Mechanistic Insight into Permeation of Plasma-Generated Species from Vacuum into Water Bulk.

Due to their potential benefits, cold atmospheric plasmas (CAPs), as biotechnological tools, have been used for various purposes, especially in medical and agricultural applications. The main effect of CAP is associated with reactive oxygen and nitrogen species (RONS). In order to deliver these RONS to the target, direct or indirect treatment approaches have been employed. The indirect method is put into practice via plasma-activated water (PAW). Despite many studies being available in the field, the permeation mechanisms of RONS into water at the molecular level still remain elusive. Here, we performed molecular dynamics simulations to study the permeation of RONS from vacuum into the water interface and bulk. The calculated free energy profiles unravel the most favourable accumulation positions of RONS. Our results, therefore, provide fundamental insights into PAW and RONS chemistry to increase the efficiency of PAW in biological applications.

Impact of Reactive Oxygen and Nitrogen Species Produced by Plasma on Mdm2-p53 Complex.

The study of protein-protein interactions is of great interest. Several early studies focused on the murine double minute 2 (Mdm2)-tumor suppressor protein p53 interactions. However, the effect of plasma treatment on Mdm2 and p53 is still absent from the literature. This study investigated the structural changes in Mdm2, p53, and the Mdm2-p53 complex before and after possible plasma oxidation through molecular dynamic (MD) simulations. MD calculation revealed that the oxidized Mdm2 bounded or unbounded showed high flexibility that might increase the availability of tumor suppressor protein p53 in plasma-treated cells. This study provides insight into Mdm2 and p53 for a better understanding of plasma oncology.

Impact of an ionic liquid on protein thermodynamics in the presence of cold atmospheric plasma and gamma rays.

Cold atmospheric plasma and gamma rays are known to have anticancer properties, even though their specific mechanisms and roles as co-solvents during their action are still not clearly understood. Despite the use of gamma rays in cancer therapy, they have oncogenic potential, whereas this has not been observed for plasma treatment (to date). To gain a better understanding, we studied the action of dielectric barrier discharge (DBD) plasma and gamma rays on the myoglobin protein. We analyzed the secondary structure and thermodynamic properties of myoglobin after both treatments. In addition, in the last few years, ammonium ionic liquids (ILs) have revealed their important role in protein folding as co-solvents. In this work, we treated the protein with ammonium ILs such as triethylammonium methanesulfonate (TEMS) and tetrabutylammonium methanesulfonate (TBMS) and later treated this IL-protein solution with DBD plasma and gamma rays. In this study, we show the chemical and thermal denaturation of the protein after plasma and gamma treatments in the presence and absence of ILs using circular dichroism (CD) and UV-vis spectroscopy. Furthermore, we also show the influence of plasma and gamma rays on the secondary structure of myoglobin in the absence and presence of ILs or ILs + urea using CD. Finally, molecular dynamic simulations were conducted to gain deeper insight into how the ILs behave to protect the protein against the hydrogen peroxide generated by the DBD plasma and gamma rays.

Effects of plasma irradiation using various feeding gases on growth of Raphanus sativus L.

In this work, we have studied the action of dielectric barrier discharge (DBD) plasma irradiation using various feeding gases on seeds of Raphanus sativus L. and analysis their growth. Our experimental data shows that Air, O2, and NO(10%)+N2 feeding gases plasma irradiation enhanced plant growth, whereas N2, He and Ar feeding gases plasma irradiation had little influence on plant growth. Moreover, humid air plasma irradiation was more effective in growth enhancement than dry one. More than 2.3 times faster growth was observed by 3 min air plasma irradiation with 40-90% relative humidity. The reactive species generated by plasma in gas phase were detected using optical emission spectroscopy and in liquid phase by electron spin resonance (ESR) spectroscopy. We concluded that OH and O radicals were key species for plant growth enhancement.

Unexpected effects of the alteration of structure and stability of myoglobin and hemoglobin in ammonium-based ionic liquids.

