Importance of bicarbonate transport in pH control during amelogenesis - need for functional studies.

Dental enamel, the hardest mammalian tissue, is produced by ameloblasts. Ameloblasts show many similarities to other transporting epithelia although their secretory product, the enamel matrix, is quite different. Ameloblasts direct the formation of hydroxyapatite crystals, which liberate large quantities of protons that then need to be buffered to allow mineralization to proceed. Buffering requires a tight pH regulation and secretion of bicarbonate by ameloblasts. Many investigations have used immunohistochemical and knockout studies to determine the effects of these genes on enamel formation, but up till recently very little functional data were available for mineral ion transport. To address this, we developed a novel 2D in vitro model using HAT-7 ameloblast cells. HAT-7 cells can be polarized and develop functional tight junctions. Furthermore, they are able to accumulate bicarbonate ions from the basolateral to the apical fluid spaces. We propose that in the future, the HAT-7 2D system along with similar cellular models will be useful to functionally model ion transport processes during amelogenesis. Additionally, we also suggest that similar approaches will allow a better understanding of the regulation of the cycling process in maturation-stage ameloblasts, and the pH sensory mechanisms, which are required to develop sound, healthy enamel.

X-ray diagnostics of ECR ion sources-Techniques, results, and challenges.

The high magnetic confinement provided by the minimum-B structure of electron cyclotron resonance ion sources (ECRIS) hosts a non-equilibrium plasma, composed of cold multi-charged ions and hot electrons whose energy can expand up to ≈1 MeV. With a very limited accessibility, the ECR plasma is difficult to study. The x-ray photons generated by the interaction of the warm and hot electron populations within the plasma, as well as the photons generated by electrons lost to the chamber wall, are a signal of great interest as it gives an insight to the properties of the ECR plasma. After an introduction presenting the mechanism generating the x rays in ECRIS, this Review presents the methodology to measure x rays both for volumetric measurements and plasma imaging. The main insights of those measurements are presented. Prospects of x-ray measurements to better understand the plasma dynamics are finally highlighted.

Two-chamber configuration of Bio-Nano electron cyclotron resonance ion source for fullerene modification.

We report on the modification of fullerenes with iron and chlorine using two individually controllable plasmas in the Bio-Nano electron cyclotron resonance ion source (ECRIS). One of the plasmas is composed of fullerene and the other one is composed of iron and chlorine. The online ion beam analysis allows one to investigate the rate of the vapor-phase collisional modification process in the ECRIS, while the offline analyses (e.g., liquid chromatography-mass spectrometry) of the materials deposited on the plasma chamber can give information on the surface-type process. Both analytical methods show the presence of modified fullerenes such as fullerene-chlorine, fullerene-iron, and fullerene-chlorine-iron.

Ion beam emittance from an ECRIS.

Simulation of ion beam extraction from an Electron Cyclotron Resonance Ion Source (ECRIS) is a fully 3 dimensional problem, even if the extraction geometry has cylindrical symmetry. Because of the strong magnetic flux density, not only the electrons are magnetized but also the Larmor radius of ions is much smaller than the geometrical dimension of the plasma chamber (Ø 64 × 179 mm). If we assume that the influence of collisions is small on the path of particles, we can do particle tracking through the plasma if the initial coordinates of particles are known. We generated starting coordinates of plasma ions by simulation of the plasma electrons, accelerated stochastically by the 14.5 GHz radio frequency power fed to the plasma. With that we were able to investigate the influence of different electron energies on the extracted beam. Using these assumptions, we can reproduce the experimental results obtained 10 years ago, where we monitored the beam profile with the help of viewing targets. Additionally, methods have been developed to investigate arbitrary 2D cuts of the 6D phase space. To this date, we are able to discuss full 4D information. Currently, we extend our analysis tool towards 5D and 6D, respectively.

X-ray pinhole camera setups used in the Atomki ECR Laboratory for plasma diagnostics.

