Natural Clinoptilolite Nanoparticles Coated with Phosphatidylcholine.

A nanocarrier was obtained by coating the natural clinoptilolite particle surface with a phosphatidylcholine layer. The shell effective size does not exceed the thickness of the phospholipid molecular layer, which was confirmed by UV spectrometry and molecular mass spectrometry. The hydrodynamic diameter of the formulated nanocarrier, which was determined by dynamic light scattering, is smaller than the clinoptilolite core size. This effect is assumed to be caused by the phospholipid shell, which reduces the aqueous medium friction. The nanosize of the formulated nanocarrier, the natural clinoptilolite core, and the phospholipid shell together allow a combination of fruitful features that can be used for the fabrication of multifunctional platforms for the delivery of biologically active substances, bioimaging, or as a basis for biosensors.

Multiphoton Microscopy and Mass Spectrometry for Revealing Metabolic Heterogeneity of Hepatocytes in vivo.

The aim of the investigation was to study the possibility of revealing the heterogeneity of normal liver hepatocytes in terms of metabolic status using the modern methods of multiphoton microscopy and mass spectrometry. Materials and Methods: Heterogeneity of hepatocytes in terms of total metabolic activity was assessed using multiphoton microscopy based on the autofluorescence intensity of intracellular cofactors NAD(P)H and FAD. Hepatocyte heterogeneity in terms of intensity of intracellular metabolic processes was determined using the fluorescence lifetime imaging (FLIM) method based on the data about fluorescence lifetime contributions of various forms of NAD(P)H. The method of time-of-flight secondary ion mass spectrometry (TоF-SIMS) was used to study the lipid and amino acid composition of hepatocytes. Results: It has been revealed using multiphoton microscopy that hepatocytes are heterogeneous in terms of general metabolic activity. Using FLIM, it was established that the heterogeneity degree was high in terms of intensity of oxidative phosphorylation, glycolysis, and synthetic processes (lipogenesis, nucleic acid synthesis, and the pentose phosphate pathway). The TоF-SIMS method revealed the presence of hepatocyte heterogeneity in terms of amino acid and lipid composition, which points to various intensities of synthetic processes in individual hepatocytes. Moreover, differences in the content of PO3 ions were revealed. The results of ToF-SIMS study correlate with the data obtained by multiphoton microscopy and FLIM, confirming the revealed heterogeneity of hepatocytes in terms of general metabolic activity and intensity of intercellular metabolic processes. Conclusion: The latest methods of fluorescence bioimaging and mass spectrometry proved to be effective in revealing hepatocyte heterogeneity in terms of metabolic status. The presence of heterogeneity should be taken into account in studying the liver tissue under various conditions with the application of fluorescence bioimaging methods.

[POPULATION AND CLUSTER APPROACH TO ASSESSMENT OF REPRODUCTIVE HEALTH OF WOMEN].

