Bactericidal Effect of Underwater Plasma Treatment on Waste Brine from Kimchi Production Process and the Evaluation of Reusability of Plasma-Treated Waste Brine in Salting Kimchi Cabbage.

Recycling waste brine from the Kimchi production process is an important environmental and industry issue. We applied an underwater plasma to reduce food-borne pathogens in the waste brine. The capillary electrodes supplied by alternating current (AC) bi-polar pulsed power were applied to treat 100 L of waste brine. The inactivation efficacy was analyzed using four different agars (Tryptic Soy Agar (TSA), Marine agar (MA), de Man Rogosa Sharpe Agar (MRS), and Yeast Extract-Peptone-Dextrose (YPD), respectively. The microbial population was reduced linearly with treatment time, irrespective of culturing medium. The inactivation followed the log-linear model (R2: 0.96-0.99). The reusability of plasma-treated waste brine (PTWB) was determined by five parameters (salinity, pH, acidity, reducing sugar, and the population of microorganisms) of salted Kimchi cabbage, and they were compared with newly made brine (NMB) and waste brine (WB), respectively. The results showed that the quality of salted Kimchi cabbage prepared by PTWB was not significantly different from that of NMB, indicating that the underwater plasma treatment is feasible to reuse waste brine in the salting process of Kimchi production.

Mechanism of (-)-epigallocatechin gallate (EGCG) dimerization by low-temperature plasma.

The efficient dimerization of (-)-Epigallocatechin gallate (EGCG), which is the major bioactive constituent isolated from the leaves of Camellia sinensis, was initially reported without changes in its stereochemistry using low-temperature plasma. The contribution of plasma during the dimerization of EGCG in a methanolic solution was quantified using a major factor, with the major factor demonstrated based on the contents of newly generated products, in this case the sum of oolonghomobisflavans A and B depending on the plasma treatment method. Samples were treated in three methods: plasma direct treatment, an indirect treatment using only reactive species, and an indirect treatment using effects other than those by reactive species. Ozone was identified as a major factor during the plasma treatment, and the operating ranges of the ozone concentration for regulated dimerization were evaluated. The mechanism by which EGCG synthesizes dimers A and B during the treatment process using low-temperature plasma was investigated using the derived major factor and prior literature. The ozone generated by the plasma reacted with methanol to form formaldehyde, and dimers A and B were synthesized by oligomers through a methylene-bridge by the formaldehyde. A plausible pathway of regulated dimerization was deduced based on these results, and the mechanism of EGCG dimerization by plasma is described using this pathway.

The Effect of Gap Distance between a Pin and Water Surface on the Inactivation of Escherichia coli Using a Pin-to-Water Plasma.

Atmospheric plasmas have been applied for the inactivation of microorganisms. Industrials demand to investigate the relation of the key reactive species induced by plasmas and the operating parameters including boundary conditions in order to control plasma treatment processes. In this study, we investigated the effect of gap distance between a pin-electrode and water surface on inactivation efficacy. When the gap distance decreased from 5 mm to 1 mm, the reduction of Escherichia coli (E. coli) was increased to more than 4 log CFU/mL. The reactive oxygen species measured optically and spectrophotometrically were influenced by gap distance. The results from electron spin resonance (ESR) analysis showed that the pin-to-water plasma generated hydroxyl radical (OH•) and singlet oxygen (1O2) in the water and superoxide radical (O2-•) served as a precursor of OH•. The inactivation of E. coli was significantly alleviated by sodium azide (1O2 scavenger), indicating that 1O2 contributes the most to bacterial inactivation. These findings provide a potentially effective strategy for bacterial inactivation using a pin-to-water plasma.

Plasma-catalytic ethylene removal by a ZSM-5 washcoat honeycomb monolith impregnated with palladium.

The effective removal of dilute ethylene in a novel honeycomb plasma reactor was investigated using a honeycomb catalyst (Pd/ZSM-5/monolith) sandwiched between two-perforated electrodes operating at ambient temperature. Herein, the dependence of catalyst performance on the binder fraction, catalyst preparation method, and catalyst loading was examined. Ethylene removal was carried out by a process comprising cycles of 30-min adsorption conjugated with 15-min plasma-catalytic oxidation. Interestingly, the performance of the cyclic process was superior to continuous plasma-catalytic oxidation and thermally activated catalyst in terms of energy conservation, i.e., ~36 compared to ~105 and ~300 J/L, respectively. Hence, the novel cyclic process can be considered advanced-oxidation technology that features room-temperature oxidation, offers low energy consumption, negligible hazardous by-products emissions such as NOx and O3. Moreover, the process operated under described conditions: low-pressure drop, ambient atmosphere, a mechanically stable system, and a simple reactor configuration, suggesting the practical applicability of this plasma process.

