DNA Chimeras as Electrochemical Biosensors for Host-Guest Measurements in Blood.

Few sensing platforms have become ubiquitous to enable rapid and convenient measurements at the point-of-care. Those, however, are "one-off" technologies, meaning that they can only detect a single target and are hardly adaptable. In response, we plan to develop a sensing platform that can be extended to detect other classes of molecules and that affords rapid, convenient, continuous measurements directly in undiluted complex matrices. For this, we decided to rely on a host molecule that presents reversible interactions toward specific guest molecules to develop a new class of sensors that we coined "Electrochemical DNA-host chimeras". As a proof-of-concept for our sensor, we decided to use cyclobis(paraquat-p-phenylene) ("blue box") that we attached on an electrode-bound DNA to allow measurements of electron-rich guests such as dopamine and aspirin. Doing so allows to promote host-guest complex that could be quantified using blue box's electrochemistry. Because of this unique sensor architecture, we achieve, to our knowledge, the first reagentless, continuous and rapid (<5 min) host-guest measurements in undiluted whole blood. We envision that given the library of electroactive host molecules that this will allow the development of a sensing platform for measurements of several classes of molecules in complex matrices at the point-of-care.

Host-Guest Formulation of Carboxylatopillar[6]arene with Ergothioneine as a New Antidote for Combinational Detoxification of Paraquat.

Paraquat (PQ) is exceptionally toxic to the human body. PQ ingestion can cause severe organ damage with a mortality rate of 50-80%, resulting from the absence of effective antidotes and detoxification solutions. Herein, a host-guest formulation is proposed, in which ergothioneine (EGT), an antioxidant drug, was encapsulated by carboxylatopillar[6]arene (CP6A) to achieve a combinational therapy for PQ poisoning. Nuclear magnetic resonance (NMR) and fluorescence titration were employed to confirm the complexation between CP6A and EGT as well as PQ with robust affinities. In vitro studies proved that EGT/CP6A significantly reduced PQ toxicity. Treatment with EGT/CP6A could effectively relieve organ damage caused by PQ ingestion and enhance the normalization of hematological and biochemical parameters. The host-guest formulation EGT/CP6A also improved the survival ratio in PQ-poisoned mice. These favorable outcomes originated from synergistic effects that PQ triggered the release of EGT to combat peroxidation damage and excess PQ was engulfed within the cavity of CP6A.

Surface molecularly imprinted-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for highly selective and sensitive direct analysis of paraquat in complicated samples.

Herein, a novel surface molecularly imprinted-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (SMI-MALDI-TOF MS) method for direct target paraquat (PQ) analysis in complicated samples is reported. Notably, a captured analyte-imprinted material can be directly detected via MALDI-TOF MS by using imprinted material as nanomatrix. Using this strategy, the molecular specific affinity performance of surface molecularly imprinted polymers (SMIPs) and the high-sensitivity detection capability of MALDI-TOF MS was integrated. The introduction of SMI endowed the nanomatrix with the capacity for rebinding the target analyte and ensuring specificity, prevented the interfering organic matrix, and enhanced the analyzing sensitivity. By using paraquat (PQ) as a template, dopamine as a monomer, and covalent organic frameworks with a carboxyl group (C-COFs) as a substrate, polydopamine (PDA) was decorated on C-COFs via a simple self-assembly procedure to generate an analyte-based surface molecularly imprinted polymer (C-COF@PDA-SMIP), which served the dual function of SMIP capturing the target analytes and high-efficiency ionization. Thus, a reliable MALDI-TOF MS detection PQ with high selectivity and sensitivity as well as an interference-free background was achieved. The synthesis and enrichment conditions of C-COF@PDA-SMIPs were optimized, and its structure and property were characterized. Under optimal experimental conditions, the proposed method achieved highly selective and ultrasensitive detection of PQ from 5 to 500 pg mL-1, and the limit of detection was as low as 0.8 pg mL-1, which is at least three orders of magnitude lower than that achieved without enrichment. In addition, the specificity of the proposed method was superior to that of C-COFs and nonimprinted polymers. Moreover, this method exhibited reproducibility, stability, and high salt tolerance. Lastly, the practical applicability of the method was successfully verified by analyzing complicated samples, such as grass and orange.

The Story of the Little Blue Box: A Tribute to Siegfried Hünig.