Imaging of the electron cyclotron resonance (ECR) plasmas by using CCD camera in combination with a pinhole is a non-destructive diagnostics method to record the strongly inhomogeneous spatial density distribution of the X-ray emitted by the plasma and by the chamber walls. This method can provide information on the location of the collisions between warm electrons and multiple charged ions/atoms, opening the possibility to investigate the direct effect of the ion source tuning parameters to the plasma structure. The first successful experiment with a pinhole X-ray camera was carried out in the Atomki ECR Laboratory more than 10 years ago. The goal of that experiment was to make the first ECR X-ray photos and to carry out simple studies on the effect of some setting parameters (magnetic field, extraction, disc voltage, gas mixing, etc.). Recently, intensive efforts were taken to investigate now the effect of different RF resonant modes to the plasma structure. Comparing to the 2002 experiment, this campaign used wider instrumental stock: CCD camera with a lead pinhole was placed at the injection side allowing X-ray imaging and beam extraction simultaneously. Additionally, Silicon Drift Detector (SDD) and High Purity Germanium (HPGe) detectors were installed to characterize the volumetric X-ray emission rate caused by the warm and hot electron domains. In this paper, detailed comparison study on the two X-ray camera and detector setups and also on the technical and scientific goals of the experiments is presented.

Evidence for Bicarbonate Secretion by Ameloblasts in a Novel Cellular Model.

Formation and growth of hydroxyapatite crystals during amelogenesis generate a large number of protons that must be neutralized, presumably by HCO3 (-)ions transported from ameloblasts into the developing enamel matrix. Ameloblasts express a number of transporters and channels known to be involved in HCO3 (-)transport in other epithelia. However, to date, there is no functional evidence for HCO3 (-)transport in these cells. To address questions related to HCO3 (-)export from ameloblasts, we have developed a polarized 2-dimensional culture system for HAT-7 cells, a rat cell line of ameloblast origin. HAT-7 cells were seeded onto Transwell permeable filters. Transepithelial resistance was measured as a function of time, and the expression of transporters and tight junction proteins was investigated by conventional and quantitative reverse transcription polymerase chain reaction. Intracellular pH regulation and HCO3 (-)transport were assessed by microfluorometry. HAT-7 cells formed epithelial layers with measureable transepithelial resistance on Transwell permeable supports and expressed claudin-1, claudin-4, and claudin-8-key proteins for tight junction formation. Transport proteins previously described in maturation ameloblasts were also present in HAT-7 cells. Microfluorometry showed that the HAT-7 cells were polarized with a high apical membrane CO2 permeability and vigorous basolateral HCO3 (-)uptake, which was sensitive to Na(+)withdrawal, to the carbonic anhydrase inhibitor acetazolamide and to H2DIDS inhibition. Measurements of transepithelial HCO3 (-)transport showed a marked increase in response to Ca(2+)- and cAMP-mobilizing stimuli. Collectively, 2-dimensional HAT-7 cell cultures on permeable supports 1) form tight junctions, 2) express typical tight junction proteins and electrolyte transporters, 3) are functionally polarized, and 4) can accumulate HCO3 (-)ions from the basolateral side and secrete them at the apical membrane. These studies provide evidence for a regulated, vectorial, basolateral-to-apical bicarbonate transport in polarized HAT-7 cells. We therefore propose that the HAT-7 cell line is a useful functional model for studying electrolyte transport by ameloblasts.

Recent developments of ion sources for life-science studies at the Heavy Ion Medical Accelerator in Chiba (invited).

With about 1000-h of relativistic high-energy ion beams provided by Heavy Ion Medical Accelerator in Chiba, about 70 users are performing various biology experiments every year. A rich variety of ion species from hydrogen to xenon ions with a dose rate of several Gy/min is available. Carbon, iron, silicon, helium, neon, argon, hydrogen, and oxygen ions were utilized between 2012 and 2014. Presently, three electron cyclotron resonance ion sources (ECRISs) and one Penning ion source are available. Especially, the two frequency heating techniques have improved the performance of an 18 GHz ECRIS. The results have satisfied most requirements for life-science studies. In addition, this improved performance has realized a feasible solution for similar biology experiments with a hospital-specified accelerator complex.

Organ perfusion with Hemopump device assistance with and without intraaortic balloon pumping.

