Репозиторий Университета

Mechanisms of homeostatic plasticity promote compensatory changes of cellular excitability in response to chronic changes in the network activity. This type of plasticity is essential for the maintenance of brain circuits and is involved in the regulation of neural regeneration and the progress of neurodegenerative disorders. One of the most studied homeostatic processes is synaptic scaling, where global synaptic adjustments take place to restore the neuronal firing rate to a physiological range by the modulation of synaptic receptors, neurotransmitters, and morphology. However, despite the comprehensive literature on the electrophysiological properties of homeostatic scaling, less is known about the structural adjustments that occur in the synapses and dendritic tree. In this study, we performed a meta-analysis of articles investigating the effects of chronic network excitation (synaptic downscaling) or inhibition (synaptic upscaling) on the dendritic spine density of neurons. Our results indicate that spine density is consistently reduced after protocols that induce synaptic scaling, independent of the intervention type. Then, we discuss the implication of our findings to the current knowledge on the morphological changes induced by homeostatic plasticity.

© 2021, The Author(s). Background: The acute phase response (APR) to CNS insults contributes to the overall magnitude and nature of the systemic inflammatory response. Aspects of this response are thought to drive secondary inflammatory pathology at the lesion site, and suppression of the APR can therefore afford some neuroprotection. In this study, we examined the APR in a mouse model of traumatic spinal cord injury (SCI), along with its relationship to neutrophil recruitment during the immediate aftermath of the insult. We specifically investigated the effect of IL-1 receptor antagonist (IL-1RA) administration on the APR and leukocyte recruitment to the injured spinal cord. Methods: Adult female C57BL/6 mice underwent either a 70kD contusive SCI, or sham surgery, and tissue was collected at 2, 6, 12, and 24 hours post-operation. For IL-1RA experiments, SCI mice received two intraperitoneal injections of human IL-1RA (100mg/kg), or saline as control, immediately following, and 5 hours after impact, and animals were sacrificed 6 hours later. Blood, spleen, liver and spinal cord were collected to study markers of central and peripheral inflammation by flow cytometry, immunohistochemistry and qPCR. Results were analysed by two-way ANOVA or student’s t-test, as appropriate. Results: SCI induced a robust APR, hallmarked by elevated hepatic expression of pro-inflammatory marker genes and a significantly increased neutrophil presence in the blood, liver and spleen of these animals, as early as 2 hours after injury. This peripheral response preceded significant neutrophil infiltration of the spinal cord, which peaked 24 hours post-SCI. Although expression of IL-1RA was also induced in the liver following SCI, its response was delayed compared to IL-1β. Exogenous administration of IL-1RA during this putative therapeutic window was able to suppress the hepatic APR, as evidenced by a reduction in CXCL1 and SAA-2 expression as well as a significant decrease in neutrophil infiltration in both the liver and the injured spinal cord itself. Conclusions: Our data indicate that peripheral administration of IL-1RA can attenuate the APR which in turn reduces immune cell infiltration at the spinal cord lesion site. We propose IL-1RA treatment as a viable therapeutic strategy to minimise the harmful effects of SCI-induced inflammation.

The family of Tribbles proteins play many critical nonenzymatic roles and regulate a wide range of key signaling pathways. Tribbles homolog 2 (Trib2) is a pseudo serine/threonine kinase that functions as a scaffold or adaptor in various physiological and pathological processes. Trib2 can interact with E3 ubiquitin ligases and control protein stability of downstream effectors. This protein is induced by mitogens and enhances the propagation of several cancer cells, including myeloid leukemia, liver, lung, skin, bone, brain, and pancreatic. Thus, Trib2 can be a predictive and valuable biomarker for the diagnosis and treatment of cancer. Recent studies have illustrated that Trib2 plays a major role in cell fate determination of stem cells. Stem cells have the capacity to self-renew and differentiate into specific cell types. Stem cells are important sources for cell-based regenerative medicine and drug screening. Trib2 has been found to increase the self-renewal ability of embryonic stem cells, the reprogramming efficiency of somatic cells, and chondrogenesis. In this review, we will focus on the recent advances of Trib2 function in tumorigenesis and stem cell fate decisions. [MediaObject not available: see fulltext.]

