Inactivation efficacy of low-pressure plasma treatment against seed-borne tomato pathogen Clavibacter michiganensis and effect of seed setting position and mesh sheet usage.

Clavibacter michiganensis, a gram-positive actinomycete, is a major seed-borne tomato pathogen. We investigated the inactivation efficacy of low-pressure plasma treatment against C. michiganensis inoculated on tomato seeds by placing them on a mesh sheet above the bottom dielectric glass plate. The 2- and 5-minute plasma treatment reduced C. michiganensis populations on the tomato seeds by 0.8 and 1.8 log cfu/seed, respectively. The reduction rates were similar to those of C. michiganensis on shirona (cruciferous) seeds, which have different shapes and surface structures. In contrast, the inactivation of C. michiganensis cells using plasma was more difficult than that of X. campestris cells. Additionally, it was found that placing seeds on a mesh sheet laid on the dielectric glass plate was remarkably effective in inactivating the pathogens on tomato seeds. Since the tomato seeds were susceptible to damage from plasma treatment, methods to reduce its damage need to be investigated.

Antibiotic-Resistant and Non-Resistant Bacteria Display Similar Susceptibility to Dielectric Barrier Discharge Plasma.

Here, we examined whether antibiotic-resistant and non-resistant bacteria show a differential susceptibility to plasma treatment. Escherichia coli DH5α were transformed with pPRO-EX-HT-CAT, which encodes an ampicillin resistance gene and chloramphenicol acetyltransferase (CAT) gene, and then treated with a dielectric barrier discharge (DBD) plasma torch. Plasma treatment reduced the viable cell count of E. coli after transformation/selection and further cultured in ampicillin-containing and ampicillin-free medium. However, there was no significant difference in viable cell count between the transformed and untransformed E. coli after 1 min- and 2 min-plasma treatment. Furthermore, the enzyme-linked immunosorbent assay (ELISA) and acetyltransferase activity assay showed that the CAT activity was reduced after plasma treatment in both transformed and selected E. coli grown in ampicillin-containing or ampicillin-free medium. Loss of lipopolysaccharide and DNA damage caused by plasma treatment were confirmed by a Limulus test and polymerase chain reaction, respectively. Taken together, these findings suggest the plasma acts to degrade components of the bacteria and is therefore unlikely to display a differential affect against antibiotic-resistant and non-resistant bacteria. Therefore, the plasma method may be useful in eliminating bacteria that are recalcitrant to conventional antibiotic therapy.

Key role of singlet oxygen and peroxynitrite in viral RNA damage during virucidal effect of plasma torch on feline calicivirus.

A dielectric barrier discharge (DBD) plasma torch has been used to evaluate the mechanism underlying inactivation of feline calicivirus (FCV) by plasma treatment. Plasma treatment of cell lysate infected with FCV F9 strain reduced the viral titer of the median tissue culture infectious dose (TCID50). The D value (treatment time required to lower the viral titer to 1/10) was 0.450 min, while the viral titer dropped below the detection limit within 2 min. FCV was not significantly inactivated by heat or UV applied at levels corresponding to those generated from the DBD plasma torch after 2 min (38.4 °C and 46.79 mJ/cm2 UV, respectively). However, TCID50 was reduced by 2.47 log after exposure to 4.62 mM ONOO-, corresponding to the concentration generated after 2 min of plasma treatment. Radical scavengers, including superoxide dismutase, dimethyl sulfoxide, and catalase, did not significantly affect viral titers; however, sodium azide, uric acid, and ascorbic acid, which are scavengers of 1O2 radicals, ONOO-, and peroxynitrous acid (ONOOH; produced from ONOO- under acidic conditions), respectively, significantly increased TCID50 and intact viral RNA. These findings suggest that ONOO- and 1O2 play an important role in FCV inactivation by attacking viral RNA during DBD plasma torch treatment.

