Interactive network pharmacology and electrochemical analysis reveals electron transport-mediating characteristics of Chinese medicine formula Jing Guan Fang.

Background: Jing Guan Fang (JGF) is an anti-COVID-19 Chinese Medicine decoction comprised of five medicinal herbs to possess anti-inflammatory and antiviral properties for treatment. This study aims to electrochemically decipher the anti-coronavirus activity of JGF and show that microbial fuel cells may serve as a platform for screening efficacious herbal medicines and providing scientific bases for the mechanism of action (MOA) of TCMs. Methods: Electrochemical techniques (e.g., cyclic voltammetry) and MFCs were adopted as the bioenergy-based platforms to assess the bioenergy-stimulating characteristics of JGF. Phytochemical analysis correlated polyphenolic and flavonoid content with antioxidant activity and bioenergy-stimulating properties. Network pharmacology on the active compounds was employed to identify anti-inflammatory and anti-COVID-19 protein targets, and molecular docking validated in silico results. Significant findings: This first-attempt results show that JGF possesses significant reversible bioenergy-stimulation (amplification 2.02 ± 0.04) properties suggesting that its antiviral efficacy is both bioenergy-steered and electron mediated. Major flavonoids and flavone glycosides identified by HPLC (e.g., baicalein and baicalin, respectively) possess electron-shuttling (ES) characteristics that allow herbal medicines to treat COVID-19 via (1) reversible scavenging of ROS to lessen inflammation; (2) inhibition of viral proteins; and (3) targeting of immunomodulatory pathways to stimulate the immune response according to network pharmacology.

Interactive deciphering electron-shuttling characteristics of Coffea arabica leaves and potential bioenergy-steered anti-SARS-CoV-2 RdRp inhibitor via microbial fuel cells.

Due to the pandemics of COVID-19, herbal medicine has recently been explored for possible antiviral treatment and prevention via novel platform of microbial fuel cells. It was revealed that Coffea arabica leaves was very appropriate for anti-COVID-19 drug development. Antioxidant and anti-inflammatory tests exhibited the most promising activities for C. arabica ethanol extracts and drying approaches were implemented on the leaf samples prior to ethanol extraction. Ethanol extracts of C. arabica leaves were applied to bioenergy evaluation via DC-MFCs, clearly revealing that air-dried leaves (CA-A-EtOH) exhibited the highest bioenergy-stimulating capabilities (ca. 2.72 fold of power amplification to the blank). Furthermore, molecular docking analysis was implemented to decipher the potential of C. arabica leaves metabolites. Chlorogenic acid (-6.5 kcal/mol) owned the highest binding affinity with RdRp of SARS-CoV-2, showing a much lower average RMSF value than an apoprotein. This study suggested C. arabica leaves as an encouraging medicinal herb against SARS-CoV-2.

Interactive deciphering electron-shuttling characteristics of agricultural wastes with potential bioenergy-steered anti-COVID-19 activity via microbial fuel cells.

Background: This first-attempt study explored indigenous herbs from agricultural waste with bioenergy and biorefinery-stimulating potentials for possible anti-COVID-19 drug development. As prior novel study revealed, medicinal herbs abundant in ortho-dihydroxyl substituents and flavonoid-bearing chemicals were likely not only electron shuttle (ES)-steered, but also virus transmission-resisted. Methods: Herbal extract preparation from agricultural wastes were implemented via traditional Chinese medicine (TCM) decoction pot. After filtration and evaporation, a crude extract obtained was used for evaluation of bioenergy-stimulating and electron-mediating characteristics via microbial fuel cells (MFCs). Combined with cyclic voltammetric analysis, MFCs provided a novel platform to distinguish electron shuttles from antioxidants with electron-transfer steered antiviral potentials of herbal extracts. Significant findings: After 50 serial cyclic voltammogram traces, considerable ES activities of herbal extracts still stably remained, indicating that possible medication-associated capabilities could be persistent. This work also extended to explore bioenergy-stimulating herbs from agricultural waste recycling for bioenergy and biorefinery applications. Water extract of Coffea arabica was more biotoxic than ethanolic extract, resulting in its lower power-generating capability. The findings revealed that water extract of Trichodesma khasianum and Euphorbia hirta could exhibit considerable bioenergy-enhancing effects. For cradle-to-cradle circular economy, agricultural waste could be specifically screened for possible regeneration of value-added anti-COVID-19 drugs via bioenergy selection.

