Comparative analysis of the metrological documents specifying chronologically the standard atmosphere parameters is completed. It was revealed, that the
GOST
4401–81—the USSR State Standard, which is ...currently in force,—contains a lot of errors. Information technology procedures are proposed to identify errors in the tables, which are the matter of the metrological documents under analysis. Examples of use of the procedures are given, the errors already identified in these standards are listed.
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•A novel electroactive secondary metabolite was discovered in medium of living yeast cells.•Pd-NPs can effectively support profiling of the formed metabolite in yeasts.•The impact of ...pH and interfering electroactive species on the obtained signal was studied.•One-pot alginate-based nanoreactor was fully cytocompatible to serve as a nanofermenter.
This study describes the development of a one-pot electrochemical miniaturized system for simultaneous cultivation and monitoring of the oxidative status of living cells. This system consisted of screen-printed electrodes modified by electroplated Pd-NPs as an electrocatalyst (i) and living yeast cells (Saccharomyces cerevisiae) (ii) immobilized on the cytocompatible alginate layer (iii).
Briefly, during the course of electrochemical investigations a novel electroactive compound methylhydrazine derivative as a secondary metabolite and result of microbial activity was found in yeast cells and used as a signaling molecule for their biochemical profiling. Under the optimized experimental conditions the signal corresponding to the found electroactive secondary metabolite formed in medium of living cells was measured without sample collecting, transport, storage or pre-treatment steps (i.e. extraction, pre-concentration, chemical derivatization or labeling). The electrochemical dependencies, which were derived by a miniaturized electroanalytical system, were fully validated in a conventional three-electrode system under inert atmosphere (Ar) and in the presence of oxygen (air, O2).
It is believed that the proposed one-pot nanoreactors serving simultaneously as nanofermenters and amperometric detectors for the quantification of secondary metabolites formed in medium of living cells can significantly enhance the understanding of ongoing fermentation processes in the future and our knowledge on the biochemistry of yeasts.
Nuclear organelles are viscous droplets, created by concentration-dependent condensation and liquid-liquid phase separation of soluble proteins. Nuclear organelles have been actively investigated for ...their role in cellular regulation and disease. However, these studies are highly challenging to perform in live cells, and therefore, their physico-chemical properties are still poorly understood. In this study, we describe a fluorescence lifetime imaging approach for real-time monitoring of protein condensation in nuclear organelles of live cultured cells. This approach unravels surprisingly large cyclic changes in concentration of proteins in major nuclear organelles including nucleoli, nuclear speckles, Cajal bodies, as well as in the clusters of heterochromatin. Remarkably, protein concentration changes are synchronous for different organelles of the same cells. We propose a molecular mechanism responsible for synchronous accumulations of proteins in the nuclear organelles. This mechanism can serve for general regulation of cellular metabolism and contribute to coordination of gene expression.
This paper considers the metallurgical processes of dissociation, ionization, oxidation, deoxidation, and dissolution of oxides during underwater wet cutting. A multiphase mechanism of underwater wet ...cutting consisting of working and idle cycles of the electrical process in a pulsating vapor gas bubble is proposed. A model of arc penetration into metal due to metal oxidation and stabilization of the arc by the inner walls of a narrow kerf is proposed. For underwater cutting of 10 KhSND, 304L steel, CuAl5, and AlMg4.5Mn0.7 alloy, we provide a principle of modeling the phase composition of the gas mixture based on high oxygen concentration, improving ionization, enthalpy, heat capacity, and thermal conductivity of plasma through the use of a mixture of KNO3, FeCO3 and aluminum. The method of improving the thermophysical properties and ionization of plasma due to the exothermic effect when introducing Fe3O4, MoO2, WO2 oxides and Al, Mg, Ti deoxidizers is proposed. Although a negative effect of refractory slag was revealed, it could be removed by using the method of reducing surface tension through the ionic dissolution of refractory oxides in Na3AlF6 cryolite. In underwater cutting of 10 KhSND and 304L, the steel welding current was 344–402 A with a voltage of 36–39 V; in cutting of CuAl5 and AlMg4.5Mn0.7 alloy, the welding current was 360–406; 240 A, with a voltage of 35–37; 38 V, respectively, with the optimal composition of flux-cored wire: 50–60% FeCO3 and KNO3, 20–30% aluminum, 20% Na3AlF6. Application of flux-cored wires of the KNO3-FeCO3-Na3AlF6-Al system allowed stable cutting of 10KhSND, AISI 304L steels, and CuAl5 bronze with kerf width up to 2.5–4.7 mm.
Abstract
Optical imaging is a most useful and widespread technique for the investigation of the structure and function of the cellular genomes. However, an analysis of immensely convoluted and ...irregularly compacted DNA polymer is highly challenging even by modern super-resolution microscopy approaches. Here we propose fluorescence lifetime imaging (FLIM) for the advancement of studies of genomic structure including DNA compaction, replication as well as monitoring of gene expression. The proposed FLIM assay employs two independent mechanisms for DNA compaction sensing. One mechanism relies on the inverse quadratic relation between the fluorescence lifetimes of fluorescence probes incorporated into DNA and their local refractive index, variable due to DNA compaction density. Another mechanism is based on the Förster resonance energy transfer (FRET) process between the donor and the acceptor fluorophores, both incorporated into DNA. Both these proposed mechanisms were validated in cultured cells. The obtained data unravel a significant difference in compaction of the gene-rich and gene-poor pools of genomic DNA. We show that the gene-rich DNA is loosely compacted compared to the dense DNA domains devoid of active genes.
