Endoscopic Hemostasis in Porcine Gastrointestinal Tract Using CO2 Low-Temperature Plasma Jet.

BACKGROUND: Recently, atmospheric low-temperature plasma (LTP) has attracted attention as a novel medical tool that might be useful for achieving hemostasis. However, conventional plasma sources are too big for use with endoscopes, and the efficacy of LTP for achieving hemostasis in cases of gastrointestinal bleeding is difficult to investigate. In this study, to solve the problem, we developed a 3D-printed LTP jet that has a diameter of 2.8 mm and metal body for endoscopic use. The characteristics, hemostasis efficacy, and safety were investigated. MATERIALS AND METHODS: On investigating the basic characteristics of the developed plasma jet, the electron densities, gas temperatures, and reactive species were measured by emission spectroscopy and thermocouple. To evaluate the efficacy of such hemostatic treatment, porcine gastrointestinal bleeding was treated with the device. In addition, to investigate the safety of such treatment, the CO2 LTP-treated tissue was compared with tissue that was treated with clipping-based or argon plasma coagulation-based hemostasis for 5 d, and hematoxylin and eosin staining was used to evaluate tissue damage in the treated regions. RESULTS: The measurement of emission spectroscopy, power, and electron density of various gas plasmas suggested that a high-density (1014 cm-3) LTP of CO2 was generated by the LTP jet, and the gas temperature was 41.5°C at 3 mm from the outlet of the LTP jet. The CO2 LTP achieved hemostasis of oozing blood by 70 ± 20 s. In addition, the CO2 LTP resulted in earlier recovery than clipping-based or argon plasma coagulation-based hemostases, and the treated regions had no damage by the CO2 LTP treatment. CONCLUSIONS: These results indicated that the developed LTP plasma jet has the potential to be used for endoscopic hemostasis.

Influence of Gas Temperature in Atmospheric Non-Equilibrium Plasma on Bactericidal Effect.

In this study, the relationship between plasma gas temperature and the bactericidal effects on five of bacteria (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis and Bacillus cereus (spore)) in liquid was investigated using a temperature-controllable plasma source. We determined that the bactericidal ability improved as the plasma gas temperature increased. Specifically, the bactericidal ability on E. coli of 80-℃ plasma was enhanced by as much as 6.3 times compared to that of 10-℃ plasma. The relationship between plasma gas temperature and the amount of hydroxyl radical, singlet oxygen, hydrogen peroxide, and ozone introduced into the solution was investigated. Our results also showed that each reactive species production increased by 2.1, 9.0, 1.6, and 17 times, respectively, with 80-℃ compared to 10-℃ plasma. The relationship between the bactericidal ability and amount of reactive species indicated that singlet oxygen and ozone introduced to the solution mostly influenced the bactericidal ability as the plasma gas temperature increased. We conclude that the plasma gas temperature is the crucial parameter for plasma sterilization.

Development of a Dual Plasma Desorption/Ionization System for the Noncontact and Highly Sensitive Analysis of Surface Adhesive Compounds.

We developed a dual plasma desorption/ionization system using two plasmas for the semi-invasive analysis of compounds on heat-sensitive substrates such as skin. The first plasma was used for the desorption of the surface compounds, whereas the second was used for the ionization of the desorbed compounds. Using the two plasmas, each process can be optimized individually. A successful analysis of phenyl salicylate and 2-isopropylpyridine was achieved using the developed system. Furthermore, we showed that it was possible to detect the mass signals derived from a sample even at a distance 50 times greater than the distance from the position at which the samples were detached. In addition, to increase the intensity of the mass signal, 0%-0.02% (v/v) of hydrogen gas was added to the base gas generated in the ionizing plasma. We found that by optimizing the gas flow rate through the addition of a small amount of hydrogen gas, it was possible to obtain the intensity of the mass signal that was 45-824 times greater than that obtained without the addition of hydrogen gas.

Investigation of blood coagulation effect of nonthermal multigas plasma jet in vitro and in vivo.

BACKGROUND: Nonthermal atmospheric pressure plasma (NTAPP) has recently received attention as a novel tool in medicine. It is thought that plasma components yield plasma effects such as sterilization, blood coagulation, and wound healing. These effects are produced without thermal damage. We investigated the blood coagulation effect of NTAPP by using a multigas plasma jet. MATERIALS AND METHODS: Multigas plasma jets can generate NTAPP by several gas species. In this study, argon, oxygen, helium, nitrogen, mock air, and carbon dioxide were used to generate NTAPP, and blood coagulation times were compared with each plasma-treated sample. The NTAPP blood coagulation effects on whole blood with four different anticoagulants were investigated. In addition, in this study, the effects of plasma treatment on porcine tissues and organs were investigated as in vivo experiment. RESULTS: A tendency to coagulate later with argon gas plasma than others was shown. There were no significant differences between oxygen, helium, nitrogen, mock air, and carbon dioxide. Whole blood with each anticoagulant demonstrated fast coagulation by NTAPP treatment. Fast control of the bleeding lesions on porcine stomach and liver by plasma treatment was observed, and no tissue damage due to the plasma treatment was detected by optical microscope. CONCLUSIONS: These experiments suggest the potential of various gas NTAPPs as a novel medical device to control bleeding lesions.

