Репозиторий Университета

Over the last decades, introduction of high-dose corticosteroids and immunosuppressive agents and later rituximab into the current algorithms for remission induction and maintenance treatment resulted in a tremendous improvement in the survival of patients with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides (AAV). However, in the recent meta-analysis of observational studies, Tan et al showed a 2.7-fold increased risk of death in patients with AAV when compared with the general population.1 Notably, there was a trend towards lower mortality in the most recent compared with the earlier cohorts. In our own study in 242 patients with granulomatosis with polyangiitis, we also found a significant decrease in mortality in the recent years (2004–2012 vs 1970–2003; p=0.04) and a shift towards a higher percentage of cardiovascular events and complications of immunosuppression as the causes of death.2

Two approaches to the synthesis of dinitrosyl iron complexes (DNIC) with glutathione and l-cysteine in aqueous solutions based on the use of gaseous NO and appropriate S-nitrosothiols, viz., S-nitrosoglutathione (GS-NO) or S-nitrosocysteine (Cys-NO), respectively, are considered. A schematic representation of a vacuum unit for generation and accumulation of gaseous NO purified from the NO2 admixture and its application for obtaining aqueous solutions of DNIC in a Thunberg apparatus is given. To achieve this, a solution of bivalent iron in distilled water is loaded into the upper chamber of the Thunberg apparatus, while the thiol solution in an appropriate buffer (рН 7.4) is loaded into its lower chamber. Further steps, which include degassing, addition of gaseous NO, shaking of both solutions and formation of the Fe²⁺-thiol mixture, culminate in the synthesis of DNIC. The second approach consists in a stepwise addition of Fe²⁺ salts and nitrite to aqueous solutions of glutathione or cysteine. In the presence of Fe²⁺ and after the increase in рН to the physiological level, GS-NO or Cys-NO generated at acid media (pH < 4) are converted into DNIC with glutathione or cysteine. Noteworthy, irrespective of the procedure used for their synthesis DNIC with glutathione manifest much higher stability than DNIC with cysteine. The pattern of spin density distribution in iron-dinitrosyl fragments of DNIC characterized by the d⁷ electronic configuration of the iron atom and described by the formula Fe⁺(NO⁺)2 is unique in that it provides a plausible explanation for the ability of DNIC to generate NO and nitrosonium ions (NO⁺) and the peculiar characteristics of the EPR signal of their mononuclear form (M-DNIC).

This study describes the development and cell culture application of nanometer thick photocrosslinkable thermoresponsive polymer films prepared by physical adsorption. Two thermoresponsive polymers, poly(N-isopropylacrylamide (NIPAm)-co-acrylamidebenzophenone (AcBzPh)) and poly(NIPAm-co-AcBzPh-co-N-tertbutylacrylamide) are investigated. Films are prepared both above and below the polymers' lower critical solution temperatures (LCSTs) and cross-linked, to determine the effect, adsorption preparation temperature has on the resultant film. The films prepared at temperatures below the LCST are smoother, thinner, and more hydrophilic than those prepared above. Human pulmonary microvascular endothelial cell (HPMEC) adhesion and proliferation are superior on the films produced below the polymers LCST compared to those produced above. Cells sheets are detached by simply lowering the ambient temperature to below the LCST. Transmission electron, scanning electron, and light microscopies indicate that the detached HPMEC sheets maintain their integrity.

Background Urethral reconstruction is one of the great surgical challenges for urologists. A cell-based tissue-engineered urethra may be an alternative for patients who have complicated long strictures and need urethral reconstruction. Here, we demonstrated the feasibility of using autologous urine-derived stem cells (USCs) seeded on small intestinal submucosa (SIS) to repair a urethral defect in a rabbit model. Methods Autologous USCs were obtained and characterized, and their capacity to differentiate into urothelial cells (UCs) and smooth muscle cells (SMCs) was tested. Then, USCs were labeled with PKH67, seeded on SIS, and transplanted to repair a urethral defect. The urethral defect model was surgically established in New Zealand white male rabbits. A ventral urethral gap was created, and the urethral mucosa was completely removed, with a mean rabbit penile urethra length of 2 cm. The urethral mucosal defect was repaired with a SIS scaffold (control group: SIS with no USCs; experimental group: autologous USC-seeded SIS; n = 12 for each group). A series of tests, including a retrograde urethrogram, histological analysis, and immunofluorescence, was undertaken 2, 3, 4, and 12 weeks after the operation to evaluate the effect of the autologous USCs on urethral reconstruction. ResultsAutologous USCs could be easily collected and induced to differentiate into UCs and SMCs. In addition, the urethral caliber, speed of urothelial regeneration, content of smooth muscle, and vessel density were significantly improved in the group with autologous USC-seeded SIS. Moreover, inflammatory cell infiltration and fibrosis were found in the control group with only SIS, but not in the experimental autologous USC-seeded SIS group. Furthermore, immunofluorescence staining demonstrated that the transplanted USCs differentiated into UCs and SMCs in vivo. Conclusions Autologous USCs can be used as an alternative cell source for cell-based tissue engineering for urethral reconstruction.

