Aggregation by lectins as an approach for exosome isolation from biological fluids: Validation for proteomic studies

Exosomes, a special type of microparticles produced by cells, are currently of considerable interest for researchers. The term “exosomes” denotes extracellular vesicles of less than 120 nm in size derived from intracellular multivesicular bodies. Multiple studies that address the distinctive features of exosome structure and biochemical composition in various pathological states imply the possibility of development of novel diagnostic techniques based on the detection of changes in the pool of proteins and nucleic acids transported by exosomes. However, methods for isolation and investigation of exosomes are rather difficult to develop because of a small size of these vesicles. A novel approach for preparative-scale isolation of exosomes based on the phenomenon of binding and aggregation of these particles in the presence of lectins has been put forward in the present study. The method developed is relatively cost-effective, allows for the isolation of exosomes from various biological fluids, and has been validated for the subsequent analysis of the protein composition of the exosomes in view of the possible clinical applications. The validation showed that the sedimentation of lectin-aggregated exosomes is a suitable approach for the isolation of these microvesicles from the complete conditioned culture medium in a research-laboratory setup.     

Activation of central orexin/hypocretin neurons by dietary amino acids

Hypothalamic orexin/hypocretin (orx/hcrt) neurons regulate energy balance, wakefulness, and reward; their loss produces narcolepsy and weight gain. Glucose can lower the activity of orx/hcrt cells, but whether other dietary macronutrients have similar effects is unclear. We show that orx/hcrt cells are stimulated by nutritionally relevant mixtures of amino acids (AAs), both in brain slice patch-clamp experiments, and in c-Fos expression assays following central or peripheral administration of AAs to mice in?vivo. Physiological mixtures of AAs electrically excited orx/hcrt cells through a dual mechanism involving inhibition of K(ATP) channels and activation of system-A amino acid transporters. Nonessential AAs were more potent in activating orx/hcrt cells than essential AAs. Moreover, the presence of physiological concentrations of AAs suppressed the glucose responses of orx/hcrt cells. These results suggest a new mechanism of hypothalamic integration of macronutrient signals and imply that orx/hcrt cells sense macronutrient balance, rather than net energy value, in extracellular fluid.     

Multi-Chord Beam Diagnostics of Plasma at the GOL-NB Device

Plasma Physics Reports - Abstract—A multi-chord beam diagnostic system for an open multiple-mirror trap GOL-NB is described. The system is based on a 10 keV ribbon beam of fast neutral...     

Experimental study of the evaporation and expansion of a solid pellet in a plasma heated by an electron beam

Results are presented from experiments on the injection of solid pellets into a plasma heated by an electron beam in the GOL-3 device. For this purpose, two pellet injectors were installed in the device. The target plasma with a density of ～10¹⁵ cm^?3 was produced in a solenoid with a field of 4.8 T and was heated by a highpower electron beam with an electron energy of ～1 MeV, a duration of ～7 s, and a total energy of 120–150 kJ. Before heating, the pellet was injected into the center of the plasma column transversely to the magnetic field. The injection point was located at a distance of 6.5 or 2 m from the input magnetic mirror. Polyethylene pellets with a mass of 0.1–1 mg and lithium-deuteride pellets with a mass of 0.02–0.5 mg were used. A few microseconds after the electron beam starts to be injected into the plasma, a dense plasma bunch is formed. In the initial stage of expansion, the plasma bunch remains spherically symmetric. The plasma at the periphery of the bunch is then heated and becomes magnetized. Next, the dense plasma expands along the magnetic field with a velocity on the order of 300 km/s. A comparison of the measured parameters with calculations by a hydrodynamic model shows that, in order to provide such a high expansion velocity, the total energy density deposited in the pellet must be ～1 kJ/cm². This value substantially exceeds the energy density yielded by the target plasma; i.e., the energy is concentrated across the magnetic field onto a dense plasma bunch produced from the evaporated particle.     

Study of the mechanism for fast ion heating in the GOL-3 multimirror magnetic confinement system

Results are presented from experimental studies of ion heating in the GOL-3 device. The experiments were carried out in a multimirror configuration with a local magnetic well. It was found that, during the injection of a relativistic electron beam, a decrease in the local density of the beam in a magnetic well, which is proportional to the decrease in the strength of the longitudinal magnetic field, results in the formation of a short plasma region with a low electron temperature. The measured longitudinal gradient of the plasma pressure corresponds to an electron temperature gradient of ～2–3 keV/m. Axially nonuniform heating of the plasma electrons gives rise to the macroscopic motion of the plasma along the magnetic field in each cell of the multimirror confinement system. The mixing of the counterpropagating plasma flows inside each cell leads to fast ion heating. Under the given experimental conditions, the efficiency of this heating mechanism is higher than that due to binary electron-ion collisions. The collision and mixing of the counterpropagating plasma flows is accompanied by a neutron and γ-ray burst. The measured ratio of the plasma pressure to the vacuum magnetic field pressure in these experiments reaches 0.2.     

