Nitrogen regulates the synthesis of hydrophobic amino acids to improve protein structural and gel properties in common buckwheat.

Nitrogen (N) fertilizer impacts the grain quality of common buckwheat, but the effects and regulatory mechanisms of N on various protein parameters of buckwheat are not fully understood. The purpose of this study was to investigate the particle morphology, structural and gel properties, and regulation mechanism of buckwheat protein under four N levels. The bulk density, surface hydrophobicity, particle size, and thermal properties of the buckwheat protein were maximized through the optimal N application (180 kg N/ha), further enhancing the thermal stability of the protein. N application increased the ß-sheet content and reduced the random coil content. Appropriate N fertilizer input enhanced the tertiary structure stability and gel elasticity of buckwheat protein by promoting hydrophobic interactions, disulfide bonds, ionic bonds, storage modulus and loss modulus. The differentially expressed proteins induced by N are primarily enriched in small ribosomal subunit and ribosome, improving protein quality mainly by promoting the synthesis of hydrophobic amino acids. Future agriculture should pay attention to the hydrophobic amino acid content of buckwheat to effectively improve protein quality. This study further advances the application of buckwheat protein in the field of food processing and provides a theoretical basis for the extensive development and utilization of buckwheat protein.

Effects of mulching types on the yield and water utilization by broomcorn millet (Panicum miliaceum): Results of a study in the Loess Plateau, China.

Mulching practices have been widely adopted to improve rainfed crop productivity. However, the major resources including water, heat, and light that influenced the yield of broomcorn millet in different dryland regions have rarely been explored. A three-season field experiment with three mulching practices i.e. traditional planting with non-mulching (TP), ridge-furrow mulching system (RF), and plastic film mulching (PFM) was conducted in three semi-arid regions in the Loess Plateau, China, i.e. Guyuan city (GY), Huining county (HN), and Yulin city (YL) between 2020 and 2022 to investigate the impacts of mulching regimes on soil hydrothermal conditions, agronomic characteristics, leaf photosynthetic properties, broomcorn millet yield, and water use efficiency (WUE). Results showed that both PFM and RF treatments increased soil temperature and moisture, and enhanced dry matter accumulation by promoting leaf photosynthetic capacity and chlorophyll content, thereby improving broomcorn millet yield and WUE. PFM and RF increased the average broomcorn millet yield by 15.08% and 24.86% at GY site, 20.86% and 25.61% at HN site, and 15.75% and 25.57% at YL site, respectively, and increased the average WUE by 16.31% and 27.48% at GY site, 23.21% and 28.80% at HN site, 15.55% and 28.57% at YL site, respectively. Partial least squares path modeling analysis revealed that soil moisture was an important environmental factor in determining broomcorn millet yield. Overall, RF practice can be taken to improve the management of agricultural climate factors and maximize yield, thereby promoting the sustainable development of dryland agriculture in the Loess Plateau.

Qualitative and quantitative analyses of chemical constituents in vitro and in vivo and systematic evaluation of the pharmacological effects of Tibetan medicine Zhixue Zhentong capsules.