Context: Stress urinary incontinence (SUI) significantly diminishes the quality of patients' lives. Currently available surgical and nonsurgical therapies remain far from ideal. At present advancements in cellular technologies have stirred growing interest in the use of autologous cell treatments aimed to regain urinary control. Objective: To conduct a review of the literature and analyze preclinical and clinical studies dedicated to various cell therapies for SUI, assessing their effectiveness, safety, and future prospects. Evidence acquisition: A systematic literature search in PubMed was conducted using the following key terms: "stem," "cell," "stress," "urinary," and "incontinence." A total of 32 preclinical studies and 15 clinical studies published between 1946 and December 2014 were included in the review. Evidence synthesis: Most preclinical trials have used muscle-derived stem cells (MDSCs) and adipose-derived stem cells (ADSCs). Yet, at present, the application of other types of cells, such as human amniotic fluid stem muscle-derived progenitor cells (hAFSCs), and bone marrow mesenchymal stromal cells (BMSCs), is becoming more extensive. While the evidence shows that these therapies are effective and safe, further work is required to standardize surgical techniques, as well as to identify indications for their use, doses and number of doses. Conclusion: Future research will have to focus on clinical applications of cell therapies; namely, it will have to determine indications for their use, doses of cells, optimal surgical techniques and methods, attractive cell sources, as well as to develop clinically relevant animal models and make inroads into understanding the mechanisms of SUI improvement by cell therapies.

Urethral strictures and anomalies remain among the difficult problems in urology, with urethroplasty procedures being the most effective treatment options. The two major types of urethroplasty are anastomotic urethroplasty and widening the urethral lumen using flaps or grafts (i.e. substitution urethroplasty). However, no ideal material for the latter has been found so far. Designing and selecting such a material is a necessary and challenging endeavour, driving the need for further bioengineered urethral tissue research. This article reviews currently available studies on the potentialities of tissue engineering in urethral reconstruction, in particular those describing the use of both acellular and recellularized tissue-engineered constructs in animal and human models. Possible future developments in this field are also discussed. Copyright © 2015 John Wiley & Sons, Ltd.

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.

Cartilaginous structures are at the core of embryo growth and shaping before the bone forms. Here we report a novel principle of vertebrate cartilage growth that is based on introducing transversally-oriented clones into pre-existing cartilage. This mechanism of growth uncouples the lateral expansion of curved cartilaginous sheets from the control of cartilage thickness, a process which might be the evolutionary mechanism underlying adaptations of facial shape. In rod-shaped cartilage structures (Meckel, ribs and skeletal elements in developing limbs), the transverse integration of clonal columns determines the well-defined diameter and resulting rod-like morphology. We were able to alter cartilage shape by experimentally manipulating clonal geometries. Using in silico modeling, we discovered that anisotropic proliferation might explain cartilage bending and groove formation at the macro-scale.

Pores are vital for functioning of avascular tissues. Laser-induced pores play an important role in the process of cartilage regeneration. The aim of any treatment for osteoarthritis is to repair hyaline-type cartilage. The aims of this study are to answer two questions: (1) How do laser-assisted pores affect the cartilaginous cells to synthesize hyaline cartilage (HC)? and (2) How can the size distribution of pores arising in the course of laser radiation be controlled? We have shown that in cartilage, the pores arise predominately near chondrocytes, which promote nutrition of cells and signal molecular transfer that activates regeneration of cartilage. In vivo laser treatment of damaged cartilage of miniature pig joints provides cellular transformation and formation of HC. We propose a simple model of pore formation in biopolymers that paves the way for going beyond the trial-anderror approach when choosing an optimal laser treatment regime. Our findings support the approach toward laser healing of osteoarthritis.