The nature of solvent-biomolecule interactions is generally weak and non-specific. The addition of ionic liquids (ILs), which have emerged as a new class of solvents, strengthen the stability of some proteins whereas the same ILs weaken the stability of some other proteins. Although ILs are commonly used for the stabilization of biomolecules, the bimolecular interactions of their stabilization-destabilization is still an active subject of considerable interest and studies on this topic have been limited. To reveal the impact of ILs on the stability of proteins, a series of protic ILs possessing a tetra-alkyl ammonium cation [R4N](+) with a hydroxide [OH](-) anion were synthesized. In this study, we report the structural stability of heme proteins such as myoglobin (Mb) and hemoglobin (Hb) in a series of ammonium-based ILs such as tetramethyl ammonium hydroxide [(CH3)4N](+)[OH](-) (TMAH), tetraethyl ammonium hydroxide [(C2H5)4N](+)[OH](-) (TEAH), tetrapropyl ammonium hydroxide [(C3H7)4N](+)[OH](-) (TPAH) and tetrabutyl ammonium hydroxide [(C4H9)4N](+)[OH](-) (TBAH) by fluorescence and circular dichroism (CD) spectroscopic studies. Our experimental results reveal that less viscous ILs carrying smaller alkyl chain such as TMAH are strong destabilizers of the heme proteins as compared to the ILs carrying bulkier alkyl chains which are more viscous ILs, such as TBAH. Therefore, our results demonstrate that the addition of these ILs to the heme proteins decreases their thermal stability allowing the protein to be in an unfolded state at lower temperatures. Further, we describe the molecular-structural interaction of the heme proteins with the ILs (molecule like a ligand) by the PatchDocking method.

Plasma modification of poly(2-heptadecyl-4-vinylthieno[3,4-d]thiazole) low bandgap polymer and its application in solar cells.

For the first time, we here propose a green methodology to modify a low bandgap polymer for highly efficient solar cells using atmospheric pressure plasma jet or soft plasma operating on different feeding gases (air, Ar and N2). The physical properties of the modified polymer were investigated using conductivity measurements, UV-visible spectroscopy, photoluminescence spectroscopy, X-ray photoelectron spectroscopy, cyclic voltammograms, atomic force microscopy, cathodoluminescence and confocal Raman spectroscopy. Further, we examined the variation of the work function of the polymer before and after plasma treatment using a γ-focused ion beam. Additionally, photovoltaic cells based on the plasma-modified polymer having ITO/PEDOT:PSS/PHVTT (with or without plasma modification):PC71BM/LiF/Al configuration were fabricated and then characterized. We found that the power conversion efficiency (PCE) of the plasma-modified polymer increased dramatically as compared to the control polymer (without plasma treatment). PCE of the control polymer was found to be 4.11%, while after air, Ar and N2 gas plasma treatment the polymer showed PCEs of 4.85%, 4.87% and 5.14% respectively. Thus, plasma treatment not only alters the surface properties, but also modifies the bulk properties (changes in HOMO and LUMO bandgap level). Hence, this work provides new dimensions to explore more about plasma and polymer chemistry.

Influence of nanosecond pulsed plasma on the non-enzymatic pathway for the generation of nitric oxide from L-arginine and the modification of graphite oxide to increase the solar cell efficiency.

In this work, we demonstrated the action of nanosecond pulsed plasma (NPP) on the generation of nitric oxide (NO) from the non-enzymatic pathway and on the modification of graphite oxide (GO) sheets to increase polymer solar cells (PSCs) efficiency. NO is an important signal and an effector molecule in animals, which is generated from the enzyme-catalyzed oxidation of L-arginine to NO and L-citrulline. Hence, L-arginine is an important biological precursor for NO formation. Therefore, we developed a new non-enzymatic pathway for the formation of NO and L-citrulline using NPP and characterized the pathway using NO detection kit, NMR, liquid chromatography/capillary electrophoresis-mass spectrometry (LC/CE-MS) for both quantitative and qualitative bioanalysis. We then synthesized and modified the functional groups of GO using NPP, and it was characterised by X-ray photoelectron spectroscopy (XPS), confocal Raman spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM) imaging, cathodoluminescence (CL) and work function using γ-FIB. Further, we also tested the power conversion efficiency of the PSCs devices with modified GO that is similar to the one obtained with poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) as HTL. This work is perceived to have great implications for inexpensive and efficient methodology for NO generation and modification of GO, which are applicable in materials from nanomaterials to biomolecules.

Optical and structural properties of nanobiomaterials.

