Репозиторий Университета

© 2018, Institute of Chemistry, Slovak Academy of Sciences. This work focuses on the behavior and comparative assessment of the antioxidant activity of several well-known antioxidants using three different approaches with the same ABTS•+/potassium persulfate radical-generating system: a decolorization assay, kinetic assay, and visual-spectrophotometric titration assay. The decolorization assay is the most common approach but gives little information on antioxidant behavior. The kinetic assay can be used for an in-depth study of the specific features of a particular antioxidant and facilitates identification of the mechanism of action. The visual-spectrophotometric titration assay is complementary to the above assays and subdivided into two stages to demonstrate the contribution of the “fast” and “slow” scavenging properties to the total antioxidant activity. The trolox equivalent antioxidant capacity (TEAC) value ranges for several flavonoids and the endogenous antioxidants derived from these assays vary from 0.7 to 5.1, 0.5–2.8, and 0.4–3.7, respectively, while the sequence of the weakest antioxidant to the strongest is similar. The analysis of the kinetic curves for some flavonoids showed that their interaction with ABTS•+ has an atypical character. Based on the results of the kinetic and visual-spectrophotometric titration assays, fast-reacting (trolox, α-tocopherol), slow-reacting (naringenin, apigenin) and moderate-reacting antioxidant (dihydroquercetin, quercetin, rutin, morin, and glutathione) compounds were distinguished.

© 2018 Background and aim: The non-glycosylated glycoform of transferrin (Tf), known as asialo-Tf, was not selected (in favor of disialo-Tf) as the measurand for the standardization of carbohydrate deficient transferrin (CDT) determination because of a lower diagnostic sensitivity provided with the currently available analytical procedures for sera. However, asialo-Tf could provide an additional value to disialo-Tf in the CDT analysis employed in forensic toxicology contexts. The present work aimed at developing an easy sample preparation based on PEG precipitation in order to improve the detectability of asialo-Tf in capillary electrophoresis (CE). Methods: Equal volumes (35 μL) of serum and of 30% PEG-8000 were mixed and briefly vortexed. After centrifugation, the supernatant was iron saturated with a ferric solution (1:1, v/v). The mixture was analyzed in CE for asialo-Tf and disialo-Tf determination. Results: PEG-8000 precipitation allowed the improvement of the baseline in the electropherograms in terms of interferences reduction particularly in the asialo-Tf migration region. The detection of asialo-Tf was possible in 89% of samples with disialo-Tf above the cut-off limit, whereas only 16% of them showed asialo-Tf by employing the traditional sample preteatment. Conclusions: Asialo-Tf represents an additional value to disialo-Tf as a biomarker of alcohol abuse in forensic toxicology.

© 2018, Springer International Publishing AG, part of Springer Nature.  Molecular oxygen (O 2 ) is a key player in cell mitochondrial function, redox balance and oxidative stress, normal tissue function and many common disease states. Various chemical, physical and biological methods have been proposed for measurement, real-time monitoring and imaging of O 2 concentration, state of decreased O 2 (hypoxia) and related parameters in cells and tissue. Here, we review the established and emerging optical microscopy techniques allowing to visualize O 2 levels in cells and tissue samples, mostly under in vitro and ex vivo, but also under in vivo settings. Particular examples include fluorescent hypoxia stains, fluorescent protein reporter systems, phosphorescent probes and nanosensors of different types. These techniques allow high-resolution mapping of O 2 gradients in live or post-mortem tissue, in 2D or 3D, qualitatively or quantitatively. They enable control and monitoring of oxygenation conditions and their correlation with other biomarkers of cell and tissue function. Comparison of these techniques and corresponding imaging setups, their analytical capabilities and typical applications are given.

© 2018 The Author(s) Deep vein thrombosis (DVT) is a major origin of morbidity and mortality. While DVT has long been considered as blood coagulation disorder, several recent lines of evidence demonstrate that immune cells and inflammatory processes are involved in DVT initiation. Here, we discuss these mechanisms, in particular, the role of immune cells in endothelial activation, and the immune cascades leading to expression of adhesion receptors on endothelial cells. We analyze the specific recruitment and functional roles of different immune cells, such as mast cells and leukocytes, in DVT. Importantly, we also speculate how immune modulation could be used for DVT prevention with a lower risk of bleeding complications than conventional therapeutic approaches.

