Chronic Ethanol Consumption by Rats before Mating Affects Working Memory in the Offspring.

Parameters of non-spatial and spatial memory were evaluated in sexually mature offspring of outbred rats (females and males F0) consuming a 10% ethanol solution for 30 weeks before mating. We found a significant increase in the recognition index in F1 males and its decrease in F1 females in the novel object recognition test. During the first days of the experiment in T-maze, a decrease in spatial memory was revealed in F1 males, which remained at the trend level until the end of testing; no significant deviations were detected in F1 females. Memory impairment in F1 females was accompanied by a decrease in BDNF level in the hippocampus, but not in the prefrontal cortex. Thus, ethanol consumption by F0 rats before mating led to impairment of long-term working memory only in female F1 offspring.

Mechanisms of Stimulation of the Growth of Mouse Mammary Adenocarcinoma by Mesenchymal Stem Cells.

Mesenchymal stem cells from the adipose tissue (AT MSC) and the bone marrow (BM MSC) stimulated migration of melanoma B16 cells, while mammary adenocarcinoma Ca755 cells stimulated migration of mesenchymal stem cells. Mesenchymal stem cells retained these properties at late terms after γ-irradiation in vitro. Tumors that developed after injection of Ca755 cells alone and in combinations with BM MSC or AT MSC had similar histological structure corresponding to breast adenocarcinoma. Only AT MSC stimulated tumor growth, which was determined by more intensive secretion of factors stimulating proliferation of tumor cells, including chemokine CCL2. The use of AT MSC in regenerative medicine requires careful monitoring of the absence of tumors in patients.

Mesenchymal Stem Cells from Mouse Adipose Tissue Stimulate Tumor Growth.

We studied the effect of mesenchymal stem cells from the bone marrow and adipose tissue on the growth rate of melanoma B16 and mammary adenocarcinoma Ca755 tumors after their co-administration with tumor cells to syngeneic mice. Stimulation of tumor growth and formation of melanoma metastases in the lungs was found under the influence of adipose tissue-derived, but not bone marrow-derived stem cells. At delayed terms after irradiation in sublethal doses, the adipose tissue-derived mesenchymal stem cells also stimulated the tumor growth. Stimulation of the tumor growth by adipose tissue-derived mesenchymal stem cells was caused by factors secreted by these cells. Transplantation of mesenchymal stem cells to humans is possible only after accurate exclusion of malignant tumors.

Proliferative and Differentiation Potential of Multipotent Mesenchymal Stem Cells Cultured on Biocompatible Polymer Scaffolds with Various Physicochemical Characteristics.

Biocompatibility of film and fibrous scaffolds from polylactide-based polymers and the relationship between their architecture and the functional characteristics of mesenchymal stem cells were studied. Cell culturing on polylactide-based film and fibrous matrixes did not deteriorate cell morphology and their proliferation and differentiation capacities. The rate of cell proliferation and penetration in microporous 3D matrices with the same porosity parameters and pore size depended on their spatial organization. The above materials can be used as scaffolds for mesenchymal stem cells for creation of tissue engineering implants. The scaffold size and structure should be determined by the defects in the organs in which the regeneration processes have to be stimulated.

CHARACTERISTICS OF TUMORS THAT DEVELOPED IN MICE AFTER TREATMENT WITH IRRADIATED SYNGENEIC MESENCHYMAL STEM CELLS OF BONE MARROW.

Mesenchymal stem cells (MSCs) are present in almost all organs and tissues of the organism. It is believed, that MSCs could be transformed into cancer stem cells spontaneously or under influence of genotoxic factors and trigger the growth of tumors. The aim of this work was to study the possibility of malignant transformation of cultured MSCs from murine bone marrow (MSCs-BM) after g-irradiation in vitro and characterize of biochemical and histological features of the tumors that developed after transplantation of MSCs-BM into syngeneic mice. Tumors were observed in 3­4 months after MSCs-BM transplantation. After administration of MSCs-BM irradiated at a dose of 1 Gy, tumors were seen in 2 of 5 mice. After transplantation of MSCs-BM irradiated at a dose of 6 Gy, tumors were found in all 5 of 5 mice. In the case of control MSCs-BM, only one tumor appeared in 6 months after transplantation. The telomerase activity was two times higher in the tumor developed from 6 Gy irradiated MSCs-BM than from 1 Gy irradiated MSCs-BM. The tumors developed from control and irradiated MSCs-BM were classified as multicomponent mesenchymomas («mixture of sarcomas¼). Histological examination showed that tumors contained tissue areas of different histogenesis. Thus, MSCs-BM g-irradiated at doses of 1 and 6 Gy and, much less frequently, control MSCs-BM can transform into tumor cells and induce development of multicomponent mesenchymomas.