In this review, the optical and structural properties of biomaterials are discussed. First, we demonstrate the optical and structural properties of natural and plasma-treated DNA, using UV-visible absorption, circular dichroism (CD), and Raman spectroscopy. Fluorescence and lasing action in the dye-doped DNA-surfactant complex are also explained. Additionally, nanomaterial-based DNA detection and DNA-templated nanomaterial growth are described. Next, we discuss protein folding studies utilizing fluorescence, CD, and nuclear magnetic resonance (NMR) spectroscopy. From the CD spectra of alpha-chymotrypsin (CT), we estimate the composition of a-helices and the beta-sheets, and random coils in the CT. 1H NMR spectroscopy is used to investigate the thermal effect on the refolding of CT in the presence of an ionic liquid. Finally, we explain the numerical simulation method used for studying the optical properties of biomaterials. Applications of the Monte-Carlo method in photodynamic therapy, skin tissue optics, and bioimaging are described.

Growth control of Marchantia polymorpha gemmae using nonthermal plasma irradiation.

Several studies have documented that treatment by cold atmospheric pressure plasma (CAPP) on plants foster seed germination and growth in recent years. However, the molecular processes that underlie the action of CAPP on the seeds and plants remain mostly enigmatic. We here introduce gemmae of Marchantia polymorpha, a basal liverwort, as a novel model plant material suitable for CAPP research. Treating the gemmae with CAPP for a constant time interval at low power resulted in consistent growth enhancement, while growth inhibition at higher power in a dose-dependent manner. These results distinctly demonstrate that CAPP irradiation can positively and negatively regulate plant growth depending on the plasma intensity of irradiation, offering a suitable experimental system for understanding the molecular mechanisms underlying the action of CAPP in plants.

A preliminary study of the effect of DBD plasma and osmolytes on T98G brain cancer and HEK non-malignant cells.

Non-thermal plasmas are emerging as a novel tool for the treatment of living tissues for biological and medical purpose. In this study, we described the effect of 4 min dielectric barrier discharge (DBD) plasma on both T98G cancer and HEK normal cell lines in the presence of different concentrations of osmolytes. This treatment strategy shows a specific inhibitory effect of a 240 s plasma exposure in the presence of osmolytes against T98G brain cancer cells only, but not on HEK normal cells. Based on these interesting properties of osmolytes, a non-thermal plasma appears to be a potential anticancer treatment strategy for different kinds of cancers in the presence of osmolytes.

Biomedical importance of indoles.

The indole nucleus is an important element of many natural and synthetic molecules with significant biological activity. This review covers some of the relevant and recent achievements in the biological, chemical and pharmacological activity of important indole derivatives in the areas of drug discovery and analysis.

Unraveling the Transport Properties of RONS across Nitro-Oxidized Membranes.

The potential of cold atmospheric plasma (CAP) in biomedical applications has received significant interest, due to its ability to generate reactive oxygen and nitrogen species (RONS). Upon exposure to living cells, CAP triggers alterations in various cellular components, such as the cell membrane. However, the permeation of RONS across nitrated and oxidized membranes remains understudied. To address this gap, we conducted molecular dynamics simulations, to investigate the permeation capabilities of RONS across modified cell membranes. This computational study investigated the translocation processes of less hydrophilic and hydrophilic RONS across the phospholipid bilayer (PLB), with various degrees of oxidation and nitration, and elucidated the impact of RONS on PLB permeability. The simulation results showed that less hydrophilic species, i.e., NO, NO2, N2O4, and O3, have a higher penetration ability through nitro-oxidized PLB compared to hydrophilic RONS, i.e., HNO3, s-cis-HONO, s-trans-HONO, H2O2, HO2, and OH. In particular, nitro-oxidation of PLB, induced by, e.g., cold atmospheric plasma, has minimal impact on the penetration of free energy barriers of less hydrophilic species, while it lowers these barriers for hydrophilic RONS, thereby enhancing their translocation across nitro-oxidized PLB. This research contributes to a better understanding of the translocation abilities of RONS in the field of plasma biomedical applications and highlights the need for further analysis of their role in intracellular signaling pathways.

Robust Cough Detection With Out-of-Distribution Detection.