The tetracationic cyclophane, cyclobis(paraquat-p-phenylene), also known as the little blue box, constitutes a modular receptor that has facilitated the discovery of many host-guest complexes and mechanically interlocked molecules during the past 35 years. Its versatility in binding small π-donors in its tetracationic state, as well as forming trisradical tricationic complexes with viologen radical cations in its doubly reduced bisradical dicationic state, renders it valuable for the construction of various stimuli-responsive materials. Since the first reports in 1988, the little blue box has been featured in over 500 publications in the literature. All this research activity would not have been possible without the seminal contributions carried out by Siegfried Hünig, who not only pioneered the syntheses of viologen-containing cyclophanes, but also revealed their rich redox chemistry in addition to their ability to undergo intramolecular π-dimerization. This Review describes how his pioneering research led to the design and synthesis of the little blue box, and how this redox-active host evolved into the key component of molecular shuttles, switches, and machines.

Azocalixarene-Based Supramolecular System for the Detection of Paraquat via an Improved Indicator Displacement Assay.

In view of the lethal toxicity of paraquat (PQ) on human health, herein, a simple indicator displacement assay (IDA) based on an azo-modified calixarene host (azoCX[4]) and a fluorophore guest (p-DPD) were elaborately constructed for PQ detection in environmental water samples and plant surfaces. The fluorescent signal of p-DPD in the probe can be quenched by azoCX[4] through a photon-induced electron transfer process and recovered upon the addition of PQ within 10 s. The detection range of the p-DPD@azoCX[4] probe was calculated to be 0.35-8 µM in the Tris-HCl buffer solutions (pH = 7.4). Moreover, this probe exhibited excellent detection selectivity toward PQ over five herbicides (glyphosate, bispyribac, atrazine, ametryn, and bensulfuron methyl), together with anti-interference abilities in the presence of inorganic ions (K+, Na+, Zn2+, Ni2+, Li+, F-, Cl-, Br-, CO32-, HCO3-, and NO3-) and amino acids (Asp, Arg, Glu, Ala, and Cys). Particularly, the probe was successfully used to detect PQ in real water samples with acceptable accuracy and showed potential applications for on-site detection with paper-based test strips and on the leaf surface. We believe that this simplified IDA-based probe provided an effective detecting tool for PQ, and the design strategy would guide the further development of new IDA sensing systems.

A Structure-Adaptive Ligand: The Conformation Changes to Fit with the Lengths and Angles of Tetravalent Bis-Paraquat Guests.

A flexible cylindrical cryptand (1b) with two m-phenylene-26-crown-8 ethers has been prepared, and its complexing properties with respect to tetravalent cationic bis-paraquat guests have been investigated. Diffusion-ordered spectroscopy nuclear magnetic resonance (NMR) and titration experiments using cold electrospray ionization mass spectrometry and 1H NMR revealed that 1b has versatile complexing properties with respect to tetravalent cationic guests used despite the guest molecules' length and shape.

Catalytic electrochemistry of the bacterial Molybdoenzyme YcbX.

Molybdenum-dependent enzymes that can reduce N-hydroxylated substrates (e.g. N-hydroxyl-purines, amidoximes) are found in bacteria, plants and vertebrates. They are involved in the conversion of a wide range of N-hydroxylated organic compounds into their corresponding amines, and utilize various redox proteins (cytochrome b5, cyt b5 reductase, flavin reductase) to deliver reducing equivalents to the catalytic centre. Here we present catalytic electrochemistry of the bacterial enzyme YcbX from Escherichia coli utilizing the synthetic electron transfer mediator methyl viologen (MV2+). The electrochemically reduced form (MV+.) acts as an effective electron donor for YcbX. To immobilize YcbX on a glassy carbon electrode, a facile protein crosslinking approach was used with the crosslinker glutaraldehyde (GTA). The YcbX-modified electrode showed a catalytic response for the reduction of a broad range of N-hydroxylated substrates. The catalytic activity of YcbX was examined at different pH values exhibiting an optimum at pH 7.5 and a bell-shaped pH profile with deactivation through deprotonation (pKa1 9.1) or protonation (pKa2 6.1). Electrochemical simulation was employed to obtain new biochemical data for YcbX, in its reaction with methyl viologen and the organic substrates 6-N-hydroxylaminopurine (6-HAP) and benzamidoxime (BA).