Introduction: Zhixue Zhentong capsules (ZXZTCs) are a Tibetan medicine preparation solely composed of Lamiophlomis rotata (Benth.) Kudo. L. rotata is the only species of the genus Laniophlomis (family Lamiaceae) that has medicinal constituents derived from the grass or root and rhizome. L. rotata is one of the most extensively used folk medicines by Tibetan, Mongolian, Naxi, and other ethnic groups in China and has been listed as a first-class endangered Tibetan medicine. The biological effects of the plant include hemostasis, analgesia, and the removal of blood stasis and swelling. Purpose: This study aimed to profile the overall metabolites of ZXZTCs and those entering the blood. Moreover, the contents of six metabolites were measured and the hemostatic, analgesic, and anti-inflammatory effects of ZXZTCs were explored. Methods: Ultra-performance liquid chromatography-tandem quadrupole time-of-flight high-resolution mass spectrometry (UPLC-Q-TOF-MS) was employed for qualitative analysis of the metabolites of ZXZTCs and those entering the blood. Six metabolites of ZXZTCs were quantitatively determined via high-performance liquid chromatography The hemostatic, analgesic, and anti-inflammatory effects of ZXZTCs were evaluated in various animal models. Results: A total of 36 metabolites of ZXZTCs were identified, including 13 iridoid glycosides, 9 flavonoids, 9 phenylethanol glycosides, 4 phenylpropanoids, and 1 other metabolite. Overall, 11 metabolites of ZXZTCs entered the blood of normal rats. Quantitative analysis of the six main metabolites, shanzhiside methyl ester, chlorogenic acid, 8-O-acetyl shanzhiside methyl ester, forsythin B, luteoloside, and verbascoside, was extensively performed. ZXZTCs exerted hemostatic effects by reducing platelet aggregation and thrombosis and shortening bleeding time. Additionally, ZXZTCs clearly had an analgesic effect, as observed through the prolongation of the latency of writhing, reduction in writhing, and increase in the pain threshold of experimental rats. Furthermore, significant anti-inflammatory effects of ZXZTCs were observed, including a reduction in capillary permeability, the inhibition of foot swelling, and a reduction in the proliferation of granulation tissue. Conclusion: Speculative identification of the overall metabolites of ZXZTCs and those entering the blood can provide a foundation for determining its biologically active constituents. The established method is simple and reproducible and can help improve the quality control level of ZXZTCs as a medicinal product. Evaluating the hemostatic, analgesic, and anti-inflammatory activities of ZXZTCs can help reveal its mechanism.

Design, synthesis, and analgesia evaluation of novel Transient Receptor Potential Vanilloid 1 (TRPV1) agonists modified from Cannabidiol (CBD).

Pain-relief is a long-term research hotspot with huge demand in clinical treatment. The analgesics currently used have several side effects, such as being addictive and causing gastrointestinal bleeding. Therefore, new drugs and targets in analgesic field are both desirable. Transient Receptor Potential Vanilloid 1 (TRPV1) plays an essential role in pain perception and regulation, providing a new strategy for the development of antinociceptive agents. Here, a series of novel TRPV1 agonists were designed and synthesized based on Cannabidiol (CBD), a widely used pain-relieving agent with weak agonistic activity on TRPV1. According to the results of systematic in vitro and in vivo biological assays, compound 10f was finally identified as a promising TRPV1 agonist, with higher target affinity, stronger analgesic activity, and weak side effect of hyperthermia. Molecular docking simulations revealed a significant hydrogen bond interaction between 10f and Arg557, an amino acid residue key to the activity of TRPV1 protein. Taken together, compound 10f can be used as a lead compound for further optimization.

Challenges for the development of mutant isocitrate dehydrogenases 1 inhibitors to treat glioma.

Glioma is one of the most common types of brain tumors, and its high recurrence and mortality rates threaten human health. In 2008, the frequent isocitrate dehydrogenase 1 (IDH1) mutations in glioma were reported, which brought a new strategy in the treatment of this challenging disease. In this perspective, we first discuss the possible gliomagenesis after IDH1 mutations (mIDH1). Subsequently, we systematically investigate the reported mIDH1 inhibitors and present a comparative analysis of the ligand-binding pocket in mIDH1. Additionally, we also discuss the binding features and physicochemical properties of different mIDH1 inhibitors to facilitate the future development of mIDH1 inhibitors. Finally, we discuss the possible selectivity features of mIDH1 inhibitors against WT-IDH1 and IDH2 by combining protein-based and ligand-based information. We hope that this perspective can inspire the development of mIDH1 inhibitors and bring potent mIDH1 inhibitors for the treatment of glioma.

The evolutionary dynamics history of canine distemper virus through analysis of the hemagglutinin gene during 1930-2020.

Canine distemper virus (CDV) is a lethal viral disease of carnivores which is considered to be a serious threat to domestic and wild species. Despite the widespread use of vaccines, CDV still occurs in vaccinated animals and current vaccines does not guarantee complete protection. In this study, a total of 286 hemagglutinin (H) gene sequences of the virus isolated in 25 countries during 90 years (1930-2020) were analyzed by Bayesian maximum likelihood analysis to estimate the population dynamics. We identified the most recent common ancestor (TMRCA) of the virus in 1868 in the USA which arrived in continental Europe in 1948, and from there, the virus spread rapidly to other continents. The Canidae family was identified as the original host as well as a source of the subsequent spread. We identified 11 lineages of geographic co-circulating strains globally. The effective population size experienced a two-phase-exponential growth between 2000-2005 and 2010-2012. Our findings provide a novel insight into the epidemic history of canine distemper virus which may facilitate more effective disease management. This study uses a large set of sequencing data on the H gene of CDV to identify distinct lineages of the virus, track its geographic spread over time, analyze its likelihood of transmission within and between animal families, and provide suggestions for improved strategies to combat the virus. Supplementary Information: The online version contains supplementary material available at 10.1007/s10344-023-01685-z.

