Heme-containing enzymes play vital functions in living organisms, including humans. Here we demonstrate two indirect effects of (electric discharge)-treated stainless steel on a model enzyme — horseradish peroxidase (HRP). The first effect is the complete loss of the enzyme’s adsorption after its incubation in grounded stainless steel chamber, which has been preliminarily subjected to electric discharge in air at atmospheric pressure. The second one is the formation of enzyme aggregates in the sample incubated in another grounded chamber two meters away from the discharge-treated one. At that, the HRP’s enzymatic activity is found to be unaffected in the both cases. These effects may be explained by the occurrence of knotted electromagnetic fields (KEMF). By using high-speed atomic force microscopy (HS-AFM), we reveal the relatively high surface mobility of cytochromes P450cam and P450 102A1 (BM3), whose isoelectric point (pI) values are acidic; at that, thymidylate synthase (TYMS) with near-neutral pI adsorbs strongly. Thus, HRP is the best model object, since its basic pI provides quite strong adsorption on mica. Since (electric discharge)-processed materials have found applications in medicine, we expect that the effects discovered will be considered in future biomedical applications of (electric discharge)-based technologies.
The synthesis of nanoparticles in the plasma of a stratified DC gas discharge was carried out under typical gas discharge conditions, i.e., at room temperature and an argon pressure of 0.11 torr, with a discharge current of 2.5 mA. The particles were formed and grown due to the sputtering of a dielectric plasma concentrator, which was used for strata stabilization. The analysis of the material collected using double-sided carbon tape placed on the glass wall of the discharge tube was performed by scanning electron microscopy and X-ray energy-dispersive microanalysis after the experiments. Three distinctive groups of particles of different shape and size were found, i.e., smooth spherical nanoparticles with a size of 10–100 nm, the main group of smooth spherical and dumbbell-like particles with a size of 200–500 nm, and micron-sized particles of complex cauliflower-like shape. EDX microanalysis of the synthesized nanoparticles revealed that the particles mainly consist of C, O, and Si, which proves that they were formed from the sputtered material of the silicone dielectric concentrator. Analysis of the particles and plasma parameters was performed, and a probable mechanism for the formation of such particles is proposed.
An analysis of lattice wave spectra in a three-dimensional dusty plasma structure formed in a direct current gas discharge with alternating polarity under microgravity conditions is reported. The spectra are determined using the Fourier transform of microparticle velocities, measured by tracking microparticles with subpixel resolution. Both longitudinal and transverse modes are detected and analyzed. The absence of a "k-gap" in the long-wavelength domain of the transverse mode strongly suggests that the microparticles form a solid structure. Therefore, the experimental spectra are compared with the spectra obtained from molecular dynamics simulations for different lattice structures and their orientation. This comparison yields important dusty plasma parameters, such as the particle charge and the plasma screening length. The measured longitudinal and transverse sound velocities allow us to estimate the elastic moduli of the particle component. These are rather small in the absolute magnitude, but when normalized by the number density and the interaction energy of the particles resemble those in conventional matter.
Fragmentation of a suspension of micron-sized plastic microparticles and their contraction into dense globules was experimentally obtained in a gas discharge plasma, when the plasma density was deliberately and abruptly increased. The globules took up spherical shapes 0.14-1.1 mm in diameters and contained from tens to thousands microparticles. The fragmentation and globule formation appears to be similar to the development of gravitational instability. This process is attributed to the Le Sage's like attraction among microparticles in a dense plasma due to the plasma losses inside a globule hypothesized theoretically in the middle of the 1990s. The key role of plasma flows in the attraction was prominently demonstrated in the same experiment by the distinctly visible disintegration of the globules when we reduced the density of the surrounding plasma to the initial one. Also molecular dynamics simulations of fragmentation of microparticle clouds and globules formation qualitatively resemble typical patterns of the fragmentation and collapse of interstellar nebulae.
