Репозиторий Университета

© 2020 Elsevier Ltd Coconut water (Cocos Nucifera) is shown to be a source of essential elements present in the form of low-molecular weight stable complexes known for their bio-availability. The total element concentrations were in the range of 0.2–2.7, 0.3–1, 3–14 and 0.5–2 ppm for Fe, Cu, Mn, and Zn, respectively, and varied as a function of the origin of the nut and its maturity. Speciation was investigated by size-exclusion chromatography - inductively coupled plasma mass spectrometry (ICP MS), and hydrophilic interaction liquid chromatography (HILIC) - electrospray-Orbitrap MS. The metal species identified included: iron complexes with citrate and malate: FeIII(Cit)3(Mal), FeIII(Cit)2(Mal)2, FeIII(Mal)2, glutamine: FeIII(Glu)2 and nicotianamine: FeII(NA); copper complexes with phenylanine: CuII(Phe)2 and CuII(Phe)3 and nicotianamine: CuII(NA); zinc complexes with citrate: ZnII(Cit)2 and nicotianamine ZnII(NA) and manganese complex with asparagine MnII(Asp)2. The contributions of the individual species to the total elements concentrations could be estimated by HILIC – ICP MS.

© 2020 Elsevier B.V. For thermoelements operating on the Peltier and Seebeck effects, including multisection ones used at temperatures up to 900 K, the physicochemical principles of creating effective thin-film multilayer contact systems obtained by magnetron ion-plasma sputtering have been developed. The formation of contact systems was carried out on thermoelectric materials based on: Bi2Te3; Sb2Te3; PbTe; GeTe with the increased thermoelectric figure of merit. A structure of contact systems consisting of contact layers providing ohmic contact, adhesion, barrier and interconnection properties of contact systems is proposed and justified. The selection criteria for materials of contact layers are substantiated. For multisection thermoelements operating on the Seebeck effect at temperatures above 500 K, the necessity of introducing diffusion-barrier layers into the structure of contact systems providing reliability and invariability of the properties of contact systems is substantiated. Based on the physicochemical analysis, the thermodynamic and kinetic factors of the stability and degradation of diffusion-barrier layers are determined. The influence of methods for preparing the surface of thermoelectric materials on the adhesion, contact resistance, and thermal stability of contact systems is established. Using Auger electron spectroscopy, the analysis of the causes of thermal stability and degradation of contact systems was carried out. The deposition modes were determined. The effective contact systems were obtained and investigated. The respective systems are based on: Ni; Mo/Ni and Ni/(Ta–W–N)/Ni having the adhesive strength of more than 12 MPa; the contact resistance not exceeding 10−9 Ω m2 and thermal stability at temperatures up to 900 K.

© 2020 Elsevier B.V. For thermoelements operating on the Peltier and Seebeck effects, including multisection ones used at temperatures up to 900 K, the physicochemical principles of creating effective thin-film multilayer contact systems obtained by magnetron ion-plasma sputtering have been developed. The formation of contact systems was carried out on thermoelectric materials based on: Bi2Te3; Sb2Te3; PbTe; GeTe with the increased thermoelectric figure of merit. A structure of contact systems consisting of contact layers providing ohmic contact, adhesion, barrier and interconnection properties of contact systems is proposed and justified. The selection criteria for materials of contact layers are substantiated. For multisection thermoelements operating on the Seebeck effect at temperatures above 500 K, the necessity of introducing diffusion-barrier layers into the structure of contact systems providing reliability and invariability of the properties of contact systems is substantiated. Based on the physicochemical analysis, the thermodynamic and kinetic factors of the stability and degradation of diffusion-barrier layers are determined. The influence of methods for preparing the surface of thermoelectric materials on the adhesion, contact resistance, and thermal stability of contact systems is established. Using Auger electron spectroscopy, the analysis of the causes of thermal stability and degradation of contact systems was carried out. The deposition modes were determined. The effective contact systems were obtained and investigated. The respective systems are based on: Ni; Mo/Ni and Ni/(Ta–W–N)/Ni having the adhesive strength of more than 12 MPa; the contact resistance not exceeding 10−9 Ω m2 and thermal stability at temperatures up to 900 K.

