Properties of essential oils absorbed on the surface of cardboard pieces after using atmospheric-pressure plasma treatments to develop long-lasting Varroa miticides in honeybees (Apis mellifera).

The ectoparasitic mite, Varroa destructor is the most serious widespread pest of managed honeybees (Apis mellifera). Several acaricide products, which include essential oils, have been proposed for mite control. In this study, we aimed to apply atmospheric-pressure plasma to modify a cardboard piece surface in order to prolong the delivery of essential oils for controlling Varroa in honeybee colonies. Absorption capacity, release rates and evaporation rates of essential oils were determined. Cardboard piece showed a higher absorption capacity of cinnamon compared to citronella and clove. Surface modification of cardboard pieces using argon plasma at different gas flow rates and treatment durations, significantly affected the absorption of clove oil. Additionally, the release rate of cinnamon, citronella and clove was significantly enhanced after argon plasma treatments. Evaporation of cinnamon was dramatically increased by plasma treatment at 6-h of incubation. The highest evaporation rate was obtained by plasma-treated cardboard piece at a gas flow rate of 0.5 Lpm for 60 s (0.2175 ± 0.0148 µl/g•h). Efficiency of plasma-treated cardboard piece, impregnated with essential oils, was also investigated for Varroa control in honeybee colonies. In the first experiment, formic acid 65% (v/v) showed the highest efficiency of 90.60% and 81.59% with the percent of mite infestation was 0.23 ± 0.13% and 0.47 ± 0.19% at 21 and 35 days, respectively after treatment. The efficacy of cardamon oil (5% (v/v)) delivered using plasma-treated cardboard pieces was 57.71% (0.70 ± 0.16% of mite infestation) at day 21 of experiment. However, the delivery of cardamon oil at the concentration of 1% and 5% (v/v) by untreated cardboard piece had 16.93% and 24.05% of efficacy to control mites. In the 2nd experiment, the application of plasma-treated cardboard pieces impregnated with 5% (v/v) clove oil induced a 38.10% reduction in the population of Varroa mites followed by 5% (v/v) of cardamon with 30% efficiency. Although, the infestation rate of Varroa in colonies was not significant different between treatments, essential oils delivered using plasma-treated cardboard pieces tended to decrease Varroa population in the treated colonies. Hence, atmospheric-pressure plasma for the modification of other materials, should be further investigated to provide alternative control treatment applications against honeybee mites.

Enhancing Bioactivity and Conjugation in Green Coffee Bean (Coffea arabica) Extract through Cold Plasma Treatment: Insights into Antioxidant Activity and Phenolic-Protein Conjugates.

Cold plasma technology is gaining attention as a promising approach to enhancing the bioactivity of plant extracts. However, its impact on green coffee bean extracts (GCBEs) still needs to be explored. In this study, an innovative underwater plasma jet system was employed to investigate the effects of cold plasma on Coffea arabica GCBEs, focusing on the conjugation reflected by the change in composition and bioactivity. The DPPH radical scavenging antioxidant activity exhibited a gradual increase with plasma treatment up to 35 min, followed by a decline. Remarkably, at 35 min, the plasma treatment resulted in a significant 66% increase in the DPPH radical scavenging activity of the GCBE. The total phenolic compound content also displayed a similar increasing trend to the DPPH radical scavenging activity. However, the phenolic profile analysis indicated a significant decrease in chlorogenic acids and caffeine. Furthermore, the chemical composition analysis revealed a decrease in free amino acids, while sucrose remained unchanged. Additionally, the SDS-PAGE results suggested a slight increase in protein size. The observed enhancement in antioxidant activity, despite the reduction in the two major antioxidants in the GCBE, along with the increase in protein size, might suggest the occurrence of conjugation processes induced by plasma, particularly involving proteins and phenolic compounds. Notably, the plasma treatment exhibited no adverse effects on the extract's safety, as confirmed by the MTT assay. These findings indicate that cold plasma treatment holds significant promise in improving the functional properties of GCBE while ensuring its safety. Incorporating cold plasma technology into the processing of natural extracts may offer exciting opportunities for developing novel and potent antioxidant-rich products.