Cough is an important defense mechanism of the respiratory system and is also a symptom of lung diseases, such as asthma. Acoustic cough detection collected by portable recording devices is a convenient way totrack potential condition worsening for patients who have asthma. However, the data used in building current cough detection models are often clean, containing a limited set of sound categories, and thus perform poorly when they are exposed to a variety of real-world sounds which could be picked up by portable recording devices. The sounds that are not learned by the model are referred to as Out-of-Distribution (OOD) data. In this work, we propose two robust cough detection methods combined with an OOD detection module, that removes OOD data without sacrificing the cough detection performance of the original system. These methods include adding a learning confidence parameter and maximizing entropy loss. Our experiments show that 1) the OOD system can produce dependable In-Distribution (ID) and OOD results at a sampling rate above 750 Hz; 2) the OOD sample detection tends to perform better for larger audio window sizes; 3) the model's overall accuracy and precision get better as the proportion of OOD samples increase in the acoustic signals; 4) a higher percentage of OOD data is needed to realize performance gains at lower sampling rates. The incorporation of OOD detection techniques improves cough detection performance by a significant margin and provides a valuable solution to real-world acoustic cough detection problems.

Health assessment of rice cultivated and harvested from plasma-irradiated seeds.

This study provides the health effects assessment of rice cultivated from plasma-irradiated seeds. The rice (Oryza sativa L.) cultivated from seeds with plasma irradiation showed a growth improvement (slope-ratios of with plasma to without plasma were 1.066, 1.042, and 1.255 for tiller, and earing, and ripening periods, respectively) and an 4% increase in yield. The cultivated rice was used for repeated oral administrations to mice for 4-week period. Distilled water and rice cultivated from seeds without plasma irradiation were also used as control. The weights of the lung, kidney, liver, and spleen, with corresponding average values of 0.22 g, 0.72 g, 2.1 g, and 0.17 g for w/ plasma group and 0.22 g, 0.68 g, 2.16 g, and 0.14 g for w/o plasma group, respectively, showing no effect due to the administration of rice cultivated from plasma-irradiated seeds. Nutritional status, liver function, kidney function, and lipid, neutral fat profiles, and glucose metabolism have no significant difference between with and without plasma groups. These results show no obvious subacute effects were observed on rice grains cultivated and harvested from the mother plant that experienced growth improvement by plasma irradiation. This study provides a new finding that there is no apparent adverse health effect on the grains harvested from the plasma-irradiated seeds.

Water and a protic ionic liquid acted as refolding additives for chemically denatured enzymes.

In this communication, we present the ability of water and a protic ionic liquid, triethyl ammonium phosphate (TEAP) to act as refolding additives for the urea-induced chemical denaturated state of the two enzymes, α-chymotrypsin and succinylated Con A. We show that the enzymatic activity is regained and in certain circumstances enhanced.

Synthesis and antiproliferative activity of ammonium and imidazolium ionic liquids against T98G brain cancer cells.

Four ammonium and imidazolium ionic liquids (ILs) have been synthesized and screened against the T98G cell line (brain cancer) and HEK normal cells. Treatment induced metabolic cell death (MTT), growth inhibition, clonogenic inhibition were studied as cellular response parameters. Treatment with ILs enhanced growth inhibition and cell death in a concentration dependent manner in both the T98G and HEK cell lines. At higher concentrations (>0.09 mg/mL) the cytotoxic effects of the ILs were highly significant. An inhibitory effect on clonogenic capacity was also observed after cell treatment. Amongst all ILs; IL 4 (BMIMCl) exhibited potent activity against T98G brain cancer cells. Despite potent in-vitro activity, all ILs exhibited less cytotoxicity against the normal human HEK cells at all effective concentrations.

Detection of NO3- introduced in plasma-irradiated dry lettuce seeds using liquid chromatography-electrospray ionization quantum mass spectrometry (LC-ESI QMS).

Discharge plasma irradiates seeds with reactive oxygen and nitrogen species (RONS). However, RONS introduced in seeds by plasma irradiation have not been successfully detected thus far. This study provides experimental evidence that nitrate ion NO3- is introduced in lettuce seeds as RONS upon irradiation with atmospheric-pressure air dielectric barrier discharge plasma. Plasma irradiation for 5 min promotes seed germination. The components of the plasma-irradiated seeds were examined using electrospray ionization quantum mass spectrometry (ESI QMS), which revealed that the plasma irradiation introduced an ion with a mass of 62 m/z in detectable amounts. This ion was identified as NO3- by liquid chromatography (LC), multiple wavelength detector (MWD), and LC-ESI QMS. A one-dimensional simulation at electron temperature Te = 1 eV, electron density Ne = 1013/m3, and gas temperature Tg = 300 K indicated the introduction of NO3-, involving nitric oxide NO. NO3- is one of the most important ions that trigger signal transduction for germination when introduced in seeds. The scanning electron microscopy (SEM) images revealed that there was no change on the surface of the seeds after plasma irradiation. Plasma irradiation is an effective method of introducing NO3- in seeds in a dry process without causing damage.