6-Shogaol as a Novel Thioredoxin Reductase Inhibitor Induces Oxidative-Stress-Mediated Apoptosis in HeLa Cells.

Inhibition of thioredoxin reductase (TrxR) is a crucial strategy for the discovery of antineoplastic drugs. 6-Shogaol (6-S), a primary bioactive compound in ginger, has high anticancer activity. However, its potential mechanism of action has not been thoroughly investigated. In this study, we demonstrated for the first time that 6-S, a novel TrxR inhibitor, promoted oxidative-stress-mediated apoptosis in HeLa cells. The other two constituents of ginger, 6-gingerol (6-G) and 6-dehydrogingerduone (6-DG), have a similar structure to 6-S but fail to kill HeLa cells at low concentrations. 6-Shogaol specifically inhibits purified TrxR1 activity by targeting selenocysteine residues. It also induced apoptosis and was more cytotoxic to HeLa cells than normal cells. The molecular mechanism of 6-S-mediated apoptosis involves TrxR inhibition, followed by an outburst of reactive oxygen species (ROS) production. Furthermore, TrxR knockdown enhanced the cytotoxic sensitivity of 6-S cells, highlighting the physiological significance of targeting TrxR by 6-S. Our findings show that targeting TrxR by 6-S reveals a new mechanism underlying the biological activity of 6-S and provides meaningful insights into its action in cancer therapeutics.

Nitrogenous Fertilizer Levels Affect the Physicochemical Properties of Sorghum Starch.

Nitrogen is a key factor affecting sorghum growth and grain quality. This experiment was designed to investigate the physicochemical properties of sorghum starch in four sorghum varieties (Liaoza 10, Liaoza 19, Jinza 31, and Jinza 34) under four nitrogen levels: 0 kg/ha urea (N1), 300 kg/ha urea as base fertilizer (N2), 300 kg/ha urea as topdressing at the jointing stage (N3), and 450 kg/ha urea as topdressing at the jointing stage (N4). The results showed that grain size and amylose content increased with increasing nitrogen fertilizer level, peaking at N3. The peak viscosity, final viscosity, gelatinization temperature, initial temperature, final temperature, and enthalpy value increased with the nitrogenous fertilizer level, peaking at N3. The application of nitrogen fertilizer at the jointing period significantly increased the above indicators. However, excess nitrogen at the jointing period (N4) can significantly reduce the above indicators, thus changing the physicochemical properties and structure of sorghum starch. Overall, nitrogen significantly affects the structure and physicochemical properties of sorghum starch.

Chronic effects of benzalkonium chlorides on short chain fatty acids and methane production in semi-continuous anaerobic digestion of waste activated sludge.

As an emerging pollutant, benzalkonium chlorides (BACs) potentially enriched in waste activated sludge (WAS). However, the microbial response mechanism under chronic effects of BACs on acidogenesis and methanogenesis in anaerobic digestion (AD) has not been clearly disclosed. This study investigated the AD (by-)products and microbial evolution under low to high BACs concentrations from bioreactor startup to steady running. It was found that BACs can lead to an increase of WAS hydrolysis and fermentation, but a disturbance to acidogenic bacteria also occurred at low BACs concentration. A noticeable inhibition to methanogenesis occurred when BAC concentration was up to 15 mg/g TSS. Metagenomic analysis revealed the key genes involved in acetic acid (HAc) biosynthesis (i.e. phosphate acetyltransferase, PTA), ß-oxidation pathway (acetyl-CoA C-acetyltransferase) and propionic acid (HPr) conversion was slightly promoted compared with control. Furthermore, BACs inhibited the acetotrophic methanogenesis (i.e. acetyl-CoA synthetase), especially BAC concentration was up to 15 mg/g TSS, thereby enhanced short chain fatty acids (SCFAs) accumulation. Overall, chronic stimulation of functional microorganisms with increasing concentrations of BACs impact WAS fermentation.