Development of a High-Density Microplasma Emission Source for a Micro Total Analysis System.

To achieve a highly sensitive and onsite analysis of a small amount samples, a microplasma-based micro total analysis systems (µ-TAS) device was developed. A dielectric barrier discharge (DBD) that can generate a stable plasma at atmospheric pressure was generated in a microchip and used as the plasma source. The use of DBD suppresses the temperature rise of the electrodes and enables operation for long times because of a reduction of the electrode damage due to suppression of the current via dielectric interposing between the electrodes. It is expected that the analytical system can be miniaturized because helium plasma is generated in the microchannel contained in the microchip. Emissions from gaseous Cl, Br, and I were analyzed using the plasma source, and it was found that the detection limits for these analytes were 0.22, 0.18, and 0.14 ppm, respectively. The calibration curves for gaseous Cl, Br, and I were also plotted obtaining correlation coefficients of 0.975, 0.955 and 0.986, respectively, and showing good linearity for the developed plasma source.

Direct protein introduction into plant cells using a multi-gas plasma jet.

Protein introduction into cells is more difficult in plants than in mammalian cells, although it was reported that protein introduction was successful in shoot apical meristem and leaves only together with a cell-penetrating peptide. In this study, we tried to introduce superfolder green fluorescent protein (sGFP)-fused to adenylate cyclase as a reporter protein without a cell-penetrating peptide into the cells of tobacco leaves by treatment with atmospheric non-thermal plasmas. For this purpose, CO2 or N2 plasma was generated using a multi-gas plasma jet. Confocal microscopy indicated that sGFP signals were observed inside of leaf cells after treatment with CO2 or N2 plasma without substantial damage. In addition, the amount of cyclic adenosine monophosphate (cAMP) formed by the catalytic enzyme adenylate cyclase, which requires cellular calmodulin for its activity, was significantly increased in leaves treated with CO2 or N2 plasma, also indicating the introduction of sGFP-fused adenylate cyclase into the cells. These results suggested that treatment with CO2 or N2 plasma could be a useful technique for protein introduction into plant tissues.

Irradiation effects of low temperature multi gas plasma jet on oral bacteria.

The purpose of this study is to evaluate the sterilization effects of a newly developed low temperature multi gas plasma jet on oral pathogenic microorganisms (Streptococcus mutans [S. mutans], Lactobacillus fermentum [L. fermentum], Aggregatibacter actinomycetemcomitans [A. actinomycetemcomitans]). Plasma gas which generated from O2, N2, Ar and 50% (O2+N2) was irradiated to the microbes. Effect of O2 plasma irradiation on S. mutans under scanning electron microscopy (SEM) was also observed. O2 plasma was directly applied to dental plaque on human extracted tooth. Then, the depth of enamel resorption area was noted by nanoscale hybrid microscope. O2 had the best sterilizing effect for all microbes. The potent bactericidal effect of plasma irradiation was also observed by SEM. Decalcification of enamel was noted significantly lower in plasma irradiated tooth surface compared to no plasma exposure group. These findings revealed that multi gas plasma jet has great potential to be used for dental treatment.

Imaging of the Staphylococcus aureus Inactivation Process Induced by a Multigas Plasma Jet.

To identify mechanisms underlying the bacterial inactivation process by atmospheric nonthermal plasma using a unique plasma jet that can generate various gas plasmas, Staphylococcus aureus were irradiated with carbon dioxide plasma, which produces a large amount of singlet oxygens, and nitrogen plasma, which produces a large amount of OH radicals. And damaged areas of plasma-treated bacteria were observed by field emission scanning electron microscopy, transmission electron microscopy, and atomic force microscopy. As a result, bacteria were damaged by both gas plasmas, but the site of damage differed according to gas species. Therefore, it suggests that singlet oxygen generated by carbon dioxide plasma or other reactive species caused by singlet oxygen contributes to the damage of internal structures of bacteria through the cell wall and membrane, and OH radicals generated by nitrogen plasma or other reactive species derived from OH radicals contribute to damage of the cell wall and membrane.

Development of Desolvation System for Single-cell Analysis Using Droplet Injection Inductively Coupled Plasma Atomic Emission Spectroscopy.