Treatment of Helicobacter pylori with dielectric barrier discharge plasma causes UV induced damage to genomic DNA leading to cell death.

Gastrointestinal endoscopy is an important tool for the indentification and treatment of disorders of the gastrointestinal tract. However, nosocomial infections of Helicobacter pylori have been linked to the use of contaminated endoscopes. Disinfectants such as glutaraldehyde, ortho-phthalaldehyde and peracetic acid are generally used in the reprocesssing of endoscopes, but these chemicals are hazardous to human health. Thus, safer reprocessing and disinfecion methods are needed. In this study, we applied a dielectric barrier discharge (DBD) plasma torch for inactivation of H. pylori to investigate a potential new methodology to disinfect endoscopes. Suspensions of H. pylori in 10% glycerol were subjected to the DBD plasma torch, which reduced the viable cell count to undetectable levels after 2 min of treatment. Furthermore, urease activity of H. pylori was eliminated after 2 min-plasma treatment, while plasma-treatment reduced the intact DNA of H. pylori in a time-dependent manner. Next, we examined several potential bactericidal factors produced by the DBD plasma torch. Two min-plasma treatment resulted in a small temperature rise (4 °C), ultraviolet radiation (UV) generation, and the production of hydrogen peroxide. H. pylori samples were then exposed to equivalent levels of each of these factors in turn. Our results showed that treatment with heat and hydrogen peroxide at the levels produced after 2-min of plasma treatment did not efficiently inactivate H. pylori, whereas exposure to UV had a significant bactericidal effect. Taken together, UV generated by the plasma torch may be crucial for efficient inactivation of H. pylori by damaging the bacterial DNA.

Low-Pressure Plasma Application for the Inactivation of the Seed-borne Pathogen Xanthomonas campestris.

The aim of this study was to investigate the effect of low-pressure plasma treatment on seed disinfection and the possible mechanisms underlying this effect. Seed-borne disease refers to plant diseases that are transmitted by seeds; seed disinfection is an important technique for prevention of such diseases. In this study, the effectiveness of low-pressure plasma treatment in the inactivation of the seed-borne plant pathogenic bacterium, Xanthomonas campestris, inoculated on cruciferous seeds, was evaluated. The highest inactivation effect was observed when the treatment voltage and argon gas flow rate were 5.5 kV and 0.5 L/min, respectively. The viable cell number of X. campestris was 6.6 log cfu/seed before plasma treatment, and decreased by 3.9 log after 5 min of treatment and by 6.6 log after 40 min. Ethidium monoazide treatment and quantitative real-time PCR results indicated that both the cell membrane and target DNA region were damaged following 5 min of plasma treatment. Although both heat and ozone were generated during the plasma treatment, the contribution of both factors to the inactivation of X. campestris was small by itself in our low-pressure plasma system. Overall, we have shown that our low-pressure plasma system has great applicability to controlling plant pathogenic bacterium contamination of seeds.

Seed disinfection effect of atmospheric pressure plasma and low pressure plasma on Rhizoctonia solani.

Gas plasma generated and applied under two different systems, atmospheric pressure plasma and low pressure plasma, was used to investigate the inactivation efficacy on the seedborne pathogenic fungus, Rhizoctonia solani, which had been artificially introduced to brassicaceous seeds. Treatment with atmospheric plasma for 10 min markedly reduced the R. solani survival rate from 100% to 3% but delayed seed germination. The low pressure plasma treatment reduced the fungal survival rate from 83% to 1.7% after 10 min and the inactivation effect was dependent on the treatment time. The seed germination rate after treatment with the low pressure plasma was not significantly different from that of untreated seeds. The air temperature around the seeds in the low pressure system was lower than that of the atmospheric system. These results suggested that gas plasma treatment under low pressure could be effective in disinfecting the seeds without damaging them.

N2 gas plasma inactivates influenza virus mediated by oxidative stress.

