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Rheum officinale, a member of the Polygonaceae family, is an important medicinal plant that is widely used in traditional Chinese medicine. Here, we report a 7.68-Gb chromosome-scale assembly of R. officinale with a contig N50 of 3.47 Mb, which was clustered into 44 chromosomes across four homologous groups. Comparative genomics analysis revealed that transposable elements have made a significant contribution to its genome evolution, gene copy number variation, and gene regulation and expression, particularly of genes involved in metabolite biosynthesis, stress resistance, and root development. We placed the recent autotetraploidization of R. officinale at ∼0.58 mya and analyzed the genomic features of its homologous chromosomes. Although no dominant monoploid genomes were observed at the overall expression level, numerous allele-differentially-expressed genes were identified, mainly with different transposable element insertions in their regulatory regions, suggesting that they functionally diverged after polyploidization. Combining genomics, transcriptomics, and metabolomics, we explored the contributions of gene family amplification and tetraploidization to the abundant anthraquinone production of R. officinale, as well as gene expression patterns and differences in anthraquinone content among tissues. Our report offers unprecedented genomic resources for fundamental research on the autopolyploid herb R. officinale and guidance for polyploid breeding of herbs., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

The detection of anthraquinone in tea leaves has raised concerns due to a potential health risk associated with this species. This led the European Union to impose a maximum residue limit (MRL) of 0.02 mg/kg for anthraquinone in dried tea leaves. As atmospheric contamination has been identified as one of the possible sources of anthraquinone residue, this study investigates the contamination resulting from the deposition of atmospheric anthraquinone using a global chemical transport model that accounts for the emission, atmospheric transport, chemical transformation, and deposition of anthraquinone on the surface. The largest contribution to the global atmospheric budget of anthraquinone is from residential combustion followed by the secondary formation from oxidation of anthracene. Simulations suggest that atmospheric anthraquinone deposition could be a substantial source of the anthraquinone found on tea leaves in several tea-producing regions, especially near highly industrialized and populated areas of southern and eastern Asia. The high level of anthraquinone deposition in these areas may result in residues in tea products exceeding the EU MRL. Additional contamination could also result from local tea production operations., (© 2023. The Author(s).)

In the present work, a two-dimensional qNMR method for the determination of sennosides was established. Using band-selective HSQC and the cross correlations of the characteristic 10-10' bonds, we quantified the total amount of the value-determining dianthranoids in five minutes, thus, rendering the method not only fast, but also specific and stability indicating. The validation of the method revealed excellent accuracy (recovery rates of 98.5 to 103%), precision (RSD values of 3.1%), and repeatability (2.2%) and demonstrated the potential of 2D qNMR in the quality control of medicinal plants. In a second method, the use of 2D qNMR for the single analysis of sennosides A, B, and A 1 was evaluated with acceptable measurement times (31 min), accuracy (93.8%), and repeatability (5.4% and 5.6%) for the two major purgatives sennoside A and B. However, the precision for sennoside B and A 1 was not satisfactory, mainly due to the low resolution of the HSQC signals of the two compounds.

In recent years, the hepatotoxicity of Polygoni Multiflora Radix (PMR) has attracted increased research interest. Some studies suggest that anthraquinone may be the main hepatotoxic component. Most of the relevant studies have focused on the mononuclear anthraquinone component rather than binuclear anthraquinones. The hepatotoxicity of dinuclear anthraquinone (dianthrone) was investigated in a cell-based model. Next, a method for the determination of six free and total dianthonones in PMR and PMR Praeparata (PMRP) was established using ultra-high-performance liquid chromatography triple quadrupole mass spectrometry (UPLC-QQQ-MS/MS), which was then used to analyze the collected samples. The data show that four binuclear anthraquinone compounds were hepatotoxic and may be potential toxicity indicators for the safety evaluation of PMR and PMRP. Herein, we provide a theoretical basis for the improvement of PMRP quality standards.

Propolis is a well-known resinous natural substance collected by honeybees ( Apis mellifera L.) from plants exudations. Variations in chemical composition of propolis are due to different sources from which it is collected and change in climate and geographical location. In this study, different propolis samples were collected from different regions of Balochistan and examined for its chemical composition, total phenolics and total flavonoid contents, and antioxidant potential by using DPPH radical scavenging assay and antimicrobial activity. Bioactive components analysis revealed the presence of steroids, carbohydrates, flavonoids, coumarins, cardiac glycosides, quinones, anthraquinones, terpenoids, tannins, and phlobatannins at different levels. The total phenolics contents were ranged from 2.9343 ± 1.247 to 6.0216 ± 2.873 mg GAE g -1 , and flavonoid contents were found to be 0.1546 ± 0.087 to 0.6586 ± 0.329 mg QE g -1, respectively. The antioxidant ability of each extract was analyzed by their concentration having 50% inhibition ( IC 50 ). The propolis sample P3 possessed lower IC 50 27.07 ± 0.73 mg mL -1 with higher % inhibition of DPPH radical, and P8 showed lower % inhibition by having IC 50 84.43 ± 2.07 mg mL -1 . The antibacterial activity of all samples was analyzed against a wide group of bacteria including Escherichia coli , Staphylococcus aureus , Pseudomonas aeruginosa , and Klebsiella pneumonia and propolis extract (P4) was highly active against Klebsiella pneumoniae with the maximum diameter of zone of inhibition 20.33 ± 1.52 mm, and propolis extract (P3) showed maximum zone of inhibition against Escherichia coli 19.06 ± 1.90, while propolis extract (P2) was found less active with minimum diameter of zone of inhibition 7.46 ± 1.50 mm. The antifungal activity of extract was considered as active against the fungal species. Propolis extract (P3) showed 82% of zone of inhibition against Aspergillus Niger , and propolis extract (P1) was highly active against Aspergillus parasiticus with 80% of zone of inhibition. By comparing the vibration frequencies in wave numbers of the sample spectrograph acquired from an FTIR spectrophotometer, the functional groups present in the extracts were identified. The presence of seven elements (Fe, Zn, Mn, Ni, Pb, Cd, and Cr) was analyzed through atomic absorption spectrophotometer. The obtained concentrations were within the permissible ranges established by the World Health Organization. The GC-MS analysis revealed the presence of 80 different compounds belonged to different classes. The obtained results confirmed the imperative potential of propolis which can be used in various biological applications., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Ali Akbar et al.)

Acacia ehrenbergiana (Hayne), also known as Salam, is a highly drought resistant shrub distributed in North and East Africa, and the Arabian Peninsula. The plant is gathered for its gum and fiber, and is an important legume species for indigenous populations. In this study, the phytochemical analysis, antibacterial, and antioxidant properties of various alcoholic and aqueous extracts of Acacia ehrenbergiana grown in Qatar were investigated. The qualitative phytochemical screening of this species exhibited the presence of glycosides, tannins, flavonoids, terpenoids, saponins, phenol, and anthraquinones in various extracts. The agar diffusion method was performed to check the antibacterial activity. The acetone and ethanol extracts showed 85% antibacterial activity of the control against Gram-negative E. coli , while the acetone extract had 65% activity against the Bacillus Gram-positive species. The highest activity against Staphylococcus aureus was 65% for the butanol extract. The antioxidant capacities were evaluated by the DPPH method. Various extracts exhibited antioxidant activities similar to or higher than standard antioxidants, with the highest percent inhibition of 95% for the acetone and ethanol extracts. The acetone extracts were further purified by reverse phase combiflash chromatography followed by HPLC. Three of the pure compounds isolated were subjected to MS, FTIR, and NMR spectral analysis and were found to be stigmasterol, spinasterol, and theogallin. In conclusion, the observed antibacterial and antioxidant activities as well as the presence of secondary metabolites with potential medicinal activities makes Acacia ehrenbergiana a potent valuable endemic medicinal plant.