Removal of ethyl acetate in air by using different types of corona discharges generated in a honeycomb monolith structure coated with Pd/γ-alumina.

A method based on the corona discharge produced by high voltage alternating current (AC) and direct current (DC) over a Pd/γ-Al2O3 catalyst supported on a honeycomb structure monolith was developed to eliminate ethyl acetate (EA) from the air at atmospheric pressure. The characteristics of the AC and DC corona discharge generated inside the honeycomb structure monolith were investigated by varying the humidity, gas hourly space velocity (GHSV), and temperature. The results showed that the DC corona discharge is more stable and easily operated at different operating conditions such as humidity, GHSV, and gas temperature compared to the AC discharge. At a given applied voltage, the EA conversion in the DC honeycomb catalyst discharge is, therefore, higher compared with that in the AC honeycomb catalyst discharge (e.g., 96% of EA conversion compared with approximately 68%, respectively, at 11.2 kV). These new results can open opportunities for wide applications of DC corona discharge combined with honeycomb catalysts to VOC treatment.

Molecular hybridization based on (-)-epigallocatechin gallate as a new class of antiglycation agents.

(-)-Epigallocatechin gallate (EGCG) and olivetol hybrid molecules 1-4 were conveniently synthesized using dielectric barrier discharge plasma irradiation. The structures of these unprecedented hybrid molecules were determined by interpretation of spectroscopic data. The unusual hybrid 1 showed improved antiglycation potency toward the advanced formation of glycation end products than the original EGCG and olivetol. The novel hybrid 1 is an interesting new class of antiglycation candidate that requires further investigation.

Growth and bioactive phytochemicals of Panax ginseng sprouts grown in an aeroponic system using plasma-treated water as the nitrogen source.

Ginseng (Panax ginseng Meyer) sprouts are grown to whole plants in 20 to 25 days in a soil-less cultivation system and then used as a medicinal vegetable. As a nitrogen (N) source, plasma-treated water (PTW) has been used to enhance the seed germination and seedling growth of many crops but has not been investigated for its effects on ginseng sprouts. This study established an in-situ system for N-containing water production using plasma technology and evaluated the effects of the PTW on ginseng growth and its bioactive phytochemicals compared with those of an untreated control. The PTW became weakly acidic 30 min after the air discharge at the electrodes because of the formation of nitrate (NO3‒) and nitrite (NO2‒) in the water. The NO3‒ and NO2‒ in the PTW, together with potassium ions (K+), enhanced the shoot biomass of the ginseng sprout by 26.5% compared to the untreated control. The ginseng sprout grown in the PTW had accumulated more free amino acids and ginsenosides in the sprout at 25 days after planting. Therefore, PTW can be used as a liquid N fertilizer for P. ginseng growth and phytochemical accumulation during sprouting under aeroponic conditions.

Novel Hybrid Molecules Based on (-)-Epigallocatechin Gallate as Potent Anti-adipogenic Agents.

A series of novel (-)-epigallocatechin gallate (EGCG)-phloroglucinol hybrid compounds 1-4 has been successfully synthesized by employing a simple and efficient methodology using a dielectric barrier discharge (DBD) plasma irradiation. The new hybrid structures were determined by interpretation of spectroscopic data, with the absolute configurations being established by analysis of the circular dichroism (CD) spectra. The novel hybrids 1 and 2 showed highly improved anti-adipogenic potencies toward both pancreatic lipase and preadipocytes differentiation in 3T3-L1 compared to the original EGCG and phloroglucinol. A novel hybrid 1 represent an interesting subclass of anti-adipogenic candidates that need further research.

Inactivation of Salmonella Typhimurium by Non-Thermal Plasma Bubbles: Exploring the Key Reactive Species and the Influence of Organic Matter.

The key reactive species generated by non-thermal plasma bubbles for the inactivation of Salmonella Typhimurium and the effects of organic matter on the inactivation efficacy were investigated. Plasma, which is primarily composed of ozone (O3), was generated by dielectric barrier discharge and injected into a solution (400 mL) as a bubble. The surviving population of S. Typhimurium decreased in proportion to the treatment time, resulting in a 5.29 log reduction after 5 min of treatment. Verification tests to specify key reactive species were conducted using an O3 destruction unit and reactive oxygen species scavengers. The results indicated that singlet oxygen (1O2) contributes substantially to the inactivation of S. Typhimurium, and that the presence of superoxide anion radicals (O2·-) from O3 is essential for the production of 1O2. When a S. Typhimurium suspension containing organic matter (final concentration: 0, 0.005, 0.05, 0.1, and 0.5 g/L), consisting of beef extract and peptone, was treated with plasma bubbles for 5, 10, 15, 20, 25, and 30 min, respectively, the potential of the plasma bubbles for inactivating S. Typhimurium successfully was verified with longer contact time, despite organic matter attenuating the inactivation efficiency in a dose-dependent manner.