Rationale: Colorectal cancer (CRC) is one of the most prevalent oncological diseases with high mortality. Invasive optical (endoscopic) colono- scopy has been recognized as a golden standard for the CRC diagnostics. A promising area is the development of non-invasive tools for CRC diagnosis with circular RNA (circRNA). One of the most sensitive non-invasive tools for detection of cancer RNA markers is considered to be the biosensor methods with the use of nanowire chips with oDNA probes (fragments of DNA oligonucleotides) immobilized on their surface. It has been previously shown that circRNA hsa_circ_0136666_CBC1, hsa_circ_0031263_CBC1, and hsa_circ_0072715_CBC1 are associated with CRC. Aim: To determine the lower limit of concentration sensitivity of detection of CRC-associated circRNA with nanowire chips with immobilized oDNA probes, to demonstrate the usability of these chips for non-invasive detection of circRNA in plasma in the CRC diagnostics, and to establish the potential for the use of nanowire chips for the early CRC diagnosis. Methods: To ensure biospecific binding of the circRNA hsa_circ_0136666_CBC1, hsa_circ_0031263_CBC1, and hsa_circ_0072715_CBC1 (the CRC markers), oDNA probes with the nucleotide sequences complementary to the target circRNA have been immobilized on the nanowire surface. At the study step 1, we detected the lower concentration limit for detection of the target molecules with the use of their analogues, i.e. synthetic model oDNA with the nucleotide sequences complementary to oDNA probes. At the study step 2, we used the nanowire chips with immobilized oDNA probes to detect the circRNA in plasma of the patients with confirmed CRC. Plasma samples from non-cancer patients were used as controls. Results: The lower concentration limit for the detection of DNA analogues of the circRNA hsa_circ_0136666_CBC1, hsa_circ_0031263_CBC1, and hsa_circ_0072715_CBC1 with nanowire chips with oDNA probes was 10-16 М. The analysis of total RNA isolated from plasma of the CRC patients showed a significant increase in the signal from the sensory elements of the nanowire chip. The analysis of plasma samples from the non-cancer patients, the nanowire signal changes were non-significant indicating the absence of detectable concentrations of the circRNA in plasma of the non-cancer patients. Conclusion: We have identified the minimal detectable concentration of the circRNA hsa_circ_0136666_CBC1, hsa_circ_0031263_CBC1, and hsa_circ_0072715_CBC1, associated to the development of CRC, with nanowire chips with immobilized oDNA probes: it was 10-16 М. The experiment showed the usability of such nanowire chips for non-invasive detection of the given circRNA markers in total RNA samples isolated from plasma of CRC patients.
Listeria monocytogenes is motile at 22°C and non-motile at 37°C. In contrast, expression of L . monocytogenes virulence factors is low at 22°C and up-regulated at 37°C. Here, we studied a character of L . monocytogenes near surface swimming (NSS) motility and its effects on adhesion patterns and invasion into epithelial cells. L . monocytogenes and its saprophytic counterpart L . innocua both grown at 22°C showed similar NSS characteristics including individual velocities, trajectory lengths, residence times, and an asymmetric distribution of velocity directions. Similar NSS patterns correlated with similar adhesion patterns. Motile bacteria, including both pathogenic and saprophytic species, showed a preference for adhering to the periphery of epithelial HEp-2 cells. In contrast, non-motile bacteria were evenly distributed across the cell surface, including areas over the nucleus. However, the uneven distribution of motile bacteria did not enhance the invasion into HEp-2 cells unless virulence factor production was up-regulated by the transient shift of the culture to 37°C. Motile L . monocytogenes grown overnight at 22°C and then shifted to 37°C for 2 h expressed invasion factors at the same level and invaded human cells up to five times more efficiently comparatively with non-motile bacteria grown overnight at 37°C. Taken together, obtained results demonstrated that (i) NSS motility and correspondent peripheral location over the cell surface did not depend on L . monocytogenes virulence traits; (ii) motility improved L . monocytogenes invasion into human HEp-2 cells within a few hours after the transition from the ambient temperature to the human body temperature.
The development of highly sensitive diagnostic systems for the early revelation of diseases in humans is one of the most important tasks of modern biomedical research, and the detection of the core antigen of the hepatitis C virus (HCVcoreAg)—a protein marker of the hepatitis C virus—is just the case. Our study is aimed at testing the performance of the nanoribbon biosensor in the case of the use of two different types of molecular probes: the antibodies and the aptamers against HCVcoreAg. The nanoribbon sensor chips employed are based on “silicon-on-insulator structures” (SOI-NR). Two different HCVcoreAg preparations are tested: recombinant β-galactosidase-conjugated HCVcoreAg (“Virogen”, Watertown, MA, USA) and recombinant HCVcoreAg (“Vector-Best”, Novosibirsk, Russia). Upon the detection of either type of antigen preparation, the lowest concentration of the antigen detectable in buffer with pH 5.1 was found to be approximately equal, amounting to ~10−15 M. This value was similar upon the use of either type of molecular probes.
Prostate cancer (PC) is one of the major causes of death among elderly men. PC is often diagnosed later in progression due to asymptomatic early stages. Early detection of PC is thus crucial for effective PC treatment. The aim of this study is the simultaneous highly sensitive detection of a palette of PC-associated microRNAs (miRNAs) in human plasma samples. With this aim, a nanoribbon biosensor system based on “silicon-on-insulator” structures (SOI-NR biosensor) has been employed. In order to provide biospecific detection of the target miRNAs, the surface of individual nanoribbons has been sensitized with DNA oligonucleotide probes (oDNA probes) complementary to the target miRNAs. The lowest concentration of nucleic acids, detectable with our biosensor, has been found to be 1.1 × 10−17 M. The successful detection of target miRNAs, isolated from real plasma samples of PC patients, has also been demonstrated. We believe that the development of highly sensitive nanotechnology-based biosensors for the detection of PC markers is a step towards personalized medicine.