Mesenchymal stem cells (MSCs) are thought to be the most attractive type of cells for bone repair. However, much still remains unknown about MSCs and needs to be clarified before this treatment can be widely applied in the clinical practice. The purpose of this study was to establish the involvement of allogeneic MSCs in the bone formation in vivo, using a model of transgenic mice and genetically labeled cells. Polylactide scaffolds with hydroxyapatite obtained by surface selective laser sintering were used. The scaffolds were sterilized and individually seeded with MSCs from the bone marrow of 5-week-old GFP(+) transgenic С57/Вl6 or GFP(−)С57/Вl6 mice. 4-mm-diameter critical-size defects were created on the calvarial bone of mice using a dental bur. Immediately after the generation of the cranial bone defects, the scaffolds with or without seeded cells were implanted into the injury sites. The cranial bones were harvested at either 6 or 12 weeks after the implantation. GFP(+) transgenic mice having scaffolds with unlabeled MSCs were used for the observation of the host cell migration into the scaffold. GFP(−) mice having scaffolds with GFP(+)MSCs were used to assess the functioning of the seeded MSCs. The obtained data demonstrated that allogeneic MSCs were found on the scaffolds 6 and 12 weeks post-implantation. By week 12, a newly formed bone tissue from the seeded cells was observed, without an osteogenic pre-differentiation. The host cells did not appear, and the control scaffolds without seeded cells remained empty. Besides, a possibility of vessel formation from seeded MSCs was shown, without a preliminary cell cultivation under controlled conditions.

It has been established that treatment of mice with sodium nitrite, S-nitrosoglutathione and the water-soluble nitroglycerine derivative isosorbide dinitrate (ISDN) as NO donors initiates in vivo synthesis of significant amounts of EPR-silent binuclear dinitrosyl iron complexes (B-DNIC) with thiol-containing ligands in the liver and other tissues of experimental mice. This effect is especially apparent if NO donors are administered to mice simultaneously with the Fe²⁺-citrate complex. Similar results were obtained in experiments on isolated liver and other mouse tissues treated with gaseous NО in vitro and during stimulation of endogenous NO synthesis in the presence of inducible NO synthase. B-DNIC appeared in mouse tissues after in vitro treatment of tissue samples with an aqueous solution of diethyldithiocarbamate (DETC), which resulted in the transfer of iron-mononitrosyl fragments from B-DNIC to the thiocarbonyl group of DETC and the formation of EPR-detectable mononitrosyl iron complexes (MNIC) with DETC. EPR-Active MNIC with N-methyl-d-glucamine dithiocarbamate (MGD) were synthesized in a similar way. MNIC-MGD were also formed in the reaction of water-soluble MGD-Fe²⁺ complexes with sodium nitrite, S-nitrosoglutathione and ISDN.

В работе обсуждаются дискуссионные аспекты статьи Дробижева М. Ю., Кикты С. В., Мачильской О. В. “Кардиопсихиатрия. Проблема перевода”, опубликованной в журнале Кардиоваскулярной терапия и профилактика 2016; 15(4): 88-97. Представленная авторами классификация кардиопсихиатрических расстройств основана лишь на анализе литературных данных и на деле представляет собой редукционистскую попытку выяснения соотношения нейрофизиологических механизмов со сложными психопатологическими и психосоматическими образованиями, реализующимися в сфере сердечно-сосудистой системы. Возражения вызывает также раздел публикации, посвященный психофармакотерапии психических расстройств в кардиологической практике, в котором авторская позиция сводится к рекомендации применения лишь одного лекарственного препарата.

In this work, we studied water solubility of ampicillin trihydrate and its solid dispersions (SD) with polyethylene glycol-1500, polyvinylpyrrolidone-10000, and β-cyclodextrin. It was found that SD formation increases the solubility by a factor of 1.34–1.73 and the rate of ampicillin dissolution by a factor of 3.43–7.40. The results of complex physicochemical studies suggest that the improved release of ampicillin from SD is due to its micronization and solubilization by the polymer.

Background: Attention-deficit/hyperactivity disorder (ADHD) is a common, highly heritable neurodevelopmental disorder with profound cognitive, behavioral, and psychosocial impairments with persistence across the life cycle. Our initial genome-wide screening approach for copy number variants (CNVs) in ADHD implicated a duplication of SLC2A3, encoding glucose transporter-3 (GLUT3). GLUT3 plays a critical role in cerebral glucose metabolism, providing energy for the activity of neurons, which, in turn, moderates the excitatory-inhibitory balance impacting both brain development and activity-dependent neural plasticity. We therefore aimed to provide additional genetic and functional evidence for GLUT3 dysfunction in ADHD. Methods: Case-control association analyses of SLC2A3 single-nucleotide polymorphisms (SNPs) and CNVs were conducted in several European cohorts of patients with childhood and adult ADHD (SNP, n = 1,886 vs. 1,988; CNV, n = 1,692 vs. 1,721). These studies were complemented by SLC2A3 expression analyses in peripheral cells, functional EEG recordings during neurocognitive tasks, and ratings of food energy content. Results: Meta-analysis of all cohorts detected an association of SNP rs12842 with ADHD. While CNV analysis detected a population-specific enrichment of SLC2A3 duplications only in German ADHD patients, the CNV + rs12842 haplotype influenced ADHD risk in both the German and Spanish cohorts. Duplication carriers displayed elevated SLC2A3 mRNA expression in peripheral blood cells and altered event-related potentials reflecting deficits in working memory and cognitive response control, both endophenotypic traits of ADHD, and an underestimation of energy units of high-caloric food. Conclusions: Taken together, our results indicate that both common and rare SLC2A3 variation impacting regulation of neuronal glucose utilization and energy homeostasis may result in neurocognitive deficits known to contribute to ADHD risk.

