Biochemical composition of tomato fruits of various colors.

Tomato (Lycopersicon esculentum Mill.) is an economically important and widely cultivated vegetable crop that is consumed both fresh and processed. The nutritional value of tomato fruits is related to the content of carotenoids, polyphenols, sugars, organic acids, minerals and vitamins. Currently, there is a growing interest in the qualitative and quantitative increase in the content of health-promoting compounds in tomato fruits. VIR Lycopersicon (Tourn.) Mill. genetic resources collection includes 7678 accessions of one cultivated and nine wild species, which in turn provides ample opportunities for searching for information on the variability of the content of biologically active substances and searching for sources with a high content of them in the gene pool. Our work presents the results of the study of 70 accessions of cultivated and wild tomato on the main biochemical characteristics: the content of dry matter, ascorbic acid, sugars, carotenoids, chlorophylls and anthocyanins. As the basis for the selection of accessions for the study, accessions with various colors of fruits, including new accessions with varying content of anthocyanin, were taken. As a result of this study, the amplitude of variability in the content of dry matter (3.72-8.88 and 9.62-11.33 %), sugars (1.50-5.65 and 2.20-2.70 %), ascorbic acid (12.40-35.56 and 23.62- 28.14 mg/100 g), titratable acidity (0.14-0.46 and 0.33-0.48 %), chlorophylls (0.14-5.11 and 2.95-4.57 mg/100 g), carotenoids (0.97-99.86 and 1.03-10.06 mg/100 g) and anthocyanins (3.00-588.86 and 84.31-152.71 mg/100 g) in the fruits of cultivated and wild tomatoes, respectively, was determined. We have determined correlations between the content of dry matter and monosaccharides (r = 0.40, p ≤ 0.05), total sugars (r = 0.37, p ≤ 0.05) and ascorbic acid (r = 0.32, p ≤ 0.05); the content of ascorbic acid and carotenoids (r = 0.25, p ≤ 0.05). A high dependence of the content of chlorophyll a and b among themselves (r = 0.89, p ≤ 0.05), as well as between the content of chlorophyll b and anthocyanins (r = 0.47, p ≤ 0.05), the content of ß-carotene (r = 0.26, p ≤ 0.05) and the content of monosaccharides (r = -0.29, p ≤ 0.05) has been noted. We have identif ied tomato accessions with a high content of individual chemical substances, as well as with a complex of traits that can be used as sources in breeding for a high content of dry matter, sugars, ascorbic acid, pigments and anthocyanins.

Conjugates of methionine γ-lyase with polysialic acid: Two approaches to antitumor therapy.

The methionine dependence is a well known phenomenon in metabolism of cancer cells. Methionine γ-lyase (EC 4.4.1.11, MGL) catalyzes the γ-elimination reaction of L-methionine and thus could effectively inhibit the growth of malignant cells. Recently we have demonstrated that the mutant form of the enzyme C115H MGL can be used as a component of the pharmacological pair enzyme/S-(allyl/alkyl)-L-cysteine sulfoxides to yield thiosulfinates in situ. Thiosulfinates were shown to be toxic to various cancer cell lines. Therefore the application of the enzyme in enzyme pro-drug therapy may be promising. The conjugates of MGL and C115H MGL with polysialic acid were obtained and their kinetic and pharmacokinetic parameters were determined. The formation of polysialic shell around the enzyme was confirmed by atomic force microscopy. The half-life of conjugated enzymes increased 3-6 times compared to the native enzyme. The cytotoxic effect of conjugated MGL against methionine dependent cancer cell lines was increased two times compared to the values for the native enzymes. The anticancer efficiency of thiosulfinates produced by pharmacological pair C115H MGL/S-(allyl/alkyl)-L-cysteine sulfoxides was demonstrated in vitro. The results indicate that the conjugates of MGL with polysialic acid could be new antitumor drugs.

[Analysis of healthcare for patients with cardiovascular diseases in Russian Federation in comparison with the practice of Europe and USA].