Synergistic deciphering of bioenergy production and electron transport characteristics to screen traditional Chinese medicine (TCM) for COVID-19 drug development.

Background: Traditional Chinese medicine (TCM) has been used as an "immune booster" for disease prevention and clinical treatment since ancient China. However, many studies were focused on the organic herbal extract rather than aqueous herbal extract (AHE; decoction). Due to the COVID-19 pandemics, this study tended to decipher phytochemical contents in the decoction of herbs and derived bioactivities (e.g., anti-oxidant and anti-inflammatory properties). As prior works revealed, the efficacy of Parkinson's medicines and antiviral flavonoid herbs was strongly governed by their bioenergy-stimulating proficiency. Methods: Herbal extracts were prepared by using a traditional Chinese decoction pot. After filtration and evaporation, crude extracts were used to prepare sample solutions for various bioassays. The phytochemical content and bioactivities of AHEs were determined via ELISA microplate reader. Microbial fuel cells (MFCs) were used as a novel platform to evaluate bioenergy contents with electron-transfer characteristics for antiviral drug development. Significant findings: Regarding 18 TCM herbal extracts for the prevention of SARS and H1N1 influenza, comparison on total polyphenol, flavonoid, condensed tannins and polysaccharides were conducted. Moreover, considerable total flavonoid contents were detected for 11 herb extracts. These AEHs were not only rich in phytonutrient contents but also plentiful in anti-oxidant and anti-inflammatory activities. Herbs with high polyphenol content had higher antioxidant activity. Forsythia suspensa extract expressed the highest inhibition against nitric oxide production for anti-inflammation. MFC bioenergy-stimulating studies also revealed that top ranking COVID-19 efficacious herbs were both bioenergy driven and electron mediated. That is, electron transfer-controlled bioenergy extraction was significant to antiviral characteristics for anti-COVID-19 drug development.

Exploring community evolutionary characteristics of microbial populations with supplementation of Camellia green tea extracts in microbial fuel cells.

This first-attempt study deciphered combined characteristics of species evolution and bioelectricity generation of microbial community in microbial fuel cells (MFCs) supplemented with Camellia green tea (GT) extracts for biomass energy extraction. Prior studies indicated that polyphenols-rich extracts as effective redox mediators (RMs) could exhibit significant electrochemical activities to enhance power generation in MFCs. However, the supplementation of Camellia GT extract obtained at room temperature with significant redox capabilities into MFCs unexpectedly exhibited obvious inhibitory effect towards power generation. This systematic study indicated that the presence of antimicrobial components (especially catechins) in GT extract might significantly alter the distribution of microbial community, in particular a decrease of microbial diversity and evenness. For practical applications to different microbial systems, pre-screening criteria of selecting biocompatible RMs should not only consider their promising redox capabilities (abiotic), but also possible inhibitory potency (biotic) to receptor microbes. Although Camellia tea extract was well-characterized as GRAS energy drink, some contents (e.g., catechins) may still express inhibition towards organisms and further assessment upon biotoxicity may be inevitably required for practice.

Deciphering Electron-Shuttling Characteristics of Neurotransmitters to Stimulate Bioelectricity-Generating Capabilities in Microbial Fuel Cells.

