This study evaluated the in vitro antioxidant capacities of extracts from Pleurotus pulmonarius via Folin-Ciocalteu, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, metal chelating, cupric ion reducing antioxidant capacity, and lipid peroxidation inhibition assays. Extract compositions were determined by phenol-sulfuric acid; Coomassie Plus (Bradford) protein; Spectroquant zinc, copper, and manganese test assays; and liquid chromatography-tandem mass spectrometry (LC/MS/MS) and gas chromatography-mass spectrometry (GC/MS). Methanol-dichloromethane extract, water fraction, hot water, aqueous extract and hexane fraction exhibited the most potent extracts in the antioxidant activities. LC/MS/MS and GC/MS showed that the extracts contained ergothioneine, ergosterol, flavonoid, and phenolic compounds. The selected potent extracts were evaluated for their inhibitory effect against oxidation of human low-density lipoproteins and protective effects against hydrogen peroxide-induced cytotoxic injury in human aortic endothelial cells. The crude aqueous extract was deemed most potent for the prevention of human low-density lipoprotein oxidation and endothelial membrane damage. Ergothioneine might be the compound responsible for the activities, as supported by previous reports. Thus, P. pulmonarius may be a valuable antioxidant ingredient in functional foods or nutraceuticals.
The application of graphene in the field of drug delivery has attracted massive interest among researchers. However, the high toxicity of graphene has been a drawback for its use in drug delivery. Therefore, to enhance the biocompatibility of graphene, a new route was developed using ternary natural deep eutectic solvents (DESs) as functionalizing agents, which have the capability to incorporate various functional groups and surface modifications. Physicochemical characterization analyses, including field emission scanning electron microscope, fourier-transform infrared spectroscopy, Raman spectroscopy, Brunauer-Emmett-Teller, X-ray diffraction, and energy dispersive X-ray, were used to verify the surface modifications introduced by the functionalization process. Doxorubicin was loaded onto the DES-functionalized graphene. The results exhibited significantly improved drug entrapment efficiency (EE) and drug loading capacity (DLC) compared with pristine graphene and oxidized graphene. Compared with unfunctionalized graphene, functionalization with DES choline chloride (ChCl):sucrose:water (4:1:4) resulted in the highest drug loading capacity (EE of 51.84% and DLC of 25.92%) followed by DES ChCl:glycerol:water (1:2:1) (EE of 51.04% and DLC of 25.52%). Following doxorubicin loading, graphene damaged human breast cancer cell line (MCF-7) through the generation of intracellular reactive oxygen species (>95%) and cell cycle disruption by increase in the cell population at S phase and G2/M phase. Thus, DESs represent promising green functionalizing agents for nanodrug carriers. To the best of our knowledge, this is the first time that DES-functionalized graphene has been used as a nanocarrier for doxorubicin, illustrating the potential application of DESs as functionalizing agents in drug delivery systems.
With the rapid development of ionic liquid analogues, termed 'deep eutectic solvents' (DESs), and their application in a wide range of chemical and biochemical processes in the past decade, the extraction of bioactive compounds has attracted significant interest. Recently, numerous studies have explored the extraction of bioactive compounds using DESs from diverse groups of natural sources, including animal and plant sources. This review summarizes the-state-of-the-art effort dedicated to the application of DESs in the extraction of bioactive compounds. The aim of this review also was to introduce conventional and recently-developed extraction techniques, with emphasis on the use of DESs as potential extractants for various bioactive compounds, such as phenolic acid, flavonoids, tanshinone, keratin, tocols, terpenoids, carrageenans, xanthones, isoflavones, α-mangostin, genistin, apigenin, and others. In the near future, DESs are expected to be used extensively for the extraction of bioactive compounds from various sources.