Simple colorimetric screening of paraquat residue in vegetables evaluated by localized surface plasmon resonance of gold nanoparticles.

The contamination of paraquat in vegetables is widely connected with human health risks, leading to the research interest in developing a paraquat sensing system. This work reports a simple detection method of paraquat based on the electrostatic interaction of paraquat and the negatively charged gold nanoparticles (AuNPs), resulting in the changes of colors from red to blue and the shifting of localized surface plasmon resonance (LSPR) peaks of AuNPs. The limit of detection concentration (CLOD ) of this system was 100 µM paraquat. Moreover, among eight cationic salts tested, NaCl was selective to enhance the detection sensitivity of the system, resulting in the reduction of CLOD to 0.10 µM. This system selectively detected paraquat, but not other tested herbicides (ametryn, atrazine, glyphosate, and 2,4-D-dimethyl ammonium). The paraquat-spiking experiment in kale demonstrated the significant recovery rate of paraquat at 96.0-103.0%, and the relative standard deviations were less than 4%. The developed system was efficient for screening contaminated paraquat in vegetables under unwashed and washed conditions. Three out of five unwashed vegetables had a significant level of paraquat as determined by LSPR values. These results suggested the potential application of this system for a simple screening of contaminated paraquat in vegetables. Simple paraquat-screening system was developed based on the negatively charged gold nanoparticles. The limit of paraquat detection of this system was 0.10 µM. This system was potentially used for a simple screening of contaminated paraquat in vegetables.

Paraquat degradation by photocatalysis: experimental desing and optimization.

This study describes the experimental design and optimization of application TiO2 catalysts doped with 0.5, 1, 1.5, 2.0% of Fe. The catalysts were prepared using the impregnation method applied in Paraquat herbicide degradation. The catalysts were characterized by the following techniques: specific surface area and volume, mean pore diameter (BET method), scanning electron microscopy and photoacoustic spectroscopy. The characterization presented results indicating that both calcination temperature and the increase nominal metallic load affected by the structure of catalysts, changing the textural properties, as well as the band gap. The catalyst that presented the best herbicide removal percentage was TiO2 calcined at 773 K with removal of 90.2%. However, according to the experimental design and optimization, both variables (calcination temperature and Fe percentage) are significant in the process. In addition, a positive effect was found in the interaction between the two variables. The values show that a third order kinetic model better described the Paraquat photocatalytic degradation.

Construction of Supramolecular Polymers with Different Topologies by Orthogonal Self-Assembly of Cryptand-Paraquat Recognition and Metal Coordination.

Recently, metal-coordinated orthogonal self-assembly has been used as a feasible and efficient method in the construction of polymeric materials, which can also provide supramolecular self-assembly complexes with different topologies. Herein, a cryptand with a rigid pyridyl group on the third arm derived from BMP32C10 was synthesized. Through coordination-driven self-assembly with a bidentate organoplatinum(II) acceptor or tetradentate Pd(BF4)2•4CH3CN, a di-cryptand complex and tetra-cryptand complex were prepared, respectively. Subsequently, through the addition of a di-paraquat guest, linear and cross-linked supramolecular polymers were constructed through orthogonal self-assembly, respectively. By comparing their proton nuclear magnetic resonance (1H NMR) and diffusion-ordered spectroscopy (DOSY) spectra, it was found that the degrees of polymerization were dependent not only on the concentrations of the monomers but also on the topologies of the supramolecular polymers.

Protein Nanobarrel for Integrating Chlorophyll a Molecules and Its Photochemical Performance.

Taking inspiration from biology's effectiveness in nanoscale organization of chlorophylls for photosynthesis, we describe here a design for chlorophyll-protein conjugates that exploits the central hydrophobic cavity of GroEL protein nanobarrel as a binding pocket for chlorophyll. We found water-soluble conjugates of chlorophyll with GroEL could be easily generated via detergent dialysis. The number of chlorophyll units bound to GroEL is tunable by varying the equilibrium concentration of chlorophyll during dialysis. Meanwhile, it is shown that an increase in the entrapped chlorophyll amount leads to an improvement of chlorophyll-GroEL photostability. Using methyl viologen as an electron acceptor, we demonstrate that chlorophyll-GroEL has photoreduction activity, which is also switchable in on/off illumination mode. Finally, it is shown that chlorophyll-GroEL-sensitized solar cells have good photoelectric properties, yielding a high photoelectric conversion efficiency of ∼0.9%. The current strategy may be adopted for integrating other photosensitizing dyes or for other photocatalytic reactions.