OBJECTIVE: Our objective was to analyze the potential advantage of combining an intraaortic balloon pump with a transthoracic Hemopump device (Medtronic, Inc., Minneapolis, Minn.) (Nimbus Medical, Inc., Rancho Cordova, Calif.). METHODS: Twelve sheep underwent implantation of a transthoracic Hemopump device and an intraaortic balloon pump. In the first series (n = 6), we analyzed the influence of the counterpulsation on the performance of the Hemopump device. In the second group (n = 6), hemodynamic changes, myocardial wall thickening, organ perfusion, and myocardial perfusion (determined with colored microspheres) were analyzed under the following conditions: (1) control situation, (2) during application of coronary stenosis, (3) during support with the Hemopump device, and (4) during support with the Hemopump device combined with intraaortic balloon pump support. RESULTS: In the first series, we found that addition of counterpulsation reduced output with the Hemopump device by 11.1% +/- 6%. In the second series, it was shown that coronary stenosis significantly reduced contractility (rate of pressure change and wall thickening) but did not cause hemodynamic collapse. Myocardial blood flow was significantly reduced in the poststenotic subendocardial regions (mean subendocardial blood flow dropped from 78 +/- 33 to 24 +/- 17 ml/min/100 gm; p = 0.0486). Support with the Hemopump device alone improved the ratio of subendocardial to subepicardial blood flow, but endocardial underperfusion remained (analysis of variance, p < 0.001). The Hemopump device with an intraaortic balloon pump completely restored perfusion in poststenotic regions. Peripheral organ perfusion did not change during ischemia or mechanical support. CONCLUSIONS: The association of balloon counterpulsation with the Hemopump device reduces the Hemopump output by 11% and increases myocardial blood flow to ischemic regions. Perfusion to peripheral organs remains unaltered. The transthoracic Hemopump device combined with an intraaortic balloon pump is an ideal support system for the ischemic, failing heart.

The intra-aortic cannula pump: A novel assist device for the acutely failing heart.

OBJECTIVE: The intra-aortic cannula pump is a catheter pump designed to support the acutely failing heart. It expels blood from the left ventricle into the ascending aorta in a pulsatile flow pattern. The aim of the study was to analyze the hemodynamic performance of this new intracardiac support system in acute heart failure. METHODS: A 24F cannula was studied in a series of 16 sheep. Hemodynamic changes were assessed in the nonfailing, the moderately failing, and the severely failing heart. Heart failure was induced by an injection of microspheres into the left anterior descending coronary artery. The cannula was inserted through the aortic arch and introduced through the aortic valve into the left ventricle. RESULTS: Cannula insertion was feasible in all animals. Flow through the intra-aortic cannula flow was increased to a maximum of 3 L/min. No hemodynamic changes were observed in the nonfailing heart. A significant increase in cardiac output was observed in the moderately and severely reduced left ventricle (2.67 +/- 0.7 L to 3.51 +/- 0.83 L; P =.001; and 1.18 +/- 0.77 L to 2.43 +/- 0.44 L; P =.001, respectively). A drop in left atrial pressure was achieved in moderate and severe heart failure (14.1 +/- 5.93 mm Hg to 9.71 +/- 2.63 mm Hg; P =.0001; and 23 +/- 7.16 mm Hg to 11.2 +/- 2.55 mm Hg; P = 0.0001, respectively). Systolic and diastolic systemic blood pressures increased in the severely failing heart (57.3 +/- 12.8 mm Hg to 75.4 +/- 11.2 mm Hg; P =.0001; and 35.6 +/- 8.2 mm Hg to 60 +/- 14.3 mm Hg; P =.0006, respectively). CONCLUSIONS: Hemodynamic data demonstrate the beneficial effects of the intra-aortic cannula pump in moderate and severe heart failure. The intra-aortic cannula pump represents a new modality for the treatment of acute heart failure.

Ascending versus descending aortic balloon pumping: organ and myocardial perfusion during ischemia.

BACKGROUND: The ICS-Supracor (Abiomed, Danvers, MA) is a preshaped ascending aorta balloon pump. We compared the effects of this catheter with the classical descending intraaortic balloon pump (IABP). The study focused on hemodynamic effects, myocardial blood flow in normal and ischemic regions, cerebral perfusion, and peripheral organ perfusion. METHODS: We placed a stenosis on the lateral branch of the coronary artery to reduce flow 50% (sheep). Measurements included hemodynamic changes, myocardial blood flow, and organ flow (colored microspheres) at baseline, after stenosis, during IABP support, and during ICS support. RESULTS: Counterpulsation with the ICS led to a significantly higher peak diastolic aortic augmentation than with the IABP (IABP, 99 +/- 14 mm Hg; ICS, 140 +/- 29 mm Hg; p = 0.003). There was no significant change in cerebral perfusion or peripheral organ perfusion. Myocardial blood perfusion was significantly increased by the IABP as well as the ICS. This effect was seen in ischemic and nonischemic regions (subendocardial and subepicardial). The ICS improved myocardial blood flow significantly more than the IABP (IABP, 0.65 +/- 0.1 mL/min/g; ICS, 0.94 +/- 0.06 mL/min/g; p = 0.0005). CONCLUSIONS: The ICS increases myocardial blood flow in ischemic regions significantly more than the IABP, without impairment of cerebral flow. Assessment of vascular complications, peripherally and in the ascending aorta, has to await results of clinical trials.