© 2018, Springer Science+Business Media, LLC, part of Springer Nature. Changes in the levels of rennin, angiotensin II, and angiotensin (1–7) were studied during normal pregnancy. The blood was taken on gestation days 140-237 and 238-280. No significant changes in renin concentration were observed during normal pregnancy (p=0.423). The level of angiotensin II increased during normal pregnancy from 9.7±1.2 to 14.7±1.9 pg/ml (p=0.019). On the contrary, angiotensin (1–7) concentration decreased from 771.1±44.2 to 390.7±13.9 pg/ml (p<0.001). The shift in the proportion between vasoconstrictor angiotensin II and vasodilaltor angiotensin (1–7) attests to high predisposition of pregnant women to hypertension-related complications.

© 2018 The Association of Coloproctology of Great Britain and Ireland.  Aim To describe the technique of a modified extraperitoneal retrotransversalis end colostomy as part of a laparoscopic abdominoperineal excision (APR). Method The colostomy site is preoperatively chosen and used intra-operatively for a trocar. After the rectum has been mobilized the descending colon is freed. The peritoneal margin is gently grasped and the parietal peritoneum and extraperitoneal together with the transversalis fascia are separated from the transverse abdominal muscle fibres upwards for 3–4 cm aiming at the trocar site to form the extraperitoneal retrotransversalis canal. The stoma site trocar is partially withdrawn and its head is turned laterally until its tip is positioned in the layer between the abdominal wall muscles and underlying transversalis and extraperitoneal fascia together with the parietal peritoneum. The CO 2 source can be attached so that the gas helps to separate the layers, after which the colostomy trephine is formed at the site of the trocar, the grasper is inserted to gently deliver the blunt end of the descending colon through the canal and the end colostomy is formed in a usual way. Results No procedure-specific complications were noted in 39 patients who had laparoscopic APR with extraperitoneal retrotransversalis end colostomy from 2009 to 2016. In 23 patients who survived for 3.7 ± 1.7 years after surgery there were no clinical or CT signs of parastomal hernia or prolapse. Conclusion This single-institution retrospective case series demonstrates that laparoscopic extraperitoneal retrotransversalis end colostomy is feasible, safe and effective in preventing parastomal hernias and stomal prolapse.

© 2018 The Authors The main aim of the study is to determine the effectiveness and safety of lifting operations in women with varying degrees of involuntary changes of facial skin, in particular when applying pharmacological conditioning, with the objectification of the role of the latter. Materials and methods: A research and surgical treatment were conducted to eliminate involutional changes of various degrees in the facial skin of 461 women aged 35–75 years. Surface lifting was performed in 20.2% of patients, SMAS-lifting – 49.0%, SMAS-lifting with a three-level endoscopic assist lift of the lower face zone was performed in 30.8% of women. Before the surgery in 13.2% of cases, I degree of involutional changes in facial skin was registered, 47.9% – grade II, 38.9% – grade III. Patients were divided into two comparable groups. With the standard preparation without additional drug correction, 299 women (64.9%) were operated on in the preoperative period, they made up a comparison group. The main group included 162 (35.1%) women who underwent therapeutic conditioning before the lifting operation (Cytoflavin, n = 86; Actovegin n = 23; Ethylmethylhydroxypyridine succinate, n = 32; Meldonium, n = 21; Pentoxifylline, n = 31; Vinpocetine n = 27). Instrumental evaluation of the skin dermal microcirculation was performed using laser Doppler flowmetry and estimation of transcutaneous oxygen tension. In the blood plasma, the parameters of free radical processes (FRP) were studied. FRP were studied in terms of generation of active oxygen forms by leukocytes – intensity of chemiluminescence basal and intensity of chemiluminescence stimulated, as well as antiperoxide plasma activity and malondialdehyde. Early postoperative complications were analyzed, the number of repeated lifting surgical corrections on the face was studied for 5 years. Results: The role of FRP in the pathogenesis of involuntary changes in the facial skin has been established. The imbalance of FRP was expressed in the intensification of the reactive oxygen species generation and products of lipid peroxidation. This correlated with disorders of cutaneous microcirculation and a decrease in the saturation of the facial tissues with oxygen, manifested by an increasing energy deficit and the severity of involutional skin changes. The obtained data justify the expediency of using pharmacological conditioning with energy correcting antioxidant medicine. Preoperative conditioning allowed to reduce the number of early postlifting complications associated with tissue trophism in a quarter, especially during surface lifting. In addition, in the preconditioning group, the scar was more cosmetic already at the seventh day after the operation. Based on the study of postoperative catamnesis, self-assessment data and laboratory-instrumental methods of skin system examination in people of different ages, it was revealed that while using SMAS-lifting with a three-level endoscopic-assisted lifting of the lower part of the face, the lowest frequency of complications and the best 5-year effectiveness were established.