Degradation of anthocyanins involves scission of the flavonoid skeleton yielding 2,4,6-trihydroxybenzaldehyde (phloroglucinaldehyde, PGA) and a phenolic acid. However, the process is not finished with the formation of PGA, as the consequent condensation of two PGA molecules providing colored hydroxylated anthraquinones was observed for the first time. This process was studied using a combination of preparative column chromatography, nuclear magnetic resonance, liquid chromatography/high resolution tandem mass spectrometry (LC/HRMS 2 ), and quantum calculations using density functional theory. 1,3,5,7-tetrahydroxyanthraquinone (anthrachrysone) and its isomers were found to rise during heating (95 °C) in a buffered PGA model solution (phosphate buffer, pH 7). These compounds were detected in heated red wine after an increase of its pH value. The concentration of the identified anthrachrysone in the red wine reached 0.01 mg·L -1 . Presence of those compounds could therefore indicate involvement of certain steps in the processing of plant materials rich in anthocyanins (e.g., utilization of a higher temperature and/or reduction of acidity) or long-term transformation of anthocyanins (potentially, for instance, in archaeological findings such as wine or fruit residues). Additionally, measurement of wine-soil suspensions proved an increase of their pH to the values suitable for anthocyanin cleavage (neutral to slightly alkaline; reached using soil from archaeologically well-known Bull Rock Cave). Although not found in artificially prepared samples (imitations) or authentic materials so far, according to our results the above mentioned conditions are suitable for the formation of tetrahydroxylated anthraquinone derivatives and their monitoring would be beneficial.

This work presents the research on the treatment of an anthraquinone derivatives of disperse dye Blue SI from aqueous solution using aluminium for the optimization of operational parameters like pH, current density, addition of electrolyte, contact time for the color removal efficiency (CRE) and the results are compared with the performance of copper electrodes in electrocoagulation (EC). The parameters for maximum CRE was found with Al at current density 40 Am -2 , time 10 min at pH 7, and for Cu at 60 Am -2 15 min, at pH 6 were optimized. The characterization of the treated water using HPLC, MS studies revealed intermediate compounds. From the XPS analysis of the sludge obtained, the mechanism of EC was deduced. Treated aqueous solution was studied for its phytotoxicity with Vigna radiata and ecotoxicity studies were conducted on Artemia salina to study the toxicity effect of the intermediatory products in the treated dye solution. Blue SI dye aqueous solution treated with aluminium electrodes shows no or lesser toxicity in plants as well as in ecotoxic study compared with copper electrodes., Competing Interests: Declaration of competing interest The authors Parameswari Kalivel, Rajkumar Pluto Singh, Subbiah Kavitha, Dhanasekaran Padmanabhan, Suresh kumar Krishnan, Jegathambal Palanichamy declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Hypericum perforatum and related species (Hypericaceae) are a reservoir of pharmacologically important secondary metabolites, including the well-known naphthodianthrone hypericin. However, the exact biosynthetic steps in the hypericin biosynthetic pathway, vis-à-vis the essential precursors and their localization in plants, remain unestablished. Recently, we proposed a novel biosynthetic pathway of hypericin, not through emodin and emodin anthrone, but skyrin. However, the localization of skyrin and its precursors in Hypericum plants, as well as the correlation between their spatial distribution with the hypericin pathway intermediates and the produced naphthodianthrones, are not known. Herein, we report the spatial distribution of skyrin and its precursors in leaves of five in vitro cultivated Hypericum plant species concomitant to hypericin, its analogs, as well as its previously proposed precursors emodin and emodin anthrone, using MALDI-HRMS imaging. Firstly, we employed HPLC-HRMS to confirm the presence of skyrin in all analyzed species, namely H. humifusum , H. bupleuroides , H. annulatum , H. tetrapterum, and H. rumeliacum . Thereafter, MALDI-HRMS imaging of the skyrin-containing leaves revealed a species-specific distribution and localization pattern of skyrin. Skyrin is localized in the dark glands in H. humifusum and H. tetrapterum leaves together with hypericin but remains scattered throughout the leaves in H. annulatum , H. bupleuroides , and H. rumeliacum . The distribution and localization of related compounds were also mapped and are discussed concomitant to the incidence of skyrin. Taken together, our study establishes and correlates for the first time, the high spatial distribution of skyrin and its precursors, as well as of hypericin, its analogs, and previously proposed precursors emodin and emodin anthrone in the leaves of Hypericum plants.

Alternative routes to degrade dyes are of crucial importance for the environment. Hence, we report the electrochemical removal of indanthrene blue by using a boron-doped diamond anode, focusing on the toxicity of the treated solutions. Different operational conditions were studied, such as current density (5, 10, and 20 mA cm -2 ) and electrolyte composition (Na 2 SO 4 , Na 2 CO 3, and NaNO 3 ). Besides, the pH was monitored throughout the experiment to consider its direct influence on the ecotoxicity effects. The highest electrochemical oxidation efficiency, measured as color removal, was seen in the 180 min condition of electrolysis in 0.033 M Na 2 SO 4 , applying 20 mA cm -2 , resulting in a color removal of nearly 91% and 40.51 kWh m -3 of energy consumption. The toxicity towards Lactuca sativa depends solely on pH variations being indifferent to color removal. While the inhibition concentration (IC 50 ) for Raphidocelis subcapitata increases 20% after treatment (in optimized conditions), suggesting that the byproducts are more toxic for this specific organism. Our data highlight the importance of analyzing the toxicity towards various organisms to understand the toxic effect of the treatment applied., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Intestinal obstruction (IO) is a kind of acute abdomen with high morbidity and mortality. Patients suffer from poor quality of life and tremendous financial pressure. Da-Cheng-Qi decoction (DCQD), a classical purgation prescription, has clinically been proven to be an effective treatment for IO., Purpose: Network pharmacology integrated with bioactive equivalence assessment was used to discover the quality marker (Q-marker) of DCQD against IO., Methods: As there is hardly any targets recorded in database, thus the collection of IO targets was conducted by searching those of alternative diseases which have similar pathological symptoms with IO. In order to improve the reliability of the obtained targets, IO metabolomics data was introduced. Active compounds combination (ACC) was focused as potential Q-markers via component-target network analysis and function query from the identified components corresponding to the common targets. Bioequivalence between ACC and DCQD was assessed from the aspects of intestine motility (somatostatin secretion), inflammation (IL-6 secretion) and injury (wound healing assay) in vitro and was further validated in ileus rat model. PPI network analysis of core targets followed by gene pedigree classification and experimental validation confirmed the potential intervention pathway., Results: A combination of 11 ingredients, including emodin, physcion, aloe-emodin, rhein, chrysophanol, gallic acid, magnolol, honokiol, naringenin, tangeretin, and nobiletin was finally confirmed bioequivalence with DQCD to some extent and could serve as Q-markers for DCQD to attenuate IO. PI3K/AKT was verified as a possible affected pathway that DCQD exerted the effectiveness against IO., Conclusion: For the disease with few recorded targets, searching those of alternative diseases which have similar pathological symptoms could be a feasible and effective approach. The proposed network pharmacology integrated bioactive equivalence evaluation paradigm is efficient to discover Q-marker of herbal formulae., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Roots of Morinda officinalis and Morinda citrifolia have been interchangeably used in traditional Chinese medicine. However, there is no experimental evidence to support this. In this study, a ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS)-based approach and a multivariate statistical analysis (MSA) were adopted to compare the difference in the chemical compounds present in the root extract of M. officinalis and M. citrifolia . There were 26 anthraquinones, 15 triterpenes, and 8 iridoid glycosides identified in the root extracts of M. officinalis , 30 anthraquinones, 1 triterpene, and 8 iridoid glycosides in the root extracts of M. citrifolia . Among these, 25 compounds presented in both plants. In addition, a principal component analysis (PCA) showed that these two herbs could be separated clearly. Furthermore, an orthogonal partial least squares-discriminant analysis (OPLS-DA) found 9 components that could be used as chemical markers to discrimination the root extracts of M. officinalis and M. citrifolia . In addition, the results of a Cell Counting Kit 8 (CCK-8) assay and cell colony formation assay indicated that methanol root extracts of M. officinalis and M. citrifolia showed no cell cytotoxicity to normal cells, even promoted the proliferation of normal liver cells. To our knowledge, this is the first time that the differences between the root extracts of M. officinalis and M. citrifolia (Hainan province) have been observed systematically at the chemistry level.