Recently, mesenchymal stem/stromal cells (MSCs) and their widespread biomedical applications have attracted great consideration from the scientific community around the world. However, reports have shown that the main populations of the transplanted MSCs are trapped in the liver, spleen, and lung upon administration, highlighting the importance of the development of cell-free therapies. Concerning rising evidence suggesting that the beneficial effects of MSC therapy are closely linked to MSC-released components, predominantly MSC-derived exosomes, the development of an MSC-based cell-free approach is of paramount importance. The exosomes are nano-sized (30–100 nm) lipid bilayer membrane vesicles, which are typically released by MSCs and are found in different body fluids. They include various bioactive molecules, such as messenger RNA (mRNA), microRNAs, proteins, and bioactive lipids, thus showing pronounced therapeutic competence for tissues recovery through the maintenance of their endogenous stem cells, the enhancement of regenerative phenotypic traits, inhibition of apoptosis concomitant with immune modulation, and stimulation of the angiogenesis. Conversely, the specific roles of MSC exosomes in the treatment of various tumors remain challenging. The development and clinical application of novel MSC-based cell-free strategies can be supported by better understanding their mechanisms, classifying the subpopulation of exosomes, enhancing the conditions of cell culture and isolation, and increasing the production of exosomes along with engineering exosomes to deliver drugs and therapeutic molecules to the target sites. In the current review, we deliver a brief overview of MSC-derived exosome biogenesis, composition, and isolation methods and discuss recent investigation regarding the therapeutic potential of MSC exosomes in regenerative medicine accompanied by their double-edged sword role in cancer.

Recently, mesenchymal stem/stromal cells (MSCs) due to their pro-angiogenic, anti-apoptotic, and immunoregulatory competencies along with fewer ethical issues are presented as a rational strategy for regenerative medicine. Current reports have signified that the pleiotropic effects of MSCs are not related to their differentiation potentials, but rather are exerted through the release of soluble paracrine molecules. Being nano-sized, non-toxic, biocompatible, barely immunogenic, and owning targeting capability and organotropism, exosomes are considered nanocarriers for their possible use in diagnosis and therapy. Exosomes convey functional molecules such as long non-coding RNAs (lncRNAs) and micro-RNAs (miRNAs), proteins (e.g., chemokine and cytokine), and lipids from MSCs to the target cells. They participate in intercellular interaction procedures and enable the repair of damaged or diseased tissues and organs. Findings have evidenced that exosomes alone are liable for the beneficial influences of MSCs in a myriad of experimental models, suggesting that MSC- exosomes can be utilized to establish a novel cell-free therapeutic strategy for the treatment of varied human disorders, encompassing myocardial infarction (MI), CNS-related disorders, musculoskeletal disorders (e.g. arthritis), kidney diseases, liver diseases, lung diseases, as well as cutaneous wounds. Importantly, compared with MSCs, MSC- exosomes serve more steady entities and reduced safety risks concerning the injection of live cells, such as microvasculature occlusion risk. In the current review, we will discuss the therapeutic potential of MSC- exosomes as an innovative approach in the context of regenerative medicine and highlight the recent knowledge on MSC- exosomes in translational medicine, focusing on in vivo researches.