The medical community, researchers and healthcare organizers are constantly challenged by comparing key indexes reflecting the effectiveness of cardiovascular care, primarily for the dynamic assessment and implementation of the world's best practices to reduce cardiovascular mortality. The analysis of health care for patients with cardiovascular diseases (CVD) is a complex, multicomponent process, the structure and key tools of which differ from country to country. Using different data sources, methodological and analytical approaches creates certain limitations and barriers to the assessment. In order to update the ideas about the modern coordinate system and tools for assessing cardiovascular care, the authors presented practices for analyzing major indexes in Russia, European countries, and the United States. The review presents sources of statistical data, principles for assessing risk factors, cardiovascular morbidity and mortality, and specific features of monitoring the availability and quality of cardiovascular care.

[Effect of retinal protective therapy on optical coherence tomography angiography (pilot study)]. / Vliyanie retinoprotektornoi terapii na pokazateli opticheskoi kogerentnoi tomografii s funktsiei angiografii (pilotnoe issledovanie).

PURPOSE: To assess the influence of the frequency of retinal protective therapy courses on the indicators of regional hemodynamics of the eye. MATERIAL AND METHODS: The study included 17 patients (34 eyes) with a diagnosis of primary open-angle glaucoma (POAG), advanced stage. The patients were divided into 2 groups: the first group received a course of retinal protective therapy with Retinalamin every 3 months, the second group received a course of retinal protective therapy with Retinalamin every 6 months. All patients underwent standard ophthalmological examination including standard automatic perimetry according to the 24-2 program, optical coherence tomography angiography (OCT-A) of the macular area and optic disc. RESULTS: The comparison of hemodynamic parameters of all vascular plexuses of the retina at the beginning and at the end of the study, as well as intergroup comparison did not reveal any statistically significant differences (p>0.05). However, when studying the density and fractal dimension of the vascular bed, multidirectional trends were observed. Specifically, in the peripapillary region, there was a decrease in the length from 19.8 (1/mm) to 19.0 (1/mm) (p=0.37) and the density from 36.6% to 35.7% (p=0.63) of the vascular bed of the peripapillary capillary plexus of the retina in patients of the first group. In the superficial vascular plexus, the trend in the density of the vascular bed in both groups slightly changed (in group 1 - 38.1% and 38.3%, p=0.97; group 2 - 37.8% and 38.7%, p=0.46). The fractal dimension of the vascular bed in the first group tended to increase during treatment from 18.8 (1/mm) to 19.1 (1/mm) (p=0.5), while in the second group, on the contrary, it had tendency to decrease from 18.6 (1/mm) to 17.9 (1/mm) (p=0.63). In the deep vascular plexus, the density of the vascular bed trended to decrease in both groups, but in group 2 (42.5% and 42.4%, p=1.0) it was more pronounced than in the first group (42.5% and 42.6%, p=0.82). However, the fractal dimension of the vascular bed increased in group 1 (21.0 (1/mm) and 21.3 (1/mm), p=0.43) and showed a slight tendency to decrease in group 2 (21.5 (1/mm) and 21.0 (1/mm), p=0.86). CONCLUSION: The general trend of changes in hemodynamic parameters demonstrates a potential positive effect, especially in the data related to the deep vascular plexus.

In Vitro Effects of Lithium Carbonate on Cell Cycle, Apoptosis, and Autophagy in Hepatocellular Carcinoma-29 Cells.

We studied the effects of lithium carbonate on the cell cycle, apoptosis, and autophagy in hepatocellular carcinoma-29 cells (HCC-29) in vitro. Flow cytofluorometry analysis revealed accumulation of G2/M-phase HCC-29 cells and increase in the number of apoptotic cells in 48 h after administration of 5 mM lithium carbonate. Induction of autophagy in HCC-29 cells was detected by transmission electron microscopy and immunofluorescence staining. Thus, lithium carbonate produces an antitumor effect by arresting cell cycle in the G2/M-phase and induction of apoptosis and autophagy in HCC-29 cells, which confirms the lithium potential as a promising drug for the treatment of hepatocellular carcinoma.

