Atmospheric-pressure plasma jet induces apoptosis involving mitochondria via generation of free radicals

The plasma jet has been proposed as a novel therapeutic method for anticancer treatment. However, its biological effects and mechanism of action remain elusive. Here, we investigated its cell death effects and underlying molecular mechanisms, using air and N₂ plasma jets from a micro nozzle array. Treatment with air or N₂ plasma jets caused apoptotic death in human cervical cancer HeLa cells, simultaneously with depolarization of mitochondrial membrane potential. In addition, the plasma jets were able to generate reactive oxygen species (ROS), which function as surrogate apoptotic signals by targeting the mitochondrial membrane potential. Antioxidants or caspase inhibitors ameliorated the apoptotic cell death induced by the air and N₂ plasma jets, suggesting that the plasma jet may generate ROS as a proapoptotic cue, thus initiating mitochondria-mediated apoptosis. Taken together, our data suggest the potential employment of plasma jets as a novel therapy for cancer.     

Diagnostics of jet structures formed in the plasma corona during the irradiation of porous targets by intense laser pulses

Jet structures formed during laser irradiation of porous targets with an average density of ?? = 1?30 mg/cm³ were studied experimentally by using the diagnostic complex of the Mishen facility. To study complicated plasma structures, the experimental data were processed using specially elaborated mathematical methods. The probability of the emergence of jet plasma structures in plane open-pore triacetate cellulose targets was studied as a function of the parameter ??d, where ?? is the average mass density and d is the target thickness. Analysis of the experimental results and their comparison with the existing data on the jet structures formed during laser irradiation of solid-density targets allowed the authors to reveal the characteristic features and mechanisms of the development of large-scale plasma jets.     

Spectroscopic measurements of the parameters of the helium plasma jets generated in the plasma focus discharge at the PF-3 facility

The spectroscopic technique used to measure the parameters of the plasma jets generated in the plasma focus discharge and those of the plasma of the immobile gas through which these jets propagate is described. The time evolution of the intensities and shapes of spectral lines in experiments carried out with helium at the PF-3 facility was studied by means of electron-optical streak cameras. The plasma electron temperature, T ≈ 4–5 eV, was determined from the intensity ratio of two spectral lines, one of which (λ₁ = 5876 Å) belongs to neutral helium, while the other (λ₂ = 4686 Å), to hydrogen-like helium ions. The plasma density at different time instants was determined from the Stark broadening of these lines in the electric fields of different nature. The plasma density is found to vary from 4 × 10¹⁴ to 2 × 10¹⁷ cm^?3.     

Study of the plasma in a preformed Z-pinch constriction

The development of a preformed constriction in cylindrical agar-agar loads at currents of up to 3 MA is studied experimentally. The loads 3–5 mm in diameter have a mass density of 0.1 g/cm³ and are filled with different materials. Due to the implosion of the constriction to a minimum size of 40–70 μm, a hot dense plasma (with the electron density n _e=10²² cm⁻³, electron temperature T _e=0.8–1.5 keV, and ion temperature T _i=3–12 keV) is produced. It is found that the ion temperature substantially exceeds the electron temperature. The lifetime of the high-temperature plasma determined from the FWHM of a soft X radiation (SXR) pulse is shorter than 5 ns, the radiation power of photons with energies of ≥1 keV is higher than 0.5×10¹⁰ W, and their total energy attains 50 J. High-speed photography in the VUV, SXR, and optical spectral regions indicates the protracted generation of the high-temperature plasma. Calculations by the two-dimensional ideal MHD model of the Z-pinch show that the most important consequence of the protracted plasma generation in the constriction region is that the current is intercepted by a freshly produced plasma. In the course of plasma generation, the current near the axis inside the region of radius 50 μm is at most one-half of the total current. After the plasma generation comes to an end, almost the entire current is concentrated in this region for several nanoseconds; this process is accompanied by a sharp increase in the plasma temperature. __________ Translated from Fizika Plazmy, Vol. 27, No. 12, 2001, pp. 1101–1110. Original Russian Text Copyright ? 2001 by Bakshaev, Blinov, Vikhrev, Gordeev, Dan’ko, Korolev, Medovshchikov, Nedoseev, Smirnova, Tumanov, Chernenko, Shashkov.     

