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Natalia M Yudintceva,1 Yulia A Nashchekina,1,2 Miralda I Blinova,1 Nadezhda V Orlova,3 Alexandr N Muraviov,3 Tatiana I Vinogradova,3 Magomed G Sheykhov,3 Elena Y Shapkova,3 Dmitriy V Emeljannikov,3 Petr K Yablonskii,3,4 Igor A Samusenko,5 Anastasiya L Mikhrina,6 Artem V Pakhomov,7 Maxim A Shevtsov1,7,8 1Department of Cell Culture, Institute of Cytology of the Russian Academy of Sciences (RAS), 2Nanotechnology and Telecommunications, Institute of Physics, Peter the Great St Petersburg Polytechnic University, 3Department of Urology, Federal State Institution Saint Petersburg Research Institute of Phthisiopulmonology, Ministry of Health of Russia, 4Faculty of Medicine, Federal State Budgetary Institute, 5Department of Pathology, Federal State Budgetary Institute “Nikiforov Russian Centre of Emergency and Radiation Medicine” of the Ministry of Health of Russia, 6Department of Pathomorphology, I.M. Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Science, 7Department of Radiology, Federal Almazov North-West Medical Research Center, 8Department of Experimental Medicine, First I.P. Pavlov State Medical University of St Petersburg, St Petersburg, Russia Abstract: In the present study, a poly-L-lactide/silk fibroin (PL-SF) bilayer scaffold seeded with allogenic bone marrow stromal cells (BMSCs) was investigated as a potential approach for bladder tissue engineering in a model of partial bladder wall cystectomy in rabbits. The inner porous layer of the scaffold produced from silk fibroin was designed to promote cell proliferation and the outer layer produced from poly-L-lactic acid to serve as a waterproof barrier. To compare the feasibility and efficacy of BMSC application in the reconstruction of bladder defects, 12 adult male rabbits were divided into experimental and control groups (six animals each) that received a scaffold seeded with BMSCs or an acellular one, respectively. For BMSC tracking in the graft in in vivo studies using magnetic resonance imaging, cells were labeled with superparamagnetic iron oxide nanoparticles. In vitro studies demonstrated high intracellular incorporation of nanoparticles and the absence of a toxic influence on BMSC viability and proliferation. Following implantation of the graft with BMSCs into the bladder, we observed integration of the scaffold with surrounding bladder tissues (as detected by magnetic resonance imaging). During the follow-up period of 12 weeks, labeled BMSCs resided in the implanted scaffold. The functional activity of the reconstructed bladder was confirmed by electromyo­graphy. Subsequent histological assay demonstrated enhanced biointegrative properties of the PL-SF scaffold with cells in comparison to the control graft, as related to complete regener­ation of the smooth muscle and urothelium tissues in the implant. Confocal microscopy studies confirmed the presence of the superparamagnetic iron oxide nanoparticle-labeled BMSCs in newly formed bladder layers, thus indicating the role of stem cells in bladder regeneration. The results of this study demonstrate that application of a PL-SF scaffold seeded with allogenic BMSCs can enhance biointegration of the graft in vivo and support bladder tissue regeneration and function. Keywords: bladder, bone marrow stromal cells, scaffold, superparamagnetic iron oxide nanoparticles, tissue engineering, stem cells

We analytically and numerically investigate surface second-harmonic generation (SHG) from a stack of dielectric layers. We develop a theoretical formalism based on the transfer matrix method for the calculation of the surface-driven second-harmonic radiation from multilayer structures and elaborate it for the case of ultrathin dielectric layers using a power series expansion to derive the effective surface nonlinear tensor for the whole stack. We show that for deeply subwavelength thicknesses of the layers the surface responses from all interfaces can efficiently sum up, leading to largely enhanced efficiency of SHG. As a result, such surface-driven nonlinearity can become comparable to the bulk nonlinearity in noncentrosymmetric semiconductors and can yield high performance for nonlinear nanophotonic applications.