© 2020 Elsevier Inc. Arecoline is a naturally occurring psychoactive alkaloid with partial agonism at nicotinic and muscarinic acetylcholine receptors. Arecoline consumption is widespread, making it the fourth (after alcohol, nicotine and caffeine) most used substance by humans. However, the mechanisms of acute and chronic action of arecoline in-vivo remain poorly understood. Animal models are a valuable tool for CNS disease modeling and drug screening. Complementing rodent studies, the zebrafish (Danio rerio) emerges as a promising novel model organism for neuroscience research. Here, we assessed the effects of acute and chronic arecoline on adult zebrafish behavior and physiology. Overall, acute and chronic arecoline treatments produced overt anxiolytic-like behavior (without affecting general locomotor activity and whole-body cortisol levels), with similar effects also caused by areca nut water extracts. Acute arecoline at 10 mg/L disrupted shoaling, increased social preference, elevated brain norepinephrine and serotonin levels and reduced serotonin turnover. Acute arecoline also upregulated early protooncogenes c-fos and c-jun in the brain, whereas chronic treatment with 1 mg/L elevated brain expression of microglia-specific biomarker genes egr2 and ym1 (thus, implicating microglial mechanisms in potential effects of long-term arecoline use). Finally, acute 2-h discontinuation of chronic arecoline treatment evoked withdrawal-like anxiogenic behavior in zebrafish. In general, these findings support high sensitivity of zebrafish screens to arecoline and related compounds, and reinforce the growing utility of zebrafish for probing molecular mechanisms of CNS drugs. Our study also suggests that novel anxiolytic drugs can eventually be developed based on arecoline-like molecules, whose integrative mechanisms of CNS action may involve monoaminergic and neuro-immune modulation.

© 2020 Elsevier Ltd Research on individual differences in the fields of chronobiology and chronopsychology mostly focuses on two – morning and evening – chronotypes. However, recent developments in these fields pointed at a possibility to extend chronotypology beyond just two chronotypes. We examined this possibility by implementing the Single-Item Chronotyping (SIC) as a method for self-identification of chronotype among six simple chart options illustrating the daily change in alertness level. Of 2283 survey participants, 2176 (95%) chose one of these options. Only 13% vs. 24% chose morning vs. evening type (a fall vs. a rise of alertness from morning to evening), while the majority of participants chose four other types (with a peak vs. a dip of alertness in the afternoon and with permanently high vs. low alertness levels throughout the day, 15% vs. 18% and 9% vs. 16%, respectively). The same 6 patterns of diurnal variation in sleepiness were yielded by principal component analysis of sleepiness curves. Six chronotypes were also validated against the assessments of sleep timing, excessive daytime sleepiness, and abilities to wake or sleep on demand at different times of the day. We concluded that the study results supported the feasibility of classification with the 6 options provided by the SIC.

Production of reactive oxygen species (ROS) in the mitochondria plays multiple roles in physiology, and excessive production of ROS leads to the development of various pathologies. ROS in the mitochondria are generated by various enzymes, mainly in the electron transporvt chain, and it is important to identify not only the trigger but also the source of free radical production. It is important to measure mitochondrial ROS in live, intact cells, because activation of ROS production could be initiated by changes in extramitochondrial processes which could be overseen when using isolated mitochondria. Here we describe the approaches, which allow to measure production of ROS in the matrix of mitochondria in live cells. We also demonstrate how to measure kinetic changes in lipid peroxidation in mitochondria of live cells. These methods could be used for understanding the mechanisms of pathology in a variety of disease models and also for testing neuro- or cardioprotective chemicals.