Non-thermal dielectric-barrier discharge plasma induces reactive oxygen species by epigenetically modifying the expression of NADPH oxidase family genes in keratinocytes.

We have previously shown that non-thermal dielectric-barrier discharge (DBD) plasma induces the generation of reactive oxygen species (ROS) in cells; however, the underlying mechanism has not been elucidated. This study aimed to identify the mechanisms through which DBD plasma induces the expression of NADPH oxidase (NOX) family members by epigenetic modification in human keratinocytes (HaCaT). Cell exposure to DBD plasma in 10% oxygen and 90% argon resulted in the generation of ROS, triggering oxidative stress that manifested in various forms, including lipid membrane peroxidation, DNA base modification, and protein carbonylation. DBD plasma upregulated the expression of NOX1, NOX5, and DUOX2 at the mRNA and protein levels; and siRNAs targeting NOX1, NOX5, and DUOX2 attenuated the generation of DBD plasma-induced ROS. DBD plasma upregulated the transcriptional activators TET1, MLL1, and HAT1 and downregulated the transcriptional repressors DNMT1, EZH2, and HDAC1. Additionally, DBD plasma increased the binding of transcriptional activators and decreased the binding of transcriptional repressors to the DUOX2 promoter. Methyl-specific polymerase chain reaction and bisulfite sequencing indicated that DBD plasma decreased methylation at the DUOX2 promoter. These results suggest that DBD plasma induces ROS generation by enhancing the expression of the NOX system through epigenetic DNA and histone modifications.

Emerging Plasma Technology That Alleviates Crop Stress During the Early Growth Stages of Plants: A Review.

Crops during their early growth stages are vulnerable to a wide range of environmental stressors; thus, earlier seed invigoration and seedling establishment are essential in crop production. As an alternative to synthetic chemical treatments, plasma technology could be one of the emerging technologies to enhance seed germination and seedling vigor by managing environmental stressors. Recent studies have shown its beneficial effects in various stress conditions, suggesting that plasma treatment can be used for early crop stress management. This paper reviewed the effects of different types of plasma treatments on plant responses in terms of the seed surface environment (seed scarification and pathogen inactivation) and physiological processes (an enhanced antioxidant system and activated defense response) during the early growth stages of plants. As a result, plasma treatment can enhance seed invigoration and seedling establishment by alleviating the adverse effects of environmental stressors such as drought, salinity, and pathogen infection. More information on plasma applications and their mechanisms against a broad range of stressors is required to establish a better plasma technology for early crop stress management.

Demethylation and microRNA differential expression regulate plasma-induced improvement of chicken sperm quality.

The sperm quality is a vital economical requisite of poultry production. Our previous study found non-thermal dielectric barrier discharge plasma exposure on fertilized eggs could increase the chicken growth and the male reproduction. However, it is unclear how plasma treatment regulates the reproductive capacity in male chickens. In this study, we used the optimal plasma treatment condition (2.81 W for 2 min) which has been applied on 3.5-day-incubated fertilized eggs in the previous work and investigated the reproductive performance in male chickens aged at 20 and 40 weeks. The results showed that plasma exposure increased sperm count, motility, fertility rate, and fertilization period of male chickens. The sperm quality-promoting effect of plasma treatment was regulated by the significant improvements of adenosine triphosphate production and testosterone level, and by the modulation of reactive oxygen species balance and adenosine monophosphate-activated protein kinase and mammalian target of rapamycin pathway in the spermatozoa. Additionally, the plasma effect suggested that DNA demethylation and microRNA differential expression (a total number of 39 microRNAs were up-regulated whereas 53 microRNAs down-regulated in the testis) regulated the increases of adenosine triphosphate synthesis and testosterone level for promoting the chicken sperm quality. This finding might be beneficial to elevate the fertilization rate and embryo quality for the next generation in poultry breeding.

Mutual Interaction between Plasma Characteristics and Liquid Properties in AC-driven Pin-to-Liquid Discharge.