We analyse possibilities to quantitatively evaluate the surface second-order optical nonlinearity in noncentrosymmetric materials based on polarization-resolved analysis of far-field radiation patterns of second-harmonic generation. We analytically demonstrate that for plane-wave illumination the contribution to the second-harmonic signal from the surface of a nonlinear medium exhibits different polarization properties and angular dependencies compared to the contribution from the bulk. In view of this, we optimize the illumination geometry in order to enable the most efficient separation and comparison of both nonlinearities. Furthermore, we consider the illumination of an AlGaAs slab by a tightly-focused linearly-polarized Gaussian beam as an alternative measurement geometry. It is found that the reliable separation of the surface nonlinearity contribution as well as a wide range of detectable values can be achieved with this geometry as well.

Up to now, full tunability of waveforms was possible only in electronics, up to radio-frequencies. Here we propose a new concept of producing few-cycle terahertz (THz) pulses with widely tunable waveforms. It is based on control of the phase delay between different parts of the THz wavefront using linear diffractive optical elements. Suitable subcycle THz wavefronts can be generated via coherent excitation of nonlinear low-frequency oscillators by few-cycle optical pulses. Using this approach it is possible to shape the electric field rather than the slow pulse envelope, obtaining, for instance, rectangular or triangular waveforms in the THz range. The method is upscalable to the optical range if the attosecond pump pulses are used.

The interaction of subcycle pulses with quantum systems is considered when the pulse duration becomes much smaller than the timescales of electron oscillations. We show analytically that the interaction process in this case is governed by the electric pulse area. The efficient nonresonant excitation of quantum systems by subcycle pulses with a high degree of unipolarity is demonstrated. The results are confirmed by direct numerical solution of multilevel Bloch equations.

Burn wounds are one of the most important causes of mortality and especially morbidity around the world. Burn wound healing and skin tissue regeneration remain thus one of the most important challenges facing the mankind. In the present study we have addressed this challenge, applying a solution-stabilized dispersion TiO 2 nanoparticles, hypothesizing that their ability to adsorb proteins will render them a strong capacity in inducing body fluid coagulation and create a protective hybrid material coating. The in vitro study of interaction between human blood and titania resulted at enhanced TiO 2 concentrations in formation of rather dense gel composite materials and even at lower content revealed specific adsorption pattern initiating the cascade response, promising to facilitate the regrowth of the skin. The subsequent in vivo study of the healing of burn wounds in rats demonstrated formation of a strongly adherent crust of a nanocomposite, preventing infection and inflammation with quicker reduction of wound area compared to untreated control. The most important result in applying the TiO 2 dispersion was the apparently improved regeneration of damaged tissues with appreciable decrease in scar formation and skin color anomalies.

Creation, erasing and ultrafast control of population density gratings using few-cycle optical pulses coherently interacting with resonant medium is discussed. In contrast to the commonly used schemes, here the pulses do not need to overlap in the medium, interaction between the pulses is mediated by excitation of polarization waves. We investigate the details of the dynamics arising in such ultrashort pulse scheme and develop an analytical theory demonstrating the importance of the phase memory effects in the dynamics.

We propose a strikingly simple method to form approximately unipolar half-cycle optical pulses via reflection of a single-cycle optical pulse from a thin flat metallic or dielectric layer. Unipolar pulses in reflection arise due to specifics of one-dimensional pulse propagation. Namely, we show that the field emitted by the layer is proportional to the velocity of the oscillating charges in the medium, instead of their acceleration. Besides, the oscillation velocity of the charges can be forced to keep a constant sign throughout the pulse duration. That is, reflection of ultrashort pulses from broad-area layers with nanometer-scale thickness can be very different from the common reflection in the case of longer pulses and thicker layers. This suggests a possibility of unusual transformations of few-cycle light pulses in completely linear optical systems.