Broad-spectrum antibacterial and pro-regenerative effects of photoactivated Photodithazine-Pluronic F127-Chitosan polymer system: In vivo study.

Emerging global danger of multidrug resistant microbes makes it essential to explore new approaches to treat infections. We studied antibacterial and pro-regenerative effects of photodynamic therapy (PDT) performed with water solutions of photodithazine and its complexes with Pluronic F127 and chitosan in rat model of full thickness wound (n = 24) infected by an associated Gram-negative and Gram-positive bacteria culture. Laboratory rats were exposed to PDT 24 and 72 h after the injury. Exudate samples were collected before and after PDT for a microbiological study. Autopsy tissues were excised and fixed in formalin on day 4 of the experiment. Fixed tissues were processed and poured into paraffin. Paraffin sections were stained with hematoxylin and eosin and studied by an experienced pathologist. Microbiological analysis revealed that the photoactivation of photodithazine and its complexes suppressed the associated microflora in vivo and inhibited suppurative inflammation in the wounds. The triple Photodithazine-Pluronic F127-Chitosan system possessed the highest antibacterial activity. The morphological study revealed that PDT with photodithazine polymer complexes accelerated wound healing, promoted restoration of microcirculation, facilitated proliferation of fibroblast and vessels and stimulated collagen synthesis. The Photodithazine-Pluronic F127-Chitosan complex may be successfully applied for PDT to prevent and treat suppurative inflammatory diseases of the skin and soft tissues.

Influence of acetic acid on the photocatalytic activity of photosensitiser-amphiphilic polymer complexes in the oxidation reaction of tryptophan.

The effect of acetic acid on the photosensitizing activity of dimegin (DMG), chlorin e6 (Ce6), and their complexes with amphiphilic polymers-Pluronic F-127 and poly-N-vinylpyrrolidone (PVP)-in the model reaction of tryptophan photo-oxidation has been established. It was shown that the photocatalytic activity of photosensitizers (PS), which is characterized by the effective constant of the tryptophan photo-oxidation rate (keff), increases by 1.4 times for DMG, while for Ce6, it decreases by 1.2 times in a weak acidic medium (pH ∼ 4). The influence of acetic acid on the effective constant keff of the tryptophan photo-oxidation rate in the presence of PS-amphiphilic polymer complexes is determined by the nature of the polymer. Thus, the photocatalytic activity of the PS-F127 system decreases at pH ∼ 4.0, and the photocatalytic activity of PS-PVP complexes is insensitive to the presence of acetic acid in the medium. It has been suggested that the observed effects in the PS-F127 system in a weak acidic medium (pH ∼ 4) are associated with the influence of acetic acid on the supramolecular structure of Pluronic F127.

Methylated flavones of the hairy root culture Scutellaria baicalensis.

Perennial plants in northern Dauria (Zabaikalsky region) grow in low temperatures in winter and in a dry hot summer. The prairies of northern Dauria are rich in a variety of medicinal herbs, including S. baicalensis, which has roots that are in demand for traditional Chinese medicine. In addition to two monomethylated flavones (wogonin and oroxylin A), determining the pharmacological significance of the root, there is also a minority of their polymethylated congeners. Little is known about their role in the plant or their connection with the conditions of growth and cultivation of their hairy root culture (HRC). Therefore, the purpose of this study was to determine whether and to what extent the biosynthesis of the latter is retained in the hairy root culture established from wild plants of Dauria. The composition of the main methylated flavones of HRC was established using LC-MS and a previously unknown pentamethylated flavone was found in the roots. This study showed a more significant accumulation of polymethylated flavones in the root of the wild plant than in HRC.

[Study of migration and distribution of bone marrow cells transplanted animals with B16 melanoma ].