Type 2 diabetes mellitus (T2DM) is a chronic disease, and dietary modification is a crucial part of disease management. Okara is a sustainable source of fibre-rich food. Most of the valorization research on okara focused more on the physical attributes instead of the possible health attributes. The fermentation of okara using microbes originated from food source, such as tea, sake, sufu and yoghurt, were explored here. The aim of this study is to investigate fermented okara as a functional food ingredient to reduce blood glucose levels. Fermented and non-fermented okara extracts were analyzed using the metabolomic approach with UHPLC-QTof-MS E . Statistical analysis demonstrated that the anthraquinones, emodin and physcion, served as potential markers and differentiated Eurotium cristatum fermented okara (ECO) over other choices of microbes. The in-vitro α-glucosidase activity assays and in-vivo mice studies showed that ECO can reduce postprandial blood glucose levels. A 20% ECO loading crispy snack prototype revealed a good nutrition composition and could serve as a fundamental formulation for future antidiabetes recipe development, strengthening the hypothesis that ECO can be used as a novel food ingredient for diabetic management.

An optimisation of extraction towards an increased antioxidant capacity and the study on the extracts' composition by HPLC-ESI-Q-TOF-MS were performed on different organs of a rarely studied plant: Rheum cordatum Losinsk (Polygonaceae) growing in Kazakhstan. More than 20 compounds from anthraquinones and phenolics were identified in an optimised method. The plant was proven to contain a wide variety of phenolic compounds (catechins, flavonoids, and their glucosides and phenolic acids) in contrast to the anthraquinone composition, which was mainly represented by emodin and its analogues. The results of the studies could determine the plant as a rich source of pharmacologically precious polyphenols. It was evidenced that the extracting solvents, the time of collection, and the organs tested affected both the chemical content and the antioxidant potential of the extracts. Ethanol : water (50 : 50 v / v ) was selected as the most beneficial extractant for all metabolites, and based on the principal component analysis of raw data, the radical scavenging potential of the plant was strictly related to the presence of epicatechin gallate (ECG), kaempferol glucoside, and epigallocatechin gallate (EGCG) occurring in this extract at the concentration of 1.69-5%, 0.16-0.47%, and 0.001-2.93%, respectively., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2019 Gulsim Zhumashova et al.)

Quinones are becoming an essential tool for refractory organics treatment, while their quantification may be not well-considered. In this paper, two kinds of potential errors in quantification were evaluated in multiple pH conditions. They were derived from the coexistence of oxidized/reduced quinone species (Type I) and pH-sensitive feature (Type II), respectively. These errors would remarkably influence the accuracy of quantification while they haven't been emphasized. Thus, to elaborate the relationship between the two types of errors and the absorbance or pH conditions, three typical quinones [Anthraquinone-1-sulfonate (α-AQS), anthraquinone-2,6-disulfonate (AQDS) and lawsone] were selected and their acid dissociation coefficients (pKa) as well as UV-Vis spectra were determined. Results revealed that, for Type I, the relative error (RE) of α-AQS concentration would exceed the limit (5%) when reduced α-AQS was below 48% of total α-AQS. Similar results were found for lawsone. However, the RE can be eliminated by the equation established in this paper. For Type II, the pH-sensitive feature was related to the pKa values of quinones. Absorbances of α-AQS and lawsone would change remarkably with pH variation. Therefore, a model for correction was established. Analog data showed high consistency with experimental data [r = 0.995 (n = 25, p < 0.01) and r = 0.997 (n = 36, p < 0.01), for lawsone and α-AQS respectively]. Especially, the determination of AQDS concentrations was noticed to be pH-independent at 437 nm under pH 4.00 to 9.18 conditions. Based on these features, a comprehensive data solution was proposed for handling these errors., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Passive air sampling was conducted in Toronto and the Greater Toronto Area from 2016 to 2017 for 6 periods, in order to investigate ambient levels of polycyclic aromatic compounds (PACs) associated with different source types. The selected sampling sites (n = 8) cover geographical areas with varying source emissions including background, traffic, urban, industrial and residential sites. Passive air samples were analyzed for PACs which include PAHs, alkylated PAHs (alk-PAHs), dibenzothiophene and alkylated dibenzothiophenes (DBTs) and results for PAHs were used to calculate inhalation cancer risks using different approaches. The samples were also characterized for PAH derivatives including nitrated PAHs (NPAHs) and oxygenated PAHs (OPAHs). Concentrations of Σalk-PAHs and DBTs, which are known to be enriched in fossil fuels, as well as ΣNPAHs, were highest at a traffic site (MECP) located adjacent to the 18-lane Highway 401 that runs across Toronto. Except for an industrial site (HH/BU), PAC compositions were similar across the sampling sites with Σalk-PAHs being the most abundant class of PACs suggesting traffic emission was a major contributor to PACs in the atmosphere of Toronto. The industrial site exhibited a distinct chemical composition with ΣPAHs dominating over Σalk-PAHs and with elevated levels of fluoranthene, 9-nitroanthracene, and 9,10-anthraquinone, which likely reflects emissions from nearby industrial sources. MECP and HH/BU exhibited higher lifetime excess inhalation cancer risks indicating an association with traffic and industrial sources. The importance of the traffic sector as a source of PACs to ambient air is further supported by strong correlations of the ΣPAHs, Σalk-PAHs, DBTs, and ΣOPAHs with NO x . This study highlights the importance of traffic as an emission source of PACs to urban air and the relevance of PAC classes other than just unsubstituted PAHs that are important but currently not included in air quality guidelines or for assessing inhalation cancer risks., (Crown Copyright © 2019. Published by Elsevier Ltd. All rights reserved.)

Electrochemical oxidation (EO) coupled with peroxydisulfate (PDS) activation as a synergistic wastewater treatment process (PDS/EO) was performed to degrade anthraquinone dye-Reactive Brilliant Blue (RBB) in aqueous solution. Introducing PDS into the EO improved the RBB removal than the sole PDS and conventional EO systems. The RBB could activate PDS to a certain degree by itself. By the comparison of various inorganic ions addition, it showed that adding NO 3 - as the background electrolyte was more effective than the systems using the Cl - and SO 4 2- , respectively. In this PDS/EO-NO 3 - system, increasing PDS concentration (1-5 mmol L -1 ) and current density (5-10 mA cm -2 ) considerably promoted the degradation of RBB. The adjustment of the solution pH displayed that the acidic and neutral condition was beneficial to the RBB removal, and the synergistic effect was inverse ratio to the RBB initial concentration. Furthermore, the scavenger experiments verified that both SO 4 ·- and HO· were the major active substances in the RBB decomposition, and other reactive oxygen species also had considerable contributions. Thereinto NO 3 - only act a catalytic agent to improve the generation of active matters in the PDS/EO-NO 3 - . Overall, the proposed synergistic process could serve as an efficient method for the degradation of anthraquinone dye., (Copyright © 2019. Published by Elsevier Inc.)

Graphene and graphene oxide can promote the adhesion, growth and differentiation of mesenchymal stem cells. Further, graphene surface coatings accelerate the differentiation of human mesenchymal stem cells acting as osteogenic inducers. Quantification of the osteogenic induction is conventionally performed with Alizarin Red S (ARS), an anthraquinone derivative used to identify calcium deposits in tissue sections and cell cultures. The ARS staining is quite versatile because the dye forms an Alizarin Red S⁻calcium complex that can be extracted from the stained monolayer of cells and readily assayed by absorbance measurements. Direct visualization of stained deposits is also feasible; however, an in-situ visualization and quantification of deposits is possible only on transparent supports and not on thick opaque materials like ceramics and graphene composites that are well-known inducers of osteogenesis. In this manuscript, the shape of the 2D-fluorescence spectra of the ARS-calcium complex is used to develop a method to detect and monitor the in-situ differentiation process occurring during the osteogenic induction mediated by opaque graphene oxide surfaces.