Recently, mesenchymal stem/stromal cells (MSCs) due to their pro-angiogenic, anti-apoptotic, and immunoregulatory competencies along with fewer ethical issues are presented as a rational strategy for regenerative medicine. Current reports have signified that the pleiotropic effects of MSCs are not related to their differentiation potentials, but rather are exerted through the release of soluble paracrine molecules. Being nano-sized, non-toxic, biocompatible, barely immunogenic, and owning targeting capability and organotropism, exosomes are considered nanocarriers for their possible use in diagnosis and therapy. Exosomes convey functional molecules such as long non-coding RNAs (lncRNAs) and micro-RNAs (miRNAs), proteins (e.g., chemokine and cytokine), and lipids from MSCs to the target cells. They participate in intercellular interaction procedures and enable the repair of damaged or diseased tissues and organs. Findings have evidenced that exosomes alone are liable for the beneficial influences of MSCs in a myriad of experimental models, suggesting that MSC- exosomes can be utilized to establish a novel cell-free therapeutic strategy for the treatment of varied human disorders, encompassing myocardial infarction (MI), CNS-related disorders, musculoskeletal disorders (e.g. arthritis), kidney diseases, liver diseases, lung diseases, as well as cutaneous wounds. Importantly, compared with MSCs, MSC- exosomes serve more steady entities and reduced safety risks concerning the injection of live cells, such as microvasculature occlusion risk. In the current review, we will discuss the therapeutic potential of MSC- exosomes as an innovative approach in the context of regenerative medicine and highlight the recent knowledge on MSC- exosomes in translational medicine, focusing on in vivo researches.

© 2021 Elsevier Ltd This review examines investigations into the functionalization of polysaccharides by substituents containing multiple (C[dbnd]C) bonds and thiol (SH) groups that are prone to (co)polymerization in the presence of thermal, redox and photoinitiators or Michael addition reactions. A comparative analysis of the approaches to grafting the mentioned substituents onto the polysaccharide macromolecules was conducted. The use of the modified polysaccharides for the design of the 3D structures, including for the development of the pore bearing matrixes of cells or scaffolds utilized in regenerative medicine was examined. These modified polymers were also examined toward the design of excipient matrixes in pharmacological compositions, including with controllable release of active pharmaceuticals, as wel as of antibacterial and antifungal agents and others. In addition, a few examples of the use of modified derivatives in other areas are given.

© 2020 Elsevier GmbH Wad Medani virus (WMV) belongs to the genus Orbivirus and is a poorly studied arbovirus with unclear medical significance. Presently, a limited number of WMV strains are characterized and available in NCBI GenBank, some isolated many years ago. A new WMV strain was isolated in 2012 from Dermacentor nuttalli ticks collected from sheep in the Tuva Republic, Russia, and sequenced using high-throughput methods. Complete coding sequences were obtained revealing signs of multiple intersegment reassortments. These point to a high variability potential in WMV that may lead to the formation of strains with novel properties. These new data on WMV can promote better understanding of: ecological features of its circulation; relationships within the genus Orbivirus; and the medical significance of the virus.

© 2020 Elsevier GmbH Wad Medani virus (WMV) belongs to the genus Orbivirus and is a poorly studied arbovirus with unclear medical significance. Presently, a limited number of WMV strains are characterized and available in NCBI GenBank, some isolated many years ago. A new WMV strain was isolated in 2012 from Dermacentor nuttalli ticks collected from sheep in the Tuva Republic, Russia, and sequenced using high-throughput methods. Complete coding sequences were obtained revealing signs of multiple intersegment reassortments. These point to a high variability potential in WMV that may lead to the formation of strains with novel properties. These new data on WMV can promote better understanding of: ecological features of its circulation; relationships within the genus Orbivirus; and the medical significance of the virus.

© 2020 Purpose: To evaluate the density and distribution of conjunctival goblet cells in mice without clinical evidence of ocular surface diseases. Methods: Immediately after euthanasia of C57BL/6 wild-type mice, the eyes including eyelids were removed and fixed in paraformaldehyde. Entire eyeballs and eyelids were cut in series along the sagittal axis from nasal to temporal on a microtome and then stained with Periodic Acid-Schiff acid to visualize the goblet cells. At each section stained in this way, the conjunctival goblet cells of the entire upper and lower lid conjunctiva were counted by light microscopy. Additional (transmission electron microscopy) (TEM)-Analysis on ultrathin sections was performed to evaluate morphological differences. Results: The total number of conjunctival goblet cells differs markedly between individual animals. Categorisation into upper eyelid (UL) and lower eyelid (LL) and into regions (nasal, middle, temporal) revealed a significant increase of goblet cells from nasal to temporal in the UL and a significant decrease in the LL. Conclusion: The distribution of conjunctival goblet cells in mice differs considerably from humans and between individual animals. Therefore, precise selection of sampling and methods are needed to obtain comparable data. We recommend to use the middle region of the conjunctiva of UL/LL for goblet cell studies in mice. These findings are of particular interest for dry eye mouse models as well as pharmacological studies on mice with influence on their goblet cells.