This first-attempt study used electrochemical methods to quantitatively assess electron-shuttling capabilities of different neurotransmitters crucial to catecholamine biosynthesis in human brain. As prior studies mentioned, aromatics bearing ortho- or para-dihydroxybenzenes could reveal promising electroactivities to stimulate bioenergy generation in microbial fuel cells (MFCs). This feasibility study extended to investigate the electrochemical characteristics of catecholamines and trace amines (e.g., 14 model compounds selected from neurotransmitters) synthesized by human brain via cyclic voltammetry methods (CVs) and MFCs. Dopamine (DA), levodopa (L-DOPA), epinephrine (EP), norepinephrine (NP), and 3,4-dihydroxyphenylacetic acid (DOPAC) would perform the electron-shuttling characteristics, and the rest would not. In particular, DA formed by decarboxylation of L-DOPA could exhibit relatively higher electrochemical activities than their precursors. In addition, carboxylic acids formed by deamination and carboxylation of trace monoamines would reveal more significant reductive potential (Epc); however, their oxidative electric currents seemed to be reduced. That is, chemical structure significantly influenced whether the electrochemical characteristics could be effectively expressed. This work also clearly revealed that neurotransmitters with ortho-dihydroxybenzenes exhibited promising stimulation to bioelectricity-generating capabilities of MFCs in the ranking of DA ~ EP > NP > L-DOPA > DOPAC. This was consistent with ES behaviors as CV analyses indicated.

Polyphenolic compounds as electron shuttles for sustainable energy utilization.

For renewable and sustainable bioenergy utilization with cost-effectiveness, electron-shuttles (ESs) (or redox mediators (RMs)) act as electrochemical "catalysts" to enhance rates of redox reactions, catalytically accelerating electron transport efficiency for abiotic and biotic electrochemical reactions. ESs are popularly used in cellular respiratory systems, metabolisms in organisms, and widely applied to support global lives. Apparently, they are applicable to increase power-generating capabilities for energy utilization and/or fuel storage (i.e., dye-sensitized solar cell, batteries, and microbial fuel cells (MFCs)). This first-attempt review specifically deciphers the chemical structure association with characteristics of ESs, and discloses redox-mediating potentials of polyphenolics-abundant ESs via MFC modules. Moreover, to effectively convert electron-shuttling capabilities from non-sustainable antioxidant activities, environmental conditions to induce electrochemical mediation apparently play critical roles of great significance for bioenergy stimulation. For example, pH levels would significantly affect electrochemical potentials to be exhibited (e.g., alkaline pHs are electrochemically favorable for expression of such electron-shuttling characteristics). Regarding chemical structure effect, chemicals with ortho- and para-dihydroxyl substituents-bearing aromatics own convertible characteristics of non-renewable antioxidants and electrochemically catalytic ESs; however, ES capabilities of meta-dihydroxyl substituents can be evidently repressed due to lack of resonance effect in the structure for intermediate radical(s) during redox reaction. Moreover, this review provides conclusive remarks to elucidate the promising feasibility to identify whether such characteristics are non-renewable antioxidants or reversible ESs from natural polyphenols via cyclic voltammetry and MFC evaluation. Evidently, considering sustainable development, such electrochemically convertible polyphenolic species in plant extracts can be reversibly expressed for bioenergy-stimulating capabilities in MFCs under electrochemically favorable conditions.

Exploring the glyphosate-degrading characteristics of a newly isolated, highly adapted indigenous bacterial strain, Providencia rettgeri GDB 1.

This study explored the characteristics of a newly isolated glyphosate (GLYP)-degrading bacterium Providencia rettgeri GDB 1, for GLYP bioremediation. Due to the serial selection pressure of high GLYP concentrations for enriched isolation, this highly tolerant GLYP biodegrader shows very promising capabilities for GLYP removal (approximately 71.4% degradation efficiency) compared to previously reported strains. High performance liquid chromatography analyses showed aminomethylphosphonic acid (AMPA) rather than sarcosine (SAR) to be the sole intermediate of GLYP decomposition via the AMPA formation pathway. Moreover, GLYP biodegradation was biochemically favorable in aerobic cultures due to its strong growth-associated characteristics. To the best of our knowledge, this is the first report to indicate that bacterial strains in the Providencia genus could demonstrate highly promising GLYP-degrading characteristics in environments with high GLYP contents.