Purpose. Reveal features migration and distribution of syngeneic bone marrow cells (BMC) and subpopulations (MSC) after transplantation into the recipient carrier B16 melanoma bodies. Methods. We used mouse male and female C57BL/6 mice. Induction of Tumor Growth: B16 melanoma cells implanted subcutaneously into right hind paw of female C57BL/6 mice at a dose of 2.5 x 105 cells / mouse. migration study in vivo distribution and BMC and MSC was performed using genetic markers - Y-chromosome specific sequence line male C57Bl/6 syngeneic intravenous transplantation in females using the polymerase chain reaction (PCR) in real time on Authorized Termal Cycler - Light Cycler 480 II / 96 (Roche). Introduction suspension of unseparated bone marrow cells, mesenchymal stem cells from donor to recipient male mice (syngeneic recipient female C57BL/6), followed by isolation of recipients of organs was performed at regular intervals, then of organ recipients isolated DNA. Results. It was shown that bone marrow cells positive for Y-chromosome in migrate lymphoid (lymph nodes, spleen, bone marrow) or in non-lymphoid organs (liver, heart, brain, skin) syngeneic recipients. In addition to the migration of cells from the bone marrow to other organs, there is a way back migration of cells from the circulation to the bone marrow. B16 melanoma stimulates the migration of transplanted MSCs and BMC in bone marrow. It is found that tumor growth enhanced migration of transplanted bone marrow cells, including populations of MSCs in the bone marrow. In the early stages of tumor formation MSC migration activity higher than the BMC. In the later stages of tumor formation undivided population of bone marrow cells migrate to the intense swelling compared with a population of MSCs. Conclusion. The possibility of using bone marrow MSCs for targeted therapy of tumor diseases, because migration of MSCs in tumor tissue can be used to effectively deliver anticancer drugs.

Surface micromorphology of cross-linked tetrafunctional polylactide scaffolds inducing vessel growth and bone formation.

In the presented study, we have developed a synthetic strategy allowing a gradual variation of a polylactide arms' length, which later influences the micromorphology of the scaffold surface, formed by a two-photon polymerization technique. It has been demonstrated that the highest number of cells is present on the scaffolds with the roughest surface made of the polylactide with longer arms (PLA760), and osteogenic differentiation of mesenchymal stem cells is most pronounced on such scaffolds. According to the results of biological testing, the PLA760 scaffolds were implanted into a created cranial defect in a mouse for an in vivo assessment of the bone tissue formation. The in vivo experiments have shown that, by week 10, deposition of calcium phosphate particles occurs in the scaffold at the defect site, as well as, the formation of a new bone and ingrowth of blood vessels from the surrounding tissues. These results demonstrate that the cross-linked microstructured tetrafunctional polylactide scaffolds are promising microstructures for bone regeneration in tissue engineering.

APOPTOSIS AND AUTOPHAGY IN HEPATOCARCINOMA CELLS INDUCED BY DIFFERENT FORMS OF LITHIUM SALTS.

Hepatocellular carcinoma (HCC) is one of the aggressive and resistant to drug therapy cancer. It is believed that the development of HCC is correlated with dysregulation of programmed cell death. So, the search for effective inducers of HCC cell death is very important. The aim of the work was to identify structural changes leading to HCC cell death when exposed to nanoscale and original forms of lithium salts. Using light and electron microscopy and flow cytometry, structural features of autophagy and apoptosis in HCC cells after their incubation with various forms of lithium salts has been revealed. It is shown that a more pronounced effect on HCC-29 cells have nanoscale forms of lithium carbonate and lithium citrate. At the same time, nanoscale lithium citrate mainly induced apoptosis, and nanosized form of lithium carbonate, along with apoptosis, induced autophagic death of HCC-29cells.

[Computerized method of objectification of dizziness and vertigo and differential diagnostic of vestibulopathies].