Biodegradation of hazardous pollutants is of immense importance for maintaining a clean environment. However, the concentration of such contaminants/pollutants can be minimized with the help of microorganisms that has the ability to degrade the toxic pollutants into non-toxic metabolites. In the current study, 23 bacterial isolates were purified from the rhizospheric soil of Sysimbrium irio, growing as a wild plant in the vicinity of gas filling stations in Peshawar city. The isolated strains were initially screened on solid nutrient agar and further purified by culturing it on anthracene amended mineral media (PNR). The bacterial growth and anthracene disappearance were observed by calculating optical density (OD). The isolates showed a concentration-dependent growth on anthracene amended PNR media at 30°C and pH7. Also, an increase in bacterial OD from 0.351 to 1.80 with increased shaking speed was noticed. On the contrary, alternate carbon sources (glucose, fructose, sucrose) or nitrogen sources (KNO3, NaNO3, NH4NO3 and CaNO3) posed inhibitory effect on bacterial growth during anthracene degradation. The recorded efficiency of anthracene degradation by the selected bacterial isolate (1.4×1023 CFUmL-1 and 1.80 OD) was 82.29%, after 120 h of incubation. The anthracene was degraded to 9, 10, dihydroxy-anthracene and anthraquinone, detected through GC-MS. The efficient bacterial isolate was identified as S13, a new strain of Bacillus cereus, using 16S rRNA analysis, showing 98% homology. The isolated bacterial strain S13 may be used as a potential tool for bioremediation of toxic hydrocarbons and to keep the environment free from PAH pollutants., Competing Interests: The authors have declared that no competing interests exist.

Hedyotis diffuse Willd. (HD) and Hedyotis corymbosa (L.) Lam. (HC), two closely related species of the same genus, are both used for health benefits and disease prevention in China. HC is also indiscriminately sold as HD in the wholesale chain and food markets. This confusion has led to a growing concern about their identification and quality evaluation. In order to further understand the molecular diversification between them, we focus on the screening of chemical components and the analysis of non-targeted metabolites. In this study, UPLC-QTOF-MS E , UNIFI platform and multivariate statistical analyses were used to profile them. Firstly, a total of 113 compounds, including 80 shared chemical constituents of the two plants, were identified from HC and HD by using the UNIFI platform. Secondly, the differences between two herbs were highlighted with the comparative analysis. As a result, a total of 33 robust biomarkers enabling the differentiation were discovered by using multivariate statistical analyses. For HC, there were 18 potential biomarkers (either the contents were much greater than in HD or being detected only in HC) including three iridoids, eight flavonoids, two tannins, two ketones, one alcohol and two monoterpenes. For HD, there were15 potential biomarkers (either the contents were much greater than in HC or being detected only in HD) including two iridoids, eight flavonoids, one tannin, one ketone, and three anthraquinones. With a comprehensive consideration of the contents or the MS responses of the chemical composition, Hedycoryside A and B, detected only in HC, could be used for rapid identification of HC. The compounds 1,3-dihydroxy-2-methylanthraquinone and 2-hydroxy-3-methylanthraquinone, detected only in HD, could be used for rapid identification of that plant. The systematic comparison of similarities and differences between two confusing Chinese herbs will provide reliable characterization profiles to clarify the pharmacological fundamental substances. HC should not be used as the substitute of HD.

Background: The massive number of ingredients in traditional Chinese medicines (TCMs) makes quality control very difficult. The concept of quality markers (Q-marker) was recently proposed to evaluate the quality of TCMs. Xuezhiling tablets (XZL) are widely used for the treatment of hyperlipidemia in China owing to its noticeable effectiveness and mild adverse effects, but there are no proper Q-markers for this Chinese patent medicine., Purpose: The aim of the present study was to determine the Q-markers of XZL against hyperlipidemia through an integration of investigations on its lipid-lowering effect, metabolomics, content determination and pharmacokinetics., Methods: XZL was prepared in accordance with the method described in the Chinese pharmacopoeia (Ch.P.). Hyperlipidemia was induced in rats through the administration of a high-fat diet (HFD). The hypolipidemic effect of XZL was investigated through the detection of the blood levels of total glyceride (TG), total cholesterol (TC), and low density lipoprotein cholesterol (LDL-C). A metabolomics study was conducted to analyze the overall effects of XZL on the regulation of lipid metabolism. The main bioactive compounds of XZL were identified and determined in the XZL preparation and the medicated plasma of hyperlipidemic rats., Results: XZL lowered the levels of TG, TC, and LDL-C through alterations of metabolic patterns. 2,3,5,4'-Tetrahydroxystilbene-2-O-β-D-glucopyranoside (THSG), chrysophanol-1-O-β-glucopyranosyl-(1→3)-O-β-D-glucopyranosy1-(1→6)-O-β-D-glucopyranosyl-(1→6)-O-β-D-glucopyranoside (SHJ), cassiaside, rubrofusarin gentiobioside, aurantio-obtusin, chryso-obtusin, and obtusinfolin were identified and determined both in the preparation and the blood of hyperlipidemic rats., Conclusion: SHJ, obtusinfolin, THSG, rubrofusarin gentiobioside, and aurantio-obtusin, which are more abundant in the preparation, leading to greater exposure in vivo, were suitable Q-markers to guarantee the medicinal quality of XZL and ensure the clinical effectiveness on hyperlipidemia., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Here we propose a new protocol for whole-mount bone staining, which allows the rapid preparation of highly cleared and nondestructive specimens. It only takes 3 days to complete whole procedure for small vertebrates, such as medaka, zebrafish, and Xenopus frogs. In this procedure, we used a newly developed fixative containing formalin, Triton X-100, and potassium hydroxide, which allows the fixation, decolorization, and transparentization of specimens at the same time. A bone staining solution containing alizarin red S with ethylene glycol and a clearing solution containing Tween 20 and potassium hydroxide also contributed the specificity and swiftness of this new system. As expected, although details of the skeletal system could be observed in specimens with high transparency, it was noteworthy that high-resolution fluorescence images acquired using zoom microscopes clearly delineated the shape of each bone. This new procedure would be expected to be widely used as a standard procedure for bone staining in the testing the developmental toxicity of chemicals and in the screening test of knockout or mutant animals.

Morinda officinalis is an important herbal medicine and functional food, and its main constituents include anthraquinone and iridoid glycosides. Quantification of the main compounds is a necessary step to understand the quality and therapeutic properties of M. officinalis , but this has not yet been performed based on liquid chromatography/tandem mass spectrometry (LC-MS/MS). Analytes were extracted from M. officinalis by reflux method. Ultrahigh-performance liquid chromatography coupled with a triple quadrupole mass spectrometry (UPLC-QqQ-MS) using multiple reaction monitoring (MRM) mode was applied for quantification. Fragmentation pathways of deacetyl asperulosidic acid and rubiadin were investigated based on UPLC with quadrupole time-of-flight tandem mass spectrometry (Q/TOF-MS) in the MS E centroid mode. The method showed a good linearity over a wide concentration range (R² ≥ 0.9930). The limits of quantification of six compounds ranged from 2.6 to 27.57 ng/mL. The intra- and inter-day precisions of the investigated components exhibited an RSD within 4.5% with mean recovery rates of 95.32⁻99.86%. Contents of selected compounds in M. officinalis varied significantly depending on region. The fragmentation pathway of deacetyl asperulosidic and rubiadin was proposed. A selective and sensitive method was developed for determining six target compounds in M. officinalis by UPLC-MS/MS. Furthermore, the proposed method will be helpful for quality control and identification main compounds of M. officinalis .