Ultrasonic-assisted extraction of polysaccharides from Auricularia auricula and effects of its acid hydrolysate on the biological function of Caenorhabditis elegans.

The present study was designed to explore the in vivo-antioxidant capacity and the probable mechanism of AAPs-H, prepared from Auricularia auricula polysaccharides with the optimal extraction conditions by Box-Behnken design and acid hydrolysis, using Caenorhabditis elegans as a model organism. The effects of AAPs-H on the locomotion behavior, life span, antioxidant-related enzymes activities, and antioxidants levels in C. elegans were studied. Furthermore, the potentials of AAPs-H in up-regulating the expression of antioxidant-related genes in C. elegans, such as skn-1, sod-3 and sir-2.1, were also discussed. AAPs-H demonstrated a highly significant protective effect against the damage caused by paraquat, could significantly increase U-Turn frequency of worms (p < 0.01), extend their lifespan, enhance antioxidant systems including GR by 63.96% (p < 0.05), GSH-Px by 71.16% (p < 0.01), SOD by 78.65% (p < 0.01) and CAT by 98.52% (p < 0.01), increase the level of GSH by 28.12% (p < 0.05), and decrease the level of MDA by 39.29% (p < 0.01). The qRT-PCR results showed that AAPs-H could up regulate mRNA expression levels of skn-1, sod-1, sod-2, sod-3 and sir-2.1 in wild-type C. elegans (>1.6 fold) when treated with the concentration of 0.4 mg/mL (p < 0.05 or p < 0.01). Our studies provide evidence that AAPs-H improves antioxidant defense system, and up-regulation of oxidative stress related genes for prevention of stress damage in C. elegans.

Selection of an artificial paraquat-specific binding protein from a ribosome display library based on a lipocalin scaffold.

In this study, an anticalin that could specifically bind paraquat (PQ), a quaternary nitrogen herbicide, as a new set of engineered receptor protein with antibody-like properties was generated to detect PQ concentration. To this end, a native and random library was constructed and engineered to allow in vitro transcription and translation using an Escherichia coli lysate system. Meanwhile, a PQ derivative that carries an active aliphatic carboxylate group at the end of an aliphatic spacer arm was synthesized. Then, this compound was coupled covalently to the carrier protein bovine serum albumin/ovalbumin and amino-functionalized paramagnetic beads. Alternating selection in solution and immobilization in microtiter wells were used to pan mRNA-ribosome-antibody complexes. After several rounds of ribosome display, three variants were selected from a random library of the bilin-binding protein. The variants that could bind complex PQ with high affinity and exhibit IC50 values as low as 14.039 ± 0.970 ng/mL were identified. Moreover, the limits of detection reached 0.083 ± 0.011 ng/mL. Our data suggest that the generation of anticalins may provide a promising alternative to recombinant antibody fragments to create a stable receptor protein against hapten with bioanalytical relevance.

Carbon tubes from biomass with prominent adsorption performance for paraquat.

Paraquat (PQ) pollution has become an increasing seriously threat to the ecological environment. Thus, it is vital to study novel adsorption materials capable of removing PQ. Modified carbon tubes (MCT-600) were prepared by carbonizing the seed hair fibers of Metaplexis japonica at 600 °C and further modifying with nitric acid. The MCT-600 was found to possess a regular morphology, a good graphitization degree and an abundance of carboxyl functional groups. Moreover, this materil exhibits excellent adsorption performance for paraquat (PQ), such as a fast adsorption rate (adsorption equilibrium within 20 min), large adsorption capacity (218.61 mg g-1) and good regeneration ability (reused more than 5 times). The adsorption process for PQ onto MCT-600 was found to be well matched to a Langmuir isotherm and pseudo-second-order kinetics model. Owing to its unique surface properties, green raw material source and simple preparation process, MCT-600 shows potential as an outstanding candidate for PQ removal from water.

Effects of Chemically-Modified Polypyridyl Ligands on the Structural and Redox Properties of Tricarbonylmanganese(I) Complexes.