Migration and Proliferative Activity of Mesenchymal Stem Cells in 3D Polylactide Scaffolds Depends on Cell Seeding Technique and Collagen Modification.

We analyzed viability of mesenchymal stem cells seeded by static and dynamic methods to highly porous fibrous 3D poly-L-lactide scaffolds with similar physical and chemical properties, but different spatial organization modified with collagen. Standard collagen coating promoted protein adsorption on the scaffold surface and improved adhesive properties of 100 µ-thick scaffolds. Modification of 600-µ scaffolds with collagen under pressure increased proliferative activity of mesenchymal stem cells seeded under static and dynamic (delivery of 100,000 cells in 10 ml medium in a perfusion system at a rate of 1 ml/min) conditions by 47 and 648%, respectively (measured after 120-h culturing by MTT test). Dynamic conditions provide more uniform distribution of collagen on scaffold fibers and promote cell penetration into 3D poly-L-lactide scaffolds with thickness >600 µ.

[Effects of Low and Sublethal Doses of γ-Radiation on Mesenchymal and Neural Stem Cells from Mouse Brain].

Mesenchymal stem cells (MSC) exist in the brain in addition to the neural stem cells (NSC). The aim of this work was to investigate the sensitivity of mouse brain MSC (MSC(BR)) to sublethal doses of γ-radiation in comparison with the sensitivity of bone marrow MSC (MSC(BM)) and NSC and to study the effects of γ-irradiation at low doses on these cells. Cells were exposed to γ-radiation (137Cs) at the doses of 10 to 200 mGy at a dose rate of 10 mGy/min; higher doses were achieved at the dose rates of 200 and 500 mGy/min (60Co). The survival of cells was assessed by counting living cells after staining with trypan blue in the Goryaev's chamber or using the MTT test for NSC growing as neurospheres. SP fraction was measured using flow cytometry after incubation with rhodamine-123. Exposure to the doses in the range of 10 to 500 mGy stimulated cell proliferation. The maximum decrease in the cells number was seen on the seventh day after irradiation and it was practically the same for the MSC(BR) and MSC(BM). NCS were more radiosensitive than MSC. Exposure to the doses of 100 to 500 mGy stimulated cells proliferation of all SCs except of MSC(BM). It was shown that the size of SP fraction of MSC(BR) was diminished after γ-irradiation at low doses. Thus, the stimulation of cell proliferation after γ-irradiation at low doses is accompanied by the redistribution of distinct cell subpopulations: the decrease in the SP fraction and the increase in the general population of cells were observed.

Suris tetrons: possible spectroscopic evidence for four-particle optical excitations of a two-dimensional electron gas.

The excitations of a two-dimensional electron gas in quantum wells with intermediate carrier density (ne∼1011 cm-2), i.e., between the exciton-trion and the Fermi-sea range, are so far poorly understood. We report on an approach to bridge this gap by a magnetophotoluminescence study of modulation-doped (Cd,Mn)Te quantum well structures. Employing their enhanced spin splitting, we analyzed the characteristic magnetic-field behavior of the individual photoluminescence features. Based on these results and earlier findings by other authors, we present a new approach for understanding the optical transitions at intermediate densities in terms of four-particle excitations, the Suris tetrons, which were up to now only predicted theoretically. All characteristic photoluminescence features are attributed to emission from these quasiparticles when attaining different final states.

[The role of adiponectin in the pathogenesis of the metabolic syndrome and approach to therapy].