© 2021 Elsevier B.V. The Astrakhan region of Russia is endemic for the number of arboviruses. In this paper, we describe the results of the detection of the list of neglected arboviruses in the Astrakhan region for the 2018 season. For the purpose of the study in-house PCR assays for detection of 18 arboviruses have been developed and validated using arboviruses obtained from Russian State Collection of Viruses. Pools of ticks (n = 463) and mosquitoes (n = 312) as well as 420 samples of human patients sera have been collected and analyzed. Using developed multiplex real-time PCR assays we were able to detect RNA of eight arboviruses (Crimean-Congo hemorrhagic fever virus, Dhori (Batken strain) virus, Batai virus, Tahyna virus, Uukuniemi virus, Inkoo virus, Sindbis virus and West Nile fever virus). All discovered viruses are capable of infecting humans causing fever and in some cases severe forms with hemorrhagic or neurologic symptoms. From PCR-positive samples, we were able to recover one isolate each of Dhori (Batken strain) virus and Crimean-Congo hemorrhagic fever virus which were further characterized by next-generation sequencing. The genomic sequences of identified Dhori (Batken strain) virus strain represent the most complete genome of Batken virus strain among previously reported.

© 2020 Elsevier Ltd COVID-19, coronavirus disease 2019, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a pandemic. At the time of writing this (October 14, 2020), more than 38.4 million people have become affected, and 1.0 million people have died across the world. The death rate is undoubtedly correlated with the cytokine storm and other pathological pulmonary characteristics, as a result of which the lungs cannot provide sufficient oxygen to the body's vital organs. While diversified drugs have been tested as a first line therapy, the complexity of fatal cases has not been reduced so far, and the world is looking for a treatment to combat the virus. However, to date, and despite such promise, we have received very limited information about the potential of nanomedicine to fight against COVID-19 or as an adjunct therapy in the treatment regimen. Over the past two decades, various therapeutic strategies, including direct-acting antiviral drugs, immunomodulators, a few non-specific drugs (simple to complex), have been explored to treat Acute Respiratory Distress Syndrome (ARDS), Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS), influenza, and sometimes the common flu, thus, correlating and developing specific drugs centric to COVID-19 is possible. This review article focuses on the pulmonary pathology caused by SARS-CoV-2 and other viral pathogens, highlighting possible nanomedicine therapeutic strategies that should be further tested immediately.

© 2020 Elsevier Ltd COVID-19, coronavirus disease 2019, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a pandemic. At the time of writing this (October 14, 2020), more than 38.4 million people have become affected, and 1.0 million people have died across the world. The death rate is undoubtedly correlated with the cytokine storm and other pathological pulmonary characteristics, as a result of which the lungs cannot provide sufficient oxygen to the body's vital organs. While diversified drugs have been tested as a first line therapy, the complexity of fatal cases has not been reduced so far, and the world is looking for a treatment to combat the virus. However, to date, and despite such promise, we have received very limited information about the potential of nanomedicine to fight against COVID-19 or as an adjunct therapy in the treatment regimen. Over the past two decades, various therapeutic strategies, including direct-acting antiviral drugs, immunomodulators, a few non-specific drugs (simple to complex), have been explored to treat Acute Respiratory Distress Syndrome (ARDS), Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS), influenza, and sometimes the common flu, thus, correlating and developing specific drugs centric to COVID-19 is possible. This review article focuses on the pulmonary pathology caused by SARS-CoV-2 and other viral pathogens, highlighting possible nanomedicine therapeutic strategies that should be further tested immediately.