© 2018, Wuhan Institute of Virology, CAS and Springer Nature Singapore Pte Ltd. Enteric viruses are the most common cause of acute gastroenteritis (AGE) in young children and a significant public health problem globally. Hospital admissions of children under 5 years of age with diarrhea are primarily associated with group A rotavirus (RVA) infection. In this retrospective study, the population structure of viruses linked to AGE etiology in young children hospitalized with AGE in Moscow was evaluated, and molecular characterization of RVA strains was performed. Fecal specimens were collected from children under 5 years old hospitalized with AGE between 2009 and 2014 in Moscow, Russia. Multiplex real-time reverse transcription PCR was used to detect enteric viruses and for G/[P]-genotyping of isolated RVAs. Sequencing of RVA VP7 and VP4 cDNA fragments was used to validate the data obtained by PCR-genotyping. The main causes for hospitalization of children with AGE were RVA (40.1%), followed by noroviruses (11.4%), while adenoviruses, astroviruses, sapoviruses, enteroviruses, and orthoreoviruses were detected in 4.7%, 1.9%, 1.4%, 1.2%, and 0.2% of samples tested, respectively. Nosocomial infections, predominantly associated with RVAs and noroviruses, were detected in 24.8% of cases and occurred significantly more frequently in younger infants. The predominant RVA genotype was G4P[8], detected in 38.7% of RVA-positive cases, whereas genotypes G1P[8], G9P[8], G3P[8], and G2P[4] were found in 11.8%, 6.6%, 4.2%, and 3.3% of cases, respectively. Together, the presence of circulating RVA strains with rare VP7 and VP4 gene variants (G6 and P[9]) highlights the need to conduct continuous epidemiological monitoring of RVA infection.

© 2018 Elsevier B.V. Over the last decade, autophagy has emerged as one of the critical cellular systems that control homeostasis. Besides management of normal homeostatic processes, autophagy can also be induced by tissue damage stress or by rapidly progressing tumors. During tumor progression, autophagy mediates a cellular reaction to the changes inside and outside of cells, which leads to tumor adaptation. Even though the regulation of autophagy seems universal and is a well-described process, its dysregulation and role in glioma progression remain an important topic of investigation. In this review, we summarize recent evidence of autophagy regulation in brain tumor tissues and possible interconnection between signaling pathways that govern cellular responses. This perspective may help to assess the qualitative differences and various outcomes in response to autophagy stimulation.

© 2018, Springer Science+Business Media, LLC, part of Springer Nature. Arbitrary and involuntary movements were studied from the position of the new chaos—self-organization theory. Analysis of organization of tappingrams and tremorograms within the framework of N. A. Bernstein “repetition without repetitions” hypothesis leads to the Eskov—Zinchenko effect. In this case, there is no statistical stability for samples of parameters of electromyograms of any movements obtained in a row, i.e. fj(xi)≠fj+1(xi) with the probability p≥0.97. In this paper, the basic problem of motion physiology, stochastic instability establishment mechanisms, is solved in the analysis of muscle electromyogram parameters under conditions of permanent static force. Statistical instability of electromyograms within the framework of the Eskov—Zinchenko effect is proven.