OBJECTIVE: To create a complex computerized method of objectification of dizziness and vertigo, and differentiation of vestibulopathies of various geneses using electrooculography approach that allows to record and analyze spontaneous, vestibular- and visually-induced eye movements, with the following classification (discriminant) analysis of the results obtained. MATERIAL AND METHODS: The study involved 69 patients of different sex and age complained of dizziness, vertigo and disequilibrium, and 64 healthy men. Based on the results of clinical examination, patients were divided into three groups: patients with peripheral vestibulopathy, patients with central vestibulopathy and patients with psychogenic vestibulopathy. Electrooculography was performed using the hardware-software complex (HSC) "OCULOSTIM-CM". RESULTS AND CONCLUSION: Significant diagnostic parameters based on the recognition and analysis of spontaneous, vestibular- and visually-induced eye movements were coefficients of efficacy and increased frequency of fixation saccades and smooth pursuit with- and without retinal optokinetic stimulation. We developed the algorithm and complex computerized method for differentiation of different types of vestibulopathy.

[NUTRITIONAL STATUS IN THE EXPERIMENT WITH 105-DAY ISOLATION AS THE FIRST PHASE OF PROJECT MARS-500].

In a 105-day experiment simulating crew life in a space exploration vehicle, shifts in the nutritional status were assessed in 6 test subjects differing in the body mass index, basal metabolism, attitude to the diet, physical exercise and workload. Results of the investigation showed that because of the hard physical work the nutritional status of test subjects was described by more intensive basal metabolism, elevated metabolism of carbohydrates and lipids, and their increased mobilization from depots. Food ration, though it was sufficient to sustain health and fairly high performance, failed to meet fully individual taste preferences and energy needs for physical activities. The heavy workloads required mobilization of lipids from adipose depots, and a decrease of the hepatic detoxification and metabolic capacities. Self-limitation of eating protein-rich desserts led to a relative deficiency of protein intake. These faults of the diet were the reason why 4 out of 6 test subjects reduced their basal metabolism and lost body mass. Recovery of metabolism and slowdown of the body mass loss were achieved by supplementing meals with available protein-containing products.

Pineal gland as an endocrine gravitational lunasensor: manifestation of moon-phase dependent morphological changes in mice.

We found that some morphological properties of the pineal gland and submandibular salivary gland of mice are significantly distinct at the new and full moon. We suppose that the differences are initiated by the displacements of the electron-dense concretions in the secretory vesicles of pinealocytes. This presumably occurs under the influence of the gravitational field, which periodically changes during different phases of the moon. It seems that the pinealocyte is both an endocrine and gravisensory cell. A periodic secretion of the pineal gland probably stimulates, in a lunaphasic mode, the neuroendocrine system that, in turn, periodically exerts influence on different organs of the body. The observed effect probably serves, within the lifelong clock of a brain, to control development and aging in time.

Effect of mitochondria-targeted antioxidant SkQ1 on programmed cell death induced by viral proteins in tobacco plants.

Programmed cell death (PCD) is the main defense mechanism in plants to fight various pathogens including viruses. The best-studied example of virus-induced PCD in plants is Tobacco mosaic virus (TMV)-elicited hypersensitive response in tobacco plants containing the N resistance gene. It was previously reported that the animal mitochondrial protein Bcl-xL, which lacks a homolog in plants, effectively suppresses plant PCD induced by TMV p50 - the elicitor of hypersensitive response in Nicotiana tabacum carrying the N gene. Our studies show that the mitochondria-targeted antioxidant SkQ1 effectively suppresses p50-induced PCD in tobacco plants. On the other hand, SkQ1 did not affect Poa semilatent virus TGB3-induced endoplasmic reticulum stress, which is followed by PCD, in Nicotiana benthamiana epidermal cells. These data suggest that mitochondria-targeted antioxidant SkQ1 can be used to study molecular mechanisms of PCD suppression in plants.

Porphyrin effect on the surface morphology of amphiphilic polymers as observed by atomic force microscopy.