The active Brownian motion of single dust particles of various types in the 3D electrostatic DC discharge trap under the action of laser radiation is studied experimentally. Spherical dust particles with a homogeneous surface, as well as Janus particles, are used in the experiment. The properties of the active Brownian motion of all types of dust particles are studied. In particular, the 3D analysis of trajectories of microparticles is carried out, well as an analysis of their root mean square displacement. The mean kinetic energy of motion of the dust particle of various types in a 3D trap is determined for different laser powers. Differences in the character of active Brownian motion in electrostatic traps with different spatial dimensions are found.
We present the results of an experimental and analytical study of the structural and dynamic properties of a monolayer consisting of dust grains in an electrostatic trap in an RF discharge plasma. The possibility of forming a monolayer with an isotropic distribution for interparticle distance and kinetic energy of particles in the structure has been experimentally shown. Isotropy has crucial importance for the study of various processes in such systems, including the kinetics of phase transitions, the formation of directed flows, wave propagation, and others.
We report an experimental study of the 2D dynamics of active particles driven by quantum vortices on the free surface of superfluid helium at T = 1.45 К. The particle motion at short times (< 25 ms) relates to anomalous diffusion mode typical for active particles, while for longer times it corresponds to normal diffusion mode. The values of the rotational and translational kinetic energies of the particle allow to determine for the first time the intensity of the particle-vortex interaction and the dissipation rate of the vortex bundle energy. Strong bonding between a particle and a vortex is explained by coupling of normal and superfluid components.
Ovarian cancer is a gynecological cancer characterized by a high mortality rate and tumor heterogeneity. Its early detection and primary prophylaxis are difficult to perform. Detecting biomarkers for ovarian cancer plays a pivotal role in therapy effectiveness and affects patients’ survival. This study demonstrates the detection of microRNAs (miRNAs), which were reported to be associated with ovarian cancer tumorigenesis, with a nanowire biosensor based on silicon-on-insulator structures (SOI-NW biosensor). The advantages of the method proposed for miRNA detection using the SOI-NW biosensor are as follows: (1) no need for additional labeling or amplification reaction during sample preparation, and (2) real-time detection of target biomolecules. The detecting component of the biosensor is a chip with an array of 3 µm wide, 10 µm long silicon nanowires on its surface. The SOI-NW chip was fabricated using the “top-down” method, which is compatible with large-scale CMOS technology. Oligonucleotide probes (oDNA probes) carrying sequences complementary to the target miRNAs were covalently immobilized on the nanowire surface to ensure high-sensitivity biospecific sensing of the target biomolecules. The study involved two experimental series. Detection of model DNA oligonucleotides being synthetic analogs of the target miRNAs was carried out to assess the method’s sensitivity. The lowest concentration of the target oligonucleotides detectable in buffer solution was 1.1 × 10−16 M. In the second experimental series, detection of miRNAs (miRNA-21, miRNA-141, and miRNA-200a) isolated from blood plasma samples collected from patients having a verified diagnosis of ovarian cancer was performed. The results of our present study represent a step towards the development of novel highly sensitive diagnostic systems for the early revelation of ovarian cancer in women.
The locomotion of droplets in emulsions is of practical significance for fields related to medicine and chemical engineering, which can be done with a magnetic field to move droplets containing magnetic materials. Here, we demonstrate a new method of droplet locomotion in the oil-in-water emulsion with the help of a nonuniform magnetic field in the case where magnetic nanoparticles (MNPs) are dispersed in the continuous phase of the emulsion. The paper analyses the motion of the droplets in a liquid film and in a capillary for various diameters of droplets, their number density, and viscosity of the continuous phase of the emulsion. It is established that the mechanism of droplet locomotion in the emulsion largely depends on the wettability of MNPs. Hydrophobic nanoparticles are adsorbed on the droplet surfaces, forming the agglomerates of MNPs with the droplets. Such agglomerates move at much higher velocities than passive droplets. Hydrophilic nanoparticles are not adsorbed at the surfaces of the droplets but form mobile magnetic clusters dispersed in the continuous phase of the emulsion. Mobile magnetic clusters set the surrounding liquid and droplets in motion. The results obtained in this paper can be used in drug delivery.