Carbonyl complexes with manganese(I) as the central metal are very attractive catalysts. The introduction of redox-active ligands, such as quinones and methyl viologen analogs into these catalysts, would be expected to lead to superior catalyst performances, since they can function as excellent electron carriers. In this study, we synthesized four tricarbonylmanganese(I) complexes containing typical bidentate polypyridyl ligands, including 1,10-phenanthroline (phen) and 2,2'-bipyridine (bpy) frameworks bound to redox-active ortho-quinone/catechol or methyl viologen-like units. The molecular structures of the resulting complexes were determined by X-ray crystallography to clarify their steric features. As expected from the infrared (IR) data, three CO ligands for each complex were coordinated in the facial configuration around the central manganese(I) atom. Additionally, the structural parameters were found to differ significantly between the quinone/catechol units. Electrochemical analysis revealed some differences between them and their reference complexes, namely [MnBr(CO)3(phen)] and [MnBr(CO)3(bpy)]. Notably, interconversions induced by two-electron/two-proton transfers between the quinone and catechol units were observed in the phenanthroline-based complexes. This work indicated that the structural and redox properties in tricarbonylmanganese(I) complexes were significantly affected by chemically modified polypyridyl ligands. A better understanding of structures and redox behaviors of the present compounds would facilitate the design of new manganese complexes with enhanced properties.

How does methylviologen cation radical supply two electrons to the formate dehydrogenase in the catalytic reduction process of CO2 to formate?

Formate dehydrogenase from Candida boidinii (EC.1.2.1.2; CbFDH) is a commercially available enzyme and can be easily handled as a catalyst for the CO2 reduction to formate in the presence of NADH, single-electron reduced methylviologen (MV+˙) and so on. It was found that the formate oxidation to CO2 with CbFDH was suppressed using the oxidized MV as a co-enzyme and the single-electron reduced MV (MV+˙) was effective for the catalytic activity of CbFDH for the CO2 reduction to formate compared with that using the natural co-enzyme of NADH [Y. Amao, Chem. Lett., 2017, 46, 780-788]. The CO2 reduction to formate catalyzed by CbFDH requires two molecules of the MV+˙. In order to clarify the two-electron reduction process using MV+˙ in the CO2 reduction to formate catalyzed with CbFDH, we attempted enzyme reaction kinetics, electrochemical and quantum chemical analyses. Kinetic parameters obtained from the enzymatic kinetic analysis metric revealed an index of affinity of MV+˙ for CbFDH in the CO2 reduction to formate. From the results of the electrochemical analysis, it was predicted that only one molecule of MV+˙ was bound to CbFDH, and the MV bound to CbFDH was to be necessarily re-reduced by the electron source outside of CbFDH to supply the second electron in the CO2 reduction to formate. From the results of docking simulation and density functional theory (DFT) calculations, it was indicated that one molecule of MV bound to the position close to CO2 in the inner part of the substrate binding pocket of CbFDH contributed to the two-electron CO2 reduction to formate.

Selective Decoating-Induced Activation of Supramolecularly Coated Toxic Nanoparticles for Multiple Applications.

In spite of the rapid emergence of numerous nanoparticles (NPs) for biomedical applications, it is often challenging to precisely control, or effectively tame, the bioactivity/toxicity of NPs, thereby exhibiting limited applications in biomedical areas. Herein, we report the construction of hyaluronic acid (HA)-laminated, otherwise toxic methylviologen (MV), NPs via ternary host-guest complexation among cucurbit[8]uril, trans-azobenzene-conjugated HA, and MV-functionalized polylactic acid NPs (MV-NPs). The high, nonspecific toxicity of MV-NPs was effectively shielded (turned off) by HA lamination, as demonstrated in cells, zebrafish, and mouse models. The supramolecular host-guest interaction-mediated HA coating offered several HA-MV-NP modalities, including hyaluronidase locally and photoirradiation remotely, to precisely remove HA lamination on demand, thereby endowing materials with the capability of selective decoating-induced activation (DIA) for applications as a user-friendly herbicide, a selective antibacterial agent, or an anticancer nanomedicine. This work offers facile supramolecular coating and DIA strategies to effectively tame and precisely control the bioactivity and toxicity of functional nanomaterials for diverse applications.