Expansion of a multicomponent current-carrying plasma jet into vacuum

An expression for the ion?ion coupling in a multicomponent plasma jet is derived for an arbitrary ratio between the thermal and relative velocities of the components. The obtained expression is used to solve the problem on the expansion of a current-carrying plasma microjet emitted from the cathode surface into vacuum. Two types of plasmas with two ion components are analyzed: (i) plasma in which the ion components of equal masses are in the charge states Z ₁= +1 and Z ₂= +2 and (ii) plasma with ions in equal charge states but with the mass ratio m ₁/m ₂ = 2. It is shown that, for such plasmas, the difference between the velocities of the plasma components remains substantial (about 10% of the average jet velocity in case (i) and 15% in case (ii)) at distances of several centimeters from the emission center, where it can be measured experimentally, provided that its initial value at the emitting cathode surface exceeds a certain threshold. This effect is investigated as a function of the mass ratio and charge states of the ion components.     

Energy balance in the interaction of intense high-temperature plasma flows with solid targets

The energy balance in the interaction of intense (W≈7 MW/cm², Q≈130 J/cm²) flows of a high-temperature (T _e+T _i≈0.7 keV deuterium plasma with targets made of different materials (graphite, tungsten, copper, etc.) is studied experimentally. It is shown that radiation plays a decisive role in the interaction energy balance: a plasma layer arising near the surface of the eroded target reemits most of the plasma-flow energy into the surrounding space. No more than 50 J/cm² reaches the surface. Then, this energy is expended primarily on the target heating and only a small fraction (less than 3 J/cm²) is spent on the evaporation of the target surface layer. It is shown that, for targets made of high-Z materials, the energy reaching the surface is transferred predominantly by radiation.     

Study of a cumulative jet in a plasma focus discharge by the method of shearing interferometry

The dynamics of the cumulative jet formed in the course of plasma compression in a plasma focus discharge is investigated by the method of differential optical interferometry. The jet propagation velocity is found to be V = (2.3?C3) × 10⁷ cm/s, which coincides with the results of calculations performed in a 2D ideal MHD model. Ejection of matter from the anode in the late stage of the discharge due to the interaction of the cumulative jet and the electron beam with the anode surface is observed.     

RF discharge-based plasma emitter

An injector of hydrogen atoms for plasma diagnostics in modern tokamaks has been developed at the Budker Institute of Nuclear Physics (Novosibirsk). The ion source of the injector produces a proton (helium ion) beam with a current of up to 2 A (1 A), an ion energy of up to 55 keV, a beam divergence of ～0.6\deg, and a pulse duration of up to 10 s. An RF discharge-based plasma emitter, which is one of the main parts of the ion source, is described. The emitter diameter is 72 mm, the ion current density is 120 mA/cm², and the inhomogeneity of the current density is ±6%. The beam is formed by a four-electrode ionoptical system with 163 round apertures. At a current of 2 A, the ion beam consists of 67% protons, 18% H ₂ ⁺ ions, and 15% H ₃ ⁺ ions, the total content of heavier ions in the beam being no higher than 2–3%.     

Charge and energy spectra of fast multicharged ions in a laser plasma

In measuring the charge and energy spectra of the ions of a single-element laser plasma, in addition to thermal ions, fast multicharged ions are recorded that are accelerated by the electric field of laser radiation in the region of the critical plasma density. The charge and energy spectra of Co ions with the charge numbers z=1–3 are measured at laser intensities of q=5×10¹¹–10¹² W/cm². The energy spectra of these ions are broad and are located on the high-energy side (z _max=3, E>5.0 keV) with respect to the thermal ions (z _max=9, E<4.0 keV). The increase in q to 10¹⁴ W/cm² results in an increase in the charge number of both thermal and fast ions.     