© 2020 Epithelial-to-mesenchymal transition (EMT) is an essential molecular and cellular process that is part of normal embryogenesis and wound healing, and also has a ubiquitous role in various types of carcinoma and glioblastoma. EMT is activated and regulated by specific microenvironmental endogenous triggers and a complex network of signalling pathways. These mostly include epigenetic events that affect protein translation-controlling factors and proteases, altogether orchestrated by the switching on and off of oncogenes and tumour-suppressor genes in cancer cells. The hallmark of cancer-linked EMT is that the process is incomplete, as it is opposed by the reverse process of mesenchymal-to-epithelial transition, which results in a hybrid epithelial/mesenchymal phenotype that shows notable cell plasticity. This is a characteristic of cancer stem cells (CSCs), and it is of the utmost importance in their niche microenvironment, where it governs CSC migratory and invasive properties, thereby creating metastatic CSCs. These cells have high resistance to therapeutic treatments, in particular in glioblastoma.

© 2020 Elsevier B.V. In this work, we study the effect of RF magnetron oblique angle deposition (OAD) on morphology, structure, and elemental composition of as-deposited and heat-treated Cu containing calcium phosphates. The control over the surface morphology and nano roughness provided by OAD is of great interest as both Mesenchymal Stem Cells and various types of bacteria respond strongly to nanoscale topography. A Cu substituted hydroxyapatite target was used to deposit coatings on the surface of titanium (Ti) and silicon (Si) substrates. The samples were placed at an oblique angle of 80° relative to the surface of the sample holder and in a normal configuration with respect to the flux direction and, therefore, parallel to the target. The dense homogeneous coatings with globular surface features deposited at normal flux incidence (NFI) configuration changed to elliptical, highly oriented structures with the direction dictated by the atomic shadowing effect when the substrate was deposited at an oblique angle. As-deposited thin films were subjected to post-deposition-heat-treatment at 700 °C in an Ar atmosphere. This led to a drastic change in the surface morphology and, namely, lost the directionality of the nanostructures. According to the X-ray diffraction data, the samples deposited obliquely showed preferential growth in the (002) plane and lower internal stress, than samples coated at NFI for both the Si and Ti substrates. The RMS roughness of the films deposited obliquely on Si was twice that of the films deposited at NFI (860 ± 80 pm and 408 ± 60 pm, respectively). However, it was not the case for the Ti substrate, the RMS roughness decreased from 42 ± 4 nm for coatings deposited at normal flux geometry to 33 ± 2 nm for coatings deposited obliquely. The heat-treatment of the samples deposited at 80° resulted in a significant increase in the surface roughness: 8 ± 0.7 nm for Si and 71 ± 4 nm for Ti substrates. The obtained results demonstrate that the oblique angle deposition can be used to fabricate nano-rough surface morphologies.

© 2020 Elsevier B.V. In this work, we study the effect of RF magnetron oblique angle deposition (OAD) on morphology, structure, and elemental composition of as-deposited and heat-treated Cu containing calcium phosphates. The control over the surface morphology and nano roughness provided by OAD is of great interest as both Mesenchymal Stem Cells and various types of bacteria respond strongly to nanoscale topography. A Cu substituted hydroxyapatite target was used to deposit coatings on the surface of titanium (Ti) and silicon (Si) substrates. The samples were placed at an oblique angle of 80° relative to the surface of the sample holder and in a normal configuration with respect to the flux direction and, therefore, parallel to the target. The dense homogeneous coatings with globular surface features deposited at normal flux incidence (NFI) configuration changed to elliptical, highly oriented structures with the direction dictated by the atomic shadowing effect when the substrate was deposited at an oblique angle. As-deposited thin films were subjected to post-deposition-heat-treatment at 700 °C in an Ar atmosphere. This led to a drastic change in the surface morphology and, namely, lost the directionality of the nanostructures. According to the X-ray diffraction data, the samples deposited obliquely showed preferential growth in the (002) plane and lower internal stress, than samples coated at NFI for both the Si and Ti substrates. The RMS roughness of the films deposited obliquely on Si was twice that of the films deposited at NFI (860 ± 80 pm and 408 ± 60 pm, respectively). However, it was not the case for the Ti substrate, the RMS roughness decreased from 42 ± 4 nm for coatings deposited at normal flux geometry to 33 ± 2 nm for coatings deposited obliquely. The heat-treatment of the samples deposited at 80° resulted in a significant increase in the surface roughness: 8 ± 0.7 nm for Si and 71 ± 4 nm for Ti substrates. The obtained results demonstrate that the oblique angle deposition can be used to fabricate nano-rough surface morphologies.