In this adiponectin-focused review, the pathophysiological role and the potential therapeutic benefits of adiponectin in metabolic syndrome (MetS) are analysed. MetS is recognized as clusters several metabolic abnormalities and the leading cause of cardiovascular diseases. Insulin resistance (IR) is a key factor in the pathogenesis MetS. Adiponectin is the most abundant and adipose-specific adipokine. Adiponectin acts through the activation of AMP-activated protein kinase and peroxisome proliferator-activated receptor-alpha (PPARalpha) pathways. The wide distribution of adiponectin receptors in various organs and tissues suggests that adiponectin has pleiotropic effects on numerous physiological processes. Its well-known insulin-sensitizing, anti-inflammatory and antiatherosclerotic properties, accumulating evidence suggests that adiponectin may have cardioprotective properties. There is an evidence that adiponectin decreases systematic IR and generally predicts cardiovascular diseases. Recent therapeutic strategies have focused on the indirect upregulation of adiponectin through the administration of various therapeutic agents and/or lifestyle modifications. Weight loss, diet, lifestyle changes and/or medications including orlistat, sibutramine, rimonabant, increase level of adiponectin. Also insulin sensitizers, including thiazolidinediones, and lipid-lowering agents, including statins and fibrates, upregulate adiponectin and may improve IR. The wider use of new treatment approaches appears to signal of a new era in the management of cardiovascular diseases, diabetes mellitus and MetS.

Thermal activation of non-radiative Auger recombination in charged colloidal nanocrystals.

Applications of semiconductor nanocrystals such as biomarkers and light-emitting optoelectronic devices require that their fluorescence quantum yield be close to 100%. However, such quantum yields have not been obtained yet, in part, because non-radiative Auger recombination in charged nanocrystals could not be suppressed completely. Here, we synthesize colloidal core/thick-shell CdSe/CdS nanocrystals with 100% quantum yield and completely quenched Auger processes at low temperatures, although the nanocrystals are negatively photocharged. Single particle and ensemble spectroscopy in the temperature range 30-300 K shows that the non-radiative Auger recombination is thermally activated around 200 K. Experimental results are well described by a model suggesting a temperature-dependent delocalization of one of the trion electrons from the CdSe core and enhanced Auger recombination at the abrupt CdS outer surface. These results point to a route for the design of core/shell structures with 100% quantum yield at room temperature.

Magneto-Stark effect of excitons as the origin of second harmonic generation in ZnO.

The magneto-Stark effect of excitons is demonstrated to be an efficient source of optical nonlinearity in hexagonal ZnO. Strong resonant second harmonic generation signals induced by an external magnetic field are observed in the spectral range of 2s and 2p excitons. The microscopic theoretical analysis shows that for excitons with a finite wave vector, exciton states of opposite parity are mixed by an effective odd parity electric field induced by the magnetic field despite its even parity. The field, spectral, and polarization dependencies of the second harmonic generation intensity validate the proposed mechanism. The observed phenomenon is not limited to a certain symmetry class and therefore must be effective in other semiconductors.

[Clinicopathogenetic characteristics of cardiorenal syndrome in non-alcoholic fatty liver disease].

AIM: To study clinicopathogenetic characteristics of cardiorenal syndrome (CRS) in non-alcoholic fatty disease of the liver (nAFDL). MATERIAL AND METHODS: Insulin resistance markers, plasma leptin and serum adiponectin concentrations. albuminuria, intima-media thickness (IMT) of the common carotid artery (CCA) were assessed in 80 patients with metabolic syndrome (60 males and 20 females, mean age 44+-11 years) and NAFDL. RESULTS: With increasing signs of affection of target organs in NAFDL there was a trend to a significant rise in detection of chronic kidney disease (CKD) and carotid artery atherosclerosis, aggravation of albuminuria and CCA IMT, lowering of glomerular filtration rate. Leptinemia was maximal in CKD patients and directly correlated with albuminuria. Patients with CCA atherosclerosis had a significant fall of serum adiponectin which negatively correlated with CCA IMT. The latter positively correlated with insulinemia, a C-peptide plasma level and HOMA index. Serum concentration of adiponectin positively correlated with DeRitis index. CONCLUSION: CRS in NAFDL is characterized with frequent development of CKD and CCA atherosclerosis defined also by unbalance of adiponectin and leptin production.