On the evaluation of the antimicrobial effect of grape seed extract and cold atmospheric plasma on the dynamics of Listeria monocytogenes in novel multiphase 3D viscoelastic models.

The demand for products that are minimally processed and produced in a sustainable way, without the use of chemical preservatives or antibiotics have increased over the last years. Novel non-thermal technologies such as cold atmospheric plasma (CAP) and natural antimicrobials such as grape seed extract (GSE) are attractive alternatives to conventional food decontamination methods as they can meet the above demands. The aim of this study was to investigate the microbial inactivation potential of GSE, CAP (in this case, a remote air plasma with an ozone-dominated RONS output) and their combination against L. monocytogenes on five different 3D in vitro models of varying rheological, structural, and biochemical composition. More specifically, we studied the microbial dynamics, as affected by 1 % (w/v) GSE, CAP or their combination, in three monophasic Xanthan Gum (XG) based 3D models of relatively low viscosity (1.5 %, 2.5 % and 5 % w/v XG) and in a biphasic XG/Whey Protein (WPI) and a triphasic XG/WPI/fat model. A significant microbial inactivation (comparable to liquid broth) was achieved in presence of GSE on the surface of all monophasic models regardless of their viscosity. In contrast, the GSE antimicrobial effect was diminished in the multiphasic systems, resulting to only a slight disturbance of the microbial growth. In contrast, CAP showed better antimicrobial potential on the surface of the complex multiphasic models as compared to the monophasic models. When combined, in a hurdle approach, GSE/CAP showed promising microbial inactivation potential in all our 3D models, but less microbial inactivation in the structurally and biochemically complex multiphasic models, with respect to the monophasic models. The level of inactivation also depended on the duration of the exposure to GSE. Our results contribute towards understanding the antimicrobial efficacy of GSE, CAP and their combination as affected by robustly controlled changes of rheological and structural properties and of the biochemical composition of the environment in which bacteria grow. Therefore, our results contribute to the development of sustainable food safety strategies.

Canal pretreatment using cold atmospheric argon plasma, chitosan and Chlorine p6 activated by photodynamic therapy with EDTA as a final irrigant on the pushout bond strength of fiber post.

AIMS: To evaluate the effect of contemporary root canal irrigants i.e., Cold atmospheric argon plasma (CAAP), Chlorine p6 (Clp6), and chitosan on the push-out bond strength (PBS) of glass fiber post (GFP) and marten hardness (MH) of root canal dentin compared to NaOCl+EDTA. MATERIALS AND METHODS: The anatomic crown of sixty human mandibular premolars was removed by transversal section till cement-enamel junction (CEJ). Root canals were prepared and canals were dried and obturated using gutta-percha points and AH plus sealer. Post space was prepared and specimens were arbitrarily divided into four groups based on the post-space irrigation. (n = 15) Group 1: 5.25% NaOCl + 17% EDTA, Group 2: CAAP + 17% EDTA, Group 3: Chitosan + 17% EDTA and Group 4: Clp6 + 17% EDTA. For MH evaluation five samples from each disinfected group were taken and MH was assessed with the help of an ultra microhardness tester. GFP was cemented using self-etch dual cure cement on the remaining 10 samples from each group. Samples were vertically planted in acrylic resin blocks 1 mm thick slices were subjected to push-out testing and failure mode analysis using a universal testing machine and Stereomicroscope. ANOVA and Tukey multiple comparison tests were used to compare the means of bond integrity attained (p = 0.05). RESULTS: The maximum values of PBS and MH were displayed in group 1 (5.25% NaOCl + 17% EDTA) specimens. However, group 3 exhibited the minimum values of bond strength and MH scores. Intergroup comparison analysis revealed that group 2, group 3, and group 4 displayed comparable outcomes of PBS and MH (p > 0.05). The cervical and middle third of group 1 specimens displayed a cohesive type of failure. However, the apical section of group 1 and other tested groups exhibited the adhesive type of failure. CONCLUSION: Canal irrigation with 5.25% NaOCl and 17% EDTA appears to be better in terms of improving the bond integrity of fiber posts with no compromise in MH of the dentin as compared to the contemporary irrigant used.