Enhanced Electrosynthetic Hydrogen Evolution by Hydrogenases Embedded in a Redox-Active Hydrogel.

Molecular hydrogen is a major high-energy carrier for future energy technologies, if produced from renewable electrical energy. Hydrogenase enzymes offer a pathway for bioelectrochemically producing hydrogen that is advantageous over traditional platforms for hydrogen production because of low overpotentials and ambient operating temperature and pressure. However, electron delivery from the electrode surface to the enzyme's active site is often rate-limiting. Here, it is shown that three different hydrogenases from Clostridium pasteurianum and Methanococcus maripaludis, when immobilized at a cathode in a cobaltocene-functionalized polyallylamine (Cc-PAA) redox polymer, mediate rapid and efficient hydrogen evolution. Furthermore, it is shown that Cc-PAA-mediated hydrogenases can operate at high faradaic efficiency (80-100 %) and low apparent overpotential (-0.578 to -0.593 V vs. SHE). Specific activities of these hydrogenases in the electrosynthetic Cc-PAA assay were comparable to their respective activities in traditional methyl viologen assays, indicating that Cc-PAA mediates electron transfer at high rates, to most of the embedded enzymes.

Micelle-dominated distribution strategy for non-matrix matched calibration without an internal standard: "Extract-and-shoot" approach for analyzing hydrophilic targets in blood and cell samples.

The analysis of trace hydrophilic targets in complex aqueous-rich matrices is considerably challenging, generally requiring matrix-matched calibration, internal standard, or time-and-labor-intensive sample preparation. To address this analytical bottleneck, a non-matrix-matched calibration strategy without using internal standard was reported for the first time to analyze complicated biosamples such as whole blood, plasma, serum, and cell samples. This strategy, termed micelle-dominated distribution, also aimed at realizing the simple "extract-and-shoot" analytical process for such complex matrices. The micelle-matrix interaction was found to efficiently eliminate the matrix effect by dominating phase separation and analyte distribution between the extraction and matrix phases. Thus, calibration linear curves prepared in water were applicable to the analysis of all the above-mentioned sample types. Rapid distribution equilibrium within 4 min was achieved. This strategy could tolerate direct large volume injection, thereby providing two-order-of-magnitude enhancement in the sensitivity of ion-pair chromatography. The analytical method integrated cell rupture, matrix cleanup, analyte extraction, and on-column preconcentration into a fast and high-throughput operation. The successful application to the determination of exogenous pesticides and endogenous glutathione exhibited low limits of detection (0.0085-0.015 µg mL-1 for pesticides; 0.52 µg mL-1 for glutathione), wide linear ranges (0.028-50 µg mL-1 and 0.049-50 µg mL-1 for pesticides; 1.7-1000 µg mL-1 for glutathione), good linearies (R2 = 0.9994-0.9999), excellent accuracy (recoveries of 91.3-105.2%), and good precision (0.7-6.2% at the levels of 0.028 (or 0.049), 0.1, 0.5, and 50 µg mL-1 for pesticides; 0.5-8.7% at 1.7, 500, and 1000 µg mL-1 for glutathione).

Hepatoprotective role of berberine against paraquat-induced liver toxicity in rat.

Paraquat (PQ) is a herbicide agent commonly used in agricultural applications. Hepatotoxicity is among clinical complications associated with PQ intoxication. Oxidative stress and its subsequent events are major mechanisms identified in PQ-induced liver toxicity. Berberine (BBR) is a natural antioxidant widely investigated for its hepatoprotective effects. The present study designed to evaluate the potential cytoprotective properties of BBR against PQ-induced cytotoxicity in primary cultured rat hepatocytes and in vivo test of liver function enzymes. Cellular and biochemical parameters including lactate dehydrogenase (LDH), cell viability, ROS formation, glutathione (GSH) content, and mitochondrial membrane potential in the PQ-treated hepatocytes were measured, and the mentioned markers were evaluated in the presence of BBR. BBR treatment caused significant decrease in PQ-induced cell death, ROS formation, and LDH release. On the other hand, it was found that BBR inhibits cellular glutathione depletion in PQ-treated hepatocytes. Also, BBR treatment significantly diminished PQ-induced the liver function enzyme elevation. These data mention the potential hepatoprotective effect of BBR with therapeutic capability against PQ-induced liver damage.