© 2020 Elsevier B.V. Alprazolam is used in the treatment of patients with anxiety disorders comorbid with alcohol use disorder. Some proportion of these patients does not respond adequately to treatment with alprazolam, while many of them experience dose-dependent adverse drug reactions. Results of the previous studies have shown that CYP3A is involved in the biotransformation of alprazolam, the activity of which is dependent, inter alia, on the polymorphism of the encoding gene. Objective: The objective of our study was to investigate the effect of 99366316G>A polymorphism of the CYP3A4 gene on the concentration/dose indicator of alprazolam in patients with anxiety disorders comorbid with alcohol use disorder, using findings on enzymatic activity of CYP3A (as evaluated by the 6-beta-hydroxy-cortisol/cortisol ratio measurement) and on CYP3A4 expression level obtained by measuring the miR-27b plasma concentration levels in patients with anxiety disorders comorbid with alcoholism. Material and methods: Our study enrolled 105 patients with anxiety disorders comorbid with alcohol use disorder (age - 37.8±14.6 years). Therapy included alprazolam in an average daily dose of 5.6±2.4 mg per day. Treatment efficacy was evaluated using the psychometric scales. Therapy safety was assessed using the UKU Side-Effect Rating Scale. For genotyping and estimation of the microRNA (miRNA) plasma levels, we performed the real-time polymerase chain reaction. The activity of CYP3A was evaluated using the HPLC-MS/MS method by the content of the endogenous substrate of the given isoenzyme and its metabolite in urine (6- beta-hydroxy-cortisol/cortisol). Therapeutic drug monitoring (TDM) has been performed using HPLC-MS/MS. Results: Our study revealed the statistically significant results in terms of the treatment efficacy evaluation (HAMA scores at the end of the treatment course): (GG) 3.0 [2.0; 5.0] and (GA) 4.0 [4.0; 5.0], p = 0.007; at the same time, the statistical significance in the safety profile was not obtained (the UKU scores): (GG) 3.0 [2.0; 3.8] and (GA) 3.0 [1.5; 4.0], p = 0.650. We revealed a statistical significance for concentration/dose indicator of alprazolam in patients with different genotypes: (GG) 1.583 [0.941; 2.301] and (GA) 2.888 [2.305; 4.394], p = 0.001). Analysis of the results of the pharmacotranscriptomic part of the study didn't show the statistically significant difference in the miR-27b plasma levels in patients with different genotypes: (GG) 25.6 [20.4; 28.8], (GA) 25.7 [19.7; 33.1], p = 0.423. At the same time, correlation analysis revealed a statistically significant relationship between the alprazolam efficacy profile evaluated by changes in HAMA scale scores and the miR-27b plasma concentration: rs = 0.20, p = 0.042. Also, we didn't reveal the correlation between the miRNA concentration and safety profile: rs = 0.15, p = 0.127. In addition, we revealed the relationship between the CYP3A enzymatic activity (as evaluated by 6-beta-hydroxycortisol/ cortisol ratio measurement) and the miR-27b plasma concentration: rs = −0.27, p = 0.006. At the same time, correlation analysis revealed a statistically significant relationship between the alprazolam concentration and the miR-27b plasma concentration: rs = 0.28, p = 0.003. Conclusion: The effect of genetic polymorphism of the CYP3A4 gene on the efficacy and safety profiles of alprazolam was demonstrated in a group of 105 patients with anxiety disorders comorbid with alcohol use disorder. At the same time, miR-27b remains a promising biomarker for assessing the level of CYP3A4 expression, because it correlates with the encoded isoenzyme's activity.