Plasma treatment effects on destruction and recovery of Porphyromonas gingivalis biofilms.

The objective of this study was to investigate the treatment effects of non-thermal atmospheric gas plasmas (NTAP) on destruction and the recovery (or re-colonization) of Porphyromonas gingivalis (P. gingivalis) in biofilms. P. gingivalis is a well-known keystone periodontal pathogen strongly associated with periodontal diseases, especially periodontitis. P. gingivalis biofilms were formed on stainless steel coupons and treated for 1, 2, and 5 minutes by NTAP of pure argon gas and argon+oxygen gas mixture. MTT assay, colony forming unit (CFU) counting assay and confocal laser scanning microscopy (CLSM) were used to assess the destruction efficiency. In addition, the plasma treated biofilms were re-cultured in the medium supplemented with antibiotics and oxidative stress sources to determine the synergy of the NTAP with other antimicrobial agents. The results showed the plasma treatment could result in 2.7 log unit reduction in bacterial load. The recovered biofilm CFU with NTAP treatment combined with sub minimal inhibition concentration of amoxicillin was 0.33 log units less than the biofilm treated with amoxicillin alone. The recovered biofilm CFU in NTAP groups was about 2.0 log units less than that in the untreated controls under H2O2 treatment. There was approximately 1.0 log unit reduction of biofilm CFU in plasma treated biofilm compared with untreated control under paraquat treatment. The plasma treated biofilms exhibited less resistance to amoxicillin and greater susceptibility to hydrogen peroxide (H2O2) and paraquat, suggesting that NTAP may enhance biofilm susceptibility to host defense. These in vitro findings suggested that NTAP could be a novel and effective treatment method of oral biofilms that cause periodontal diseases.

Cold Atmospheric Plasma Activates Selective Photothermal Therapy of Cancer.

Due to the body's systemic distribution of photothermal agents (PTAs), and to the imprecise exposure of lasers, photothermal therapy (PTT) is challenging to use in treating tumor sites selectively. Striving for PTT with high selectivity and precise treatment is nevertheless important, in order to raise the survival rate of cancer patients and lower the likelihood of adverse effects on other body sections. Here, we studied cold atmospheric plasma (CAP) as a supplementary procedure to enhance selectivity of PTT for cancer, using the classical photothermic agent's gold nanostars (AuNSs). In in vitro experiments, CAP decreases the effective power of PTT: the combination of PTT with CAP at lower power has similar cytotoxicity to that using higher power irradiation alone. In in vivo experiments, combination therapy can achieve rapid tumor suppression in the early stages of treatment and reduce side effects to surrounding normal tissues, compared to applying PTT alone. This research provides a strategy for the use of selective PTT for cancer, and promotes the clinical transformation of CAP.

Effect of atmospheric cold plasma (ACP) on chlorine-adapted Salmonella enterica on spring onion.