Role of the plasma membrane in the mechanism of cholesterol efflux from cells

In order to investigate the role of the plasma membrane in determining the kinetics of removal of cholesterol from cells, the efflux of [³H]cholesterol from intact cells and plasma membrane vesicles has been compared. The release of cholesterol from cultures of Fu₅AH rat hepatoma and WIRL-3C rat liver cells to complexes of egg phosphatidylcholine (1 mg / ml) and human high-density apolipoprotein is first order with respect to concentration of cholesterol in the cells, with half-times ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$) for at least one-third of the cell cholesterol of 3.2 ± 0.6 and 14.3 ± 1.5 h, respectively. Plasma membrane vesicles (0.5–5.0 μm diameter) were produced from both cell lines by incubating the cells with 50 mM formaldehyde and 2 mM dithiothreitol for 90 min. The efflux of cholesterol from the isolated vesicles follows the same kinetics as the intact, parent cells: the $\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ values for plasma membrane vesicles of Fu₅AH and WIRL cells are 3.9 ± 0.5 and 11.2 ± 0.7 h, respectively. These $\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ values reflect the rate-limiting step in the cholesterol efflux process, which is the desorption of cholesterol molecules from the plasma membrane into the extracellular aqueous phase. The fact that intact cells and isolated plasma membranes release cholesterol at the same rate indicates that variations in the plasma membrane structure account for differences in the kinetics of cholesterol release from different cell types. In order to investigate the role of plasma membrane lipids, the kinetics of cholesterol desorption from small unilamellar vesicles prepared from the total lipid isolated from plasma membrane vesicles of Fu₅AH and WIRL cells were measured. Half-times of cholesterol release from plasma membrane lipid vesicles of Fu₅AH and WIRL cells were the same, with values of 3.1 ± 0.1 and 2.9 ± 0.2 h, respectively. Since bilayers formed from isolated plasma membrane lipids do not reproduce the kinetics of cholesterol efflux observed with the intact plasma membranes, it is likely that the local domain structure, as influenced by membrane proteins, is responsible for the differences in $\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}$ values for cholesterol efflux from these cell lines.     

Flow visualisation and high speed video analysis of water jets in the snapping shrimp (Alpheus heterochaelis)

Snapping shrimp (Alpheus heterochaelis) produce a fast, well-focused water jet by rapid closure of their specialised snapper claw. As shown previously, water jets may injure the opponent in interspecific encounters (e.g. with small crabs) although no damage was observed in intraspecific encounters. For conspecific receivers the jet represents a potential hydrodynamic signal and can be analysed with the help of mechanosensory hairs. To gain more insight in the biophysical characteristics of the water jet we visualised and analysed jets of tethered snapping shrimp using standard and high speed video recordings. Water jet width increases with increasing distance from the snapper claw tip, and both width and distance increase with increasing snapper claw size. Water jet distances do not increase with increasing claw cocking duration (building up muscle tension) but medium cocking durations of about 550 ms result in longest distances. Mean water jet velocity is 6.5 m s⁻¹ shortly after claw closure but rapidly decreases subsequently. At the mean distance between snapping conspecifics (9 mm) water jet velocities produced by snapping shrimp with larger snapper claws are significantly higher than those of animals with smaller claws. Interestingly, males with equal snapper claw size as females produce significantly faster water jets. Accepted: 31 March 1999     

Excitation of the isomeric states of silver isotope nuclei in the plasma produced by a subrelativistic picosecond laser pulse

Arrangement and results of experiments on the excitation of the (E _m = 93.125 keV, J ^p = 7/2⁺, T _1/2 = 44.3 s) and (E _m = 88.034 keV, J ^p = 7/2⁺, T _1/2 = 39.6 s) isomeric states of Ag¹⁰⁷ and Ag¹⁰⁹ nuclei under the action of X-ray emission in a hot (T _e ∼0.5 keV) dense plasma produced by a laser pulse with the energy ∼9 J, intensity ∼1.2 × 10¹⁸ W/cm², and duration 0.82 ps on the SOKOL-P facility are described. The experimentally determined half-life of the isomeric states agrees satisfactorily with the half-life of the Ag^107m and Ag^109m isomers, and their number N _m ∼ 6.9 × 10⁴ agrees with the qualitative estimate N _m ∼ 2.8 × 10⁴, obtained within the refined model of the physical processes in laser plasma.     