Emissions from burning of biomass in the Amazon region have adverse effects on the environment and human health. Herein, particulate matter (PM) emitted from biomass burning in the Amazon region during two different periods, namely intense and moderate, was investigated. This study focused on: i) organic characterization of nitro- and oxy-polycyclic aromatic hydrocarbons (PAHs); ii) assessment of the excess lifetime cancer risk (LCR); and iii) assessment of the in vitro mutagenic effects of extractable organic matter (EOM). Further, we compared the sensitivity of two mutagenicity tests: Salmonella/microsome test and cytokinesis-block micronucleus (CBMN) with human lung cells. Among the nitro-PAHs, 2-nitrofluoranthene, 7-nitrobenz[a]anthracene, 1-nitropyrene, and 3-nitrofluoranthene showed the highest concentrations, while among oxy-PAHs, 2-metylanthraquinone, benz[a]anthracene-7,12-dione, and 9,10-anthraquinone were the most abundant. The LCR calculated for nitro-PAH exposure during intense biomass burning period showed a major contribution of 6-nitrochrysene to human carcinogenic risk. The EOM from intense period was more mutagenic than that from moderate period for both TA98 and YG1041 Salmonella strains. The number of revertants for YG1041 was 5-50% higher than that for TA98, and the most intense responses were obtained in the absence of metabolic activation, suggesting that nitroaromatic compounds with direct-acting frameshift mutagenic activity are contributing to the DNA damage. Treatment of cells with non-cytotoxic doses of EOM resulted in an increase in micronuclei frequencies. The minimal effective dose showed that Salmonella/microsome test was considerably more sensitive in comparison with CBMN mainly for the intense burning period samples. This was the first study to assess the mutagenicity of EOM associated with PM collected in the Amazon region using Salmonella/microsome test. The presence of compounds with mutagenic effects, particularly nitro- and oxy-PAHs, and LCR values in the range of 10 -5 indicate that the population is potentially exposed to an increased risk of DNA damage, mutation, and cancer., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The aim of this study was to select optimal conditions (C and N sources, initial pH and temperature) for biodecolorization of 0.03% anthraquinone dye Alizarin Blue Black B (ABBB) by microscopic fungi: Haematonectria haematococca BwIII43, K37 and Trichoderma harzianum BsIII33. The phenolic compounds, phytotoxicity (Lepidium sativum L.), biotoxicity (Microtox), cytotoxicity and yeast viability assay were performed to determine the extent of ABBB detoxification. Biodecolorization and detoxification of 0.03% ABBB in H. haematococca BwIII43 and T. harzianum BsIII33 cultures was correlated with extracellular oxidoreductases activity. In turn, secondary products, toxic to human fibroblasts and respiring sod1 Saccharomyces cerevisiae cells, were formed in H. haematococca K37 strain cultures, despite efficient decolorization., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Our previous study revealed that mesoporous Ca-Si-based materials exhibited excellent osteoconduction because dissolved ions could form a layer of hydroxycarbonate apatite on the surface of the materials. However, the biological mechanisms underlying bone regeneration were largely unknown. The main aim of this study was to evaluate the osteogenic ability of large-pore mesoporous Ca-Si-based bioceramics (LPMSCs) by alkaline phosphatase assay, real-time PCR analysis, von Kossa, and alizarin red assay. Compared with large-pore mesoporous silica (LPMS), LPMSCs had a better effect on the osteogenic differentiation of dental pulp cells. LPMSC-2 and LPMSC-3 with higher calcium possessed better osteogenic abilities than LPMSC-1, which may be related to the calcium-sensing receptor pathway. Furthermore, the loading capacity for recombinant human platelet-derived growth factor-BB was satisfactory in LPMSCs. In vivo, the areas of new bone formation in the calvarial defect repair were increased in the LPMSC-2 and LPMSC-3 groups compared with the LPMSC-1 and LPMS groups. We concluded that LPMSC-2 and LPMSC-3 possessed both excellent osteogenic abilities and satisfactory loading capacities, which may be attributed to their moderate Ca/Si molar ratio. Therefore, LPMSCs with moderate Ca/Si molar ratio might be potential alterative grafts for craniomaxillofacial bone regeneration., Competing Interests: Disclosure The authors report no conflicts of interest in this work.

Semen cassiae is the ripe seed of Cassia obtusifolia L. or Cassia tora L. of the family Leguminosae. In traditional Chinese medicine, the two forms of Semen cassiae are raw Semen cassiae (R-SC) and parched Semen cassiae (P-SC). To clarify the processing mechanism of Semen cassiae, the pharmacokinetics of R-SC and P-SC extracts were examined. A simple, rapid, sensitive ultra-high-performance liquid chromatography-tandem mass spectroscopy (UPLC-MS/MS) method was developed and validated for the simultaneous determination of seven anthraquinone aglycones of Semen cassiae (aurantio-obtusin, obtusifolin, questin, 2-hydroxyemodin-1-methyl-ether, rhein, emodin, 1,2,7-trimethoxyl-6,8-dihydroxy-3-methylanthraquinone) to compare the pharmacokinetics of raw and parched Semen cassiae in rat plasma. Compared with the R-SC group, C max and AUC 0-12 tended to be higher in the P-SC group. In particular, C max values for aurantio-obtusin, obtusifolin, questin, 2-hydroxyemodin-1-methyl-ether and rhein were significantly higher in the P-SC group ( p < 0.05). Meanwhile, T max and MRT 0-12 tended to be lower in the P-SC group. Specifically, T max for aurantio-obtusin and 2-hydroxyemodin-1-methyl-ether and MRT 0-12 for obtusifolin and rhein were significantly higher in the P-SC group ( p < 0.05)., Competing Interests: The authors declare no conflict of interest.

Background: In this study, Aloe vera samples were collected from different climatic regions of India. Quantitative HPTLC (high performance thin layer chromatography) analysis of important anthraquinones aloin and aloe-emodin and antiplasmodial activity of crude aqueous extracts was done to estimate the effects of these constituents on antiplasmodial potential of the plant., Methods: HPTLC system equipped with a sample applicator Linomat V with CAMAG sample syringe, twin rough plate development chamber (20 x 10 cm), TLC Scanner 3 and integration software WINCATS 1.4.8 was used for analysis of aloin and aloe-emodin amount. The antiplasmodial activity of plant extracts was assessed against a chloroquine (CQ) sensitive strain of P. falciparum (MRC-2). Minimum Inhibitory Concentration (MIC) of aqueous extracts of selected samples was determined according to the World Health Organization (WHO) recommended method that was based on assessing the inhibition of schizont maturation in a 96-well microtitre plate. EC (effective concentration) values of different samples were observed to predict antiplasmodial potential of the plant in terms of their climatic zones., Results: A maximum quantity of aloin and aloe-emodin i.e. 0.45 and 0.27 mg/g respectively was observed from the 12 samples of Aloe vera. The inhibited parasite growth with EC 50 values ranging from 0.289 to 1056 μg/ml. The antiplasmodial EC 50 value of positive control Chloroquine was observed 0.034 μg/ml and EC 50 values showed by aloin and aloe-emodin was 67 μg/ml and 22 μg/ml respectively. A positive correlation was reported between aloin and aloe-emodin. Antiplasmodial activity was increased with increase in the concentration of aloin and aloe-emodin. The quantity of aloin and aloe-emodin was decreased with rise in temperature hence it was negatively correlated with temperature., Conclusions: The extracts of Aloe vera collected from colder climatic regions showed good antiplasmodial activity and also showed the presence of higher amount of aloin and aloe-emodin in comparison to collected from warmer climatic sites. Study showed significant correlation between quantities of both the anthraquinones used as marker compounds and EC 50 values of the different Aloe vera extracts. Although, both the anthraquinones showed less antiplasmodial potential in comparison to crude extracts of different Aloe vera samples. Diverse climatic factors affect the quantity of tested compounds and antiplasmodial potential of the plant in different Aloe vera samples.

Objective: To develop a reliable and easy to use expression system for antibiotic production improvement of Streptomyces., Results: A two-compound T7 RNA polymerase-dependent gene expression system was developed to fulfill this demand. In this system, the T7 RNA polymerase coding sequence was optimized based on the codon usage of Streptomyces coelicolor. To evaluate the functionality of this system, we constructed an activator gene overexpression strain for enhancement of actinorhodin production. By overexpression of the positive regulator actII-ORF4 with this system, the maximum actinorhodin yield of engineered strain was 15-fold higher and the fermentation time was decreased by 48 h., Conclusion: The modified two-compound T7 expression system improves both antibiotic production and accelerates the fermentation process in Streptomyces. This provides a general and useful strategy for strain improvement of important antibiotic producing Streptomyces strains.