© 2018, Pleiades Publishing, Ltd. Fibroblast growth factor (FGF) plays an important role in human embryogenesis, angiogenesis, cell proliferation, and differentiation. Carcinogenesis is accompanied by aberrant constitutive activation of FGF receptors (FGFRs) resulting from missense mutation in the FGFR1-4 genes, generation of chimeric oncogenes, FGFR1-4 gene amplification, alternative splicing shift toward formation of mesenchymal FGFR isoforms, and FGFR overexpression. Altogether, these alterations contribute to auto-and paracrine stimulation of cancer cells and neoangiogenesis. Certain missense mutations are found at a high rate in urinary bladder cancer and can be used for non-invasive cancer recurrence diagnostics by analyzing urine cell pellet DNA. Chimeric FGFR1/3 and amplified FGFR1/2 genes can predict cell response to the targeted therapy in various oncological diseases. In recent years, high-throughput sequencing has been used to analyze exomes of virtually all human tumors, which allowed to construct phylogenetic trees of clonal cancer evolution with special emphasis on driver mutations in FGFR1-4 genes. At present, FGFR blockers, such as multi-kinase inhibitors, specific FGFR inhibitors, and FGF ligand traps are being tested in clinical trials. In this review, we discuss current data on the functioning of the FGFR family proteins in both normal and cancer cells, mutations in the FGFR1-4 genes, and mechanisms underlying their oncogenic potential, which might be interesting to a broad range of scientists searching for specific tumor markers and targeted anti-cancer drugs.

© 2018, Pleiades Publishing, Ltd. Abstract: Mono- and diethylene oxide derivatives of tetra(p-hydroxyphenyl)porphyrin (THPP) with different conformation states are synthesized. These compounds exhibit high photosensitization activity in the generation of singlet oxygen in organic and aqueous (in the solubilized state) phases. It is shown that introducing ethylene oxide moieties into the hydroxyphenyl substituents of THPP increases its solubility in chloroform. In addition, the activity of singlet oxygen 1Δg generation in the reaction of anthracene photooxidation by THPP tetra derivatives in chloroform, where the ethylene oxide fragments are introduced into two phenyl rings and hexadecyl fragments are introduced into another two, is higher than the activity of mono- and di-modified THPP molecules with enthylene oxide molecules at one hydroxyphenyl cycle. The activity of tetra-substituted porphyrin in chloroform, however, is comparable to that of nonsubstituted tetraphenylporphyrin, considered to be one of the most active photosensitizers. The produced ethylene oxide derivatives of THPP are solubilized with pluronic F-127 (triblock copolymer of ethylene- and propylene-oxides)—one of the least toxic and effective polymeric detergents—forming water-soluble forms of the respective porphyrins. It is established that the pluronic solubilization capability (the lowest molar concentration of pluronic required for the complete transfer of porphyrin of a particular molar concentration dissolved in organic phase to the water-soluble form) is higher for asymmetrical mono- and di-derivatives of the THPP than for symmetric tetra-substituted THPP. It is shown that the activity of the solubilized water-soluble form of mono- and tetra-derivatives in tryptophan photoxidation is higher than that of unsubstituted THPP and is comparable to the activity of tetraphenylporphyrin.

© 2018 Elsevier B.V. The complex of mitochondrial protein cytochrome c (CytC) with anionic phospholipid cardiolipin (CL) plays a crucial role in the initiation of apoptosis by catalyzing lipid peroxidation in mitochondrial membranes. In our previous papers, we found that CytC and CL mixed in millimolar concentrations form a sediment showing microcrystals composed of nanospheres (Cyt-CL) of 11–12 and 8 nm in diameter. The hypothesis was proposed that Cyt-CL, having hydrophobic shell, may appear inside the membrane lipid bilayer in mitochondria and peroxidase membrane phospholipids so initiating the apoptotic cascade. In this work, Cyt-CL complex dissolved in chloroform or hexane was investigated as a model of the complex in mitochondrial membranes. We used dynamic light scattering method to measure the size of the particles. The analysis of particles size distribution of Cyt-CL in chloroform allows to reveal three dominant diameters of 12.1 ± 1.4, 7.8 ± 1.0, and 4.7 ± 0.7 nm. The first two values are closed to those, earlier obtained with small-angle X-ray scattering method in Cyt-CL microcrystals, 11.1 ± 1.0 and 8.0 ± 0.7 nm. CL extracted in chloroform-methanol forms a real solution of particles with diameter of 0.7 ± 0.1 nm. In methanol-water phase, CL and CL + CytC mixture form particles of 83.7 ± 9.8 and 71.3 ± 11.6 nm, respectively. Apparently, cardiolipin in 50% methanol forms single-layer liposomes regardless of the presence of CytC in the medium. Partial unfolding of CytC in the complex was evidenced by (a) appearance of fluorescence of tyrosine and tryptophan residues and (b) disappearance of the absorption band at 699 nm due to breakdown of heme iron – methionine bond > F⋯S(Met80). In hydrophobic solvent Cyt-CL exhibited quasi-lipoperoxidase and lipoxygenase activity as was shown in kinetic measurements of chemiluminescence enhanced by coumarin C-525, a selective sensitizer of chemiluminescence, associated with reactions of lipid peroxyl radicals. Our data in this model system do not contradict the hypothesis (Vladimirov, Y.A. et al. Biochemistry (Mosc) 78, 1086–1097) that nanospheres of Cyt-CL complex, embedded into the lipid phase of mitochondrial membrane, catalyze lipid peroxidation, thereby initiating apoptosis.

