Growth-Stimulating Effect of Platelet Preparations Obtained by Different Methods on Human Fibroblast Culture.

We studied the growth-stimulating effect of platelet-based preparations obtained by different methods on cultured human fibroblasts M-22. Platelet lysates prepared from platelet-rich plasma, platelet-poor plasma, concentrated suspension of platelets washed from plasma, and platelet-rich plasma activated with calcium chloride (ActPRP) were used. The volume of the platelet preparations was 10-500 µl per 104 cells. The most effective dose of platelet-rich and platelet-poor plasma in cell culture was 20 µl, whereas for ActPRP, the most effective dose was 500 µl. Lysates of platelet-rich and platelet-poor plasma in doses of 100-500 µl inhibited fibroblast growth and disturbed their structural integrity. At the same time, lysates of washed platelets in doses of 10-500 µl stimulated cell growth and preserved their viability. An inverse correlation was found between the number of cells in culture and the level of proinflammatory cytokines in the platelet preparations.

New Reactive Force Field for Simulations of MoS2 Crystallization.

We present a new reactive force field (ReaxFF) parameter set for simulations of Mo-S structures. We compare our parameterization to the state-of-the-art ones in their performance against density functional theory (DFT) benchmarks and MoS2 crystallization simulations. Our new force field matches DFT data significantly better than any previously published force fields and provides a realistic layered MoS2 structure in crystallization simulations. It significantly improves the state-of-the-art force fields, which tend to crystallize in the experimentally unknown rock-salt MoS structure. Therefore, our new force field is a good candidate for further development and inclusion of other practically relevant elements, such as O, C, N, and H, which can be used to study the formation and tribological or catalytical properties of molybdenum disulfide.

Experimental Study of the Effect of Biological Matrixes with Stabilized and Non-Stabilized Platelets on Reparative Process in the Wound Equivalent to Deep Burn.

We studied reparative effect of platelet-filled biological matrixes in the treatment of mice with wounds equivalent to deep burn. The wound coatings were based on decellularized dermal matrix without platelets (control), with native platelets, and with platelets stabilized with 2.5 µM nanosilver. In 3 days, the epithelial layer and derma were absent in all groups and extensive scab was formed. Dermal matrix with platelets simulated intensive migration of macrophages and fibroblasts to the wound bottom; in the control group, this migration was absent. In 14 days, granulation tissue appeared in the wound bottom in animals of all groups; in the experimental groups, the number of vessels was 2-4-fold higher than in the control, though the number of inflammatory cells in experimental groups remained high. On day 21, the scab on the most of the wound area was absent in all animals of the experimental groups and epithelialization and hair growth were pronounced, comparing to control. Nevertheless, in experiment dermal layer was not already completed, inflammation reaction remained.

Interaction of Magnesium-Based Materials with Human Blood Cells and Culture of Human Diploid Cells In Vitro.

We studied morphofunctional properties of human blood cells and diploid culture cells exposed to different types of magnesium materials: pure magnesium (Mg), magnesium-yttrium-neodymium-zirconium alloy (Mg-Y-Nd-Zr) and magnesium-zinc-calcium alloy (Mg-Zn-Ca). The materials were incubated with donor blood and mesenchymal multipotent stromal cells over 3 days. The studied materials did not induce massive lysis of human erythrocytes and leukocytes in vitro, but gradually impaired their structural integrity. In all cases, spontaneous platelet aggregation was observed in 6 h. In the presence of pure Mg and Mg-Zn-Ca alloy, this was accompanied by a decrease in the number of platelets with granules. In 24 h, substantial platelet degranulation occurred in all cases and in 72 h, the platelets did not contain granules. In parallel, the formation of large aggregates (60 µ) was observed. In the culture of stromal cells, all Mg-based materials reduced structural integrity of cells in 24 h, but did not significantly inhibit cell proliferation. Structural integrity of stromal cells partially recovered by day 3 in culture. The studied materials (Mg, Mg-Y-Nd-Zr, and Mg-Zn-Ca) seemed to be low-toxic for human cells during short-term contact, but could stimulate platelet aggregation and spontaneous degranulation and reduced the viability of diploid cells in vitro.

Growth-Stimulating Effect of Human Platelets Stabilized with Silver Nanoparticles.