An efficient strategy for digging protein-protein interactions for rational drug design - A case study with HIF-1α/VHL.

The ubiquitination of the hypoxia-inducible factor-1α (HIF-1α) is mediated by interacting with the von Hippel-Lindau protein (VHL), and is associated with cancer, chronic anemia, and ischemia. VHL, an E3 ligase, has been reported to degrade HIF-1 for decades, however, there are few successful inhibitors currently. Poor understanding of the binding pocket and a lack of in-depth exploration of the interactions between two proteins are the main reasons. Hence, we developed an effective strategy to identify and design new inhibitors for protein-protein interaction targets. The hydroxyproline (Hyp564) of HIF-1α contributed the key interaction between HIF-1α and VHL. In this study, detailed information of the binding pocket were explored by alanine scanning, site-directed mutagenesis and molecular dynamics simulations. Interestingly, we found the interaction(s) between Y565 and H110 played a key role in the binding of VHL/HIF-1α. Based on the interactions, 8 derivates of VH032, 16a-h, were synthesized by introducing various groups bounded to H110. Further assay on protein and cellular level exhibited that 16a-h accessed higher binding affinity to VHL and markable or modest improvement in stabilization of HIF-1α or HIF-1α-OH in HeLa cells. Our work provides a new orientation for the modification or design of VHL/HIF-1α protein-protein interaction inhibitors.

Efficient methane production from waste activated sludge and Fenton-like pretreated rice straw in an integrated bio-electrochemical system.

Integrated microbial electrolysis cell-anaerobic digestion (MEC-AD) systems have demonstrated potential advantages for methane production in the presence of small amounts of residual inhibitors. In this study, a series of tests were conducted to analyse the acidification and methanogenesis performance of pretreated rice straw (RS) in anaerobic digestion (AD) and MEC-AD systems after the addition of Fenton-like reagents. The results indicated that the short-chain acids (SCFAs) accumulations reached 2284.64 ± 21.57 mg COD/L with a dosage ratio of 1/4 (g RS/g VSS sludge) in the MEC-AD system and that methane production increased by 63.8% compared with that of an individual AD system. In the interim, the net energy output reached 1.09 × 103 J/g TCOD, which was 1.23 times higher than that of the AD system. The residual Fe3+/Fe2+ in the pretreatment reagent was capable of promoting acidification and methanogenesis in sludge and RS fermentation. The RS hydrolysis products could constrain methanogenesis, which can be mitigated by introducing an MEC. The microbiological analyses revealed that the MEC strongly increased the enrichment of hydrogenotrophic methanogens, especially Methanobacterium (61.16%). Meanwhile, the Syntrophomonas and Acetobacterium abundances increased to 2.81% and 2.65%, respectively, which suggested the reinforcement of acetogenesis and methanogenesis. Therefore, the enhanced hydrogenotrophic methanogens might have served as the key for enhancing the efficiency of methanogenesis due to the introduction of an MEC.

Pseudoephedrine and its derivatives antagonize wild and mutated severe acute respiratory syndrome-CoV-2 viruses through blocking virus invasion and antiinflammatory effect.

The coronavirus disease 2019 has infected over 150 million people worldwide and led to over 3 million deaths. Severe acute respiratory syndrome (SARS)-CoV-2 lineages B.1.1.7, B.1.617, B.1.351, and P.1 were reported to have higher infection rates than that of wild one. These mutations were noticed to happen in the receptor-binding domain of spike protein (S-RBD), especially mutations N501Y, E484Q, E484K, K417N, K417T, and L452R. Currently, there is still no specific medicine against the virus; moreover, cytokine storm is also a dangerous factor for severe infected patients. In this study, potential S-RBD-targeted active monomers from traditional Chinese medicine Ephedra sinica Stapf (ephedra) were discovered by virtual screening. NanoBiT assay was performed to confirm blocking activities of the screened compounds against the interaction between SARS-CoV-2 S-RBD and angiotensin-converting enzyme 2 (ACE2). We further analyzed the blocking effect of the active compounds on the interactions of mutated S-RBD and ACE2 by computational studies. Moreover, antiinflammatory activities were evaluated using qRT-PCR, enzyme-linked immune sorbent assay, and Western blot analysis. As a result, pseudoephedrine (MHJ-17) and its derivative (MHJ-11) were found as efficient inhibitors disrupting the interactions between ACE2 and both wild and mutated S-RBDs. In addition, they also have antiinflammatory activities, which can be potential drug candidates or lead compounds for further study.