[The role of leptin, adiponectin and insulin-resistance markers in development of early stages of chronic kidney disease and atherosclerosis of carotid arteries in obese patients].

AIM: To characterize clinicopathogenetically factors influencing development of early chronic kidney disease (CKD) and impairment of other target organs in obese patients. MATERIAL AND METHODS: The examination of 86 obese patients (64 males and 22 females, mean age 44 +/- 11 years) included standard clinical tests, test for albuminuria, calculation of glomerular filtration rate (GFR) by MDRD formula, ultrasound investigation of the carotid arteries to detect atherosclerotic lesion of the carotid arteries, assessment of insulin resistance - IR (plasma concentration of insulin before meal and blood C-peptide, HOMA-index), test for plasma adipokinins (leptin, adiponectin). RESULTS: Significant direct correlations were found between blood plasma leptin concentration, body mass index (BMI), plasma concentration of insulin and C-peptide, HOMA index, adiponectinemia and albuminuria. CKD patients have significantly higher than patients free of CKD levels of IR markers, waist circumference, BMI, leptinemia (38.2 +/- 28.8 and 21.6 +/- 19.8 ng/ml, respectively; p < 0.01). Obstructive sleep apnea syndrome was associated with higher IR and albuminuria, significantly lower estimated GFR (81 +/- 2 and 95 +/- 2 ml/min/1.73 m2, respectively; p < 0.05). Ultrasound evidence for atherosclerotic lesions of the carotid arteries was associated with a significant increase in blood plasma concentration of C-peptide, reduction of adiponectinemia (14.9 +/- 10.8 and 32.5 +/- 22.5 mcg/ml; p < 0.01), a rise in proportion fasting insulinemia/adiponectinemia (1.6 +/- 1.2 and 0.6 +/- 0.8, respectively; p < 0.05) and reduction of estimated GFR (86 +/- 19 and 102 +/- 25 ml/min/1.73 m2, respectively; p = 0.001). CONCLUSION: In obesity, CKD at early stages develops in parallel with atherosclerotic lesion of the carotid arteries, which correlates with progression of leptinemia, IR and attenuation of organ-protecting properties of adiponectin.

[Urokinase-type plasminogen activator and its inhibitor type 1 in the diagnosis of evolving complications in coronary heart disease].

Fibrinolytic system components are important in the regulation of thrombogenesis therefore the aim of the investigation was to compare the level of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor 1 (PAI-1) in the monocytes of patients with stable coronary heart disease (CHD) and acute coronary syndrome (ACS) to reveal an association of the content of these proteins with the severity of disease, by applying two different techniques: immunocytochemistry and flow cytofluorimetry. The counts of uPA- and PAI-1-expressing monocytes were equal in each case and accounted for 81.9-99.9% in all groups. At the same time, the level of PAI-1 was higher than that of uPA and significantly higher in patients with ACS than in those without ACS and in the controls. No significant differences were found in uPA levels between the ACS and stable CHD groups; however, it was significantly higher in the patient groups than in the control one. The detection of the higher expression of PAI-1 in the peripheral blood monocytes of patients with ACS suggests that it can be used as a marker of disease severity.

[Prognostic, diagnostic, and therapeutic prospects of using adiponectine as a biomarker in cardiobascular diseases].

The role of adipose tissue as an important endocrine organ is today beyond doubt. The adipose tissue is known to be a source of many biologically active substances, adipocitokines, one of them being the adiponectine possessing anti-inflammatory, antiatherogenic and cardioprotective properties; it is believed to be one of prospective biomarkers for risk assessment, for diagnostics and, possibly, for therapy of cardiovascular diseases.

Production of biologically active human myelocytokines in plants.

An effective system for expression of human granulocyte and granulocyte macrophage colony-stimulating factors (hG-CSF and hGM-CSF) in Nicotiana benthamiana plants was developed using viral vector based on tobacco mosaic virus infecting cruciferous plants. The genes of target proteins were cloned into the viral vector driven by actin promoter of Arabidopsis thaliana. The expression vectors were delivered into plant cells by agroinjection. Maximal synthesis rate was detected 5 days after injection and was up to 500 and 300 mg per kg of fresh leaves for hG-CSF and hGM-CSF, respectively. The yield of purified hG-CSF and hGM-CSF was 100 and 50 mg/kg of fresh leaves, respectively. Recombinant plant-made hG-CSF and hGM-CSF stimulated proliferation of murine bone marrow and human erythroleucosis TF-1 cells, respectively, at the same rate as the commercial drugs.