Identification of aldosterone secretion inhibitory factor in bovine adrenal medulla.

We report the first demonstration of an Aldosterone Secretion Inhibitory Factor (ASIF) in acid extracts of bovine adrenal medulla. Following separation from catecholamines and enkephalins, this factor leads to an 80% inhibition of PGE1-stimulated secretion of aldosterone from bovine adrenal zona glomerulosa. ASIF is retained on cation exchange gels and behaves as a small 5K-dalton peptide on Sephadex G-50. This factor cross-reacts in a radio-receptor assay for [125I] atrial natriuretic factor (ANF). ASIF is distinct from all neuropeptides formerly detected in the adrenal medulla, e.g. somatostatin, enkephalin, neuropeptide Y, dynorphin, neurotensin. In the adrenal gland, this ANF-like factor is predominantly found in the medulla (4 pmol/mg protein), with only trace amounts in the cortex (0.1 pmol/mg protein). ASIF might perhaps correspond to the endogenous ligand for the receptor sites that we have previously identified with [125I]ANF in bovine adrenal cortex and could contribute to the formerly reported attenuating influence of the adrenal medulla on mineralocorticoid production.

Effect of transmyocardial laser revascularization on chronic ischemic hearts in sheep.

BACKGROUND: We investigated the effect of transmyocardial laser revascularization (TMR) on myocardial function and regional blood flow in an animal model of ischemic heart disease. METHODS: Chronic ischemia was induced in 11 sheep by the application of coronary stenosis on the left anterior descending (LAD) and circumflex coronary artery (LCX). Ten weeks later, in six of them, transmyocardial channels were created in the anterior free wall and in the posterior wall of the left ventricle. Five animals served as controls. The myocardial function was assessed by echocardiography taken at baseline and every 2 weeks after coronary stenosis and after TMR. Myocardial perfusion was measured by colored microspheres, injected at baseline, immediately after coronary stenosis, before and after TMR, and at 20 weeks after coronary stenosis. The hearts were retrieved at 20 weeks for light microscopic examination. RESULTS: The left ventricular end-diastolic and end-systolic cavity area was elevated 20 weeks after coronary stenosis in the control and TMR groups. There was no difference between groups (analysis of variance; ANOVA, non-significant). The wall thickening fraction (WTF) decreased progressively and significantly after coronary stenosis in both groups. The WTF was further acutely reduced by TMR, and recovered gradually to the pre-TMR level. No significant difference in WTF was observed between the TMR and control groups. The resting myocardial blood flow was significantly increased by TMR at 20 weeks (P=0.03). Light microscopic examination revealed channel patency in 49% of the laser scars at 10 weeks post-TMR. A dense capillary network was observed at the edges of the surrounding scar. CONCLUSIONS: In an experimental model of ischemic heart disease, TMR developed angiogenesis in the lased channels, but, however, failed to improve myocardial function.

Calcification characteristics of porcine stentless valves in juvenile sheep.

OBJECTIVE: To compare calcification characteristics of two porcine stentless valves (Toronto SPV and Freestyle) with different designs, fixation and antimineralization techniques using a juvenile sheep model of valve implantation inside the circulation. METHODS: The stentless valves (n = 2 x 6) were implanted in juvenile sheep in the pulmonary artery as an interposition, while the circulation was maintained with a right ventricular assist device. The model was validated by the implantation of, clinically well-known, porcine (Hancock II) and pericardial (Pericarbon) valves. Half of the valves were explanted after 3 months, the rest after 6 months. Valves were examined macroscopically, by X-ray, light microscopy (HE, Masson, Von Giesson, Von Kossa, PTAH stains), and transmission electron microscopy. Quantitative determination of the calcium content of the cusps was performed with atomic absorption spectrometry. RESULTS: After 3 months, the Freestyle had an extensively calcified aortic wall, most prominent at the outflow side of the porcine valve. After 6 months, calcification increased transmurally, but the valve cusps were free of calcification, and the inflow side was only slightly calcified. The Toronto SPV valve also started to calcify at the inflow side of the valve after 3 months with increased calcification after 6 months. The base of the Toronto SPV valve cusps showed slight calcification after 6 months of implantation. CONCLUSIONS: The pattern of calcification of the porcine aortic wall differs between the two studied stentless valves, with calcification located predominantly at the outflow side in the Freestyle valve, but also at the inflow side in the Toronto SPV valve. The cusps of the Freestyle valve were less prone to calcification than those from the Toronto SPV valve.