Time-lapse resistivity measurements and groundwater geochemistry were used to study salinity effect on groundwater aquifer at the ex-promontory-land of Carey Island in Malaysia. Resistivity was measured by ABEM Terrameter SAS4000 and ES10-64 electrode selector. Relationship between earth resistivity and total dissolved solids (TDS) was derived, and with resistivity images, used to identify water types: fresh (ρ ( e ) > 6.5 Ω m), brackish (3 Ω m < ρ ( e ) < 6.5 Ω m), or saline (ρ ( e ) < 3 Ω m). Long-term monitoring of the studied area's groundwater quality via measurements of its time-lapse resistivity showed salinity changes in the island's groundwater aquifers not conforming to seawater-freshwater hydraulic gradient. In some aquifers far from the coast, saline water was dominant, while in some others, freshwater 30 m thick showed groundwater potential. Land transformation is believed to have changed the island's hydrogeology, which receives saltwater pressure all the time, limiting freshwater recharge to the groundwater system. The time-lapse resistivity measurements showed active salinity changes at resistivity-image bottom moving up the image for two seasons' (wet and dry) conditions. The salinity changes are believed to have been caused by incremental tide passing through highly porous material in the active-salinity-change area. The study's results were used to plan a strategy for sustainable groundwater exploration of the island.
Mosquito collections were carried out from May to June 1992 and from September to December 1992 in an area where a case of Japanese encephalitis was confirmed. A total of 40,072 mosquitoes belonging to 35 species and 8 genera were collected. The dominant species in that locality were Culex vishnui, Culex tritaeniorhynchus, Culex pseudovishnui, Culex gelidus, Aedes butleri, and Mansonia uniformis.
The applications of eutectic systems, including deep eutectic solvents (DESs), in diverse sectors have drawn significant interest from researchers, academicians, engineers, medical scientists, and pharmacists. Eutecticity increases drug dissolution, improves drug penetration, and acts as a synthesis route for drug carriers. To date, DESs have been extensively explored as potential drug delivery systems on account of their unique properties such as tunability and chemical and thermal stability. This review discusses two major topics: first, the application of eutectic mixtures (before and after the introduction of DES) in the field of drug delivery systems, and second, the most promising examples of DES pharmaceutical activity. It also considers future prospects in the medical and biotechnological fields. In addition to the application of DESs in drug delivery systems, they show greatly promising pharmaceutical activities, including anti-fungal, anti-bacterial, anti-viral, and anti-cancer activities. Eutecticity is a valid strategy for overcoming many obstacles inherently associated with either introducing new drugs or enhancing drug delivery systems.
One of the recently developed metaheuristic algorithms, the coyote optimization algorithm (COA), has shown to perform better in a number of difficult optimization tasks. The binary form, BCOA, is used in this study as a solution to the descriptor selection issue in classifying diverse antifungal series. Z-shape transfer functions (ZTF) are evaluated to verify their efficiency in improving BCOA performance in QSAR classification based on classification accuracy (CA), the geometric mean of sensitivity and specificity (G-mean), and the area under the curve (AUC). The Kruskal-Wallis test is also applied to show the statistical differences between the functions. The efficacy of the best suggested transfer function, ZTF4, is further assessed by comparing it to the most recent binary algorithms. The results prove that ZTF, especially ZTF4, significantly improves the performance of the original BCOA. The ZTF4 function yields the best CA and G-mean of 99.03% and 0.992%, respectively. It shows the fastest convergence behaviour compared to other binary algorithms. It takes the fewest iterations to reach high classification performance and selects the fewest descriptors. In conclusion, the obtained results indicate the ability of the ZTF4-based BCOA to find the smallest subset of descriptors while maintaining the best classification accuracy performance.
Enzymatic treatment on lignocellulosic biomass has become a trend in preparing nanocellulose (NC), but the process must be optimized to guarantee high production yield and crystallinity. This study offers insights into an innovative protocol using cultivated fungal cellulase and xylanase to improve NC production from raw oil palm leaves (OPL) using five-factor-four-level Taguchi orthogonal design for optimizing parameters, namely substrate and enzyme loading, surfactant concentration, incubation temperature and time. Statistical results revealed the best condition for producing NC (66.06 % crystallinity, 43.59 % yield) required 10 % (w/v) substrate, 1 % (v/v) enzyme, 1.4 % (w/v) Tween-80, with 72-h incubation at 30 °C. Likewise, the highest sugar yield (47.07 %) was obtained using 2.5 % (w/v) substrate, 2.0 % (v/v) enzyme, 2.0 % (w/v) Tween-80, with 72-h incubation at 60 °C. The auxiliary enzymes used in this study, i.e., xylanase, produced higher crystallinity NC, showing widths between 8 and 12 nm and lengths >1 μm and sugars at 47.07 % yield. Thus, our findings proved that optimizing the single-step enzymatic hydrolysis of raw OPL could satisfactorily produce relatively crystalline NC and sugar yield for further transformation into bio-nanocomposites and biofuels. This study presented a simple, innovative protocol for NC synthesis showing characteristics comparable to the traditionally-prepared NC, which is vital for material's commercialization.