One of the main drawbacks of chlorine disinfectants is the emergence of chlorine adapted (CA) or resistant microbial cells. This research aimed to investigate the effect of chlorine adaptation on resistance of Salmonella enterica upon atmospheric cold plasma (ACP) application at different voltages (6, 8 and 11 kV) and times (5, 10 and 15 min). Due to higher conversion efficiency and reduced dielectric barrier discharge power consumption, this method was used for cold plasma generation in this study. A higher lethality effect was observed from a higher voltage and longest times (11 kV-15 min) on CA S. enterica than on non-CA (P < 0·05). Still, it induced higher percentages of injured cells in CA (58·77%) than on non-CA (0·61%) (P < 0·05). The highest ACP effect on the inactivation of the indigenous natural flora of onion leaves was observed at the lowest voltage (P < 0·05). More than 3 log CFU/g reduction (P < 0·05) was observed at 6 kV after 5 and 10 min. ACP reduced CA and non-CA S. enterica cells on onion leaf surface to a lower extent than pure-treated cells in broth media. Nevertheless, similar to broth media, a high percentage of injury (61·03%) was induced on CA cells at higher voltage (11 kV-10 min) compared to non-CA (2·15%) (P < 0·05). Biofilm results revealed ACP application (6 kV-5 min) reduced average ODs in CA and non-CA cells (P < 0·05). Chlorine adaptation and ACP treatment influenced the antibiotic resistance pattern according to applied voltage, time and antibiotic type. The finding showed despite the highest lethality of high voltages and long times (11 kV-15 min), given the high percentages of injured cells, lower voltages may offer acceptable inactivation of pathogenic bacteria with lower injury induction. In conclusion, ACP has the potential ability to eliminate CA cells of S. enterica, which are predominant in fresh-cut vegetable outbreaks.

Molecular Mechanisms for Anti-aging of Low-Vacuum Cold Plasma Pretreatment in Caenorhabditis elegans.

Cold plasma pretreatment has the potential of anti-aging. However, its molecular mechanism is still not clear. Here, cold plasma pretreatment was firstly used to investigate the anti-aging effects of Caenorhabditis elegans using transcriptomic technique. It showed that the optimal parameters of discharge power, processing time, and working pressure for cold plasma pretreatment were separately 100 W, 15 s, and 135 Pa. The released 0.32 mJ/cm2 of the moderate apparent energy density was possibly beneficial to the strong positive interaction between plasma and C. elegans. The longest lifespan (13.67 ± 0.50 for 30 days) was obviously longer than the control (10.37 ± 0.46 for 23 days). Furthermore, compared with the control, frequencies of head thrashes with an increase of 26.01% and 37.31% and those of body bends with an increase of 33.37% and 34.51% on the fourth and eighth day, respectively, indicated movement behavior was improved. In addition, the variation of the enzyme activity of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) hinted that the cold plasma pretreatment contributed to the enhanced anti-aging effects in nematodes. Transcriptomics analysis revealed that cold plasma pretreatment resulted in specific gene expression. Anatomical structure morphogenesis, response to stress, regulation of biological quality, phosphate-containing compound metabolic process, and phosphorus metabolic process were the most enriched biological process for GO analysis. Cellular response to heat stress and HSF1-dependent transactivation were the two most enriched KEGG pathways. This work would provide the methodological basis using cold plasma pretreatment and the potential gene modification targets for anti-aging study.

Cold atmospheric plasma treatment improves the γ-aminobutyric acid content of buckwheat seeds providing a new anti-hypertensive functional ingredient.

Buckwheat (BW) is rich in γ-aminobutyric acid (GABA), which has great potential as a functional anti-hypertensive food ingredient. This study utilized cold atmospheric plasma (CAP) to promote GABA accumulation in BW during germination. The effect of this approach on GABA enrichment and anti-hypertensive activity of BW along with its processing properties were investigated. The results indicated that CAP stress treatment (50 W for 40 s) of BW seeds followed by germination at 30 °C for 60 h resulted in a GABA content of 2.22 ± 0.06 mg/g, which was a 2.64-fold greater than that of pristine BW. Cracking of the seed coat, faster germination, and the activation of glutamate decarboxylase might be responsible for the GABA enrichment. Compared with pristine BW, the GABA-enriched BW powder showed a greater inhibitory effect on the angiotensin-converting enzyme, an increased degree of pre-gelatinization and elasticity when it was formed into a dough.

Characterisation of Flavour Attributes in Egg White Protein Using HS-GC-IMS Combined with E-Nose and E-Tongue: Effect of High-Voltage Cold Plasma Treatment Time.