Measurement of resistance versus flow in assessing efficiency of aortocoronary bypass grafts.

Measurement of flow in saphenous bypass grafts with an electromagnetic flowmeter is complicated and poorly reproducible. Since coronary flow is largely dependent on variable factors the stable value of resistance seems more appropriate for comparison. A simple method has been developed for intraoperative measurement of resistance in the respective coronary bed. Pressure is recorded in the saphenous graft by an electromanometer during continuous flushing with known amounts of blood, and resistance is calculated instantaneously. The procedure is very simple and takes less than one minute. The quality of the saphenous vein itself can be assessed simultaneously by the same method. Resistances were measured during coronary surgery in over 500 saphenous grafts. The results were highly reproducible and comparable. Excellent flows can be expected if resistance is below 200 Peripheral Resistance Units (PRU); if this is over 800 PRU flow is very poor.

The enabler cannula pump: a novel circulatory support system.

BACKGROUND: The enabler circulatory support system is a catheter pump which expels blood from the left or right ventricular cavity and provides pulsatile flow in the ascending aorta or pulmonary artery. It is driven by a bedside installed pulsatile driving console. The device can easily be implanted by a minimal invasive approach, similar to the Hemopump. PURPOSE: To demonstrate the hemodynamic performance of this new intracardiac support system. METHODS: In a series of 9 sheep, hemodynamic evolutions were recorded in various conditions of myocardial contractility (the non-failing, the moderately failing and the severely failing heart). Heart failure was induced by injection of microspheres in the coronary arteries. RESULTS: Introduction of the cannula through the aortic valve was feasible in all cases. Pump flow by the enabler was gradually increased to a maximum of 3.5 L/min. Diastolic (and mean) aortic blood pressure is significantly increased in the non-failing and moderately failing condition (counterpulsation mode). In heart failure, cardiac output is significantly increased by the pump (p < 0.0001). A drop in left atrial pressure (indicating unloading) is achieved in all conditions but reaches significant levels only during heart failure (p=0.0068). CONCLUSIONS: This new circulatory support system contributes to stabilization of the circulation in the presence of cardiac unloading. In heart failure it actually supports the circulation by increasing cardiac output and perfusion pressure.

Fast camera studies at an electron cyclotron resonance table plasma generator.

A simple table-size ECR plasma generator operates in the ATOMKI without axial magnetic trap and without any particle extraction tool. Radial plasma confinement is ensured by a NdFeB hexapole. The table-top ECR is a simplified version of the 14 GHz ATOMKI-ECRIS. Plasma diagnostics experiments are planned to be performed at this device before installing the measurement setting at the "big" ECRIS. Recently, the plasma generator has been operated in pulsed RF mode in order to investigate the time evolution of the ECR plasma in two different ways. (1) The visible light radiation emitted by the plasma was investigated by the frames of a fast camera images with 1 ms temporal resolution. Since the visible light photographs are in strong correlation with the two-dimensional spatial distribution of the cold electron components of the plasma it can be important to understand better the transient processes just after the breakdown and just after the glow. (2) The time-resolved ion current on a specially shaped electrode was measured simultaneously in order to compare it with the visible light photographs. The response of the plasma was detected by changing some external setting parameters (gas pressure and microwave power) and was described in this paper.

Experiments with biased side electrodes in electron cyclotron resonance ion sources.