Exploring optimal supplement strategy of medicinal herbs and tea extracts for bioelectricity generation in microbial fuel cells.

This first-attempt study used extracts of appropriate antioxidant abundant Camellia and non-Camellia tea and medicinal herbs as model ESs to stably intensify bioelectricity generation performance in microbial fuel cells (MFCs). As electron shuttles (ESs) could stimulate electron transport phenomena by significant reduction of electron transfer resistance, the efficiency of power generation for energy extraction in microbial fuel cells (MFCs) could be appreciably augmented. Using environmentally friendly natural bioresource as green bioresource of ESs is the most promising to sustainable practicability. As comparison of power-density profiles indicated, supplement of Camellia tea extracts would be the most appropriate, then followed non-Camellia Chrysanthemum tea and medicinal herbs. Antioxidant activities, total phenolic contents and power stimulating activities were all electrochemically associated. In particular, the extract of unfermented Camellia tea (i.e., green tea) was the most promising ESs to augment bioenergy extraction compared to other refreshing medicinal herb extracts.

Kinetic study of Reactive Black 5 degradation by Fe2+/S2O82- process via interactive model-based response surface methodology.

This study aimed to kinetically discover optimal conditions on characteristics of Reactive Black 5 decolorization/degradation via ferrous (Fe2+)-activated potassium persulfate (PS). Monod-like kinetics and interactive model-based response surface methodology (RSM) were applied to fitting and predict optimize treatment. Biodegradability of the intermediates was also tested by shaking culture with two species (Proteus hauseri ZMd44 and Shewanella sp. WLP72). Results showed that the optimal degradation efficiency was predicted (through RSM) as pH 3.72, (PS) = 0.39 mM, and (Fe2+) = 0.29 mM. The transformation products (dl-4-hydroxymandelic acid, benzoic acid, benzene, formic acid, oxalic acid and acetic acid) were less toxic than the original dye solution. According to those results, clean-up of dye pollutants by the Fe2+/S2O82- process is feasible as a pre-processing for the biodegradation, and the predicted optimal conditions are meaningful for further industry utilization.

Exploring redox-mediating characteristics of textile dye-bearing microbial fuel cells: thionin and malachite green.

Prior studies indicated that biodecolorized intermediates of azo dyes could act as electron shuttles to stimulate wastewater decolorization and bioelectricity generation (WD&BG) in microbial fuel cells (MFCs). This study tended to explore whether non-azo textile dyes (i.e., thionin and malachite green) could also own such redox-mediating capabilities for WD&BG. Prior findings mentioned that OH and/or NH2 substitute-containing auxochrome compounds (e.g., 2-aminophenol and 1,2-dihydroxybenzene) could effectively mediate electron transport in MFCs for simultaneous WD&BG. This work clearly suggested that the presence of electron-mediating textile dyes (e.g., thionin and malachite green (MG)) in MFCs is promising to stimulate color removal and bioelectricity generation. That is, using MFCs as operation strategy for wastewater biodecolorization is economically promising in industrial applications due to autocatalytic acceleration of electron-flux for WD&BG in MFCs.

Deciphering characteristics of bicyclic aromatics--mediators for reductive decolorization and bioelectricity generation.

This first-attempt study quantitatively assessed electron-mediating characteristics of bicyclic aromatics - 1-amino-2-naphthol, 4-amino-1-naphthol (i.e., decolorized intermediates of azo dyes - orange I and II) for color removal and power generation in MFCs. According to cyclic-voltammetric profiles, the presence of reduction and oxidation peak potentials clearly suggested a crucial role of these intermediates as electron-shuttling mediators. Shake-flask cultures also showed that appropriate accumulation of 1A2N, 4A1N apparently enhanced color-removal efficiencies of bacterial decolorization. This study clearly suggested that suitable supplementation of electrochemically active electron shuttle(s) to dye-bearing MFCs is a promising strategy to stimulate reductive decolorization and bioelectricity generation.