© 2017 Elsevier B.V. Background: A major role of the innate immunity in the defence of mucosal tissue is well established. However, a balance between the main components of the immunity such as toll-like receptors (TLRs) and defensins in the pathology of upper respiratory tract in children has not been addressed yet. Our aim was to investigate the gene expression of some TLRs as well as alpha and beta-defensins in children suffered from adenoid hyperthrophy in comparison with healthy children. Methods: Samples (nasal epithelium and adenoids) from patients with hypertrophic adenoids (n = 77) and control group (n = 33) were investigated. Quantification of HBD-1 and 2 mRNA, alpha-defensin-HNP1 and toll-like receptors (TLR) 2, 4 and 9 mRNA expression was performed by real-time polymerase chain reaction (PCR). The detection of TLR4 and TLR9 was performed by immunohistochemistry. Results: The main finding of the study is a dramatic up-regulation of TLR2 and TLR4 expression (but down-regulation of TLR9) along with a significant reduction in the expression of the defensins in children with adenoid hyperthrophy. Conclusion: The data suggest that one of the mechanisms of mucosal involvement in the pathogenesis of upper respiratory tract infection might by a disbalance between TLRs and defensins revealed in our study.

© 2018, Pleiades Publishing, Inc. The COI gene fragment was first examined in representatives of the family Celyphidae. The presence of a considerable hiatus between interspecific and intraspecific distances was demonstrated, which enabled using the barcoding method to distinguish species of the family Celyphidae. The results of the analysis showed that different species of Celyphidae clustered together. Within the Celyphus (Hemiglobus) porosus cluster, some haplotype heterogeneity, which was, however, within the limits of intraspecific variability, was observed.

©2018 by the authors. Licensee MDPI, Basel, Switzerland. Engineering of a “smart” drug delivery system to specifically target tumour cells has been at the forefront of cancer research, having been engineered for safer, more efficient and effective use of chemotherapy for the treatment of cancer. However, selective targeting and choosing the right cancer surface biomarker are critical for a targeted treatment to work. Currently, the available delivery systems use a two-dimensional monolayer of cancer cells to test the efficacy of the drug delivery system, but designing a “smart” drug delivery system to be specific for a tumour in vivo and to penetrate the inner core remains a major design challenge. These challenges can be overcome by using a study model that integrates the three-dimensional aspect of a tumour in a culture system. Here, we tested the efficacy of a functionalized folic acid-conjugated amphiphilic alternating copolymer poly(styrene-alt-maleic anhydride) (FA-DABA-SMA) via a biodegradable linker 2,4-diaminobutyric acid (DABA) to specifically target and penetrate the inner core of three-dimensional avascular human pancreatic and breast tumour spheroids in culture. The copolymer was quantitatively analyzed for its hydrophobic drug encapsulation efficiency using three different chemical drug structures with different molecular weights. Their release profiles and tumour targeting properties at various concentrations and pH environments were also characterized. Using the anticancer drug curcumin and two standard clinical chemotherapeutic hydrophobic drugs, paclitaxel and 5-fluorouracil, we tested the ability of FA-DABA-SMA nanoparticles to encapsulate the differently sized drugs and deliver them to kill monolayer pancreatic cancer cells using the WST-1 cell proliferation assay. The findings of this study revealed that the functionalized folic acid-conjugated amphiphilic alternating copolymer shows unique properties as an active “smart” tumor-targeting drug delivery system with the ability to internalize hydrophobic drugs and release the chemotherapeutics for effective killing of cancer cells. The novelty of the study is the first to demonstrate a functionalized “smart” drug delivery system encapsulated with a hydrophobic drug effectively targeting and penetrating the inner core of pancreatic and breast cancer spheroids and reducing their volumes in a dose-and time-dependent manner.