Production of reactive oxygen species (ROS) in the mitochondria plays multiple roles in physiology, and excessive production of ROS leads to the development of various pathologies. ROS in the mitochondria are generated by various enzymes, mainly in the electron transporvt chain, and it is important to identify not only the trigger but also the source of free radical production. It is important to measure mitochondrial ROS in live, intact cells, because activation of ROS production could be initiated by changes in extramitochondrial processes which could be overseen when using isolated mitochondria. Here we describe the approaches, which allow to measure production of ROS in the matrix of mitochondria in live cells. We also demonstrate how to measure kinetic changes in lipid peroxidation in mitochondria of live cells. These methods could be used for understanding the mechanisms of pathology in a variety of disease models and also for testing neuro- or cardioprotective chemicals.

© 2020 Elsevier B.V. Over the last two decades, the scientific community and industry have made huge efforts to develop environmental protection technologies. In particular, the scarcity of drinking water has prompted the investigation of several physico-chemical treatments, and synergistic effects have been observed in hyphenated techniques. Herein, we report the first example of water treatment under simultaneous hydrodynamic cavitation and plasma discharge with the intense generation of radicals, UV light, shock waves and charged particles. This highly reactive environment is well suited to the bulk treatment of polluted water (i.e. E. coli disinfection and organic pollutant degradation). We have developed a new prototype and have efficiently applied this hybrid technology to water disinfection and the complete degradation of methanol in water with the aim of demonstrating its scalability. We have analyzed the mechanisms of water disinfection under the abovementioned conditions and verified them by measuring cavitation noise spectra and plasma emission spectra. We have also used the degradation of textile dyes and methanol solutions as an indicator for the formation of radicals.

© 2020 Elsevier B.V. Over the last two decades, the scientific community and industry have made huge efforts to develop environmental protection technologies. In particular, the scarcity of drinking water has prompted the investigation of several physico-chemical treatments, and synergistic effects have been observed in hyphenated techniques. Herein, we report the first example of water treatment under simultaneous hydrodynamic cavitation and plasma discharge with the intense generation of radicals, UV light, shock waves and charged particles. This highly reactive environment is well suited to the bulk treatment of polluted water (i.e. E. coli disinfection and organic pollutant degradation). We have developed a new prototype and have efficiently applied this hybrid technology to water disinfection and the complete degradation of methanol in water with the aim of demonstrating its scalability. We have analyzed the mechanisms of water disinfection under the abovementioned conditions and verified them by measuring cavitation noise spectra and plasma emission spectra. We have also used the degradation of textile dyes and methanol solutions as an indicator for the formation of radicals.

© 2020 Elsevier B.V. Over the last two decades, the scientific community and industry have made huge efforts to develop environmental protection technologies. In particular, the scarcity of drinking water has prompted the investigation of several physico-chemical treatments, and synergistic effects have been observed in hyphenated techniques. Herein, we report the first example of water treatment under simultaneous hydrodynamic cavitation and plasma discharge with the intense generation of radicals, UV light, shock waves and charged particles. This highly reactive environment is well suited to the bulk treatment of polluted water (i.e. E. coli disinfection and organic pollutant degradation). We have developed a new prototype and have efficiently applied this hybrid technology to water disinfection and the complete degradation of methanol in water with the aim of demonstrating its scalability. We have analyzed the mechanisms of water disinfection under the abovementioned conditions and verified them by measuring cavitation noise spectra and plasma emission spectra. We have also used the degradation of textile dyes and methanol solutions as an indicator for the formation of radicals.