The output of highly charged ions from an electron cyclotron resonance ion source (ECRIS) consists of ionic losses from a highly confined plasma. Therefore, an increase of the output of the ions of interest always is a compromise between an increase in the confinement and an increase of the losses. One route towards a solution consists of attacking the losses in directions - i.e., radial directions - that do not contribute to the required output. This was demonstrated in an experiment (using the Kei ECRIS at NIRS, Japan) where radial losses were electrostatically reduced by positively biasing one set of six "side" electrodes surrounding the plasma in side-ward directions attached (insulated) to the cylindrical wall of the plasma chamber. Recently new studies were performed in two laboratories using two essentially different ion sources. At the BioNano ECRIS (Toyo University, Japan) various sets of electrodes were used; each of the electrodes could be biased individually. At the Atomki ECRIS (Hungary), one movable, off-axis side electrode was applied in technically two versions. The measurements show indeed a decrease of ionic losses but different effectivities as compared to the biased disk.

Chronic hibernating myocardium in sheep can occur without degenerating events and is reversed after revascularization.

INTRODUCTION: Our goal was to show that blunting of myocardial flow reserve is mainly involved in adaptive chronic myocardial hibernation without apparent cardiomyocyte degeneration. METHODS AND RESULTS: Sheep chronically instrumented with critical multivessel stenosis and/or percutaneous transluminal coronary angioplasty (PTCA)-induced revascularization were allowed to run and feed in the open for 2 and 5 months, respectively. Regional myocardial blood flow (MBF) with colored microspheres, regional and global left ventricular function and dimensions (2D echocardiography), and myocardial structure were studied. In sheep with a critical stenosis, a progressive increase in left ventricular end-diastolic and end-systolic cavity area and a decrease in fractional area change were found. Fraction of wall thickness decreased in all left ventricular wall segments. MBF was slightly but not significantly decreased at rest at 2 months. Morphological quantification revealed a rather small but significant increase in diffusely distributed connective tissue, cardiomyocyte hypertrophy, and presence of viable myocardium of which almost 30 % of the myocytes showed depletion of sarcomeres and accumulation of glycogen. The extent of myolysis in the transmural layer correlated with the degree of left ventricular dilation. Structural degeneration of cardiomyocytes was not observed. Balloon dilatation (PTCA) of one of the coronary artery stenoses at 10 weeks revealed recovery of fraction of wall thickness and near normalization of global subcellular structure at 20 weeks. CONCLUSION: These data indicate that chronic reduction of coronary reserve by itself can induce ischemic cardiomyopathy characterized by left ventricular dilatation, depressed regional and global function, adaptive chronic myocardial hibernation, reactive fibrosis and cardiomyocyte hypertrophy in the absence of obvious degenerative phenomena. SUMMARY: Reduction of myocardial flow reserve due to chronic coronary artery stenosis in sheep induces adaptive myocardial hibernation without involvement of degenerative phenomena.

Two-frequency heating technique at the 18 GHz electron cyclotron resonance ion source of the National Institute of Radiological Sciences.

The two-frequency heating technique was studied to increase the beam intensities of highly charged ions provided by the high-voltage extraction configuration (HEC) ion source at the National Institute of Radiological Sciences (NIRS). The observed dependences on microwave power and frequency suggested that this technique improved plasma stability but it required precise frequency tuning and more microwave power than was available before 2013. Recently, a new, high-power (1200 W) wide band-width (17.1-18.5 GHz) travelling-wave-tube amplifier (TWTA) was installed. After some single tests with klystron and TWT amplifiers the simultaneous injection of the two microwaves has been successfully realized. The dependence of highly charged ions (HCI) currents on the superposed microwave power was studied by changing only the output power of one of the two amplifiers, alternatively. While operating the klystron on its fixed 18.0 GHz, the frequency of the TWTA was swept within its full limits (17.1-18.5 GHz), and the effect of this frequency on the HCI-production rate was examined under several operation conditions. As an overall result, new beam records of highly charged argon, krypton, and xenon beams were obtained at the NIRS-HEC ion source by this high-power two-frequency operation mode.

Fullerene-rare gas mixed plasmas in an electron cyclotron resonance ion source.

A synthesis technology of endohedral fullerenes such as Fe@C60 has developed with an electron cyclotron resonance (ECR) ion source. The production of N@C60 was reported. However, the yield was quite low, since most fullerene molecules were broken in the ECR plasma. We have adopted gas-mixing techniques in order to cool the plasma and then reduce fullerene dissociation. Mass spectra of ion beams extracted from fullerene-He, Ar or Xe mixed plasmas were observed with a Faraday cup. From the results, the He gas mixing technique is effective against fullerene destruction.