Egg white protein (EWP) is susceptible to denaturation and coagulation when exposed to high temperatures, adversely affecting its flavour, thereby influencing consumers' decisions. Here, we employ high-voltage cold plasma (HVCP) as a novel nonthermal technique to investigate its influence on the EWP's flavour attributes using E-nose, E-tongue, and headspace gas-chromatography-ion-mobilisation spectrometry (HS-GC-IMS) due to their rapidness and high sensitivity in identifying flavour fingerprints in foods. The EWP was investigated at 0, 60, 120, 180, 240, and 300 s of HVCP treatment time. The results revealed that HVCP significantly influences the odour and taste attributes of the EWP across all treatments, with a more significant influence at 60 and 120 s of HVCP treatment. Principal component analyses of the E-nose and E-tongue clearly distinguish the odour and taste sensors' responses. The HS-GC-IMS analysis identified 65 volatile compounds across the treatments. The volatile compounds' concentrations increased as the HVCP treatment time was increased from 0 to 300 s. The significant compounds contributing to EWP characterisation include heptanal, ethylbenzene, ethanol, acetic acid, nonanal, heptacosane, 5-octadecanal, decanal, p-xylene, and octanal. Thus, this study shows that HVCP could be utilised to modify and improve the EWP flavour attributes.

Induced genetic and chemical changes in medicinally important plant Catharanthus roseus (L.) G. Don: cold plasma and phytohormones.

BACKGROUND: Catharanthus roseus (L.) G. Donis a medicinal plant species belonging to the Apocynaceae family, which produces vinblastine and vincristine along with 100 other monoterpenoid indole alkaloids. The process of biosynthesis of C. roseus alkaloids is complex, in which many genes, enzymes, and regulators are involved. Induced mutations may be considered as a potential source for producing a higher amount of vinblastine and vincristine in this plant species. Therefore, the objective of the present study was to examine the effects of different treatments utilized on the induced genetic changes in C. roseus plants and enzyme activities. METHODS AND RESULTS: Spermine, jasmonic acid, methyjasmonate, putrescine, and cold plasma treatments were used for seed treatments. Different molecular markers, namely inter simple sequence repeat, inter retrotransposon amplified polymorphism, and retrotransposon microsatellite amplified polymorphism were employed to reveal the induced genetic changes. Antioxidant enzyme activities were also studied. The treated plants showed genetic variability and a significant increase in antioxidant enzyme activity compared to the control plants. The putrescine treatment resulted in the highest level of activity in superoxidase. A significant positive correlation occurred between the molecular markers data and antioxidant enzyme activities in treated plants. CONCLUSION: Our data revealed that the different phytohormones and cold plasma treatments could induce both genetic and chemical content changes in C. roseus plants.

[Molecular Mechanisms Underlying Cellular Responses to the Loading of Non-thermal Atmospheric Pressure Plasma-activated Solutions].

Plasma medicine is a rapidly expanding new field of interdisciplinary research that combines physics, chemistry, biology, and medicine. Non-thermal atmospheric pressure plasma (NTAPP) has recently been applied to living cells and tissues, and has emerged as a novel technology for medical applications, such as wound healing, blood coagulation, and cancer treatment. NTAPP was found to affect cells indirectly through the treatment of cells with previously prepared medium irradiated by NTAPP, termed plasma-activated medium (PAM). The treatment of culture media with NTAPP results in the generation of a large amount of reactive oxygen species and reactive nitrogen species, and their derived species. We found that PAM triggered a spiral apoptotic cascade in the mitochondrial-nuclear network in A549 cancer cells. This process induced the depletion of total cellular NAD+ and elevations in intracellular calcium ion, ultimately leading to cell death. We also detected the production of hydroxyl radical and elevations in intracellular ferrous ions in PAM-treated cells. The elevations observed in ferrous ions may have been due to their release from the intracellular iron store, ferritin. However, difficulties are associated with applying PAM to the clinical phase because culture media cannot be used for medical treatments. The anti-tumor activity of plasma-activated Ringer's solution was significantly stronger than that of PAM. At the end, we herein demonstrated the advantages of the combined application of plasma-activated acetate Ringer's solution and hyperthermia, a heat treatment at 42℃, for A549 cancer cell death and elucidated the underlying mechanisms.