© 2020, The Author(s). In the original version of this Article, Ghulam Md Ashraf was incorrectly affiliated with ‘Enzymoics and Novel Global Community Educational Foundation, Hebersham, NSW, Australia’. The correct affiliation is listed below. King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia. Additionally, within the Supplementary Information file originally published with this Article, authors Vijaya Rao Pidugu, Mohammad A. Kamal, Ghulam Md Ashraf, Vadim V. Tarasov, Vladimir N. Chubarev, Sergey G. Klochkov, George E. Barreto and Sergey O. Bachurin were omitted. These errors have now been corrected in the PDF and HTML versions of the Article, and in the accompanying Supplementary material.

© 2020, The Author(s). Somatic mutation profiling in gastric cancer (GC) enables main driver mutations to be identified and their clinical and prognostic value to be evaluated. We investigated 77 tumour samples of GC by next-generation sequencing (NGS) with the Ion AmpliSeq Hotspot Panel v2 and a custom panel covering six hereditary gastric cancer predisposition genes (BMPR1A, SMAD4, CDH1, TP53, STK11 and PTEN). Overall, 47 somatic mutations in 14 genes were detected; 22 of these mutations were novel. Mutations were detected most frequently in the CDH1 (13/47) and TP53 (12/47) genes. As expected, somatic CDH1 mutations were positively correlated with distant metastases (p = 0.019) and tumours with signet ring cells (p = 0.043). These findings confirm the association of the CDH1 mutations with diffuse GC type. TP53 mutations were found to be significantly associated with a decrease in overall survival in patients with Lauren diffuse-type tumours (p = 0.0085), T3-T4 tumours (p = 0.037), and stage III-IV tumours (p = 0.013). Our results confirm that the detection of mutations in the main driver genes may have a significant prognostic value for GC patients and provide an independent GC-related set of clinical and molecular genetic data.

© 2020, The Author(s). Machine learning techniques have been previously applied for classification of tumors based largely on morphological features of tumor cells recognized in H&E images. Here, we tested the possibility of using numeric data acquired from software-based quantification of certain marker proteins, i.e. key autophagy proteins (ATGs), obtained from immunohistochemical (IHC) images of renal cell carcinomas (RCC). Using IHC staining and automated image quantification with a tissue microarray (TMA) of RCC, we found ATG1, ATG5 and microtubule-associated proteins 1A/1B light chain 3B (LC3B) were significantly reduced, suggesting a reduction in the basal level of autophagy with RCC. Notably, the levels of the ATG proteins expressed did not correspond to the mRNA levels expressed in these tissues. Applying a supervised machine learning algorithm, the K-Nearest Neighbor (KNN), to our quantified numeric data revealed that LC3B provided a strong measure for discriminating clear cell RCC (ccRCC). ATG5 and sequestosome-1 (SQSTM1/p62) could be used for classification of chromophobe RCC (crRCC). The quantitation of particular combinations of ATG1, ATG16L1, ATG5, LC3B and p62, all of which measure the basal level of autophagy, were able to discriminate among normal tissue, crRCC and ccRCC, suggesting that the basal level of autophagy would be a potentially useful parameter for RCC discrimination. In addition to our observation that the basal level of autophagy is reduced in RCC, our workflow from quantitative IHC analysis to machine learning could be considered as a potential complementary tool for the classification of RCC subtypes and also for other types of tumors for which precision medicine requires a characterization.

© 2020, The Author(s). Semiconductor quantum dots (QDs) embedded into polymer microbeads are known to be very attractive emitters for spectral multiplexing and colour encoding. Their luminescence lifetimes or decay kinetics have been, however, rarely exploited as encoding parameter, although they cover time ranges which are not easily accessible with other luminophores. We demonstrate here the potential of QDs made from II/VI semiconductors with luminescence lifetimes of several 10 ns to expand the lifetime range of organic encoding luminophores in multiplexing applications using time-resolved flow cytometry (LT-FCM). For this purpose, two different types of QD-loaded beads were prepared and characterized by photoluminescence measurements on the ensemble level and by single-particle confocal laser scanning microscopy. Subsequently, these lifetime-encoded microbeads were combined with dye-encoded microparticles in systematic studies to demonstrate the potential of these QDs to increase the number of lifetime codes for lifetime multiplexing and combined multiplexing in the time and colour domain (tempo-spectral multiplexing). These studies were done with a recently developed novel luminescence lifetime flow cytometer (LT-FCM setup) operating in the time-domain, that presents an alternative to reports on phase-sensitive lifetime detection in flow cytometry.

Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) causes a severe respiratory disease in humans. The MERS-CoV spike (S) glycoprotein mediates viral entry into target cells. For this, MERS-CoV S engages the host cell protein dipeptidyl peptidase 4 (DPP4, CD26) and the interface between MERS-CoV S and DPP4 has been resolved on the atomic level. Here, we asked whether naturally-occurring polymorphisms in DPP4, that alter amino acid residues required for MERS-CoV S binding, influence cellular entry of MERS-CoV. By screening of public databases, we identified fourteen such polymorphisms. Introduction of the respective mutations into DPP4 revealed that all except one (Δ346-348) were compatible with robust DPP4 expression. Four polymorphisms (K267E, K267N, A291P and Δ346-348) strongly reduced binding of MERS-CoV S to DPP4 and S protein-driven host cell entry, as determined using soluble S protein and S protein bearing rhabdoviral vectors, respectively. Two polymorphisms (K267E and A291P) were analyzed in the context of authentic MERS-CoV and were found to attenuate viral replication. Collectively, we identified naturally-occurring polymorphisms in DPP4 that negatively impact cellular entry of MERS-CoV and might thus modulate MERS development in infected patients.

© 2020, The Author(s). Machine learning techniques have been previously applied for classification of tumors based largely on morphological features of tumor cells recognized in H&E images. Here, we tested the possibility of using numeric data acquired from software-based quantification of certain marker proteins, i.e. key autophagy proteins (ATGs), obtained from immunohistochemical (IHC) images of renal cell carcinomas (RCC). Using IHC staining and automated image quantification with a tissue microarray (TMA) of RCC, we found ATG1, ATG5 and microtubule-associated proteins 1A/1B light chain 3B (LC3B) were significantly reduced, suggesting a reduction in the basal level of autophagy with RCC. Notably, the levels of the ATG proteins expressed did not correspond to the mRNA levels expressed in these tissues. Applying a supervised machine learning algorithm, the K-Nearest Neighbor (KNN), to our quantified numeric data revealed that LC3B provided a strong measure for discriminating clear cell RCC (ccRCC). ATG5 and sequestosome-1 (SQSTM1/p62) could be used for classification of chromophobe RCC (crRCC). The quantitation of particular combinations of ATG1, ATG16L1, ATG5, LC3B and p62, all of which measure the basal level of autophagy, were able to discriminate among normal tissue, crRCC and ccRCC, suggesting that the basal level of autophagy would be a potentially useful parameter for RCC discrimination. In addition to our observation that the basal level of autophagy is reduced in RCC, our workflow from quantitative IHC analysis to machine learning could be considered as a potential complementary tool for the classification of RCC subtypes and also for other types of tumors for which precision medicine requires a characterization.

© 2020, The Author(s). Semiconductor quantum dots (QDs) embedded into polymer microbeads are known to be very attractive emitters for spectral multiplexing and colour encoding. Their luminescence lifetimes or decay kinetics have been, however, rarely exploited as encoding parameter, although they cover time ranges which are not easily accessible with other luminophores. We demonstrate here the potential of QDs made from II/VI semiconductors with luminescence lifetimes of several 10 ns to expand the lifetime range of organic encoding luminophores in multiplexing applications using time-resolved flow cytometry (LT-FCM). For this purpose, two different types of QD-loaded beads were prepared and characterized by photoluminescence measurements on the ensemble level and by single-particle confocal laser scanning microscopy. Subsequently, these lifetime-encoded microbeads were combined with dye-encoded microparticles in systematic studies to demonstrate the potential of these QDs to increase the number of lifetime codes for lifetime multiplexing and combined multiplexing in the time and colour domain (tempo-spectral multiplexing). These studies were done with a recently developed novel luminescence lifetime flow cytometer (LT-FCM setup) operating in the time-domain, that presents an alternative to reports on phase-sensitive lifetime detection in flow cytometry.

Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) causes a severe respiratory disease in humans. The MERS-CoV spike (S) glycoprotein mediates viral entry into target cells. For this, MERS-CoV S engages the host cell protein dipeptidyl peptidase 4 (DPP4, CD26) and the interface between MERS-CoV S and DPP4 has been resolved on the atomic level. Here, we asked whether naturally-occurring polymorphisms in DPP4, that alter amino acid residues required for MERS-CoV S binding, influence cellular entry of MERS-CoV. By screening of public databases, we identified fourteen such polymorphisms. Introduction of the respective mutations into DPP4 revealed that all except one (Δ346-348) were compatible with robust DPP4 expression. Four polymorphisms (K267E, K267N, A291P and Δ346-348) strongly reduced binding of MERS-CoV S to DPP4 and S protein-driven host cell entry, as determined using soluble S protein and S protein bearing rhabdoviral vectors, respectively. Two polymorphisms (K267E and A291P) were analyzed in the context of authentic MERS-CoV and were found to attenuate viral replication. Collectively, we identified naturally-occurring polymorphisms in DPP4 that negatively impact cellular entry of MERS-CoV and might thus modulate MERS development in infected patients.

© 2020, The Author(s). Semiconductor quantum dots (QDs) embedded into polymer microbeads are known to be very attractive emitters for spectral multiplexing and colour encoding. Their luminescence lifetimes or decay kinetics have been, however, rarely exploited as encoding parameter, although they cover time ranges which are not easily accessible with other luminophores. We demonstrate here the potential of QDs made from II/VI semiconductors with luminescence lifetimes of several 10 ns to expand the lifetime range of organic encoding luminophores in multiplexing applications using time-resolved flow cytometry (LT-FCM). For this purpose, two different types of QD-loaded beads were prepared and characterized by photoluminescence measurements on the ensemble level and by single-particle confocal laser scanning microscopy. Subsequently, these lifetime-encoded microbeads were combined with dye-encoded microparticles in systematic studies to demonstrate the potential of these QDs to increase the number of lifetime codes for lifetime multiplexing and combined multiplexing in the time and colour domain (tempo-spectral multiplexing). These studies were done with a recently developed novel luminescence lifetime flow cytometer (LT-FCM setup) operating in the time-domain, that presents an alternative to reports on phase-sensitive lifetime detection in flow cytometry.

Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) causes a severe respiratory disease in humans. The MERS-CoV spike (S) glycoprotein mediates viral entry into target cells. For this, MERS-CoV S engages the host cell protein dipeptidyl peptidase 4 (DPP4, CD26) and the interface between MERS-CoV S and DPP4 has been resolved on the atomic level. Here, we asked whether naturally-occurring polymorphisms in DPP4, that alter amino acid residues required for MERS-CoV S binding, influence cellular entry of MERS-CoV. By screening of public databases, we identified fourteen such polymorphisms. Introduction of the respective mutations into DPP4 revealed that all except one (Δ346-348) were compatible with robust DPP4 expression. Four polymorphisms (K267E, K267N, A291P and Δ346-348) strongly reduced binding of MERS-CoV S to DPP4 and S protein-driven host cell entry, as determined using soluble S protein and S protein bearing rhabdoviral vectors, respectively. Two polymorphisms (K267E and A291P) were analyzed in the context of authentic MERS-CoV and were found to attenuate viral replication. Collectively, we identified naturally-occurring polymorphisms in DPP4 that negatively impact cellular entry of MERS-CoV and might thus modulate MERS development in infected patients.