Thawing in the water bath is often considered as a standard procedure. The thermal history of samples thawed in this way is poorly controlled, but cryopreservation and banking of cell-based products require standardization, automation and safety of all the technological stages including thawing. The programmable freezers allow implementation of the controlled cooling as well as the controlled thawing. As the cell damage occurs during the phase transformation that takes place in the cryoprotectant medium in the process of freezing-thawing, the choice of warming rates within the temperature intervals of transformations is very important. The goal of the study was to investigate the influence of warming rates within the intervals of the phase transformations in the DMSO-based cryoprotectant medium on the cell recovery and to develop a cryopreservation protocol with controlled cooling and warming rates. The temperature intervals of phase transformations such as melting of the eutectic mixture of the cryoprotectant solution (MEMCS), melting of the eutectic salt solution (MESS), melting of the main ice mass (MMIM), recrystallization before MEMCS, recrystallization before MESS and recrystallization before MMIM were determined by thermo-mechanical analysis. The biological experiments were performed on the rat testicular interstitial cells (TIC). The highest levels of the cell recovery and metabolic activity after cryopreservation were obtained using the protocol with the high (20 °C/min) warming rate in the temperature intervals of crystallization of the eutectics as well as recrystallizations and the low (1 °C/min) warming rate in the temperature intervals of melting of the eutectics as well as MMIM. The total cell recovery was 65.3 ± 2.1 %, the recovery of the 3-beta-HSD-positive (Leydig) cells was 82.9 ± 1.8 %, the MTT staining was 32.5 ± 0.9 % versus 42.1 ± 1.7 %; 57.4 ± 2.1 % and 24.0 ± 1.1 % respectively, when compared to the thawing in the water bath.

Takayasu arteritis is well-known rare form of large vessel vasculitis. Pulmonary involvement is observed in 50% of cases. However, pulmonary hypertension is a rare complication of pulmonary artery stenosis. In this article we present a rare clinical case of Takayasus arteritis with typical stenosis of subclavian, carotid arteries, and involvement of pulmonary arteries with development of severe pulmonary hypertension. The article also contains discussion of current methods of diagnosis and treatment of pulmonary artery stenosis in Takayasus arteritis with special focus on effectiveness and complications of angioplasty and surgery.

Background: The mucopolysaccharidoses (MPSs) are rare genetic disorders caused by mutations in lysosomal enzymes involved in the degradation of glycosaminoglycans (GAGs). In this study, we analyzed a total of 48 patients including MPSI (n=6), MPSII (n=18), MPSIIIA (n=11), MPSIVA (n=3), and MPSVI (n=10)., Methods: In MPS patients, urinary GAGs were colorimetrically assayed. Enzyme activity was quantified by colorimetric and fluorimetric assays. To find mutations, all IDUA, IDS, SGSH, GALNS, and ARSB exons and intronic flanks were sequenced. New mutations were functionally assessed by reconstructing mutant alleles with site-directed mutagenesis followed with expression of wild-type and mutant genetic variants in CHO cells, measuring enzymatic activity, and Western blot analysis of protein expression of normal and mutated enzymes in cell lysates., Results: A total of five novel mutations were found including p.Asn348Lys (IDUA) in MPSI, p.Tyr240Cys (GALNS) in MPSIVA, and three ARSB mutations (p.Gln110*, p.Asn262Lysfs*14, and pArg315*) in MPSVI patients. In case of mutations p.Asn348Lys, p.Asn262Lysfs*14, and p.Gln110*, no mutant protein was detected while activity of the mutant protein was <1% of that of the normal enzyme. For p.Tyr240Cys, a trace of mutant protein was observed with a remnant activity of 3.6% of the wild-type GALNS activity. For pArg315*, a truncated 30-kDa protein that had 7.9% of activity of the normal ARSB was detected., Conclusions: These data further enrich our knowledge of the genetic background of MPSs., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The authors conducted histological, immunohistochemical, and morphometric studies of 47 endomyocardial biopsies and 33 hearts with a high morphological criterion for the diagnosis of arrhythmogenic right ventricular dysplasia after F.I. Marcus et al. (2010), as well as intravital and postmortem cardiac magnetic resonance imaging (MRI). The average residual cardiomyocyte area was 39.7 +/- 12.8%, and the average cardiomyocyte diameter was 10.4 +/- 1.5 microm. Muscle fiber atrophy, lipomatosis, and fibrosis in the right ventricle were detected in all cases, myocarditis in 44%, mitral valve prolapse in 42%, and muscle fiber dyscomplexation in the right ventricular in 33%. The most informative signs for the diagnosis of arrhythmogenic right ventricular dysplasia using MRI are right ventricular contraction dyssynchrony and those using a morphological study are a residual cardiomyocyte area and muscle fiber atrophy in the right ventricle. A decrease in or focal absence of gamma-catenin expression in the muscle fibers was observed only in 60% of the patients.