Antimicrobial mechanism and the effect of atmospheric pressure N2 plasma jet on the regeneration capacity of Staphylococcus aureus biofilm

Abstract

This study systematically assessed the inactivation mechanism on Staphylococcus aureus biofilms by a N₂ atmospheric-pressure plasma jet and the effect on the biofilm regeneration capacity from the bacteria which survived, and their progenies. The total bacterial populations were 7.18?±?0.34 log10 CFU ml^?1 in biofilms and these were effectively inactivated (>5.5-log10 CFU ml^?1) within 30?min of exposure. Meanwhile, >80% of the S. aureus biofilm cells lost their metabolic capacity. In comparison, ～20% of the plasma-treated bacteria entered a viable but non-culturable state. Moreover, the percentage of membrane-intact bacteria declined to ～30%. Scanning electron microscope images demonstrated cell shrinkage and deformation post-treatment. The total amount of intracellular reactive oxygen species was observed to have significantly increased in membrane-intact bacterial cells with increasing plasma dose. Notably, the N₂ plasma treatment could effectively inhibit the biofilm regeneration ability of the bacteria which survived, leading to a long-term phenotypic response and dose-dependent inactivation effect on S. aureus biofilms, in addition to the direct rapid bactericidal effect.     

Anion channels in the sea urchin sperm plasma membrane

Ionic fluxes in sea urchin sperm plasma membrane regulate cell motility and the acrosome reaction (AR). Although cationic channels mediate some of the ionic movements, little is known about anion channels in these cells. The fusion of sperm plasma membranes into lipid bilayers allowed identification of a 150 pS anion channel. This anion channel was enriched from detergent-solubilized sperm plasma membranes using a wheat germ agglutinin Sepharose column. Vesicles formed from this preparation were fused into black lipid membranes (BLM), yielding single channel anion-selective activity with the properties of those found in the sperm membranes. The following anion selectivity sequence was found: NO₃^? > CNS^? > Br^? > CI^?. This anion channel has a high open probability at the holding potentials tested, it is partially blocked by 4,4′-diisothiocyano-2,2′ -stilbendisulfonic acid (DIDS), and it often displays substates. The sperm AR was also inhibited by DIDS. © 1993 Wiley-Liss, Inc.     

Isolation and characterization of plasma membranes from the adrenal medulla.

Abstract– The isolation of a plasma membrane fraction from the bovine adrenal medulla and its characterization are described. The plasma membranes are enriched 13-fold in AChE, a plasma membrane marker, and represent 0.7% of the homogenate membrane protein. The yield of these membranes is typically 10-12% by the criterion of the percentage of total membrane bound AChE in the homogenate. The membranes were characterized as to their polypeptide, phospholipid and cholesterol content and compared with chromaffin vesicle, mitochondrial and microsomal membranes by these parameters. Two enzymatic components of the plasma membranes, ATPase and adenylate cyclase, were also studied. Calcium ATPase activity is 2.5-fold higher than magnesium ATPase activity, appears to be the result of a single enzyme, and is a genuine component of the plasma membranes. The magnesium stimulated activity appears to have at least two enzymatic components, one of which may be identical to the calcium ATPase. Adenylate cyclase is a plasma membrane component, but may not be uniquely localized there, as it is rather unstable throughout the fractionation procedure. It is stimulated by GTP (0.7-fold at 10^?5M), GPP(NH)P (4.8-fold at 10^?5M) and sodium fluoride (4.6-fold at 10^?2M). It is refractory to stimulation by all other compounds tested.     

Targeting Cancer Cells with Reactive Oxygen and Nitrogen Species Generated by Atmospheric-Pressure Air Plasma

The plasma jet has been proposed as a novel therapeutic method for cancer. Anticancer activity of plasma has been reported to involve mitochondrial dysfunction. However, what constituents generated by plasma is linked to this anticancer process and its mechanism of action remain unclear. Here, we report that the therapeutic effects of air plasma result from generation of reactive oxygen/nitrogen species (ROS/RNS) including H₂O₂, Ox, OH⁻, •O_2, NOx, leading to depolarization of mitochondrial membrane potential and mitochondrial ROS accumulation. Simultaneously, ROS/RNS activate c-Jun NH₂-terminal kinase (JNK) and p38 kinase. As a consequence, treatment with air plasma jets induces apoptotic death in human cervical cancer HeLa cells. Pretreatment of the cells with antioxidants, JNK and p38 inhibitors, or JNK and p38 siRNA abrogates the depolarization of mitochondrial membrane potential and impairs the air plasma-induced apoptotic cell death, suggesting that the ROS/RNS generated by plasma trigger signaling pathways involving JNK and p38 and promote mitochondrial perturbation, leading to apoptosis. Therefore, administration of air plasma may be a feasible strategy to eliminate cancer cells.     