With a view to enhance the sensitivity of analytical instruments used in the measurement of trace elements contained in a single cell, we have now equipped the previously reported micro-droplet injection system (M-DIS) with a desolvation system. This modified M-DIS was coupled to inductively coupled plasma atomic emission spectroscopy (ICP-AES) and evaluated for its ability to measure trace elements. A flow rate of 100 mL/min for the additional gas and a measurement point -7.5 mm above the load coil (ALC) have been determined to be the optimal parameters for recording the emission intensity of the Ca(II) spectral lines. To evaluate the influence of the desolvation system, we recorded the emission intensities of the Ca(I), Ca(II), and H-ß spectral lines with and without inclusion of the desolvation system. The emission intensity of the H-ß spectral line reduces and the magnitude of the Ca(II)/Ca(I) emission intensity ratio increases four-fold with inclusion of the desolvation system. Finally, the elements Ca, Mg, and Fe present in a single cell of Pseudococcomyxa simplex are simultaneously determined by coupling the M-DIS equipped with the desolvation system to ICP-AES.

Development of a gas-cylinder-free plasma desorption/ionization system for on-site detection of chemical warfare agents.

A gas-cylinder-free plasma desorption/ionization system was developed to realize a mobile on-site analytical device for detection of chemical warfare agents (CWAs). In this system, the plasma source was directly connected to the inlet of a mass spectrometer. The plasma can be generated with ambient air, which is drawn into the discharge region by negative pressure in the mass spectrometer. High-power density pulsed plasma of 100 kW could be generated by using a microhollow cathode and a laboratory-built high-intensity pulsed power supply (pulse width: 10-20 µs; repetition frequency: 50 Hz). CWAs were desorbed and protonated in the enclosed space adjacent to the plasma source. Protonated sample molecules were introduced to the mass spectrometer by airflow through the discharge region. To evaluate the analytical performance of this device, helium and air plasma were directly irradiated to CWAs in the gas-cylinder-free plasma desorption/ionization system and the protonated molecules were analyzed by using an ion-trap mass spectrometer. A blister agent (nitrogen mustard 3) and nerve gases [cyclohexylsarin (GF), tabun (GA), and O-ethyl S-2-N,N-diisopropylaminoethyl methylphosphonothiolate (VX)] in solution in n-hexane were applied to the Teflon rod and used as test samples, after solvent evaporation. As a result, protonated molecules of CWAs were successfully observed as the characteristic ion peaks at m/z 204, 181, 163, and 268, respectively. In air plasma, the limits of detection were estimated to be 22, 20, 4.8, and 1.0 pmol, respectively, which were lower than those obtained with helium plasma. To achieve quantitative analysis, calibration curves were made by using CWA stimulant dipinacolyl methylphosphonate as an internal standard; straight correlation lines (R(2) = 0.9998) of the peak intensity ratios (target per internal standard) were obtained. Remarkably, GA and GF gave protonated dimer ions, and the ratios of the protonated dimer ions to the protonated monomers increased with the amount of GA and GF applied.

Fundamental properties of a touchable high-power pulsed microplasma jet and its application as a desorption/ionization source for ambient mass spectrometry.

Plasma-based ambient desorption/ionization mass spectrometry (ADI-MS) has attracted considerable attention in many fields because of its capacity for direct sample analyses. In this study, a high-power pulsed microplasma jet (HPPMJ) was developed and investigated as a new plasma desorption/ionization source. In an HPPMJ, a microhollow cathode discharge is generated in a small hole (500 µm in diameter) using a pulsed high-power supply. This system can realize a maximum power density of 5 × 10(8) W/cm(3). The measured electron number density, excitation temperature and afterglow gas temperature of the HPPMJ were 3.7 × 10(15) cm(-3), 7000 K at maximum and less than 60 °C, respectively, which demonstrate that the HPPMJ is a high-energy, high-density plasma source that is comparable with an argon inductively coupled plasma while maintaining a low gas temperature. The HPPMJ causes no observable damage to the target because of its low gas temperature and electrode configuration; thus, we can apply it directly to human skin. To demonstrate the analytical capacity of ADI-MS using an HPPMJ, the plasma was applied to direct solid sample analysis of the active ingredients in pharmaceutical tablets. Caffeine, acetaminophen, ethenzamide, isopropylantipyrine and ibuprofen were successfully detected. Application to living tissue was also demonstrated, and isopropylantipyrine on a finger was successfully analyzed without damaging the skin. The limits of detection (LODs) for caffeine, isopropylantipyrine and ethenzamide were calculated, and LODs at the picogram level were achieved. These results indicate the applicability of the HPPMJ for high-sensitivity analysis of materials on a heat-sensitive surface.

Decomposition of tetrodotoxin using multi-gas plasma jet.

In this study, non-thermal multi-gas plasma treatments were performed for Tetrodotoxin (TTX) solution, and TTX decomposition was analyzed by liquid chromatography coupled with electrospray time-of-flight mass spectrometry. The TTX mass spectrum signal was reduced by plasma irradiations to different levels by using various gas species. Nitrogen plasma exhibited the optimal capability for TTX decomposition, followed by oxygen, argon, and carbon dioxide plasmas. The TTX concentration decreased 100-fold by nitrogen plasma treatment for 10 min.

