate. A negative correlation was detected between the levels of serum folate and homocysteine concentration (r=-0.262, p<0.05). A positive correlation has been revealed between the concentration of folic acid and the level of HDL-C (r=0.356, p<0.01), and iron (r=0.378, p<0.05).

[The study of the peculiarities of metabolism in individuals with rs9939609 polymorphism of FTO gene].

A total of 104 people, among them were 18 men and 86 women aged 18 to 67 years--people in the Moscow region of the Russian Federation, in the sanatorium treatment. Association embodiment rs9939609 FTO obesity was studied using allele-specific amplification, detection results in real time using TaqMan-probes complementary DNA polymorphisms. The frequency of the mutant allele in individuals was 47.0%, while the TT genotype was detected in 25.0% of cases, AT--in 56.0%, AA.0%. In men, prevalence rates of obesity risk allele (A) is higher than in women. In individuals with obesity (BMI over 30 kg/m2) indicated the presence of more AA genotypes (2 7.8%) compared to individuals with a body mass index of less than 30 kg/m2 (14.1%) and a high frequency of occurrence (54.2 versus 43.0%) risk allele A. Compared with individuals with genotype TT, the examinees with the AA genotype rs9939609 FTO gene was significantly higher BMI, waist circumference, hip circumference, fat mass. The same patients energy expenditure at rest value, calculated per kg of lean body mass or muscle, was slightly less than with TT genotype. In individuals with the AA genotype rs9939609 FTO gene compared with the TT genotype had significantly lower levels of HDL cholesterol and serum iron in the background significantly elevated concentrations of uric acid. Thus, the evaluation of patients with the use of innovative technologies in a spa treatment allows their personal alimentary correction for the prevention and treatment of non-communicable diseases.

[The role of the Consultative and Diagnostic Centre "Healthy Nutrition" in the diagnosis and nutritional prevention of non-communicable diseases].

In a consultative and diagnostic center "Healthy Nutrition" of Institute of Nutrition the nutritional status of 3500 patients (mean age 48.4 ± 0.3 years) liv- ing in the Moscow region, using a system Nutritest IP-3, including genomic analysis has been examined. In the analysis of dietary intake by an average review, increased energy intake due to excess intake of the total (44.2% energy) and saturated fat (13.6%) has been shown. 30.0% of patients were overweight and 34.1% were obese. Osteopenia was detected in 31.0% of men and 25.0% women, osteoporosis--20.9% and 30.3%, respectively. Analysis of the results of biochemical studies revealed increased cholesterol in 68.7% of patients, LDL cholesterol--at 63.9%, triglycerides-- at 22.5%, glucose--at 29.4%. The frequency of the occurrence of risk alleles of genes associated with the development of obesity and type 2 diabetes mellitus was: 47.8%--for the polymorphism rs9939609 (FTO gene), 8.3%--for polymorphism rs4994 (gene ADRB3), 60.2%--for the polymorphism rs659366 (gene UCP2), 36.6%--for the rs5219 polymorphism in the gene of ATP-dependent potassium channel.

[Complex approach to evaluation of natural quality of coloured juices]. / Kompleksnyi podkhod k otsenke natural'nosti okrashennykh sokov.

A novel multidimensional approach to estimation of fruit and berry colored juice authenticity is proposed. It includes determination of quality (determination of 5-hydroxymethylfurfurol) and authenticity (determination of organic acids, hydrocarbons, anthocyanin pigments and artificial colorants) parameters. Both experimental and literature data on chemical composition of 12 colored fruits and berries are summarized. New composition characteristics of pomegranate, bilberry, red currant, blackberry and lingonberry are presented.

Diagnosis of external root resorption using TACT (tuned-aperture computed tomography).

This study compared the ability of the TACT system of imaging and conventional D-speed film to detect simulated resorptive defects in cadaver teeth and jaws. Mandibular human jaw blocks were prepared and then split through the teeth in a mesial to distal direction so that the mandibular halves could be opened and reassembled when necessary. One half of each of 42 teeth was extracted, and areas on the tooth and corresponding bone identified to be studied. The jaws were opened and reassembled after no defects or after defects of different sizes (0.25, 0.5, 1.0, 2.0 mm) were placed in the tooth and the corresponding bone at predetermined evaluation areas. The teeth were imaged after each stage with conventional D-speed film using a standard paralleling technique, and with a modified orthopantomograph OP100 machine using a Schick #2 size CCD sensor as the image receptor. The source images were registered and TACT slices were generated using TACT Workbench Software. Three observers were asked to identify the presence of resorptive defects in the conventional film group and the TACT image group using specific criteria. TACT imaging was statistically superior to conventional radiographs in detecting defects that were present (Wilcoxon matched pairs signed ranks test P = 0.003). This was true for all sizes of defects examined. There was no difference between the two modalities when a defect was not present.