This study investigated the mutual interaction between the plasma and plasma treated water (PTW). Many works have shown that the plasma treatment decreases the pH of PTW due to nitric oxide electrolyte ion but the interactions between PTW and the plasma are still largely unknown. We investigated the effect of PTW on a plasma as well as the effect of a plasma on PTW using a pin-to-liquid discharge system. It is found that PTW affects not only the chemical properties but also the physical properties of the plasma such as breakdown voltage and concentration of plasma column. The decrease of the liquid surface tension of PTW due to nitric oxide electrolyte ion from the plasma results in the increase of plasma current onto the surface of PTW and vice versa. The feedback process will be continued until the transition from normal discharge to abnormal discharge. These results can be basic data for the development of plasma sources to treat liquids.

Innovative Approach of Non-Thermal Plasma Application for Improving the Growth Rate in Chickens.

As an innovative technology in biological applications-non-thermal plasma technique-has recently been applied to living cells and tissues. However, it is unclear whether non-thermal plasma treatment can directly regulate the growth and development of livestock. In this study, we exposed four-day-incubated fertilized eggs to plasma at 11.7 kV for 2 min, which was found to be the optimal condition in respect of highest growth rate in chickens. Interestingly, plasma-treated male chickens conspicuously grew faster than females. Plasma treatment regulated the reactive oxygen species homeostasis by controlling the mitochondrial respiratory complex activity and up-regulating the antioxidant defense system. At the same time, growth metabolism was improved due to the increase of growth hormone and insulin-like growth factor 1 and their receptors expression, and the rise of thyroid hormones and adenosine triphosphate levels through the regulation of demethylation levels of growth and hormone biosynthesis-related genes in the skeletal muscles and thyroid glands. To our knowledge, this study was the first to evaluate the effects of a non-thermal plasma treatment on the growth rate of chickens. This safe strategy might be beneficial to the livestock industry.

MicroRNA-7450 regulates non-thermal plasma-induced chicken Sertoli cell apoptosis via adenosine monophosphate-activated protein kinase activation.

Non-thermal plasma treatment is an emerging innovative technique with a wide range of biological applications. This study was conducted to investigate the effect of a non-thermal dielectric barrier discharge plasma technique on immature chicken Sertoli cell (SC) viability and the regulatory role of microRNA (miR)-7450. Results showed that plasma treatment increased SC apoptosis in a time- and dose-dependent manner. Plasma-induced SC apoptosis possibly resulted from the excess production of reactive oxygen species via the suppression of antioxidant defense systems and decreased cellular energy metabolism through the inhibition of adenosine triphosphate (ATP) release and respiratory enzyme activity in the mitochondria. In addition, plasma treatment downregulated miR-7450 expression and activated adenosine monophosphate-activated protein kinase α (AMPKα), which further inhibited mammalian target of rapamycin (mTOR) phosphorylation in SCs. A single-stranded synthetic miR-7450 antagomir disrupted mitochondrial membrane potential and decreased ATP level and mTOR phosphorylation by targeting the activation of AMPKα, which resulted in significant increases in SC lethality. A double-stranded synthetic miR-7450 agomir produced opposite effects on these parameters and ameliorated plasma-mediated apoptotic effects on SCs. Our findings suggest that miR-7450 is involved in the regulation of plasma-induced SC apoptosis through the activation of AMPKα and the further inhibition of mTOR signaling pathway.

DUOX2-mediated production of reactive oxygen species induces epithelial mesenchymal transition in 5-fluorouracil resistant human colon cancer cells.

The therapeutic benefits offered by 5-fluorouracil (5-FU) are limited because of the acquisition of drug resistance, the main cause of treatment failure and metastasis. The ability of the cancer cells to undergo epithelial-mesenchymal transition (EMT) contributes significantly to cancer metastatic potential and chemo-resistance. However, the underlying molecular mechanisms of 5-FU-resistance have remained elusive. Here, we show that reactive oxygen species (ROS), produced by dual oxidase 2 (DUOX2), promote 5-FU-induced EMT. First, we showed that 5-FU-resistant SNUC5 colon cancer cells (SNUC5/FUR cells) undergo EMT by analyzing the expression of EMT markers such as N-cadherin, vimentin and E-cadherin. In addition, we found that the resistant cells expressed higher levels of Snail, Slug, Twist and Zeb1, which are all critical EMT regulators and had enhanced migratory and invasive capabilities. Furthermore, SNUC5/FUR cells had increased level of DUOX2, resulting in increased ROS level. This effect was due to the enhanced binding of the ten eleven translocation 1 (TET1) demethylase to the DUOX2 promoter in the SNUC5/FUR cells. Importantly, silencing of TET1 reversed the effects of 5-FU on the cells. Finally, the antioxidant N-acetylcysteine attenuated the effects of 5-FU on EMT and metastasis. Our study demonstrates the existence of a TET1/DUOX2/ROS/EMT axis that could play a role in colon cancer chemo-resistance and the aggressiveness of this cancer.

Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels.

The quality of avian semen is an important economic trait in poultry production. The present study examines the in vitro effects of non-thermal dielectric barrier discharge plasma on chicken sperm to determine the plasma conditions that can produce the optimum sperm quality. Exposure to 11.7 kV of plasma for 20 s is found to produce maximum sperm motility by controlling the homeostasis of reactive oxygen species and boosting the release of adenosine triphosphate and respiratory enzyme activity in the mitochondria. However, prolonged exposure or further increase in plasma potential impairs the sperm quality in a time- and dose-dependent manner. Optimal plasma treatment of sperm results in upregulated mRNA and protein expression of antioxidant defense-related and energetic metabolism-related genes by increasing their demethylation levels. However, 27.6 kV of plasma exerts significant adverse effects. Thus, our findings indicate that appropriate plasma exposure conditions improve chicken sperm motility by regulating demethylation levels of genes involved in antioxidant defense and energetic metabolism.

Exposure of keratinocytes to non­thermal dielectric barrier discharge plasma increases the level of 8­oxoguanine via inhibition of its repair enzyme.

Oxidative stress enhances cellular DNA oxidation and may cause mutations in DNA bases, including 8­oxoguanine (8­oxoG). Our recent study reported that exposure of cells to non­thermal dielectric barrier discharge (DBD) plasma generates reactive oxygen species and damages DNA. The present study investigated the effect of non­thermal DBD plasma exposure on the formation of 8­oxoG in HaCaT human keratinocytes. Cells exposed to DBD plasma exhibited increased level of 8­oxoG. In addition, mRNA and protein expression levels of 8­oxoguanine glycosylase 1 (OGG1), an 8­oxoG repair enzyme, were reduced in plasma­exposed cells. Furthermore, the expression level of nuclear factor erythroid 2­related factor 2 (Nrf2), a transcription factor that regulates OGG1 gene expression, was reduced following exposure to DBD plasma. Pretreatment of cells with an antioxidant, N­acetyl cysteine (NAC), prior to plasma exposure suppressed the formation of 8­oxoG and restored the expression levels of OGG1 and Nrf2. In addition, phosphorylation of protein kinase B (Akt), which regulates the activation of Nrf2, was reduced following plasma exposure. However, phosphorylation was restored by pretreatment with NAC. These findings suggested that non­thermal DBD plasma exposure generates 8­oxoG via inhibition of the Akt­Nrf2­OGG1 signaling pathway in HaCaT cells.

Non-thermal gas plasma-induced endoplasmic reticulum stress mediates apoptosis in human colon cancer cells.

Colorectal cancer is a common type of tumor among both men and women worldwide. Conventional remedies such as chemotherapies pose the risk of side­effects, and in many cases cancer cells develop chemoresistance to these treatments. Non­thermal gas plasma (NTGP) was recently identified as a potential tool for cancer treatment. In this study, we investigated the potential use of NTGP to control SNUC5 human colon carcinoma cells. We hypothesized that NTGP would generate reactive oxygen species (ROS) in these cells, resulting in induction of endoplasmic reticulum (ER) stress. ROS generation, expression of ER stress­related proteins and mitochondrial calcium levels were analyzed. Our results confirmed that plasma­generated ROS induce apoptosis in SNUC5 cells. Furthermore, we found that plasma exposure resulted in mitochondrial calcium accumulation and expression of unfolded protein response (UPR) proteins such as glucose­related protein 78 (GRP78), protein kinase R (PKR)­like ER kinase (PERK), and inositol­requiring enzyme 1 (IRE1). Elevated expression of spliced X­box binding protein 1 (XBP1) and CCAAT/enhancer­binding protein homologous protein (CHOP) further confirmed that ROS generated by NTGP induces apoptosis through the ER stress signaling pathway.

Design and characterization of 2.45 GHz electron cyclotron resonance plasma source with magnetron magnetic field configuration for high flux of hyperthermal neutral beam.

A 2.45 GHz electron cyclotron resonance (ECR) source with a magnetron magnetic field configuration was developed to meet the demand of a hyperthermal neutral beam (HNB) flux on a substrate of more than 1x10(15) cm(-2) s(-1) for industrial applications. The parameters of the operating pressure, ion density, electron temperature, and distance between the neutralization plate and the substrate for the HNB source are specified in a theoretical analysis. The electron temperature and the ion density are measured to characterize the ECR HNB source using a Langmuir probe and optical emission spectroscopy. The parameters of the ECR HNB source are in good agreement with the theoretically specified parameters.