Recent developments in Raman spectroscopy instrumentation and data processing algorithms have led to the emergence of Ramanomics - an independent discipline with unprecedented capabilities to map the ...distribution of distinct molecular groups in live cells. Here, we introduce a method for probing the absolute concentrations of proteins, RNA and lipids in single organelles of live cultured cells by biomolecular component analysis using microRaman data. We found significant cell-to-cell variations in the molecular profiles of organelles, thus providing a physiologically relevant set of markers of cellular heterogeneity. At the same cell the molecular profiles of different organelles can strongly correlate, reflecting tight coordination of their functions. This correlation was significant in WI-38 diploid fibroblasts and weak in HeLa cells, indicating profound differences in the regulation of biochemical processes in these cell lines.
Biodistribution of photosensitizer (PS) in photodynamic therapy (PDT) can be assessed by fluorescence imaging that visualizes the accumulation of PS in malignant tissue prior to PDT. At the same ...time, excitation of the PS during an assessment of its biodistribution results in premature photobleaching and can cause toxicity to healthy tissues. Combination of PS with a separate fluorescent moiety, which can be excited apart from PS activation, provides a possibility for fluorescence imaging (FI) guided delivery of PS to cancer site, followed by PDT.
In this work, we report nanoformulations (NFs) of core-shell polymeric nanoparticles (NPs) co-loaded with PS 2-(1-hexyloxyethyl)-2-devinyl pyropheophorbide-a, HPPH and near infrared fluorescent organic dyes (NIRFDs) that can be excited in the first or second near-infrared windows of tissue optical transparency (NIR-I, ~ 700-950 nm and NIR-II, ~ 1000-1350 nm), where HPPH does not absorb and emit. After addition to nanoparticle suspensions, PS and NIRFDs are entrapped by the nanoparticle shell of co-polymer of N-isopropylacrylamide and acrylamide poly(NIPAM-co-AA), while do not bind with the polystyrene (polySt) core alone. Loading of the NIRFD and PS to the NPs shell precludes aggregation of these hydrophobic molecules in water, preventing fluorescence quenching and reduction of singlet oxygen generation. Moreover, shift of the absorption of NIRFD to longer wavelengths was found to strongly reduce an efficiency of the electronic excitation energy transfer between PS and NIRFD, increasing the efficacy of PDT with PS-NIRFD combination. As a result, use of the NFs of PS and NIR-II NIRFD enables fluorescence imaging guided PDT, as it was shown by confocal microscopy and PDT of the cancer cells in vitro. In vivo studies with subcutaneously tumored mice demonstrated a possibility to image biodistribution of tumor targeted NFs both using HPPH fluorescence with conventional imaging camera sensitive in visible and NIR-I ranges (~ 400-750 nm) and imaging camera for short-wave infrared (SWIR) region (~ 1000-1700 nm), which was recently shown to be beneficial for in vivo optical imaging.
A combination of PS with fluorescence in visible and NIR-I spectral ranges and, NIR-II fluorescent dye allowed us to obtain PS nanoformulation promising for see-and-treat PDT guided with visible-NIR-SWIR fluorescence imaging.
To advance an understanding of cellular regulation and function it is crucial to identify molecular contents in cellular organelles, which accommodate specific biochemical processes. Toward ...achievement of this goal, we applied micro-Raman-Biomolecular Component Analysis assay for molecular profiling of major organelles in live cells. We used this assay for comparative analysis of proteins 3D conformation and quantification of proteins, RNA, and lipids concentrations in nucleoli, endoplasmic reticulum, and mitochondria of WI 38 diploid lung fibroblasts and HeLa cancer cells. Obtained data show substantial differences in the concentrations and conformations of proteins in the studied organelles. Moreover, differences in the intraorganellar concentrations of RNA and lipids between these cell lines were found. We report the biological significance of obtained macromolecular profiles and advocate for micro-Raman BCA assay as a valuable proteomics tool.
By the finite-element method, we study the stress-strain state of an interface crack in a piezoelectric bimaterial polarized in the direction perpendicular to the crack faces and subjected at ...infinity to the action of an electric field parallel to the crack and stresses parallel to the axis of polarization. On the crack faces, we consider the main versions of electrostatic boundary conditions, namely, impermeability (insulation), electric permeability, electric conductivity, and also a version of mixed boundary conditions, i.e., electric conductivity of a middle part of the top face and insulation of the remaining part of the crack. The formation of zones of mechanical contact between the faces is possible. The problem is considered in the statement of plane deformation. The energy release rates are found with the help of the virtual crack closure integral method. It is shown that the type of boundary conditions strongly affects the parameters of fracture.
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