Artemisia capillaries Thunb, Gardenia jasminoides Ellis, and Rheum officinale Baill have been combined to treat jaundice for thousands of years. Studies have revealed that these herbs induce anti-hepatic fibrosis and anti-hepatic apoptosis and alleviate hepatic oxidative stress. This study aims to determine the quality and quantity of an herbal formulation (Chinese name: Yin-Chen-Hao-Tang) using physical and chemical examinations. Physical examination of Yin-Chen-Hao-Tang in pharmaceutical herbal products, raw fiber powders, and decoction preparations was performed using Congo red and iodine-potassium staining. A sensitive and validated method employing ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) was developed to simultaneously quantify the bioactive compounds scoparone, geniposide, and rhein in the Yin-Chen-Hao-Tang formulation in different preparations. Physical examination indicated that cellulose fibers with irregular round shapes were present in the pharmaceutical herbal products. The developed UHPLC-MS/MS method showed good linearity and was well validated. The quantification results revealed that the decoction preparations had the highest amounts of geniposide and rhein. Scoparone appeared in pharmaceutical herbal products from two manufacturers. This experiment provides a qualitative and quantitative method using physical and chemical examinations to test different preparations of herbal products. The results provide a reference for clinical herbal product preparations and further pharmacokinetic research.

Anthraquinones (AQs) and phenolic compounds are important phytochemicals that are biosynthesized in cell suspension cultures of Polygonum multiflorum . We wanted to optimize the effects of plant growth regulators (PGRs), media, sucrose, l-glutamine, jasmonic acid (JA), and salicylic acid (SA) for the production of phytochemicals and biomass accumulation in a cell suspension culture of P. multiflorum . The medium containing Murashige and Skoog (MS) salts and 4% sucrose supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid, 0.5 mg/L thidiazuron, and 100 µM l-glutamine at 28 days of cell suspension culture was suitable for biomass accumulation and AQ production. Maximum biomass accumulation (12.5 and 12.35 g fresh mass (FM); 3 and 2.93 g dry mass (DM)) and AQ production (emodin 295.20 and 282 mg/g DM; physcion 421.55 and 410.25 mg/g DM) were observed using 100 µM JA and SA, respectively. JA- and SA-elicited cell cultures showed several-fold higher biomass accumulation and AQ production than the control cell cultures. Furthermore, the cell suspension cultures effectively produced 23 phenolic compounds, such as flavonols and hydroxycinnamic and hydroxybenzoic acid derivatives. PGR-, JA-, and SA-elicited cell cultures produced a higher amount of AQs and phenolic compounds. Because of these metabolic changes, the antioxidant, antimicrobial, and anticancer activities were high in the PGR-, JA-, and SA-elicited cell cultures. The results showed that the elicitors (JA and SA) induced the enhancement of biomass accumulation and phytochemical (AQs and phenolic compounds) production as well as biological activities in the cell suspension cultures of P. multiflorum . This optimized protocol can be developed for large-scale biomass accumulation and production of phytochemicals (AQs and phenolic compounds) from cell suspension cultures, and the phytochemicals can be used for various biological activities., Competing Interests: The authors declare no conflict of interest.

Ingestion of soot present in soil or other environmental particles is expected to be an important route of exposure to nitro and oxygenated derivatives of polycyclic aromatic hydrocarbons (PAHs). We measured the apparent bioaccessibility (B app ) of native concentrations of 1-nitropyrene (1N-PYR), 9-fluorenone (9FLO), anthracene-9,10-dione (ATQ), benzo[a]anthracene-7,12-dione (BaAQ), and benzanthrone (BZO) in a composite fuel soot sample using a previously-developed in vitro human gastrointestinal model that includes silicone sheet as a third-phase absorptive sink. Along with B app , we determined the 24-h sheet-digestive fluid partition coefficient (K s,24h ), the soot residue-fluid distribution ratio of the labile sorbed fraction after digestion (K r,lab ), and the maximum possible (limiting) bioaccessibility, B lim . The B app of PAH derivatives was positively affected by the presence of the sheet due to mass-action removal of the sorbed compounds. In all cases B app increased with imposition of fed conditions. The enhancement of B app under fed conditions is due to increasingly favorable mass transfer of target compounds from soot to fluid (increasing bile acid concentration, or adding food lipids) or transfer from fluid to sheet (by raising small intestinal pH). Food lipids may also enhance B app by mobilizing contaminants from nonlabile to labile states of the soot. Compared to the parent PAH, the derivatives had larger K r,lab , despite having lower partition coefficients to various hydrophobic reference phases including silicone sheet. The B lim of the derivatives under the default conditions of the model ranged from 65.5% to 34.4%, in the order, 1N-PYR > ATQ > 9FLO > BZO > BaAQ, with no significant correlation with hydrophobic parameters, nor consistent relationship with B lim of the parent PAH. Consistent with earlier experiments on a wider range of PAHs, the results suggest that a major determinant of bioaccessibility is the distribution of chemical between nonlabile and labile states in the original solid., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

An anaerobic sludge (AS), capable of decolorizing a variety of synthetic dyes, was acclimated and is reported here. The sludge presented a much better dye decolorizing ability than that of different individual strains. A broad spectrum of dyes could be decolorized by the sludge. Continuous decolorization tests showed that the sludge exhibited the ability to decolorize repeated additions of dye. The chemical oxygen demand (COD) removal rate of the dye wastewater reached 52% after 12 h of incubation. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) profiles revealed that the microbial community changed as a result of varying initial concentrations of dyes. Phylogenetic analysis indicated that microbial populations in the sludge belonged to the phyla Acidobacteria, Firmicutes, Bacteroidetes, Chloroflexi and Proteobacteria. The degradation products of the three types of dye were identified. For azo dyes, the anaerobic sludge converted Methyl Orange to N , N -dimethylbenzene-1,4-diamine and 4-aminobenzenesulfonic acid; for triphenylmethane dyes, after Malachite Green was decolorized, the analyzed products were found to be a mixture of N , N -dimethylbenzenamine, 3-dimethyl-aminophenol and 4-dimethylaminobenzophenone; for anthraquinone dyes, two products (acetophenone and 2-methylbenzoic acid) were observed after Reactive Blue 19 decolorization. Together, these results suggest that the anaerobic sludge has promising potential for use in the treatment of industrial wastewater containing various types of dyes., Competing Interests: The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

This study investigated the degradation of a model organic compound, reactive blue (RB-19), in aqueous solution using a needle-plate non-thermal plasma (NTP) reactor, which was operated using three gas atmospheres (Ar, air, O2) at room temperature and atmospheric pressure. The relative discharge and degradation parameters, including the peak to peak applied voltage, power, ozone generation, pH, decolorization rates, energy density and the total organic carbon (TOC) reduction were analyzed to determine the various dye removal efficiencies. The decolorization rate for Ar, air and O2 were 59.9%, 49.6% and 89.8% respectively at the energy density of 100 kJ/L. The best TOC reduction was displayed by Ar with about 8.8% decrease, and 0% with O2 and air atmospheres. This phenomenon could be explained by the formation of OH• and O3 in the Ar and O2 atmospheres, which are responsible for increased mineralization and efficient decolorization. A one-dimension model was developed using software COMSOL to simulate the RB-19-ozone reaction and verify the experiments by comparing the simulated and experimental results. It was determined that ozone plays the most important role in the dye removal process, and the ozone contribution rate ranged from 0.67 to 0.82., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Non-reducing iterative type I polyketide synthase genes, pnk1 and pnk2, were cloned from the fungus Phoma sp. BAUA2861, which produces the topoisomerase I inhibitors, topopyrones A to D. Heterologous expression of these polyketide synthase genes under the α-amylase promoter in Aspergillus oryzae was carried out to identify their functions. The pnk2 transformant produced topopyrones C, D, and haematommone. Therefore, the pnk2 gene was found to encode for the topopyrone nonaketide synthase.