Формативная фабрикация — новый способ производства трехмерных конструкций из неорганических и органических веществ, включая живые клетки, путем их направленной и программируемой сборки в различных физических полях, которые обеспечивают функцию временной и удаляемой бесконтактной поддержки. В качестве физических полей могут выступать магнитные, акустические, электрические и другие, а также их комбинации. В обзоре сформулированы главные физические принципы технологии трехмерной сборки и фабрикации, приведены наглядные примеры ее успешного практического применения (в частности, в тканевой инженерии), описаны остающиеся пока еще нерешенными научно-технологические проблемы и изложены основные направления ее будущего развития. Отдельно подчеркнута роль отечественных ученых в разработке этой новой перспективной технологии, которая, по сути, может стать производственной основой следующей индустриальной революции.
We analyze the experimental data on the motion of active Brownian micrograins in RF discharge plasmas. In the experiments, two types of microparticles were used: first—plastic grains fully covered with metal, and second—Janus particles with a thin metal cap. We have tracked the trajectories of the separate grains and plotted the pair correlation functions of the observed structures. To examine the motion of the grains, we studied the dependencies of the MFPT dynamic entropy on the coarsening parameter, the fractal dimension of the system on its mean kinetic temperature, and the mean localization area of the grain on its mean kinetic temperature. Based on the obtained results, we conclude that the character of motion of our active Brownian systems changes as the power of an illuminating laser (and, therefore, the mean kinetic temperature of the grains) increases. Janus particles change their trajectories from more chaotic to spiral-like ones; in the case of fully covered particles, we observe the dynamical phase transition from the more ordered structure to the less ordered one.
The nonlinear dust-acoustic instability in the condensed submicron fraction of dust particles in the low-pressure glow discharge at ultra-low temperatures is experimentally and theoretically investigated. The main discharge parameters are estimated on the basisof the dust-acoustic wave analysis. In particular, the temperature and density of ions, as well as the Debye radius, are determined. It is shown that the ion temperature exceeds the temperature of the neutral gas. The drift characteristics of all plasma fractions are estimated. The reasons for the instability excitation are considered.
The systems of active Brownian grains can be considered as open systems, in which there is an exchange of energy and matter with the environment. The collective phenomena of active Brownian grains can demonstrate analogies with ordinary phase transitions. We study the active Brownian motion of light-absorbing and strongly interacting grains far from equilibrium suspended in gas discharge under laser irradiation when the nature and intensity of the active motion depend on the effect of radiation. Active Brownian motion is caused by photophoresis, i.e., absorption of laser radiation at the metal-coated surface of the grain creates radiometric force, which in turn drives the grains. We experimentally observed the active Brownian motion of charged grains in the transition of the grain monolayer from the solid to liquid state. An analysis of the character of motion, including the mean-square and linear displacement and persistence length at various values of the randomization (coupling parameter) of the grain structure, was presented.
Complex structures, consisting of a large number of interacting subsystems, have the ability to self-organize and evolve, when the scattering of energy coming from the outside ensures the maintenance of stationary ordered structures with an entropy less than the equilibrium entropy. One of the fundamental problems here is the role of quantum phenomena in the evolution of macroscopic objects. We provide experimental evidence for the active Brownian motion and evolution of structures driven by quantum effects for micron-sized grains levitating in superfluid helium. The active Brownian motion of grains was induced by quantum turbulence during the absorption of laser irradiation by grains. The intensity of Brownian motion associated with quantum vortices increased by 6–7 orders of magnitude compared to the values from the Einstein formula. We observed the grain structures in a state far from thermodynamic equilibrium and their evolution to more complex organized structures with lower entropy due to the quantum mechanism of exceedingly high entropy loss in superfluid helium.
Self-organization of dust grains into stable filamentary dust structures (or "chains ") largely depends on dynamic interactions between individual charged dust grains and complex electric potential arising from the distribution of charges within a local plasma environment. Recent studies have shown that the positive column of the gas discharge plasma in the Plasmakristall-4 (PK-4) experiment at the International Space Station supports the presence of fast-moving ionization waves, which lead to variations of plasma parameters by up to an order of magnitude from the average background values. The highly variable environment resulting from ionization waves may have interesting implications for the dynamics and self-organization of dust particles, particularly concerning the formation and stability of dust chains. Here, we investigate the electric potential surrounding dust chains in the PK-4 experiment by employing a molecular dynamics model of the dust and ions with boundary conditions supplied by a particle-in-cell with Monte Carlo collision simulation of the ionization waves. The model is used to examine the effects of the plasma conditions within different regions of the ionization wave and compare the resulting dust structure to that obtained by employing the time-averaged plasma conditions. The comparison between simulated dust chains and experimental data from the PK-4 experiment shows that the time-averaged plasma conditions do not accurately reproduce observed results for dust behavior, indicating that more careful treatment of plasma conditions in the presence of ionization waves is required. It is further shown that commonly used analytic forms of the electric potential do not accurately describe the electric potential near charged dust grains under these plasma conditions.