© 2020 Elsevier B.V. Alprazolam is used in the treatment of patients with anxiety disorders comorbid with alcohol use disorder. Some proportion of these patients does not respond adequately to treatment with alprazolam, while many of them experience dose-dependent adverse drug reactions. Results of the previous studies have shown that CYP3A is involved in the biotransformation of alprazolam, the activity of which is dependent, inter alia, on the polymorphism of the encoding gene. Objective: The objective of our study was to investigate the effect of 99366316G>A polymorphism of the CYP3A4 gene on the concentration/dose indicator of alprazolam in patients with anxiety disorders comorbid with alcohol use disorder, using findings on enzymatic activity of CYP3A (as evaluated by the 6-beta-hydroxy-cortisol/cortisol ratio measurement) and on CYP3A4 expression level obtained by measuring the miR-27b plasma concentration levels in patients with anxiety disorders comorbid with alcoholism. Material and methods: Our study enrolled 105 patients with anxiety disorders comorbid with alcohol use disorder (age - 37.8±14.6 years). Therapy included alprazolam in an average daily dose of 5.6±2.4 mg per day. Treatment efficacy was evaluated using the psychometric scales. Therapy safety was assessed using the UKU Side-Effect Rating Scale. For genotyping and estimation of the microRNA (miRNA) plasma levels, we performed the real-time polymerase chain reaction. The activity of CYP3A was evaluated using the HPLC-MS/MS method by the content of the endogenous substrate of the given isoenzyme and its metabolite in urine (6- beta-hydroxy-cortisol/cortisol). Therapeutic drug monitoring (TDM) has been performed using HPLC-MS/MS. Results: Our study revealed the statistically significant results in terms of the treatment efficacy evaluation (HAMA scores at the end of the treatment course): (GG) 3.0 [2.0; 5.0] and (GA) 4.0 [4.0; 5.0], p = 0.007; at the same time, the statistical significance in the safety profile was not obtained (the UKU scores): (GG) 3.0 [2.0; 3.8] and (GA) 3.0 [1.5; 4.0], p = 0.650. We revealed a statistical significance for concentration/dose indicator of alprazolam in patients with different genotypes: (GG) 1.583 [0.941; 2.301] and (GA) 2.888 [2.305; 4.394], p = 0.001). Analysis of the results of the pharmacotranscriptomic part of the study didn't show the statistically significant difference in the miR-27b plasma levels in patients with different genotypes: (GG) 25.6 [20.4; 28.8], (GA) 25.7 [19.7; 33.1], p = 0.423. At the same time, correlation analysis revealed a statistically significant relationship between the alprazolam efficacy profile evaluated by changes in HAMA scale scores and the miR-27b plasma concentration: rs = 0.20, p = 0.042. Also, we didn't reveal the correlation between the miRNA concentration and safety profile: rs = 0.15, p = 0.127. In addition, we revealed the relationship between the CYP3A enzymatic activity (as evaluated by 6-beta-hydroxycortisol/ cortisol ratio measurement) and the miR-27b plasma concentration: rs = −0.27, p = 0.006. At the same time, correlation analysis revealed a statistically significant relationship between the alprazolam concentration and the miR-27b plasma concentration: rs = 0.28, p = 0.003. Conclusion: The effect of genetic polymorphism of the CYP3A4 gene on the efficacy and safety profiles of alprazolam was demonstrated in a group of 105 patients with anxiety disorders comorbid with alcohol use disorder. At the same time, miR-27b remains a promising biomarker for assessing the level of CYP3A4 expression, because it correlates with the encoded isoenzyme's activity.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

© 2019 Elsevier B.V. The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.