Thermodynamic characterization of the biocompatible ionic liquid effects on protein model compounds and their functional groups.

The stability of proteins under co-solvent conditions is dependant on the nature of the co-solvent; the co-solvent can alter a protein's properties and structural effects through bimolecular interactions between its functional groups and co-solvent particles. Ionic liquids (ILs) represent a rather diverse class of co-solvents that are combinations of different ions, which are liquids at or close to room temperature. To quantify the bimolecular interactions of protein functional groups with biocompatible ILs, we report the systematic and quantitative apparent transfer free energies (ΔG'(tr)) of a homologous series of cyclic dipeptides (CDs) from water to aqueous solutions of ILs through solubility measurements, as a function of IL concentration at 25 °C under atmospheric pressure. The materials investigated in the present work included the CDs of cyclo(Gly-Gly), cyclo(Ala-Gly), cyclo(Ala-Ala), cyclo(Leu-Ala), and cyclo(Val-Val). The ILs used such as diethylammonium acetate ([Et(2)NH][CH(3)COO], DEAA), triethylammonium acetate ([Et(3)NH][CH(3)COO], TEAA), diethylammonium dihydogen phosphate ([Et(3)NH][H(2)PO(4)], DEAP), triethylammonium dihydogen phosphate ([Et(3)NH][H(2)PO(4)], TEAP), diethylammonium sulfate ([Et(3)NH][HSO(4)], DEAS) and triethylammonium sulfate ([Et(3)NH][HSO(4)], TEAS). We observed positive values of ΔG'(tr) for CDs from water to ILs, indicating that interactions between ILs and CDs are unfavourable, which leads to stabilization of the native structure of CDs. The experimental results were further used for estimating the transfer free energies (Δg'(tr)) of the peptide bond (-CONH-), the peptide backbone unit (-CH(2)C=ONH-), and various functional groups from water to IL solutions. Our results explicitly elucidate that a series of all ammonium ILs act as stabilizers for tested model compounds through the exclusion of ILs from CDs surface.

Activity and stability of α-chymotrypsin in biocompatible ionic liquids: enzyme refolding by triethyl ammonium acetate.

In view the of wide scope of structural information of biomolecules in biocompatible ionic liquids (ILs) in various applications including chemical and biochemical, it is essential to study the productive preferential interactions between biological macromolecules and biocompatible ILs. We have therefore explored the stability and activity of α-chymotrypsin (CT) in the presence of five ILs from different families, such as triethyl ammonium acetate (TEAA), triethyl ammonium phosphate (TEAP) from ammonium salts, 1-benzyl-3-methylimidazolium chloride ([Bzmim][Cl]), 1-benzyl-3-methylimidazolium tetrafluoroborate ([Bzmim][BF(4)]) from imidazolium salts and tetra-butyl phosphonium bromide (TBPBr) from phosphonium families. Circular dichroism (CD) and UV-vis spectrophotometer experiments were used to study CT stabilization by ILs, related to the associated structural changes and enzyme activity studies, respectively. We observed that all ILs have a dominant contribution to the stabilization of CT. The stability and activity of CT depends on the structural arrangement of the ions of ILs. Our experimental results explicitly elucidate that more hydrophobic imidazolium and phosphonium cations carrying longer alkyl chains of ILs ([Bzmim][Cl], [Bzmim][BF(4)] and TBPBr) were weak stabilizers for CT, while small alkyl chain molecules of triethyl ammonium salts (TEAA and TEAP) are strong stabilizers and therefore more biocompatible for CT stability. Our CD and NMR measurements reveal that TEAA is a refolding additive for CT from a quenched thermal unfolded enzyme structure.

A protic ionic liquid attenuates the deleterious actions of urea on α-chymotrypsin.

We present a biocompatible ionic liquid, triethyl ammonium acetate (TEAA), that attenuates the denaturation action of a non-ionic chaotrope, urea, on the industrially relevant proteolytic enzyme α-chymotrypsin.