Identification of root canals in molars by tuned-aperture computed tomography.

AIM: To compare the tuned-aperture computed tomography system of imaging to conventional D-speed film for their ability to identify root canals in extracted human molars. METHODOLOGY: Thirteen maxillary and six mandibular human molars were mounted in acrylic blocks to simulate clinical conditions by surrounding the teeth with a radiodense structure. The teeth were then imaged with conventional D-speed film using a standard paralleling technique, and with a modified orthopantomograph OP100 machine using a Schick no. 2 size CCD sensor as the image receptor. The source images were registered and TACT slices were generated using TACT Workbench Software. Three observers were asked to identify the number of canals in the conventional film group and the TACT image group using specific criteria. Ground truth was established by cross-sectioning the teeth at the coronal, middle, and apical thirds of the roots and directly visualizing the root canal morphology. RESULTS: TACT imaging detected 36% of 4th canals in maxillary molars and 80% of third canals in mandibular molars. Conventional film detected 0% of fourth canals in maxillary molars and 0% of third canals in mandibular molars. The differences in canal detection between the two techniques were statistically significant (Wilcoxon matched pair sign rank test, P = 0.001). CONCLUSIONS: In this study, the TACT system of digital imaging was superior to conventional film in the detection of root canals in human molars and may be useful for the detection of root canals that will probably be missed upon conventional X-ray examination.

Expression of IL-8 by cells of the odontoblast layer in vitro.

Due to their peripheral location in the dental pulp and their cellular extension into dentin, odontoblasts are the first pulpal cells to encounter dental pathogens. The association of odontoblasts with immunoglobulins and dendritic cells during microbial invasion of dentin implies that these cells may possess a role in the innate and adaptive pulpal immune responses, however this has not been examined. A pivotal step in the innate immune response is the detection of foreign antigen and the recruitment of immune effector cells to the area. IL-8 is a potent chemotactic cytokine that plays an important role in the inflammatory response. The purpose of this study was to determine if odontoblasts are capable of expressing the pro-inflammatory chemokine IL-8. Human odontoblasts from intact, noncarious third molars were maintained in culture and exposed to Escherichia coli lipopolysaccharide (LPS) (serotype 055:B5) on day 4 for 8-10 h in a humidified 5% CO2 incubator. Control and experimental samples were assayed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot for the production of IL-8 mRNA and protein. Analysis of the PCR products revealed that cells of the odontoblast layer maintained in this culture model constitutively expressed low levels of IL-8, which were increased in response to E. coli LPS exposure. Western blotting confirmed that the mRNA was translated into protein. These results imply that odontoblasts are capable of producing of pro-inflammatory mediators, thereby actively participating in the recruitment of neutrophils in response to bacterial by-products.

TNF-alpha release in monocytes after exposure to calcium hydroxide treated Escherichia coli LPS.

Lipopolysaccharide (LPS), a cell wall component of Gram negative anaerobic bacteria, has been implicated in the pathogenesis of periapical disease resulting from infected root canals. Calcium hydroxide [Ca(OH)2] has been shown to be an effective medicament in such infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)2 may also be the result of direct inactivation of LPS. The aim of this study was to investigate whether the toxic potential of an Escherichia coli LPS could be reduced or eliminated by Ca(OH)2. Four concentrations of E. coli LPS ranging from 1-1000 ng/ml sterile water were incubated in duplicate either with 25 mg Ca(OH)2 or sterile water alone. Controls consisted of Ca(OH)2 without LPS or sterile water only. Monocytes were collected from peripheral blood by centrifuging through a gradient and plated to a specific density. Adherent monocytes were incubated for 4 days at 37 degrees C with 5% CO2 in M199 medium with 10% autologous serum. The different LPS solutions were added to the wells on day 5. After 4 h the supernatants were collected and quantitatively assayed for TNF-alpha using a commercial ELISA kit. Statistical analysis was performed with ANOVA. Results indicated that Ca(OH)2 is able to eliminate the ability of an E. coli LPS to stimulate TNF-alpha production in peripheral blood monocytes (P < 0.0001).