We studied proliferative activity of multipotent mesenchymal stromal cells on collagen matrices containing platelets stabilized with silver nanoparticles. Dose-dependent stimulation of the growth of stromal cells without their structural damage was observed in the presence of stabilized platelets in proportion of 20-120 mln per 105 multipotent mesenchymal stromal cells. The same doses of non-stabilized platelets produced less pronounced stimulation effect. In higher doses (≥180 mln) stabilized platelets inhibited cell proliferation and induced their damage. Stabilized platelets enhanced migration of multipotent mesenchymal stromal cells; after formation of the monolayer, they actively colonized deep layer of the collagen matrix. The formed monolayer of multipotent mesenchymal stromal cells survived over 14 days without appreciable cell damage.

Giant crystals inside mitochondria of equine chondrocytes.

The present study reports for the first time the presence of giant crystals in mitochondria of equine chondrocytes. These structures show dark contrast in TEM images as well as a granular substructure of regularly aligned 1-2 nm small units. Different zone axes of the crystalline structure were analysed by means of Fourier transformation of lattice-resolution TEM images proving the crystalline nature of the structure. Elemental analysis reveals a high content of nitrogen referring to protein. The outer shape of the crystals is geometrical with an up to hexagonal profile in cross sections. It is elongated, spanning a length of several micrometres through the whole cell. In some chondrocytes, several crystals were found, sometimes combined in a single mitochondrion. Crystals were preferentially aligned along the long axis of the cells, thus appearing in the same orientation as the chondrocytes in the tissue. Although no similar structures have been found in the cartilage of any other species investigated, they have been found in cartilage repair tissue formed within a mechanically stimulated equine chondrocyte construct. Crystals were mainly located in superficial regions of cartilage, especially in joint regions of well-developed superficial layers, more often in yearlings than in adult horses. These results indicate that intramitochondrial crystals are related to the high mechanical stress in the horse joint and potentially also to the increased metabolic activity of immature individuals.

Alcohol consumption induces global gene expression changes in VTA dopaminergic neurons.

Alcoholism is associated with dysregulation in the neural circuitry that mediates motivated and goal-directed behaviors. The dopaminergic (DA) connection between the ventral tegmental area (VTA) and the nucleus accumbens is viewed as a critical component of the neurocircuitry mediating alcohol's rewarding and behavioral effects. We sought to determine the effects of binge alcohol drinking on global gene expression in VTA DA neurons. Alcohol-preferring C57BL/6J × FVB/NJ F1 hybrid female mice were exposed to a modified drinking in the dark (DID) procedure for 3 weeks, while control animals had access to water only. Global gene expression of laser-captured tyrosine hydroxylase (TH)-positive VTA DA neurons was measured using microarrays. A total of 644 transcripts were differentially expressed between the drinking and nondrinking mice, and 930 transcripts correlated with alcohol intake during the last 2 days of drinking in the alcohol group. Bioinformatics analysis of alcohol-responsive genes identified molecular pathways and networks perturbed in DA neurons by alcohol consumption, which included neuroimmune and epigenetic functions, alcohol metabolism and brain disorders. The majority of genes with high and specific expression in DA neurons were downregulated by or negatively correlated with alcohol consumption, suggesting a decreased activity of DA neurons in high drinking animals. These changes in the DA transcriptome provide a foundation for alcohol-induced neuroadaptations that may play a crucial role in the transition to addiction.

[Behavioral features of the imago of malaria mosquitoes (Diptera, Culicidae, Anopheles) in uzbekistan].

Morphological, cytogenetic, and molecular genetic analyses made in the Fergana, Chirchik-Akhangaran, Mirzachul, and Zarafshan physicogeographical districts of Uzbekistan revealed the closely related species An. artemievi malaria mosquito from the An. maculipennis complex. In the human settlements and natural biotopes under their canopy of 7 physicogeographical districts of Uzbekistan, there were 6 Anopheles mosquito species (An. artemievi, An. claviger, An. hyrcanus, An.martinius, An. pulcherrimus, and An. superpictus); An. superpictus is a dominant species in the human settlements and An. artemievi in subdominant. An.pulcherrimus was dominant and An. superpictus was subdominant under natural canopy conditions. The latter is of widespread occurrence in the mountain and piedmont areas of Uzbekistan. It is encountered in all the physicogeographical districts. An. artemievi is distributed in the river valleys in the Fergana, Chirchik-Akhangaran, Mirzachul, and Zarafshan physicogeographical districts. An. pulcherrimus is common in the plain river valleys, except in the Qashqadaryo physicogeographical district. An. martinius is found in the Qashqadaryo and Nizhneamudryo physicogeographical districts. Livestock houses are the most attractive day's rests for mosquitoes; utility rooms rank next in mosquito density. Housing premises are slightly occupied by mosquitoes. The maximum size of aggressive mosquitoes is noted in July, August, and early September.