Cold Atmospheric Pressure Plasma Treatment of Maize Grains-Induction of Growth, Enzyme Activities and Heat Shock Proteins.

Zea mays L. is one of the most produced crops, and there are still parts of the world where maize is the basic staple food. To improve agriculture, mankind always looks for new, better methods of growing crops, especially in the current changing climatic conditions. Cold atmospheric pressure plasma (CAPP) has already showed its potential to enhance the culturing of crops, but it still needs more research for safe implementation into agriculture. In this work, it was shown that short CAPP treatment of maize grains had a positive effect on the vitality of grains and young seedlings, which may be connected to stimulation of antioxidant and lytic enzyme activities by short CAPP treatment. However, the prolonged treatment had a negative impact on the germination, growth, and production indexes. CAPP treatment caused the increased expression of genes for heat shock proteins HSP101 and HSP70 in the first two days after sowing. Using comet assay it was observed that shorter treatment times (30-120 s) did not cause DNA damage. Surface diagnostics of plasma-treated grains showed that plasma increases the hydrophilicity of the surface but does not damage the chemical bonds on the surface.

Cold atmospheric pressure He-plasma jet and plasma ball interactions with the Venus flytrap: Electrophysiology and side effects.

Cold atmospheric pressure radio frequency plasma (CAPP) can play an important role in agriculture, medicine, biophysical and bioelectrochemical applications, disinfection and sterilization, synthesis of different compounds, nitrogen fixation, and treatment of surfaces. Here we found that reactive oxygen and nitrogen species, UV-Vis photons, and high-frequency strong electromagnetic fields with an amplitude of a few kV produced by a cold plasma jet can interact with bio-tissue and damage it if the plasma treatment is long enough. The electrophysiological effects of CAPP treatment of bio-tissue and electrical signals transmission were measured in the Venus flytrap. The plasma ball does not produce any visible side effects on the Venus flytrap, but induces electrical signals in bio-tissue with very high amplitude. Plasma (Kirlian) photography shows the existence of a blue aura around the plasma ball due to a corona discharge. Understanding the mechanisms of interactions between CAPP and bio-tissue and preventing side effects can contribute to the application of plasma technology in medicine and agriculture. The use of cold plasma in medicine and agriculture should be monitored for side effects from strong high-frequency electro-magnetic fields, UV photons, and reactive oxygen and nitrogen species to protect against undesirable consequences.

The synergistic effect of Canady Helios cold atmospheric plasma and a FOLFIRINOX regimen for the treatment of cholangiocarcinoma in vitro.

Cholangiocarcinoma (CCA) is a rare biliary tract cancer with a low five-year survival rate and high recurrence rate after surgical resection. Currently treatment approaches include systemic chemotherapeutics such as FOLFIRINOX, a chemotherapy regimen is a possible treatment for severe CCA cases. A limitation of this chemotherapy regimen is its toxicity to patients and adverse events. There exists a need for therapies to alleviate the toxicity of a FOLFIRINOX regimen while enhancing or not altering its anticancer properties. Cold atmospheric plasma (CAP) is a technology with a promising future as a selective cancer treatment. It is critical to know the potential interactions between CAP and adjuvant chemotherapeutics. In this study the aim is to characterize the efficacy of FOLFIRINOX and CAP in combination to understand potential synergetic effect on CCA cells. FOLFIRINOX treatment alone at the highest dose tested (53.8 µM fluorouracil, 13.7 µM Leucovorin, 5.1 µM Irinotecan, and 3.7 µM Oxaliplatin) reduced CCA cell viability to below 20% while CAP treatment alone for 7 min reduced viability to 3% (p < 0.05). An analysis of cell viability, proliferation, and cell cycle demonstrated that CAP in combination with FOLFIRINOX is more effective than either treatment alone at a lower FOLFIRINOX dose of 6.7 µM fluorouracil, 1.7 µM leucovorin, 0.6 µM irinotecan, and 0.5 µM oxaliplatin and a shorter CAP treatment of 1, 3, or 5 min. In conclusion, CAP has the potential to reduce the toxicity burden of FOLFIRINOX and warrants further investigation as an adjuvant therapy.