Bombesin-induced cytosolic Ca2+ spiking in pancreatic acinar cells depends on cyclic ADP-ribose and ryanodine receptors

Different hormones and neurotransmitters, using Ca2+ as their intracellular messenger, can generate specific cytosolic Ca2+ signals in different parts of a cell. In mouse pancreatic acinar cells, cytosolic Ca2+ oscillations are triggered by activation of acetylcholine (ACh), cholecystokinin (CCK) and bombesin receptors. Low concentrations of these three agonists all induce local Ca(2+)spikes, but in the case of bombesin and CCK these spikes can also trigger global Ca2+ signals. Here we monitor cytosolic Ca2+ oscillations induced by low (2-5 pM) concentrations of bombesin and show that, like ACh- and CCK-induced oscillations, the bombesin-elicited responses are inhibited by ryanodine(50 microM). We then demonstrate that, like CCK- but unlike ACh-induced oscillations, the responses to bombesin are abolished by intracellular infusion of the cyclic ADP ribose (cADPr) antagonist 8-NH2-cADPr (20 microM). We conclude that in mouse pancreatic acinar cells, bombesin, CCK and ACh all produce local Ca2+ spikes by recruiting common oscillator units composed of ryanodine and inositol trisphosphate receptors. However, bombesin and CCK also recruit cADPr receptors, which may account for the global Ca2+ signals that can be evoked by these two agonists. Our new results indicate that each Ca2+ -mobilizing agonist, acting on mouse pancreatic acinar cells, recruits a unique combination of intracellular Ca2+ channels.     

Study of plasma rotation in the GOL-3 facility

The MHD activity of plasma in the GOL-3 facility was studied experimentally. The complicated azimuthal structure of the profile of the longitudinal current is revealed by magnetic measurements and high-speed imaging. The azimuthal mode composition of magnetic perturbations, as well as the frequency of their rotation in different regimes, is determined. The signs of differential plasma rotation in the facility are described. Possible mechanisms responsible for the relation of differential rotation to current filamentation and magnetic reconnection in the GOL-3 plasma are proposed. These mechanisms explain the previously observed experimental evidences of these processes.     

Experiments on the transportation of a magnetized plasma stream in the GOL-3 facility

The program of the deep upgrade of the GOL-3 multiple-mirror trap is presented. The upgrade is aimed at creating a new GOL-NB open trap located at the GOL-3 site and intended to directly demonstrate the efficiency of using multiple-mirror magnetic cells to improve longitudinal plasma confinement in a gasdynamic open trap. The GOL-NB device will consist of a new central trap, adjoint cells with a multiple-mirror magnetic field, and end tanks (magnetic flux expanders). Plasma in the central trap will be heated by neutral beam injection with a power of up to 1.5 MW and duration of 1 ms. At present, physical experiments directed at developing plasma technologies that are novel for this facility are being carried out using the 6-m-long autonomous part of the GOL-3 solenoid. The aim of this work was to develop a method for filling the central trap with a low-temperature start plasma. Transportation of a plasma stream from an arc source over a distance of 3 m in a uniform magnetic field with an induction of 0.5–4.5 T is demonstrated. In these experiments, the axial plasma density was (1–4) × 10²⁰ m^–3 and the mirror ratio varied from 5 to 60. In general, the experiments confirmed the correctness of the adopted decisions for the start plasma source of the GOL-NB device.     

Design of an experiment on wakefield acceleration on the VEPP-5 injection complex

Relativistic beams produced by the VEPP-5 injection complex (Budker Institute of Nuclear Physics, Siberian Division, Russian Academy of Sciences) can be used to generate plasma waves with a longitudinal electric field of 1 GV/m. A part of the electron (or positron) driver bunch is accelerated by this field over a distance of up to 1 m. The main advantage of the proposed design over the previous wakefield acceleration experiments is the beam preparation system capable of compressing bunches to a length of σ_z = 0.1 mm in the longitudinal direction and producing an optimal longitudinal profile of the beam density. The main parameters of the planned device are as follows: the electron energy at the entrance to the plasma is 510 MeV, the number of particles in the bunch is 2 × 10¹⁰, the plasma density is up to 10¹⁶ cm^?3, the number of accelerated particles is up to 3 × 10⁹, and their energy spread is less than 10%. The physical project of the experiment is presented, and the results of computer simulations of the beam-plasma interaction are described.