Effect of 3D chondrocyte culturing conditions on the formation of extracellular matrix in cartilage tissue-engineering constructs.

We studied biochemical, morphological, and histological parameters of the extracellular matrix in scaffold-free tissue engineering chondrotransplants prepared from chondrocytes isolated from knee joint cartilage biopsy specimens of Haflinger horses (age 3.5-14 years) and in transplants prepared on the basis of commercial matrixes of Ethisorb® and Chondro-Gide(®). A total of 50 tissue-engineering constructs were designed and analyzed. Passage 2 cells populations were used. Mechanical stimulation during culturing of scaffold-free constructs considerably activated synthesis of the basic components of the cartilage matrix (proteoglycan concentration was 35% of the content in the native tissue, and the content of collagen-specific amino acids hydroxyproline and hydroxylysine attained 29.3 and 12.7%, respectively). The architecture of these cartilage constructs was morphologically and histologically similar to the native cartilage tissue. Insufficient support of the chondrogenesis by scaffold-based chondrotransplants and no differences between these constructs by the studied parameters were noted despite different chemical nature and structure of these scaffolds.

Equine articular chondrocytes on MACT scaffolds for cartilage defect treatment.

Treatment of cartilage defects poses challenging problems in human and veterinary medicine, especially in horses. This study examines the suitability of applying scaffold materials similar to those used for human cartilage regeneration on equine chondrocytes. Chondrocytes gained from biopsies of the talocrural joint of three horses were propagated in 2D culture and grown on two different scaffold materials, hyaluronan (HYAFF®) and collagen (BioGide®), and evaluated by light and electron microscopy. The equine chondrocytes developed well in both types of materials. They were vital and physiologically highly active. On the surface of the scaffolds, they formed cell multilayers. Inside the hyaluronan web, the chondrocytes were regularly distributed and spanned the large scaffold fibre distances by producing their own matrix sheath. Half-circle-like depressions occasionally found in the cell membrane were probably related to movement on the flexible matrix sheath. Inside the dense collagen scaffold, only single cells were found. They passed through the scaffold strands by cell shape adaptation. This study showed that the examined scaffold materials can be used for equine chondrocyte cultivation. Chondrocytes tend to form multilayers on the surface of both, very dense and very porous scaffolds, and have strategies to span between and move in large gaps.

[Detection of Anopheles artemievi (Diptera, Culicidae) in Uzbekistan].

An. artemievi was diagnosed by morphological and molecular genetic analyses in the Fergana valley, Uzbekistan, in 2008-2009. Four species of the genus Anopheles: An. superpictus, An. artemievi, An. hyrcanus, and An. claviger inhabiting the irrigated oases with the bulk of population were detected in the Fergana valley. An. artemievi and An. superpictus were prevalent in the foci of malaria. An. artemievi larvae inhabited the rice fields, springs, collection-drainage systems, and water reservoirs enriched in oxygen. The most attractive day's rest for mosquitoes was a cattle house where their bulk (on average 41.8%) was present. Next was cattle sheds, with the mosquitoes averaging 26.4%. An. artemievi, attacked human beings in the field conditions. This species showed a high susceptibility to test insecticides, such as propoxur, cipermethrin, deltamethrin, and lambda-Cyhalothrin.

Testing the silence of mutations: Transcriptomic and behavioral studies of GABA(A) receptor α1 and α2 subunit knock-in mice.