Complexes of porphyrin photosensitizers (PPS) with triblock copolymers of ethylene- and propylene oxide - Pluronics(®) - exhibit markedly increased activity in the generation of singlet oxygen in aqueous media, as compared to pure porphyrins. Pluronics are amphiphilic polymers with surfactant properties suitable for a number of medical applications. PPS-Pluronic systems are considered as promising agents for photodynamic therapy which implies generation of singlet oxygen in the water-based human tissue. Importantly, Pluronics are capable of solubilization of not only water-soluble, but also hydrophobic PPS providing their transfer into the aqueous phase. It has been shown earlier that specific interactions of PPS with Pluronics must play a primary role for the photocatalytic properties of PPS-Pluronic systems. In the process of solubilization of a hydrophobic porphyrin by a Pluronic, both components are dissolved in an organic solvent, which is then removed, and the dry film is re-dissolved in water. Apparently, the initial binding between the porphyrin and the lipophilic part of the polymer takes place already at the stage of the film formation. We applied atomic force microscopy (AFM) to visualize structures formed by Pluronics upon their interactions with meso-tetraphenylporphyrin (TPP). We studied the surface structure of Pluronics(®) F87, F108 and F127 crystallized alone or together with TPP on silicon substrates from chloroform solutions. We found Pluronics to form similar dendritic structures independently of their molecular weight and degree of hydrophobicity. In the presence of TPP, though, we observed formation of distinct convex structures on top of the Pluronic dendrites. These structures appeared to consist of multiple flat layers placed on top of each other. Their sizes varied among the three Pluronics. We believe that TPP aggregates interact with the hydrophobic units of Pluronics causing the polymer chains to pack themselves in a distinct manner around those TPP-containing "cores". These interactions apparently direct formation of complexes between the porphyrin and the polymer upon their dissolution in water, thus resulting in the encapsulation of TPP aggregates inside a Pluronic micelle. A single mechanism for the TPP solubilization by Pluronics is consistent with the same catalytic activity of the three TPP-Pluronic systems observed in the photooxidation of tryptophan.

The effect of porphyrin supramolecular structure on singlet oxygen photogeneration.

This work is to show that activity of porphyrins in singlet oxygen generation (SOG) is determined not only by their molecular structure but also by supramolecular structure of porphyrin containing systems. This is demonstrated by results of studies of SOG in the gas phase by vacuum deposited tetraphenylporphyrin (TPP) layers under photoexcitation conditions. The structure of the layers was studied by atomic force microscopy (AFM), TEM in diffraction regime (TED) and by the flicker-noise spectroscopy (FNS) method. It was shown that substrates affect the deposited layers structure only at earlier stages of the layer formation. AFM and TED data, together with FNS quantitative parameters of TPP layers, show directly that the chemical activity of solid-phase systems can be determined not only by their chemical nature but also by their supramolecular structure.

Experimental evaluation on the transmission optical microscopy for the diagnosis of lymphedema.

The mechanisms of disturbance of lymph microcirculation during development postmastectomy lymphedema (PML) are little known. The study of these mechanisms on human subjects has many difficulties: time of clinical onset of PML is indefinite and unpredictable and the methods for imaging of microlymphatics are limited. The advantages of the optical methods, especially, light microscopy are the high quality of lymphocytes dynamic imaging, possibility of the determination of absolute lymph flow velocity in a wide range, capability for simultaneous registration of blood microvessel function around lymphatics. The experimental lymphedema is created on rat mesentery by ligation of the collecting vein. It was found that after such ligation edema was clearly developed, and microcirculation was significantly disturbed including reduction of lymph flow velocity, constriction of lymphatics, inhibition of phasic activity on the background of pathological increasing of blood vessels permeability and stasis of blood flow.

Induction of interferon in mice and cell cultures by Mycoplasma pneumoniae.

Inoculation of mice and L and human embryonic lung (HEL) cell cultures with Mycoplasma pneumoniae failed to induce the production of interferon. M. pneumoniae multiplied in these cell cultures without a marked cytopathic effect. M. pneumoniae induced interferon in human peripheral blood leukocytes with maximum titres of 32 units/ml at 48 hours after infection.