The purpose of this study was to explore the possibility of using a population approach to assessing the risk of reproductive health disorders in women of childbearing age. We observed 240 clinically healthy women aged 20 to 43 years, half of them lived in the Middle Black Earth region of Russia, and 120 women lived in Tajikistan. The study identified population differences in women with different ethnic backgrounds and regions of residence according to a set of genetic, hormonal, and immune signs. All women underwent venous blood sampling for the purpose of HLA typing by molecular genetic analysis and determining the range of normative values ​​of hormonal and immune status parameters. DNA samples were obtained from peripheral blood lymphocytes using reagent kits and a protocol for isolating DNA from various biological materials from DLAtomTM DNAPrep 100 (Russia). Hormonal status was studied in terms of follicle-stimulating hormone, luteinizing hormone, prolactin, estradiol, progesterone, 17-OH-progesterone; Thyroid hormones - thyroid-stimulating hormone, total triiodothyronine, total thyroxine; Androgens - testosterone, dihydroepiandrosterone; Steroid hormone-cortisol. In order to exclude the variability of the data, the examination was carried out on the 3-5th days of the menstrual cycle: luteinizing hormone, follicle-stimulating hormone, estradiol, prolactin, testosterone and on the 20-22nd Day of the progesterone cycle. Statistical data processing was carried out on the basis of SPSS programs and included descriptive and comparative nonparametric statistics, discriminant, regression analysis, one - factorial analysis of variance, calculation of 95% confidence intervals, construction of ROC curves. The study included the determination of allelic variants at the three loci of the HLA-D genes (HLA-DRB1, HLA-DQA1, HLA-DQB1) controlling the immune response. Population differences in the locus of the HLA-DRB1 gene were determined. The HLA-DRB1 * 08 and HLA-DRB1 * 12 alleles are found in the Russian women's population, and the HLA-DRB1 * 04 and HLA-DRB1 * 17 alleles are more reliably detected in the Tajik women's population, while the HLA-DRB1 * 04 allele Is associated with a higher incidence of miscarriage. The population characteristics of the HLA-DQA1 gene locus were also established. In the group of women of the Russian population, the incidence of HLA-DQA1 * 0101 and HLA-DQA1 * 0103 alleles is significantly higher, of which the former is associated with protective properties for reproductive pathology, and the latter, on the contrary, with miscarriage. At the same time, the alleles HLA-DQA1 * 0201 and HLA-DQA1 * 0301 were significantly more often detected in the compared with the population of Tajik women. As in the previous case, for the HLA-DQA1 * 0201 allele reproductive health disorders are not characteristic, and in the case of the HLA-DQA1 * 0301 allele they accompany it. Our studies have revealed that there are differences between the populations of Russian and Tajik women in a number of parameters of the hormonal and immune status, as well as at the level of allelic variants of genes that control the immune response. The population approach, based on the use of discriminant analysis, is a highly effective way of grouping women according to their reproductive health status and the risk factors that caused reproductive damage. The risk factors that caused the reproductive failure are different in combination and manifestation in the populations of Russian and Tajik women, except for the adverse allelic variants of the HLA-DRB1 * 04 and HLA-DQA1 * 0103 genes, which are the same in both populations. The obtained data also show that in different populations in the evaluation of reproductive health a differentiated approach is needed both to establish physiological norms in these categories of parameters and to assess the reproductive health of women.

Femtosecond Nanostructuring of Glass with Optically Trapped Microspheres and Chemical Etching.

Laser processing with optically trapped microspheres is a promising tool for nanopatterning at subdiffraction-limited resolution in a wide range of technological and biomedical applications. In this paper, we investigate subdiffraction-limited structuring of borosilicate glass with femtosecond pulses in the near-field of optically trapped microspheres combined with chemical postprocessing. The glass surface was processed by single laser pulses at 780 nm focused by silica microspheres and then subjected to selective etching in KOH, which produced pits in the laser-affected zones (LAZs). Chemical postprocessing allowed obtaining structures with better resolution and reproducibility. We demonstrate production of reproducible pits with diameters as small as 70 nm (λ/11). Complex two-dimensional structures with 100 nm (λ/8) resolution were written on the glass surface point by point with microspheres manipulated by optical tweezers. Furthermore, the mechanism of laser modification underlying selective etching was investigated with mass spectrum analysis. We propose that the increased etching rate of laser-treated glass results from changes in its chemical composition and oxygen deficiency.

[Evaluation of functional adaptation level in air specialists according to biochemical indexes of saliva secretion].

It was examined a capability of evaluation of functional condition of air staff by indexes of natrium, kalium, cortisol and glucose in saliva. There were realized 5 series of examinations with participations of 71 airplane pilot of the same level in conditions of realizing flies of different difficultness. Saliva sampling was effectuated before and after the flies not later then 10-15 minutes after landing. On pre-flight medical examination and after performance of task of air relay there was registration of systolic, diasystolic blood pressure and cardiac rate. It was posed the correlation of physiological indexes with percentage of examined ingredients in saliva in different flight loads. The results of examinations speak for capability of using of indexes of percentage of natrium, kalium, cortisol and glucose in saliva for evaluation of functional condition of airplane pilots during effectuating the flies and rating of value of flight load with account of individual peculiarities.

[Complex evaluation of parachutists' stress-endurance by biochemical characteristics of saliva].

Variations in saliva biochemical characteristics of parachuting sportsmen were analyzed after flight duty. The investigation revealed three types of saliva biochemical reactions to the stresses of parachuting. Our data point to the possibility to judge about the level of tension in organism of people engaged in difficult and extreme activities by saliva concentrations of Na+, K+, cortisol and glucose along with the physiological and psychological investigations traditionally employed by aviation and sport medicine.