Knock-in mice were constructed with mutations in the α1 (H(270), A(277)) and α2 (H(270), A(277)) subunits of the GABAA receptor, which resulted in receptors that lacked modulation by ethanol but retained normal responses to GABA in vitro. A key question is whether these mutant receptors also function normally in vivo. Perturbation of brain function was evaluated by gene expression profiling in the cerebral cortex and by behavioral pharmacology experiments with GABAergic drugs. Analysis of individual transcripts found only six transcripts that were changed in α1 knock-in mice and three in the α2 mutants (p<0.05, corrected for multiple comparisons). Two transcripts that are sensitive to neuronal activity, Arc and Fos, increased about 250% in the α2 mutants, and about 50% in the α1 mutants. Behavioral effects (loss of righting reflex, rotarod) of flurazepam and pentobarbital were not different between α2 mutants and wild-type, but they were enhanced for α1 knock-in mice. These results indicate that introduction of these mutations in the α2 subunit of the GABAA receptor does not produce marked perturbation of brain function, as measured by gene expression and GABAergic behavioral responses, but the same mutations in the α1 subunit produce more pronounced changes, especially in GABAergic function.

Alcohol trait and transcriptional genomic analysis of C57BL/6 substrains.

C57BL/6 inbred mice have been widely used as research models; however, widespread demand has led to the creation of several B6 substrains with markedly different phenotypes. In this study, we report that two substrains of C57BL/6 mice, C57BL/6J (B6J) and C57BL/6NCrl (B6C), separated over 50 years ago at two different breeding facilities differ significantly in alcohol consumption and alcohol preference. The genomes of these two substrains are estimated to differ by only 1-2% of all gene loci, providing a unique opportunity to extract particular expression signatures between these substrains that are associated with quantifiable behavioral differences. Expression profiling of the cortex and striatum, hippocampus, cerebellum and the ventral brain region from alcohol-naïve B6C and B6J mice showed intervals on three chromosomes that are enriched in clusters of coregulated transcripts significantly divergent between the substrains. Additional analysis identified two genomic regions containing putative copy number differences between the substrains. One such region on chromosome 14 contained an estimated 3n copy number in the B6J genome compared with B6C. Within this interval, a gene of unknown function, D14Ertd449e, was found to be both associated with alcohol preference and vary in copy number across several inbred strain lineages. H2afz, Psen1, Wdfy1 and Clu were also identified as candidate genes that may be involved in influencing alcohol consumption.

Photoluminescence spectroscopy of the molecular biexciton in vertically stacked InAs-GaAs quantum dot pairs.

We present photoluminescence studies of the molecular neutral biexciton-exciton spectra of individual vertically stacked InAs/GaAs quantum dot pairs. We tune either the hole or the electron levels of the two dots into tunneling resonances. The spectra are described well within a few-level, few-particle molecular model. Their properties can be modified broadly by an electric field and by structural design, which makes them highly attractive for controlling nonlinear optical properties.

Defining the dopamine transporter proteome by convergent biochemical and in silico analyses.

Monoamine transporters play a key role in neuronal signaling by mediating reuptake of neurotransmitters from the synapse. The function of the dopamine transporter (DAT), an important member of this family of transporters, is regulated by multiple signaling mechanisms, which result in altered cell surface trafficking of DAT. Protein-protein interactions are likely critical for this mode of transporter regulation. In this study, we identified proteins associated with DAT by immunoprecipitation (IP) followed by mass spectrometry. We identified 20 proteins with diverse cellular functions that can be classified as trafficking proteins, cytoskeletal proteins, ion channels and extracellular matrix-associated proteins. DAT was found to associate with the voltage-gated potassium channel Kv2.1 and synapsin Ib, a protein involved in regulating neurotransmitter release. An in silico analysis provided evidence for common transcriptional regulation of the DAT proteome genes. In summary, this study identified a network of proteins that are primary candidates for functional regulation of the DAT, an important player in mechanisms of mental disorders and drug addiction.

Electrically tunable g factors in quantum dot molecular spin states.

We present a magnetophotoluminescence study of individual vertically stacked InAs/GaAs quantum dot pairs separated by thin tunnel barriers. As an applied electric field tunes the relative energies of the two dots, we observe a strong resonant increase or decrease in the g factors of different spin states that have molecular wave functions distributed over both quantum dots. We propose a phenomenological model for the change in g factor based on resonant changes in the amplitude of the wave function in the barrier due to the formation of bonding and antibonding orbitals.