Here we show that N2 gas plasma, produced by applying a short high-voltage pulse using a static induction (SI) thyristor power supply inactivates influenza virus. N2 gas plasma treatment of influenza A and B viruses induced the degradation of viral proteins, including nucleoprotein, hemagglutinin, and neuraminidase. The injury of viral RNA genome and the inactivation of hemagglutination were also observed after N2 gas plasma treatment. These changes were possibly due to changes in the viral envelope, because modification of the lipid content was also suggested by Fourier-transformed infrared spectroscopy. At least three major mechanisms of action (heat, UV-A, and oxidative stress (i.e. hydrogen peroxide-like molecules)) were found in this system. Among them, oxidative stress appeared to be the main factor in the inactivation of influenza virus. In addition, there was an increase in the nitrotyrosine content of viral proteins, suggesting that oxidative stress produced by N2 gas plasma generation oxidized proteins. As a result, oxidation may be the most important factor in the inactivation, degradation, and modification of influenza virus by N2 gas plasma.

Comparison of MR images for age determination; T1 weighted images (T1WI) versus T2* weighted images (T2*WI).

PURPOSE: T1WI (T1 weighted image) was acquired in order to grade bone fusion following the studies by FIFA (Federation Internationale de Football Associations). Research using images other than T1WI has not been reported. The aim of this study is to evaluate the grade of epiphyseal fusion by T2* weighted images (T2*WI) and to investigate new findings on T2*WI as compared with T1WI. METHODS: A total of 87 subjects, all junior football players between the ages of 12 and 17 years old, were examined. T1 and T2* WI were obtained using a 1.2T Open type MR system. The T1WI and T2*WI were rated twice randomly by four radiologists using the FIFA grading system. RESULTS: The intra-rater reliability for grading was higher in T1WI (The Intraclass Correlation Coefficient (ICC)=0.949-0.985) than in T2*WI (ICC=0.917-0.943). The inter-rater reliability for grading was also higher in T1WI (ICC=0.923) than in T2*WI (ICC=0.867). CONCLUSIONS: This research showed that T1WI is a better sequence than T2*WI to evaluate bone fusion following FIFA protocol. It was speculated that the reason for this is that T1WI has higher tissue contrast resolution and enables clearer images of the epiphyseal fusion than T2*WI and the grading system by T1WI was not suitable for T2*WI.

A pilot trial on kinematic magnetic resonance imaging using a superconducting, horizontally opened, 1.2 T magnetic resonance system.

PURPOSE: This study was performed to introduce and evaluate the potential of kinematic magnetic resonance imaging (KMRI) using a high-field open-magnet magnetic resonance (MR) system. METHODS: We attempted to perform KMRI of healthy volunteers' lumbar spine and knee in the lateral position and ankle in the supine position utilizing the superconducting, horizontally opened, 1.2 T MR system (OASIS, HITACHI, Tokyo, Japan). For the KMRI of the lumbar spine, the volunteer had to lie on one side while maintaining maximally anteflexed, neutral, and maximally retroflexed positions and remain still for the duration of the acquisition time for each posture. In the same way, KMRI of the knee was performed with the volunteer's knee flexed at 0°, 30°, 60°, 90°, and 120° in the lateral position, and KMRI of the ankle was performed with the volunteer's ankle in maximally dorsiflexed, neutral, and maximally plantarflexed positions while lying in the supine position. RESULTS: We could acquire higher quality kinematic MR images than those acquired using low-field MR systems. The spinal canal, intervertebral discs and foramina, and facet joints in lumbar spine KMRI; the ligaments, menisci and patellofemoral joint in knee KMRI; and the tibiotalar articulation and peroneal tendon in ankle KMRI were clearly depicted. CONCLUSION: The results of our pilot trial indicated that a superconducting horizontally opened, 1.2 T MR system offers high-quality KMRI images and can be utilized for the kinematic diagnosis and evaluation of sports injuries.