Combined Antimicrobial Effect of Bio-Waste Olive Leaf Extract and Remote Cold Atmospheric Plasma Effluent.

A novel strategy involving Olive Leaf Extract (OLE) and Cold Atmospheric Plasma (CAP) was developed as a green antimicrobial treatment. Specifically, we reported a preliminary investigation on the combined use of OLE + CAP against three pathogens, chosen to represent medical and food industries (i.e., E. coli, S. aureus and L. innocua). The results indicated that a concentration of 100 mg/mL (total polyphenols) in OLE can exert an antimicrobial activity, but still insufficient for a total bacterial inactivation. By using plain OLE, we significantly reduced the growth of Gram positive S. aureus and L. innocua, but not Gram-negative E. coli. Instead, we demonstrated a remarkable decontamination effect of OLE + CAP in E. coli, S. aureus and L. innocua samples after 6 h. This effect was optimally maintained up to 24 h in S. aureus strain. E. coli and L. innocua grew again in 24 h. In the latter strain, OLE alone was most effective to significantly reduce bacterial growth. By further adjusting the parameters of OLE + CAP technology, e.g., OLE amount and CAP exposure, it could be possible to prolong the initial powerful decontamination over a longer time. Since OLE derives from a bio-waste and CAP is a non-thermal technology based on ionized air, we propose OLE + CAP as a potential green platform for bacterial decontamination. As a combination, OLE and CAP can lead to better antimicrobial activity than individually and may replace or complement conventional thermal procedures in food and biomedical industries.

The amino acid metabolism is essential for evading physical plasma-induced tumour cell death.

BACKGROUND: Recent studies have emphasised the important role of amino acids in cancer metabolism. Cold physical plasma is an evolving technology employed to target tumour cells by introducing reactive oxygen species (ROS). However, limited understanding is available on the role of metabolic reprogramming in tumour cells fostering or reducing plasma-induced cancer cell death. METHODS: The utilisation and impact of major metabolic substrates of fatty acid, amino acid and TCA pathways were investigated in several tumour cell lines following plasma exposure by qPCR, immunoblotting and cell death analysis. RESULTS: Metabolic substrates were utilised in Panc-1 and HeLa but not in OVCAR3 and SK-MEL-28 cells following plasma treatment. Among the key genes governing these pathways, ASCT2 and SLC3A2 were consistently upregulated in Panc-1, Miapaca2GR, HeLa and MeWo cells. siRNA-mediated knockdown of ASCT2, glutamine depletion and pharmacological inhibition with V9302 sensitised HeLa cells to the plasma-induced cell death. Exogenous supplementation of glutamine, valine or tyrosine led to improved metabolism and viability of tumour cells following plasma treatment. CONCLUSION: These data suggest the amino acid influx driving metabolic reprogramming in tumour cells exposed to physical plasma, governing the extent of cell death. This pathway could be targeted in combination with existing anti-tumour agents.

Effects of in-package atmospheric cold plasma treatment on the qualitative, metabolic and microbial stability of fresh-cut pears.

BACKGROUND: The greatest hurdle to commercial marketing of fresh-cut fruits and vegetables is limited shelf life due to microbial hazards and quality deterioration. Atmospheric cold plasma (ACP) is an emerging non-thermal technology with significant potential to improve the safety and storability of fresh products. The objective of this study was to evaluate the effects of ACP, generated in sealed packaging, on the qualitative, metabolic and microbial stability of fresh-cut pears during simulated cold storage. RESULTS: ACP treatments were effective in inhibiting the growth of mesophilic aerobic bacteria, yeast and mold, particularly CP3 (65 kV, 1 min), which could prolong shelf life to the greatest extent. While decontamination was not always associated with an increase in plasma intensity. Moreover, at 65 kV for 1 min, ACP treatment had the potential to retard respiration, and maintain organoleptic properties and other quality attributes. Additionally, peroxidase and pectin methylesterase (PME) activities were reduced immediately after treatments. These effects were dependent on treatment voltage and time, while a subsequent recovery in activity was only observed for PME. CONCLUSION: The results obtained from this study will contribute to an understanding of the effects of in-package ACP treatments on the storability and microbial safety of fresh-cut pears. This knowledge could be beneficial in reducing quality losses for fresh-cut pears and the preservation of other products. © 2021 Society of Chemical Industry.