Mixed antihypertensive drug intoxication poses a significant risk for patient mortality. In tandem to antihypertensives, hypolipidemic medicines (especially statins) are often prescribed. Among their well-known adverse effects belongs rhabdomyolysis. We report a case of fatal multi-drug overdose in a 65-year-old female alcoholic. The patient was unconscious at admission. Empty blister packs indicated the abuse of 250 tablets of urapidil, 42 tablets of verapamil/trandolapril, 50 tablets of moxonidin, 80 tablets of atorvastatin and 80 tablets of diacerein. Standard measures (gastric lavage, activated charcoal, mechanical ventilation, massive doses of vasopressors, volume expansion, diuretics and alkalinization) failed to provide sufficient drug elimination and hemodynamic support and the sufferer deceased on the fourth day. Dramatic elevations of serum myoglobin (34,020 μg/L) and creatine kinase (219 μkat/L) were accompanied by rise in cardiac troponin I and creatinine. Gas chromatography revealed ethanol 1.17 g/kg (blood) and 2.81 g/kg (urine). Thin layer chromatography and gas chromatography of gastric content and urine verified verapamil, moxonidin and urapidil fragment (diacerein method was unavailable). Atorvastatin and trandolapril concentrations (LC-MS(n)) equaled 277.7 μg/L and 57.5 μg/L, resp. (serum) and 8.15 μg/L and 602.3 μg/L, resp. (urine). Histology confirmed precipitates of myoglobin with acute necrosis of proximal renal tubules in association with striated muscle rhabdomyolysis and myocardial dystrophy. Cardiogenic-distributive shock in conjunction with acute renal failure due to the combined self-poisoning with vasoactive agents and atorvastatin were determined to be this decedent's immediate cause of death. The manner of death was assigned to be suicidal., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Background: The use of herbal medicines as complements or alternatives to orthodox medicines has been on the increase. There has been the erroneous belief that these medicines are free from adverse effects. Schefflera barteri is popularly used in the West region of Cameroon for the treatment of various diseases such as diarrhea, spasm, pneumonia and animals bite. Considering the ethnopharmacological relevance of this plant, this study was designed to investigate the possible toxic effects of the stem bark extract of S. barteri., Methods: The extract was prepared by maceration of stem bark dry powder in methylene chloride/methanol mixture. Phytochemical analysis was performed by chemical reaction method. Oral acute toxicity study was carried out by administering single geometric increasing doses (2 to 16 g/kg body weight) of plant extract to Swiss albino mice. For sub-acute toxicity study, repeated doses (100, 200, 400 and 800 mg/kg bw) of plant extract were given to Wistar albino rats for 28 consecutive days by oral route. At the end of the treatment period, hematological and biochemical parameters were assessed, as well as histopathological studies., Results: Phytochemical analysis of stem bark extract of S. barteri revealed the presence of anthocyanins, anthraquinons and saponins. Acute toxicity results showed that the LD50 was greater than 16000 mg/kg. Sub-acute treatment significantly (P < 0.05) increased the level of serum transaminase, proteins and HDL cholesterol. On the other hand, the extract significantly (P < 0.05) reduced the level of leucocytes as well as neutrophils, basophils and monocytes in female. No significant variation of serum creatinine, LDL cholesterol, serum triglycerides as well as liver, spleen, testicles and ovaries proteins was noted. Histopathological analysis of organs showed vascular congestion, inflammation of peri-portal and vacuolization of hepatocytes at the level of the liver. Leucocytes infiltration of peri-portal veins were noticed on lungs and liver cells as well as inflammatory peri-bronchial and basal membranes seminar tube merely joined on lungs and testis respectively., Conclusion: The results suggest that acute administration of the stem bark extract of S. barteri is associated with signs of toxicity, administration over a long duration provokes hepatotoxicity, testes and lungs toxicities.

The objective of this study was to develop a novel analytical chemistry method, comprised of a coupled high-performance liquid chromatography-gas chromatography/mass spectrometry system (LC-GC/MS) with low detection limits and high selectivity, for the identification and determination of oxygenated polycyclic aromatic hydrocarbons (OPAHs) and polycyclic aromatic hydrocarbons (PAHs) in urban air and diesel particulate matter. The linear range of the four OPAHs, which include 9,10-anthraquinone, 4H-cyclopenta[def]phenanthrene-4-one, benzanthrone, and 7,12-benz[a]anthraquinone, was 0.7 pg-43.3 ng with limits of detection (LODs) and limits of quantification (LOQs) on the order of 0.2-0.8 and 0.7-1.3 pg, respectively. The LODs in this study are generally lower than values reported in the literature, which can be explained by using large-volume injection. The recoveries of the OPAHs spiked onto glass fiber filters using two different pressurized liquid extraction (PLE) methods were in the ranges of 84-107 and 67-110 %, respectively. The analytical protocols were validated using the following National Institute of Standards and Technology standard reference materials: SRM 1649a (Urban Dust), SRM 1650b (Diesel Particulate Matter), and SRM 2975 (Diesel Particulate Matter, Industrial Forklift). The measured mass fractions of the OPAHs in the standard reference materials (SRMs) in this present study are higher than the values from the literature, except for benzanthrone in SRM 1649a (Urban Dust). In addition to the OPAHs, 44 PAHs could be detected and quantified from the same particulate extract used in this protocol. Using data from the literature and applying a two-sided t test at the 5 % level using Bonferroni correction, significant differences were found between the tested PLE methods for individual PAHs. However, the measured mass fractions of the PAHs were comparable, similar to, or higher than those previously reported in the literature.

Lianhua-Qingwen capsule (LQC) is a commonly used Chinese medical preparation to treat viral influenza and especially played a very important role in the fight against severe acute respiratory syndrome (SARS) in 2002-2003 in China. In this paper, a rapid ultraperformance liquid chromatography coupled with diode-array detector and quadrupole time-of-flight mass spectrometry (UPLC-DAD-QTOF-MS) method was established for qualitative and quantitative analysis of the major constituents of LQC. A total of 61 compounds including flavonoids, phenylpropanoids, anthraquinones, triterpenoids, iridoids, and other types of compounds were unambiguously or tentatively identified by comparing the retention times and accurate mass measurement with reference compounds or literature data. Among them, twelve representative compounds were further quantified as chemical markers in quantitative analysis, including salidroside, chlorogenic acid, forsythoside E, cryptochlorogenic acid, amygdalin, sweroside, hyperin, rutin, forsythoside A, phillyrin, rhein, and glycyrrhizic acid. The UPLC-DAD method was evaluated with linearity, limit of detection (LOD), limit of quantification (LOQ), precision, stability, repeatability, and recovery tests. The results showed that the developed quantitative method was linear, sensitive, and precise for the quality control of LQC.

The present study concerns the identification of nine thus-far unknown derivatives of carminic acid extracted from pre-Columbian Peruvian textiles dyed with American cochineal-these derivatives are not found in commercially available preparations of the dye. These compounds probably represent a unique fingerprint of dyed textiles from this region, as they have never been reported to occur in other fabrics of historical value. They were separated by reversed-phase high-performance liquid chromatography (phenyl column) and detected using a UV/vis spectrophotometer and two tandem mass spectrometers. Peaks observed in chromatograms registered at 450 and 500 nm were further identified by ESI QqQ MS (mainly in the negative ion mode), supported by high-resolution ESI QIT/ToF MS data. The characteristic fragmentation pathways of isolated carminic acid and its derivatives provided additional information concerning lost neutrals and thus the functional groups and substituents present in the parent molecules. This information mainly related to multiple cleavages of the hexoside moiety (initially cross-ring cleavage), which are characteristic of C-glucosides (loss of 90, 120, and 148 Da). This is accompanied by the elimination of H2O as well as the further loss of 60 Da from the hexoside moiety. Moreover, other losses from the carbonyl groups (44 Da from CO2 loss, 62 Da from ethylene glycol loss, 32 Da from O2 loss, 138 Da from hydroxybenzoic acid, and 120 Da from oxomethylene cyclohexadienone) provided more specific information about structures of the identified derivatives of carminic acid.