Chronic spontaneous urticaria (CSU) is a mast-cell driven skin disease, characterized by the recurrence of transient wheals, angioedema, or both for more than 6 weeks. Autoimmunity is thought to be one of the most frequent causes of CSU. Type I and type II autoimmunity, i.e. IgE to autoallergens and IgG autoantibodies to IgE or its receptor, respectively, have been implicated in the etiology and pathogenesis of CSU. We analyzed the relevant literature and assessed the existing evidence in support of a role for type I and II autoimmunity in CSU with the help of Hill’s criteria of causality. For each of these criteria, i.e. strength of association, consistency, specificity, temporality, biological gradient, plausibility, coherence, experiment and analogy, we categorized the strength of evidence as “insufficient”, “low”, “moderate” or “high” and then assigned levels of causality for type I and II autoimmunity in CSU, from level 1 (causal relationship) to level 5 (causality not likely). Based on the evidence in support of Hill’s criteria, type I autoimmunity in CSU has level 3 causality (causal relationship suggested) and type II autoimmunity has level 2 causality (causal relationship likely). There are still many aspects of the pathologic mechanisms of CSU that need to be resolved, but it is becoming clear that there are at least two distinct pathways, type I and type II autoimmunity, that contribute to the pathogenesis of this complex disease.

Physical adsorption was used to produce nanometer thick thermoresponsive films with a view to nonenzymatic cell detachment. Two polymers were investigated, poly-(N-isopropylacrylamide) and poly (N-isopropylacrylamide-co-N-tertbutylacrylamide). Substrates were prepared above and below the polymers’ LCST to investigate the effect of polymer conformation on the prepared substrates. Endothelial cells were seeded on the prepared films; cell proliferation was higher on the films produced below the polymers’ LCST than on those prepared above and cells detached from the surfaces upon temperature reduction. Physical adsorption of poly-(N-isopropylacrylamide)–based films is a viable approach to produce substrates compliant with cell growth and temperature modulated detachment.

In recent years, engineering of blood vessels, which can provide the effective transport of nutrients and various metabolites, is one of the major challenges in tissue reconstruction. Many researches are carried out to develop cell-seeded bioconstructs based on natural polymers, particularly on PEGylated fibrin. Therefore, the aim of this study was to reveal the optimal component ratio for modified fibrin hydrogels in order to provide favorable conditions for vascular development of endothelial and mesenchymal stem cell co-culture. It has been found out that the PEGylated fibrin hydrogels can support 3D cell growth in HUVECs and hASCs co-culture. The microporous filamentous hydrogel prepared from PEGylated 5 : 1 fibrinogen and using the 1 : 0.2 protein to thrombin ratio had the most favorable microenvironment for cell distribution, growth and development in the studied co-culture that resulted in high levels of expression of proteins required for angiogenesis.
(PDF) Evaluation of the Vasculogenic Potential of Hydrogels Based on Modified Fibrin.

Conducting, mechanically durable, elastic nanocomposite films were prepared with chitosan (CS) as the polymer matrix, graphene obtained from highly exfoliated graphite as the nanofiller, and poly(vinyl pyrrolidone) (PVP) as the stabilizer of the graphene sheets. The maximum graphene content in the composites without a loss of uniformity and other useful properties increased up to 4.0 wt %. The resulting composites were characterized by scanning electron microscopy, Raman spectroscopy, X-ray diffraction analysis, mechanical testing, and electrical conductivity testing to determine the effects of the addition of graphene on the morphology and mechanical and electrical properties of the CS–PVP–graphene nanocomposite films. In this study, we took an approach to making nanocomposites from the perspectives of green chemistry, environmental protection, regenerative medicine, and low cost. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017, 134, 45038. © 2017 Wiley Periodicals, Inc.

The discovery and development of effective antihypoxic drugs is an important issue in modern experimental and clinical pharmacology. Complexes of zinc containing imidazole cycles as ligands are promising candidates for the design of novel drugs, including pharmacotherapeutic tools for patient’s protection under conditions of oxygen deficiency. Herein, we report antihypoxic activity of zinc complexes with N-alkenylimidazoles, and, specifically, a novel drug Acyzol, which can be used as efficient antidote after poisoning by carbon monoxide and other combustion products.