A long course of anticancer therapy may lead to testicular steroidogenesis destruction. Cryopreservation of testicular interstitial cells (TIC) would be a strategy to protect hormonal and fertile potential of pre-pubertal boys treated with chemo - or radiotherapy. The aim of this research was to optimize protocols for freezing of TIC. Essential physical processes associated with the presence of dimethyl sulphoxide (Me(2)SO) in the cryoprotectant solution take place at the temperatures below -60°С. These processes are the eutectic crystallization at the stage of freezing and the recrystallization before the melting of the eutectic mixture at the stage of heating. Both of the processes affect the viability of the cells subjected to cryopreservation. Temperature intervals when these processes take place were determined by the method of thermoplastic deformation for 10% Me(2)SO selected for cryopreservation of TIC. Rat TIC were cryopreserved using five different protocols which varied in cooling rates within the chosen temperature intervals. Post-thaw cell viability and metabolic activity were evaluated by Trypan Blue and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) staining assays. Leydig cell recovery after cryopreservation was measured by 3-beta-hydroxysteroid dehydrogenase reaction. Based on the obtained results, the authors developed a cryopreservation protocol for TIC which makes it possible to achieve great cell viability due to using controlled cooling rates within the temperature intervals below -60°С., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Biolistic transformation of soybean by construction carrying a mutant alpha-tubulin gene as selective marker gene conferring resistance to dinitroaniline herbicides, and by additional construction pAHTUBI carrying a full-length barley beta-tubulin gene for correct co-expression of exogeneous tubulin in cells of transgenic soybean lines has been realized. It was established that 10 microM trifluralin is the most optimal selective concentration to pick up transformed soybean lines. Transgenic nature of offained regenerants was confirmed by Southern blotting hybridization using specific probe to selective alpha-tubulin gene.

Embryogenic potential of 19 soybean cultivars originated from different eco-geographic regions (Europe, China, America and Far East) has been established and analyzed. Cultivars TSZ-14, Amurskaja-111, Gribovskaja mestnaja were emphasized as highly embryogenic genotypes. Cultivar Mantherova biela vel. revealed the highest embryogenic potential (80%). The comparison of embryogenic potential of the Ukrainian cultivars with given data indicated various parameters which could affect the induction of soybean somatic embryogenesis. It had also been supposed that cultivars from the Far East and Europe could have the potential for further screening of new highly embryogenic genotypes. The data obtained enlarge the data base pool of the soybean somatic embryogenesis research and enable using the most appropriate cultivars for further genetic improvement.

Seven soybean (Glycine max (L.) Merr.) cultivars zoned in forest-steppe and marshy woodlands of Ukraine: "Chernuatka", "Vasylkivska", "Kyivska-91", "Kyivska-27", "Marjana", "Chernobura" and "Altair" were established in vitro for somatic embryogenesis inducing. Cultivars "Marjana" (88%) and "Vasylkivska" (86%) demonstrated the highest embryogenic capacity among all the tested cultivars. During the further plant regeneration cultivars "Marjana" and "Kyivska-91" showed the best capacity to form adult plants despite the fact that embryogenic capability of the cultivar "Kyivska-91" was 71%. According to the obtained data three genotypes were selected for the further investigation of effective methods of biolictic transformation of ukrainian cultivars.