Kelvin-Helmholtz instability of a cylindrical plasma flow with an arbitrary temperature

The dispersion relation for Kelvin-Helmholtz magnetohydrodynamic instability of a cylindrical plasma flow in a longitudinal magnetic field is studied with allowance for plasma compressibility. Stability of the system in a wide range of plasma parameters is thoroughly analyzed in the incompressible plasma approximation. Using the results obtained, a diagram of the system stability is constructed in terms of the magnetic field and the ratio between the plasma densities in the flow and the ambient space. It is shown by numerically solving the dispersion relation for the case of a compressible plasma that perturbations with scale lengths on the order of the flow diameter and larger can develop even at a zero temperature. For low ion-sound velocities, c _S ²/U ₀² < 0.25, the growth rate of the axisymmetric mode with m = 0 is much smaller than that of non-axisymmetric modes. It is shown that, in an incompressible plasma, the eigenmodes are damped monotonically with distance from the flow. In plasma with a finite temperature, the character of damping is oscillatory; in this case, the lower the plasma temperature, the larger the distance at which the ambient plasma is perturbed.     

Experimental study of the excitation of rhodium isomer in a plasma produced by a picosecond laser pulse

Estimates and first experimental results on the excitation of a long-lived isomer state (E _m = 39.756 keV, J ^p = 9/2⁻, and T _1/2 = 56.114 min) of Rh¹⁰³ nuclei under the action of X radiation in a hot solid-state-density rhodium plasma produced by a picosecond laser pulse in the SOKOL-P laser facility are presented.     

Calcium pump activity of the renal plasma membrane and renal microsomes

ATP-dependent Ca²⁺ uptake distinct from that of the mitochondria is found in both plasma membrane and microsomal membranes of rat kidney. Activity attributed to these fractions is enhanced by ammonium oxalate and is apparently insensitive to NaN₃. In contrast, rat kidney mitochondrial Ca²⁺ uptake is blocked by NaN₃. The pH of optimal activity is significantly higher for the mitochondrial fraction. Microsomal membrane Ca²⁺ uptake differs from that of the plasma membrane. Microsomal membranes are four times as active as the plasma membrane at high (5 mM) ATP levels. Apparent K_m values for Mg²⁺-ATP differ in the two preparations with a higher affinity for Mg²⁺-ATP found in the plasma membrane Ca²⁺ uptake activity of the plasma membrane preparation is readily inhibited by Na⁺. Sucrose gradient density fractionation indicates that the observed microsomal membrane Ca²⁺ pump activity is associated with membrane vesicles derived from the endoplasmic reticulum. Ca²⁺ pump activity of both plasma membrane and microsomal fraction is depressed din the adrenalectomized rat. This activity is not restored by a single natriuretic dose of aldosterone.     

Diffusion coefficient of fluorescent phosphatidylinositol 4,5-bisphosphate in the plasma membrane of cells

Phosphatidylinositol 4,5-bisphosphate (PIP₂) controls a surprisingly large number of processes in cells. Thus, many investigators have suggested that there might be different pools of PIP₂ on the inner leaflet of the plasma membrane. If a significant fraction of PIP₂ is bound electrostatically to unstructured clusters of basic residues on membrane proteins, the PIP₂ diffusion constant, D, should be reduced. We microinjected micelles of Bodipy TMR-PIP₂ into cells, and we measured D on the inner leaflet of fibroblasts and epithelial cells by using fluorescence correlation spectroscopy. The average ± SD value from all cell types was D = 0.8 ± 0.2 μm²/s (n = 218; 25°C). This is threefold lower than the D in blebs formed on Rat1 cells, D = 2.5 ± 0.8 μm²/s (n = 26). It is also significantly lower than the D in the outer leaflet or in giant unilamellar vesicles and the diffusion coefficient for other lipids on the inner leaflet of these cell membranes. The simplest interpretation is that approximately two thirds of the PIP₂ on inner leaflet of these plasma membranes is bound reversibly.