Muscone derivative ZM-32 inhibits breast tumor angiogenesis by suppressing HuR-mediated VEGF and MMP9 expression.

Inhibition of tumor angiogenesis is a highly effective strategy for cancer treatment. Human antigen R (HuR), an RNA-binding protein, is overexpressed in many cancers and regulates the mRNAs of multiple angiogenic factors by binding to the adenylate-uridylate-rich element in their 3' untranslated region. HuR protein has been demonstrated to be an important regulatory factor in macrophage-mediated angiogenesis, a process in which macrophages are critical for tumor progression. Muscone is a synthetic equivalent of musk, and recent studies have shown that it has a regulatory effect on angiogenesis. In this study, we synthesized five series of muscone derivatives and discovered that compound ZM-32 was effective in preventing HuR RRM1/2-Vegf-a mRNA complex formation. ZM-32 bound to HuR RRM1/2 protein with a KD value of 521.7 nmol/L. Furthermore, ZM-32 inhibited endothelial cell proliferation, migration, and tubule formation, and suppressed the VEGF/VEGFR2/ERK1/2 signaling axis mediated by macrophages in vitro. We also demonstrated that ZM-32 effectively prevented the proliferation and migration of breast cancer cells and inhibited the growth and angiogenesis of MDA-MB-231 xenograft tumors without any obvious toxicity in vivo. Mechanistically, exposure to ZM-32 influenced the mRNA stability of Vegf-a and Mmp9 in a HuR-dependent manner in both macrophages and MDA-MB-231 cells. Thus, in this study we identified a new muscone derivative, ZM-32, with anti-angiogenesis effects mediated via targeting HuR in breast cancer, that may become a potentially valuable lead compound for anti-cancer angiogenesis.

Optimization of a novel single air-lift sequencing bioreactor for raw piggery wastewater treatment: Nutrients removal and microbial community structure analysis.

In this study, a sequencing batch and intermittent air-lift bioreactor (SBIAB) was evaluated under the three independent variables to treat raw piggery wastewater. The effects of hydraulic retention time (HRT), air flow rate and sludge retention time (SRT) on the nutrients removal of SBIAB were researched. The optimum values of HRT, air flow rate and SRT were 8 d, 2 l/min and 20 d, respectively. Meanwhile, the removal rates of chemical oxygen demand (COD), NH4+-N, total nitrogen (TN) and total phosphorus (TP) were up to 89.5%, 93.5%, 61.1% and 57.3%, respectively. Generally, the nutrients removal performance could be enhanced with increasing HRT from 6 to 10 d, while it was inhibited at air flow rate of 3 l/min. Higher air flow rate caused the alkaline pH and high free ammonia concentration, which imposed restrictions on the process of wastewater treatment. In the SBIAB, a coupling of aerobic/anoxic/anaerobic zone was formed according to the changes of oxidation-reduction potential (ORP) values at the optimum condition. Microbial community structure analysis indicated that the functional microbes including Brachymonas, Prokaryote, Giesbergeria, Comamonadaceae bacterium, Clostridiales bacterium, Comamonas, Tissierella, Aequorivita were enriched in the SBIAB, which played a significant role in the removal of complex organics and nitrogen.

Glycyrrhizic acid exerts inhibitory activity against the spike protein of SARS-CoV-2.

Coronavirus causes a disease with high infectivity and pathogenicity, especially SARS in 2003, MERS in 2012, and COVID-2019 currently. The spike proteins of these coronaviruses are critical for host cell entry by receptors. Thus, searching for broad-spectrum anti-coronavirus candidates, such as spike protein inhibitors, is vital and desirable due to the mutations in the spike protein. In this study, a combination of computer-aided drug design and biological verification was used to discover active monomers from traditional Chinese medicine. Surface plasmon resonance (SPR) assays and NanoBit assays were used to verify the predicated compounds with their binding activities to spike proteins and inhibitory activities on the SARS-CoV-2 RBD/ACE2 interaction, respectively. Furthermore, an MTT assay was used to evaluate the cell toxicities of active compounds. As a result, glycyrrhizic acid (ZZY-44) was found to be the most efficient and nontoxic broad-spectrum anti-coronavirus molecule in vitro, especially, the significant effect on SARS-CoV-2, which provided a theoretical basis for the study of the pharmacodynamic material basis of traditional Chinese medicine against SARS-CoV-2 and offered a lead compound for further structural modification in order to obtain more effective candidate drugs against SARS-CoV-2.