High voltage atmospheric cold plasma treatment inactivates Aspergillus flavus spores and deoxynivalenol toxin.

Fungal contamination is a concern for the food industry. Fungal spores resist food sterilization treatments and produce mycotoxins that are toxic for animals and humans. Technologies that deactivate spores and toxins without impacting food quality are desirable. This study demonstrates the efficiency of a high voltage atmospheric cold plasma (HVACP) technology using air to generate reactive oxygen (ROS) and nitrogen (RNS) species for the degradation of Aspergillus flavus cultures and the deoxynivalenol (DON) mycotoxin. Optical emission and absorption spectroscopy demonstrate ionization of hydroxyl groups, atomic oxygen and nitrogen, and confirm production of ROS and RNS, e.g. O3, NO2, NO3, N2O4, and N2O5. Fungal cultures show a depletion in pigmentation and an ~50% spore inactivation after 1-min treatments. Treated spores show surface ablation and membrane degradation by scanning electron microscopy. Twenty-minute direct HVACP treatments of 100 µg of DON in one mL aqueous suspensions resulted in a greater than 99% reduction in DON structure and rescued over 80% of Caco-2 cell viability; however, the same treatment on 100 µg of powdered DON toxin only showed a 33% reduction in DON and only rescued 15% of cell viability. In summary, HVACP air treatment can inactivate both fungal spores and toxins in minutes.

Effect of cold plasma and elicitors on bioactive contents, antioxidant activity and cytotoxicity of Thai rat-tailed radish microgreens.

BACKGROUND: Raphanus sativus var. caudatus or Thai rat-tailed radish (RTR) contains glucosinolates and isothiocyanates with chemopreventive effects; however, only mature plants have been investigated to date. Thus, the present study aimed to determine isothiocyanates, phenolic compounds and flavonoid compounds, antioxidant activity, cytotoxicity, and antiproliferative activity of RTR microgreens grown from seeds treated with cold plasma (21 kV for 5 min), organic elicitor (160 mmol L-1 NaCl, 10 mmol L-1 CaCl2 or 176 mmol L-1 sucrose) or both in combination. Seeds were germinated on vermiculite and sprayed with deionized water or elicitor for 7 days before harvest. RESULTS: Cold plasma had insignificant effect on growth, whereas NaCl and CaCl2 increased fresh weight. Plasma with CaCl2 led to the highest total isothiocyanate (ITC) content [1.99 g kg-1 dry weight (DW)] in RTR microgreens containing raphasatin as the only ITC detected. Plasma treatment gave the highest total phenolic content (7.56 mg gallic acid equivalents g-1 DW), antioxidant activity from a 2,2-diphenyl-1-picrylhydrazyl assay (7.70 mg trolox equivalents g-1 DW) and ferric reducing antioxidant power assay (21.72 mg Fe2+ g-1 DW). Microgreen extracts from plasma showed an IC50 value of 29.28 and 13.83 µg mL-1 towards MCF-7 and HepG2, respectively, with inhibitory properties on matrix metalloproteinase (MMP)-2 and MMP-9 proteins. Plasma enhanced Bax and Caspase-3 gene expression but reduced Bcl-2 and MMP-9 expression, indicating activation of apoptosis. CONCLUSION: Cold plasma shows promise as an innovative tool to enhance bioactive compounds with chemopreventive benefits in microgreens. © 2020 Society of Chemical Industry.