A new air-cooled argon/helium-compatible inductively coupled plasma torch.

A new inductively coupled plasma (ICP) torch with an air-cooling system has been designed and developed for both argon and helium plasma. The same torch and impedance-matching network could be used to generate stable Ar- and He-ICP. The torch consists of three concentric quartz tubes. The carrier gas, plasma gas, and cooling gas flow through the intervals between each tube. In an experiment, it was found that Ar-ICP could form a stable plasma under the following conditions: RF power of 1 kW, plasma gas flow rate of 11 L min(-1), and cooling gas flow rate of 20 L min(-1). For He-ICP, an input RF power of 2 kW, which is two-times higher than that of a conventional He-ICP, could be constantly applied to the plasma with plasma gas and cooling gas flow rates of 15 and 20 L min(-1), respectively. Using this torch, it is possible to realize lower plasma gas consumption for Ar- and He-ICP and a high-power drive for He-ICP. It has been found that the air-cooling gas stabilizes the shape of the plasma due to the pressure difference between the cooling gas and the plasma gas.

Settlement of planulae of the Moon jellyfish Aurelia aurita onto hydrophilic polycarbonate plates modified by atmospheric plasma treatment.

It has been reported that planula larvae of some jellyfish prefer artificial substrates for settlement. This research focused on the relationship between the settlement of planulae and the wettability of artificial substrate surfaces. We used atmospheric plasmas to change the wettability of the surfaces of polycarbonate (PC) plates because plasma treatment has no chemical side effects. The treatment made the surfaces hydrophilic, as evidenced by the decrease of contact angle from 85° to 35°. X-ray photoelectron spectroscopy revealed that the change of wettability of the PC plates could be attributed to N2, which was probably ionized in the air above the plates. Scanning electron microscopy revealed no difference in the surface morphology of the plates before and after plasma treatment. Results of bioassays using treated PC plates showed that planulae tended to preferentially settle on hydrophobic surfaces.

Development of the atmospheric plasma soft-ablation method (APSA) for elemental analysis of materials on heat-sensitive substrates.

To achieve a highly sensitive and prompt elemental analysis of materials on heat-sensitive substrates, like living tissues, the atmospheric plasma soft-ablation method (APSA) was developed. The damage-free plasma, which has room temperature and no risk of electrical shock, was used as a sampling medium for materials, and the sampled materials were introduced to inductively coupled plasma mass spectrometer (ICP-MS). By using APSA, a mass signal of molybdenum, using a thin molybdenum-grease layer, was successfully obtained without damaging the subjacent glass substrate. The use of a hydrogen admixture to generate the plasma was examined in order to achieve more effective sampling by utilizing chemical reactions between radicals in the plasma and the sample material. As a result, the sensitivity of all measured elements contained in tablet supplements increased by up to 20 times upon the addition of 1% hydrogen, even though the plasma-gas temperature did not change significantly.

Development of injection gas heating system for introducing large droplets to inductively coupled plasma.

We developed an injection gas heating system for introducing large droplets, because we want to effectively to measure elements in a single cell. This system was applied to ICP-atomic emission spectrometry (ICP-AES), to evaluate it performance. To evaluate the effect of the emission intensity, the emission intensity of Ca(II) increased to a maximum of tenfold at 147°C and the peak was shifted upstream of the plasma. To investigate in detail the effect of an injection gas heating system, we studied different conditions of the injection gas temperature and droplet volume. When the injection gas temperature was 89°C, smaller droplets were easily ionized. At 147°C, the emission intensity ratio and the absolute amount of the sample including the droplet exhibited close agreement. These results show the advantages of the injection gas heating system for large droplet introduction, and the sufficient reduction in the solvent load. The solvent load could be reduced by heating to 147°C using the system.

Wide-bore capillary hydrodynamic chromatography with ICP-MS detection for evaluation of lanthanide uptake by molecular aggregates.

Wide-bore capillary hydrodynamic chromatography (W-HDC) resolves analytes on the basis of a difference in the extent of radial diffusion simply by their passage through an empty capillary. The combination of this method with ICP-MS proves efficient for the evaluation of the interaction of metal ions with molecular aggregates. Lecithin vesicles are suitable molecular aggregates for the uptake of the lanthanide ions in the presence of the first row transition metal ions, suggesting that the present method is applicable to the screening of the molecular aggregate system suitable for selective extraction of a particular targeted small molecule. The visual inspection of the elution profiles gives us qualitative but useful information on the interaction between the vesicle and metal ions. In addition, studying the slope of the front edge of an elution curve provides more quantitative implications.