This study explores an eco-friendly delignification technique for raw oil palm leaves (OPL), highlighting the optimized conditions of choline chloride-lactic acid deep eutectic solvent (DES)-mediated ball milling pretreatment to maximize the co-production yields of highly crystalline cellulose and lignin. Our five-level-four-factor Taguchi design identified the optimal reaction settings for cellulose production (85.83 % yield, 47.28 % crystallinity) as 90-minute milling, 1500 rpm, mill-ball size ratio of 30:10, ball-to-sample mass ratio of 20:1, DES-to-sample mass ratio of 3:1. Conversely, the maximal lignin extraction yield (35.23 %) occurred optimally at 120-minute milling, 600 rpm, mill-ball size ratio of 25:5, ball-to-sample mass ratio of 20:1 and DES-to-sample mass ratio of 9:1. Statistical results showed that milling frequency (p-value ≤ 0.0001) was highly significant in improving cellulose crystallinity and yield, while DES-to-sample mass ratio (p-value ≤ 0.0001) was the most impacting on lignin yield. The thermogravimetric method affirmed the elevated cellulose thermal stability, corroborating the enhanced cellulose content (40.14 % to 73.67 %) alongside elevated crystallinity and crystallite size (3.31 to 4.72 nm) shown by X-ray diffractograms. The increased surface roughness seen in micrographs mirrored the above-said post-treatment changes. In short, our optimized one-pot dual-action pretreatment effectively delignified the raw OPL to produce cellulose-rich material with enhanced crystallinity and lignin solidity.
Dehalogenase E (DehE) is a non-stereospecific enzyme produced by the soil bacterium, Rhizobium sp. RC1. Till now, the catalytic mechanism of DehE remains unclear although several literature concerning its structure and function are available. Since DehE is non-stereospecific, the enzyme was hypothesized to follow a 'direct attack mechanism' for the catalytic breakdown of a haloacid. For a molecular insight, the DehE modelled structure was docked in silico with the substrate 2-chloropropionic acid (2CP) in the active site. The ideal position of DehE residues that allowed a direct attack mechanism was then assessed via molecular dynamics (MD) simulation. It was revealed that the essential catalytic water was hydrogen bonded to the 'water-bearer', Asn114, at a relatively constant distance of ∼2.0 Å after 50 ns. The same water molecule was also closely sited to the catalytic Asp189 at an average distance of ∼2.0 Å, signifying the imperative role of the latter to initiate proton abstraction for water activation. This reaction was crucial to promote a direct attack on the α-carbon of 2CP to eject the halide ion. The water molecule was oriented favourably towards the α-carbon of 2CP at an angle of ∼75°, mirrored by the formation of stable enzyme-substrate orientations throughout the simulation. The data therefore substantiated that the degradation of a haloacid by DehE followed a 'direct attack mechanism'. Hence, this study offers valuable information into future advancements in the engineering of haloacid dehalogenases with improved activity and selectivity, as well as functionality in solvents other than water.
A novel greener MNC/PES membrane was developed through an electrospinning technique for lipase immobilization to catalyze the synthesis of ethyl valerate (EV). In this study, the covalent immobilization of Aspergillus oryzae lipase (AOL) onto an electrospun nanofibrous membrane consisting of magnetic nanocellulose (MNC) and polyethersulfone (PES) to produce EV was statistically optimized. Raman spectroscopy, Fourier-transform infrared spectroscopy: attenuated total reflection, field emission scanning electron microscopy, energy dispersive X-ray spectroscopy, thermal gravimetric analysis (TGA), and differential thermal gravimetric (DTG) of MNC/PES-AOL demonstrated that AOL was successfully immobilized onto the fibers. The Taguchi design-assisted immobilization of AOL onto MNC/PES fibers identified that 1.10 mg/mL protein loading, 4 mL reaction volume, 250 rpm stirring rate, and 50 °C were optimal to yield 72.09% of EV in 24 h. The thermal stability of MNC/PES-AOL was improved by ≈20% over the free AOL, with reusability for up to five consecutive esterification cycles while demonstrating an exceptional half-life of 120 h. Briefly, the electrospun MNC/PES fibers that immobilized AOL showed promising applicability in yielding relatively good EV levels. This study suggests that using MNC as fillers in a PES to improve AOL activity and durability for a longer catalytic process could be a viable option.