[Cranberries: chemical composition, nutritional and medicinal properties]. / Iagody brusniki: khimicheskii sostav, pishchevye i tselebnye svoistva.

The contents of protein, fats, carbohydrates (glucose, fructose, sucrose, dietary cellulose), some vitamin (ascorbic acid, riboflavin, beta-carotene, vitamin E), polyphenolic compounds, organic acids, pectin, glycosides, macro- and microelements in red bilberry are presented. Antimicrobial effect and other effect of red bilberry and its use in traditional medicine and dietetics are also outlined.

[Content of toxic substances in breast milk after timely and premature labor]. / Soderzhanie toksichnykh veshchestv v zhenskom moloke posle srochnykh i prezhdevremennykh rodov.

Data on the contents of protein, fats and toxic compounds (PCB, DDT, gamma-Lindane) in breast milk Russian and Vietnamese women after timely and premature childbed are resulted. The contents of protein in transitive milk was higher than in mature milk, both timely childbed and premature one. The contents of protein in nature milk of the Vietnamese women was lower, than in mature milk of the Russian women (12 g/l against 16.5 g/l accordingly). The contents of fat in breast milk of the Russian women in lactation increased. Vietnamese women had very low fat level in mature milk (29 g/l against 42 g/l at the Russian women). The contents of toxic compounds (PCB, DDT, gamma-Lindane) in breast milk both, Vietnamese women and Russian women was rather high and significant frequency of detection of these compounds in milk was marked. The correlation contents of basic food components in breast milk and level of toxic compounds was absent.

Induction of human factor VIII inhibitors in rats 2: Fine mapping of rat anti-human rFVIII antibodies.

Inhibiting antibodies in patients with hemophilia A pose a significant therapeutic dilemma in the treatment of bleeding episodes. The genetic factors which predispose hemophiliacs to inhibitors and the optimal method for inhibitor suppression remain obscure. Hence, an animal model of the human FVIII inhibitor response is of potential value. Sprague-Dawley rats immunized with human recombinant FVIII (rFVIII) subsequently developed abnormal coagulation parameters coincident with the development of an immune response to the human protein. The epitopes for the resultant rat anti-rFVIII antibodies were mapped using a random fragment expression library constructed from the FVIII cDNA. Antigenic regions located within the A1, First and Second Acidic and B domains were mapped. Rat immunoglobulins reactive with the individual epitopes were immunoaffinity purified and assayed for inhibitory activity. Several of the epitopes mapped using the rat antibodies were similar to regions previously shown to be antigenic for human inhibitors. By contrast, no epitopes were mapped to the A2 domain with the techniques used. This may be due to the possible presence of conformational epitopes in this area which cannot undergo fragmentation and still retain antigenicity or the presence of relatively low concentrations of antibodies to this region. The rat model shares some similarity with both the auto- and alloimmune human response to FVIII and therefore may be a valuable model for studies on the induction and suppression of the inhibitor response.

Induction of human factor VIII inhibitors in rats by immunization with human recombinant factor VIII: a small animal model for humans with high responder inhibitor phenotype.

Hemophilia A is a clotting disorder that is due to reduced or absent coagulation factor VIII (FVIII) activity. In approximately 25% of people with severe hemophilia A, standard treatment with intravenous plasma-derived or recombinant FVIII (rFVIII) induces anti-FVIII antibodies that inhibit FVIII activity (inhibitors). We describe the development of a rat model to study the formation of inhibitors. Immunization of rats with human rFVIII in adjuvant induced an anti-human rFVIII antibody response characteristic of an anti-FVIII inhibitor response in hemophilia A patients. The rats exhibited a rapid, polyclonal secondary antibody response to human rFVIII. These antibodies were reactive against epitopes located in the heavy and light chains. All the rFVIII-immunized rats developed antibodies against the FVIII C2 domain, a region of major reactivity in hemophilia A patients with inhibitors. Furthermore, competition ELISAs demonstrated that rat and human anti-FVIII antibodies recognized identical or overlapping epitopes of the FVIII molecule. The rat anti-FVIII antibodies also functioned as human FVIII inhibitors with titers ranging from 120 to 2048 Bethesda Units (B.U.). We propose that this rat model may be useful to investigate immune responses to FVIII and may lead to better therapies for FVIII inhibitors.