Deciphering mediating characteristics of decolorized intermediates for reductive decolorization and bioelectricity generation.

As decolorized intermediates could play a role of electron-shuttling mediator to enhance the performance of dye decolorization and bioelectricity generation, this study selected model compounds with auxochromes (e.g., benzene-1,2-diol, 1,2-diaminobenzene) to explore how chemical structure(s) affected color removal and power producing capabilities in microbial fuel cells (MFCs). According to cyclic voltammetry, respiratory testing and MFC data, promising electron-shuttling capabilities of aforementioned compounds were revealed using Proteus hauseri ZMd44, Aeromonas sp. C78, Acinetobacter johnsonii NIUx72 bearing MFCs. These findings clearly indicated that chemical structure(s) of decolorized mediators directly affected characteristics of simultaneous reductive decolorization and bioelectricity generation in MFCs, suggesting feasible operation strategy of MFCs for industrial applications.

Exploring new strains of dye-decolorizing bacteria.

This study unveiled a new strategy to explore new indigenous strains with excellent decolorization capabilities from freshwaters and seawaters. Two new bacterial decolorizers DX2b and SH7b, which have the capability to decolorize textile dyes, were isolated from Cross-Strait Taiwan and China. According to PCR-augmented 16S rRNA gene analyses for strain identification, >99% of nucleotide sequences in isolated strains were identical to type strains Rahnella aquatilis, Acinetobacter guillouiae, Microvirgula aerodenitrificans, and Pseudomonas sp. Time-series inspection upon azoreductase activity assay and generation of decolorized intermediates all confirmed in parallel with reductive decolorization of new decolorizers DX2b and SH7b. The result also showed that bacterial decolorization of these new strains was mainly catalyzed via the enzymatic expression of azoreductase and riboflavin reductase, and biosorption seemed not to play a crucial role color removal (approximately <10%).

Assessment upon azo dye decolorization and bioelectricity generation by Proteus hauseri.

This study explored dye decolorization and bioelectricity generation of indigenous Proteus hauseri ZMd44 for dye-bearing wastewater treatment. Chemical structures of azo dyes apparently affected the performance of dye biodecolorization. Additions of diazo dye C.I. reactive blue 160 (RBu160) stimulated simultaneous dye decolorization and bioelectricity generation of ZMd44 in single chamber microbial fuel cells (MFCs). However, high-level additions of RBu160 repressed capabilities of power production in MFC due to competition of electrons used for reductive decolorization. Decolorized intermediates of RBu160-phenyl methadiamine and 5-sulfoanthranilic acid as electron shuttles might mediate electron transport for current generation in MFC.

Comparative study on characteristics of azo dye decolorization by indigenous decolorizers.

This study provides a first attempt to explore indigenous strains with excellent decolorization capability from the most biodiverse region in Taiwan for dye-bearing wastewater treatment. Bacterial isolates were obtained via serial selections under selection pressure of the fungicide nystatin and model textile dye(s). According to profiles of protein expression and PCR-augmented 16S rRNA gene analyses for strain identification, >99% of nucleotide sequences in isolated strains were identical to type strains Aeromonas hydrophila, Klebsiella pneumoniae, Enterobacter cancerogenus, Proteus hauseri, Acinetobacter johnsonii. This first-attempt study not only explored most abundant decolorizers in Taiwan, but also compared their color removal performance for further applications.

Cost-effective biostimulation strategy for wastewater decolorization using immobilized-cell systems.