Design of a novel interleukin-13 antagonist from analysis of informational structure.

Interleukin-13 (IL-13) is one of the cytokines involved in the development of Th2-type immune response. It plays an important role in the pathogenesis of asthma and other allergic diseases. Two deletion forms of IL-13 were constructed on a basis of informational structure analysis and expressed in E. coli cells. They were found to differ in ability to stimulate proliferation of TF-1 cell line. Deletion variant 146 (DV146) completely lacks such activity, whereas DV148 provides about 50% of the proliferation stimulation. The simultaneous addition of DV146 with full-length IL-13 suppresses proliferation depending on the concentration of the deletion form. Thus, the designed protein acts as an antagonist of IL-13.

Localization of Mullerian inhibiting substance receptors in various human cancer cell lines.

Recombinant human MIS (rhMIS) produced in transfected Chinese hamster ovary cells has been purified by immunoaffinity chromatography. In the absence of reducing agents, 140 kD homodimer and several oligomers with molecular masses from 280 to 1000 kD are present. Homodimer, tetramer, and higher-molecular-weight rhMIS fractions reduced survival of tumor cells. For these experiments, FITC-labeled rhMIS was used for binding and endocytosis studies by flow cytometry. Flow cytometry performed on MIS-sensitive cancer cell lines demonstrated specific binding of rhMIS. The majority of rhMIS receptors have cytosolic localization. Thus, the level of MIS receptors on the cell membrane was proportional to the content of MIS-binding proteins in the whole cell and defines a level of receptor-mediated endocytosis. The immunopurified rhMIS caused significant growth inhibition of ovarian and prostate adenocarcinoma and melanoma human cell lines in inhibition assays.

[Inactivation and sensitization of the tumor cells by the gene Bax transfection].

After the transfection of the gene Bax into the cultured tumor cells of human ovary adenocarcinoma SKOV3 and uterus carcinoma HeLa in vitro the high sensitivity of the cells SKOV3 to the protein Bax produced after the gene Bax transfection was found. The sensitivity of the cells HeLa to the gene Bax transfection was much smaller. The hyperexpression of gene Bax and hypersensitivity to doxorubicin were seen in HeLa cells received as a result of the gene Bax transfection and subsequent selection. All cells of the line SKOV3 with the increased expression of the transfected gene Bax died. In the cell line SKOV3 the mutation in a gene Bax was found which has a genotype G7/G9 against a native type of a gene Bax--G8/G8. It was concluded that the found in the exone 3 of the gene Bax mutation G7/G9 in cells SKOV3 results in an inactivation of proapoptotic activity of the protein Bax.

[Deletion mutants of human granulocyte-macrophage colony-stimulating factor]. / Deletsionnye mutanty granulotsitarno-makrofagal'nogo koloniestimuliruiushchego faktora cheloveka.

To study the structure-function relationship of the human granulocyte-macrophage colony-stimulating factor (GM-CSF), genes were constructed that encode its three deletion mutants: D1, a mutant with the deletion of six amino acid residues (37-42) some of which are a part of a beta-structural region; D2, a mutant with the deletion of the unstructured six-aa sequence of a loop (45-50); and D3, a mutant with the deletion of 14 aa residues (37-50) corresponding to the A-B loop and encoded by the second exon of the gmcsf gene. The expression products of these genes in E. coli were accumulated in a fraction of insoluble proteins. The secondary structures of the mutant proteins were similar to that of the full-size GM-CSF, but the biological activity of the deletion mutants was 130 times lower than that of the GM-CSF: they stimulated the proliferation of the TF-1 cell line at 3 ng/ml concentration. The resulting proteins displayed antagonistic properties toward the full-size GM-CSF, with the inhibition degree of its colony-stimulating activity being 27%. A decrease in the mutant activity in the row D2 > D1 > D3 implies the importance of the conserved hydrophobic residues involved in the formation of the beta-structure for the formation of the GM-CSF functional conformation.