Osteoblasts increase their rate of division and align in response to cyclic, mechanical tension in vitro.

Bone adapts to physical deformation in vivo, yet the mechanism of the adaptive process remains unknown. One reason for this perplexity has been the difficulty in examining the effects of a well-defined deformation regimen on individual bone cells. With the utilization of novel, flexible-bottomed cell culture plates, one can study the effects of cyclic strain on the morphologic and biochemical adaptations of individual osteoblasts in vitro. Avian, calvarial osteoblast-like cells, from passes 2-5, responded to cyclic strain, by increasing their rates of DNA synthesis and cell division during the first 72 h after initiation of a continuous deformation regimen comprised of 3 cycles per min of 0-24% elongation. In addition, within hours after initiation of the deformation regimen, cells oriented 90 degrees to the applied strain field at the periphery of the culture plate in the region of maximum strain and elongation.

Mechanical stress stimulates aortic endothelial cells to proliferate.

The effects of applied cyclic tensional deformation and relaxation on cultured bovine aortic endothelial cells were examined. Endothelial cells from passages 3 to 9 were seeded in flexible-bottomed plates and allowed to attach for 24 hours. Endothelial cells in the experimental group (n = 6 wells per time point) were placed in a vacuum-operated stress-providing instrument that exerted an average elongation of 10% at maximum downward deflection of the culture plate bottom. The stretched endothelial cells were subjected to repeating cycles of 10 seconds elongation and 10 seconds relaxation from days 1 through 7 in culture. Endothelial cells in the control group (n = 6 wells per time point) were subjected to similar incubation conditions as the experimental group but without tensional deformation. Tritiated thymidine was added to cells 24 hours before harvesting. On days 0, 1, 3, 5, and 7 cells were counted and analyzed for trichloroacetic acid-precipitable tritiated thymidine incorporation. The results showed that 3 cycles/min mechanical stretching stimulated deoxyribonucleic acid synthesis and endothelial cell division. We conclude that cyclic tensional deformation may stimulate endothelial cell proliferation. It is possible that naturally occurring cyclic mechanical deformation in vivo, such as the repetitive stretching and relaxation of aortic tissue by the heart, may invoke a particular pattern of synthesis and division in endothelial cells.

[Effect of alimentary alpha-tocopherol deficiency on the fatty acid composition of phospholipids in the subcellular membranes of the liver cells and high density lipoproteins in the blood plasma]. / Vliianie alimentarnoi nedostatochnosti alpha-tokoferla na zhirno-kislotnyi sostav fosfolipidov subkletochnykh membran pechenochnykh kletok i lipoproteidov vysokoi plotnosti plazmy krovi.

In experiments on rats a positive correlation was demonstrated between the changes in the fatty-acid composition of hepatocytic intracellular membrane phospholipids and blood plasma high density lipoproteins. The data obtained evidence the existence in the blood of several mechanisms of alpha-tocopherol deficiency action on hepatocytic phospholipids and blood plasma protein-lipid complexes.

[Effect of dietary fats and carbohydrates on protein restoration in the myocardium]. / Vliianie zhirov i uglevodov pishchi na obnovlenie belkov miokarda.

A study was made of the effect of high-fatty and high-carbohydrate diets on the rate of the renewal of individual rat myocardial proteins, particularly on the half-life and the constant of degradation of proteins of sarcoplasm, myosin and actin. Unlike the control diet, the high-carbohydrate diet increased the rate of myosin renewal by 25%, while the fatty diet retarded 20% the rate of the renewal of the main contractile myocardial protein. Similar changes were also found for proteins of sarcoplasma: with the high-carbohydrate diet the rate of the renewal increased 10%, whereas with the high-fatty one it diminished 47%. The rate of actin renewal dropped 11% with the high-carbohydrate diet and 30% with the high-fatty diet. Unbalance in carbohydrate and fatty supply of the body produces appreciable changes in the rate of the renewal of individual proteins of the rat myocardium.