This study tended to evaluate threshold operation criteria of biostimulation for optimal biodecolorization in immobilized-cell systems (ICSs) using Porites corals as packing matrices. Indigenous Aeromonas hydrophila with high efficiency for decolorization isolated from Northeast Taiwan was used for study. As maximal treatment performance of ICS could only be achieved with maximal absorbed biomass with highest color removal capability. Maintaining optimal attached cells for cost-effective color removal efficiency inevitably required essential nutrients provided from rich media for biostimulation. With consideration of efficient cell attachment and maximal dye biodecolorization, our proposed method of "graphical reconstruction" quantitatively revealed the most economically-feasible strategy of medium stimulation for color removal. Our findings pointed out the maximal allowable inlet concentration and treatment capacity using our prediction of constant-slope isoclines of ICSs at cultures fed with different concentrations of nutrient sources. The method of isoclines upon transient dynamics of ICSs also provided a technically viable assessment for on-site professionals to quantitatively determine maximal biotreatment thresholds in biostimulation.

Understanding effects of chemical structure on azo dye decolorization characteristics by Aeromonas hydrophila.

This novel comparative study tended to disclose how the molecular structures present in seven azo dyes including two types of azo dyes (i.e., naphthol type azo dyes--Reactive Black 5 (RB 5), Reactive Blue 171 (RB 171), Reactive Green 19 (RG19), Reactive Red 198 (RR198), Reactive Red 141 (RR141), and non-naphthol type azo dyes--Direct Yellow 86 (DY86), Reactive Yellow 84 (RY84)) affected color removal capability of Aeromonas hydrophila. Generally speaking, the decolorization rate of naphthol type azo dye with hydroxyl group at ortho to azo bond was faster than that of non-naphthol type azo dye without hydroxyl group, except of RG19. The azo dyes with electron-withdrawing groups (e.g., sulfo group in RR198, RB5 and RR141) would be easier to be decolorized than the azo dyes with the electron-releasing groups (e.g., -NH-triazine in RB171 and RG19). In addition, the azo dyes containing more electron-withdrawing groups (e.g., RR198, RB5 and RR141) showed significantly faster rate of decolorization. The azo dyes with electron-withdrawing groups (e.g., sulfo group) at para and ortho to azo bond (e.g., RR198, RB5 and RR141) could be more preferred for color removal than those at meta (e.g., DY86 and RY84). The former azo dyes with para and ortho sulfo group provided more effective resonance effects to withdraw electrons from azo bond, causing azo dyes to be highly electrophilic for faster rates of reductive biodecolorization. However, since the ortho substituent caused steric hindrance near azo linkage(s), azo dyes with para substituent could be more favorable (e.g., SO(2)(CH(2))(2)SO(4)(-) in RR198 and RB5) than those with ortho substituent (e.g., sulfo group at RR141) for decolorization. Thus, the ranking of the position for the electron-withdrawing substituent in azo dyes to escalate decolorization was para>ortho>meta. This study suggested that both the positions of substituents on the aromatic ring and the electronic characteristics of substituents in azo dyes all significantly affected the performance of biodecolorization of A. hydrophila.

Comparative assessment upon dye removal capability of indigenous bacterial strains from Lanyang Plain in northeast Taiwan.

This study provides a first attempt from a geological and ecological perspective to look forward isolations of indigenous strains with the decolorization capability from the most biodiverse region in Taiwan for dye-laden wastewater treatment. Serial selections were conducted by a specific use of the fungicide nystatin and model azo dye C.I. reactive red 141 (RR141) during isolation. Several bacterial strains with the excellent capability of azo dye decolorization were predominantly isolated from river water and mud samples of Lanyang River Basin. Phase-curve profiles indicated that azo dye decolorization was found to be non-growth associated for both mixed cultures and isolated pure strains. The color removal efficiency of the mixed culture was nearly 10-fold to that of Pseudomonas luteola at ca. 600mgL(-1) RR141, indicating a promising feasibility of isolated cultures to be applicable for practical treatments. The decolorization performance of unacclimated and acclimated pure cultures was at most 20% and 70-80% to that of the mixed cultures, respectively. It might suggest that combined interactions among decolorizers were crucial for the optimal color removal. According to the results of physiological and 16S rRNA gene sequence examinations, the isolated strains should belong to Aeromonas species (very likely A. hydrophila).