Over the past decade, deep eutectic systems (DES) have become popular, yet their potential toxicity to living organisms is not well understood. This study fills this gap by examining the toxicity, antibacterial activity and biodegradability of p-toluenesulfonic acid monohydrate (PTSA)-based DESs prepared from ammonium or phosphonium salts. Brine shrimp assays revealed varying toxicity levels of PTSA and salts. Allyltriphenylphosphonium bromide showing the longest survival time among all tested salts while PTSA exhibited a significantly longer duration of cell survival compared to other hydrogen bond donors. PTSA and ammonium salts (N,N-diethylethanolammonium chloride and choline chloride) as individual components showed non-toxic behavior for Gram-negative and Gram-positive bacteria while different PTSA-based DESs showed significant inhibition zones. Fish acute ecotoxicity tests indicated moderately toxicity for individual components and DESs, though higher concentrations increased fish mortality, highlighting the need for careful handling and disposal of PTSA-based DESs to the environment. Biodegradability analyses found all tested DESs to be readily biodegradable and it was reported that, DES 3 prepapred form PTSA and choline chloride has the highest biodegradability level. Notably, all tested DESs showed over 60 % biodegradability after 28 days. This groundbreaking study explores PTSA-based DESs, highlighting their biodegradability and potential use as antibacterial agents. Results revealed that PTSA as individual component is much better from toxicity point of view in comparison with PTSA-based DESs for any further industrial applications.
The widespread use of disposable face masks as a preventative strategy to address transmission of the SARS-CoV-2 virus has been a key environmental concern since the pandemic began. This has led to an unprecedented new form of contamination from improperly disposed masks, which liberates significant amounts of heavy metals and toxic chemicals in addition to volatile organic compounds (VOCs). Therefore, this study monitored the liberation of heavy metals, VOCs, and microfibers from submerged disposable face masks at different pH (4, 7 and 12), to simulate distinct environmental conditions. Lead (3.238% ppb), cadmium (0.672 ppb) and chromium (0.786 ppb) were found in the analyzed leachates. By pyrolysis, 2,4-dimethylhept-1-ene and 4-methylheptane were identified as the VOCs produced by the samples. The chemically degraded morphology in the FESEM images provided further evidence that toxic heavy metals and volatile organic compounds had been leached from the submerged face masks, with greater degradation observed in samples submerged at pH 7 and higher. The results are seen to communicate the comparable danger of passively degrading disposable face masks and the release of micro- or nanofibers into the marine environment. The toxicity of certain heavy metals and chemicals released from discarded face masks warrants better, more robust manufacturing protocols and increased public awareness for responsible disposal to reduce the adverse impact on ecology and human health.
There has been a growing emphasis on developing extraction methods that are not only efficient but also environmentally friendly and sustainable. One promising avenue is the exploration of deep eutectic solvents (DESs) as neoteric extraction media. This study aims to investigate the potential of DESs as neoteric extraction media for phenolics-rich flower clove extracts. Two DESs were synthesised by mixing choline chloride with glycerol and lactic acid at a molar ratio of 1:2. The thermal profiles of the mixture were analysed using differential scanning calorimetry, and the viscosity and density were measured at different temperatures. The phenolic compounds were quantitatively characterised for all of the extractants using high-performance liquid chromatography. The total phenolic content and the antioxidant activities of the extracts were determined. The results showed that DESs significantly improved the extraction of antioxidant compounds from clove, especially for the case of phenolic compounds, and also considerably enhanced the antioxidant activity of the extracts. The use of DESs offers a green, efficient method for extracting value-added products from natural sources.