Polygonum perfoliatum L., an Excellent Herbal Medicine Widely Used in China: A Review.

Polygonum perfoliatum L. (synonym: Polygonum knotweed L.; Persicaria perfoliata; family: Polygonaceae) is a kind of folk traditional Chinese medicine with a long history of wide use in the treatment of ancient internal, surgical, and gynecological diseases. At present, 80 chemical constituents have been isolated from P. perfoliatum, including flavonoids, anthraquinones, terpenoids, phenolic acids, phenylpropanoids, and alkaloids, among which flavonoids are the main active components. Modern studies have shown that P. perfoliatum has pharmacological activities such as anti-inflammatory, anti-bacterial, antiviral, anti-liver fibrosis, antitussive and expectorant, anti-tumor, anti-oxidation, and so on. By consulting and sorting out a large number of related literatures at home and abroad in recent years, this paper systematically reviewed the botany, traditional uses, phytochemistry, pharmacological activities, and quality control of P. perfoliatum, and discussed its development potential in new drug research and clinical application in the future, in order to provide a reference basis for further research and promote the in-depth development and utilization.

Novel probiotic-bound oxidized Bletilla striata polysaccharide-chitosan composite hydrogel.

As a new strategy for wound management, probiotics are highly sensitive to the external environment during use. In this study, an attempt is made to apply oxidized Bletilla striata polysaccharide (OBSP) hydrogel as a delivery system to put up a physical barrier to probiotics. Bletilla striata, as a famous traditional Chinese medicine, has been widely used for over 1500 years to promote wound healing. Firstly, probiotic-bound OBSP-Chitosan composite hydrogel (OBSP-CS-LP hydrogel) was prepared by a simple and environmentally friendly approach. Subsequently, the SEM image, swelling and rheological properties of the OBSP-CS-LP hydrogel were investigated. Notably, composite probiotics to wounds are an attractive novel intervention that significantly improves the antibacterial properties of the hydrogel and avoids the pitfalls of many antibiotic therapies. Furthermore, the full-thickness skin defect model in vivo indicated an outstanding wound healing efficacy. Therefore, OBSP-CS-LP hydrogel could be applied as a promising dressing in the wound healing.

Alkaline aided thermophiles pretreatment of waste activated sludge to increase short chain fatty acids production: Microbial community evolution by alkaline on hydrolysis and fermentation.

Adding alkaline into an anaerobic waste activated sludge (WAS) fermentation with thermophilic bacteria pretreatment could efficiently improve short-chain fatty acids (SCFAs) accumulation to 3550 ± 120 mg COD/L. The acidification rate in combined test was 21.2%, while that was 15.6% and 10.7% in sole thermophilic bacteria pretreatment and control tests respectively. Four distinct groups of microbes could be identified with noticeable shifts using the combined pretreatments, and tremendous effects were analyzed on organic content especially of the soluble proteins and SCFAs concentrations. Particularly, alkaline addition would significantly change the functional microbial structures, including the decrease of Caloramator with the function of thermophilic proteolytic and the increase of Acidobacteria TM7 and Petrimonas sp. The results above suggested that alkaline addition could decrease the hydrolytic substances consume by thermotolerance bacteria and final improve SCFAs accumulation in fermentation process.

Design, synthesis, and biological activity evaluation of BACE1 inhibitors with antioxidant activity.

The proteolytic enzyme ß-secretase (BACE1) plays a central role in the synthesis of the pathogenic ß-amyloid peptides (Aß) in Alzheimer's disease (AD), antioxidants could attenuate the AD syndrome and prevent the disease progression. In this study, BACE1 inhibitors (D1-D18) with free radical-scavenging activities were synthesized by molecular hybridization of 2-aminopyridine with natural antioxidants. The biological activity evaluation showed that D1 had obvious inhibitory activity against BACE1, and strong antioxidant activity in 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS+• ) assay, which could be used as a lead compound for further study.