© 2018 Astro Ltd. Lasers offer new possibilities in the treatment of such widespread diseases as osteoarthritis, with both direct and indirect effects on cell metabolism. Cyclic hydrostatic pressure is one of the main natural stimuli of cartilage chondrocytes. The present work shows that hydrostatic stimulation with magnitudes of up to 20 MPa can be realized locally through infrared impact on the neighboring media of chondrocytes. We compare indirect (thermomechanical, λ = 1560 nm) and direct (photo-modulation, λ1 = 1560 nm, λ2 = 670 nm) laser effects on the synthetic activity of chondrocytes in cultures within a 1 min exposure time limit, to study separately the photo-modulation and thermomechanical components of laser impact. The chondrocyte activity was monitored by immunohistochemical analysis in normoxic and hypoxic conditions. Collagen II and proteoglycan accumulation increased significantly (up to 70%) after a pulsed thermomechanical laser impact. Thermomechanical laser irradiation showed the more pronounced stimulation in both normoxic and hypoxic conditions, while the effect of photo-modulation was inhibited by oxygen concentration increase. Theoretical calculations of the laser-induced temperature and stress fields show that the spreading of the stress field with a maximum at 19.2 MPa is approximately three times greater than that of appreciable (>1 °C) heating. Thus, thermomechanical infrared stimulation of chondrocytes can be a perspective method for the restoration of hyaline-type cartilage.

© 2018 Elsevier Inc. Growing evidence demonstrates that the serotonin system influences punishment behavior in social decision-making and that individual differences in the propensity to punish are, at least in part, due to genetic variation. However, the specific genes and their mechanisms by which they influence punishment behavior are not yet fully characterized. Here, we examined whether serotonin system-related gene variation impacts on altruistic punishment in the ultimatum game by using a longitudinal approach with three time points, covering a time frame up to four months in young adults (N = 106). Specifically, we investigated additive effects of 5-HTTLPR and TPH2 G-703T genotypes by using a composite score. This composite score was significantly associated with altruistic punishment, with individuals carrying both the S-allele and the G-allele demonstrating less punishment behavior. The results suggest that serotonin system-related gene variation contributes to individual differences in altruistic punishment. Furthermore, comparably high test-retest correlations suggest that punishment behavior in the ultimatum game represents a relatively stable, trait-like behavior.

© 2018 American Society for Microbiology. The tick-borne pathogen Borrelia burgdorferi is responsible for approximately 300,000 Lyme disease (LD) cases per year in the United States. Recent increases in the number of LD cases, in addition to the spread of the tick vector and a lack of a vaccine, highlight an urgent need for designing and developing an efficacious LD vaccine. Identification of protective epitopes that could be used to develop a second-generation (subunit) vaccine is therefore imperative. Despite the antigenicity of several lipoproteins and integral outer membrane proteins (OMPs) on the B. burgdorferi surface, the spirochetes successfully evade antibodies primarily due to the VlsE-mediated antigenic variation. VlsE is thought to sterically block antibody access to protective epitopes of B. burgdorferi. However, it is highly unlikely that VlsE shields the entire surface epitome. Thus, identification of subdominant epitope targets that induce protection when they are made dominant is necessary to generate an efficacious vaccine. Toward the identification, we repeatedly immunized immunocompetent mice with live-attenuated VlsE-deleted B. burgdorferi and then challenged the animals with the VlsE-expressing (host-adapted) wild type. Passive immunization and Western blotting data suggested that the protection of 50% of repeatedly immunized animals against the highly immune-evasive B. burgdorferi was antibody mediated. Comparison of serum antibody repertoires identified in protected and nonprotected animals permitted the identification of several putative epitopes significantly associated with the protection. Most linear putative epitopes were conserved between the main pathogenic Borrelia genospecies and found within known subdominant regions of OMPs. Currently, we are performing immunization studies to test whether the identified protection-associated epitopes are protective for mice.