Background: Polygonum hydropiper L decoctions are traditionally used in the treatment of various ailments including inflammation, dyspepsia, diarrhea, menorrhagia, hemorrhoids, helminthiasis and CNS disorders. Present study was undertaken to investigate P. hydropiper L. for heavy metals content, phytoconstituents, Phytotoxic and anthelmintic activities to explore its toxicological and pharmacological potentials and rationalize its ethnomedicinal uses., Methods: Plant crude powder, methanolic extract, fractions and soil samples were analyzed for heavy metals using atomic absorption spectrophotometer. Qualitative phytochemical analysis of the plant extracts was carried out for the existence of alkaloids, flavonoids, glycosides, anthraquinones, saponins, terpenoids, sterols and tannins. Radish seeds phytotoxicity assay was used to study phytotoxic action of plant extracts. Pheretima posthuma and Ascaridia galli were used to study anthelmintic potential of the plant using albendazole and levamisole HCl as standard drugs., Results: Plant crude powder, methanolic extract (Ph.Cr), its subsequent fractions; n-hexane (Ph.Hex), chloroform (Ph.Chf), ethyl acetate (Ph.EtAc), n-Butanol (Ph.Bt), aqueous (Ph.Aq), saponins (Ph.Sp) and soil samples were found to contain copper (Cu), iron (Fe), chromium (Cr), zinc (Zn), lead (Pb), nickel (Ni), cadmium (Cd) and lead (Pb) in different concentrations. In crude powder of the plant, heavy metals concentrations were within WHO specified limits, whereas different fractions and soil samples exhibited high metals content. Ph.Cr was tested positive for the presence of alkaloids, flavonoids, saponins, tannins, triterpenoids and anthraquinone glycosides. Among different fractions Ph.EtAc, Ph.Sp, Ph.Chf and Ph.Bt were most effective causing 89.32, 89.25, 86.68 and 85.32% inhibition of seeds in phytotoxicity assay, with IC50 values of 50, 60, 35 and 100 μg/ml respectively. In anthelmintic study, Ph.Sp, Ph.Chf, Ph.EtAc and Ph.Cr were most effective against P. posthuma at 10 mg/ml concentration with an average death time of 50, 64.67, 68.67 and 71 minutes respectively. Ph.EtAc, Ph.Chf and Ph.Aq were most effective against A. galli with average death time of 7, 9 and 10 min respectively at 1 mg/ml concentration., Conclusions: Our findings indicate that P. hydropiper contains different heavy metals and secondary metabolites. Different fractions exhibited phytotoxic and anthelmintic activites comparable to control drugs, thus provide pharmacological basis for ethnomedicinal uses of this plant.

Background: Isodon rugosus is used traditionally in the management of hypertension, rheumatism, tooth-ache and pyrexia. Present study was arranged to investigate I. rugosus for phytoconstituents, phytotoxic and cytotoxic activities to explore its toxicological, pharmacological potentials and to rationalize its ethnomedicinal uses. Briefly, qualitative phytochemical analysis of the plant extracts were carried out for the existence of alkaloids, flavonoids, saponins, oils, glycosides, anthraquinones, terpenoids, sterols and tannins. Plant crude methanolic extract (Ir.Cr), its subsequent fractions; n-hexane (Ir.Hex), chloroform (Ir.Chf), ethyl acetate (Ir.EtAc), aqueous (Ir.Aq) and saponins (Ir.Sp) in different concentrations were tested for phytotoxic and cytotoxic activities using radish seeds and brine shrimps (Artemia salina) respectively. The phytotoxic activity was determined by percent root length inhibition (RLI) and percent seeds germination inhibition (SGI) while the cytotoxicity was obtained with percent lethality of the brine shrimps., Results: Ir.Cr was tested positive for the presence of alkaloids, glycosides, flavonoids, oils, terpenoids, saponins, tannins and anthraquinones. Among different fractions Ir.Sp, Ir.Chf, Ir.EtAc, and Ir.Cr were most effective causing 93.55, 89.32, 81.32 and 58.68% inhibition of seeds in phytotoxicity assay, with IC50 values of 0.1, 0.1, 0.1 and 52 μg/ml respectively. Similarly, among all the tested samples, Ir.Sp exhibited the highest phytotoxic effect causing 91.33% root length inhibition with IC50 of 0.1 μg/ml. Ir.Sp and Ir.Chf were most effective against brine shrimps showing 92.23 and 76.67% lethality with LC50 values of 10 and 12 μg/ml respectively., Conclusions: It may be inferred from the current investigations that I. rugosus contains different secondary metabolites and is a potential source for the isolation of natural anticancer and herbicidal drug molecules. Different fractions exhibited phytotoxic and cytotoxic activities, thus providing pharmacological basis for ethnomedicinal uses of this plant.

A cDNA encoding for a laccase was isolated from the white-rot fungus Lenzites gibbosa by RT-PCR and expressed in the Pichia pastoris. The laccase native signal peptide efficiently directed the secretion of the recombinant laccase in an active form. Factors influencing laccase expression, such as pH, cultivation temperature, copper concentration and methanol concentration, were optimized. The recombinant enzyme was purified to electrophoretic homogeneity, and was estimated to have a MW of ~61.5 kDa. The purified enzyme behaved similarly to the native laccase produced by L. gibbosa and efficiently decolorized Alizarin Red, Neutral Red, Congo Red and Crystal Violet, without the addition of redox mediators. The decolorization capacity of this recombinant enzyme suggests that it could be a useful biocatalyst for the treatment of dye-containing effluents. This study is the first report on the synthetic dye decolorization by a recombinant L. gibbosa laccase.

Oxidative degradation of Alizarin Red S (ARS) in aqueous solutions by using electro-Fenton was studied. At first, effect of operating parameters such as current density, aeration rate and initial pH on the degradation of ARS were studied by using UV-vis spectrum, respectively. Then, under the optimal operating conditions (current density: 10.0mAcm(-2), aeration rate: 1000mLmin(-1), initial pH: 2.8), the identification of degradation products of ARS was carried out by using GC-MS and HPLC, meanwhile its degradation pathway was proposed according to the intermediates. Considering the location, intensity and intensity ratio of fluorescence center peak of the ARS in aqueous solution, a convenient and quick monitoring method by using excitation-emission matrix fluorescence spectrum technology was developed to monitor the degradation degree of ARS through electro-Fenton process. Furthermore, it is suggested that the developed method would be promising for the quick analysis and evaluation of the degradation degree of the pollutants with π-conjugated system., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) have recently received much attention in discussions regarding the negative impacts of particulate matter (PM) on human health and the environment. The National Institute of Standards and Technology provides several environmental matrix standard reference materials (SRMs) with certified and reference values for polycyclic aromatic hydrocarbons (PAHs) and nitrated PAHs. In this study, the concentrations of oxygenated PAHs are determined in three air PM SRMs (1649b, 1648a, and 2786) and three diesel PM SRMs (1650b, 2975, and 1975) using two independent gas chromatography-mass spectrometry methods. Concentrations of oxy-PAHs were at the milligrams per kilogram level with higher overall concentrations in diesel PM (up to 50 mg/kg for 9,10-anthraquinone). One of the highest oxy-PAH concentrations (up to 5 mg/kg) measured in the air particulate SRMs was for 7,12-benz[a]anthracenquinone. These results suggest that oxygenated PAHs should not be neglected in the analysis of PM as their concentrations can be as high as those of some PAHs and are one to two orders of magnitude higher than those for nitro-PAHs.

Acetylene black (AB) nanoparticles were readily dispersed into water in the presence of dihexadecyl hydrogen phosphate. After evaporation of water, the surface of glassy carbon electrode (GCE) was coated with AB nanoparticles as confirmed from the scanning electron microscopy measurements. The transmission electron microscopy images indicated that AB nanoparticles possessed porous structure. Electrochemical behavior of chrysophanol was studied, and a sensitive oxidation peak was observed in pH 3.6 acetate buffer solution. Compared with the bare GCE, the AB nanoparticles-modified GCE greatly increased the oxidation peak current of chrysophanol, showing remarkable signal enhancement effect. The influences of pH value, amount of AB, accumulation potential and time on the signal enhancement of chrysophanol were studied. As a result, a novel electrochemical method was developed for the determination of chrysophanol. The linear range was from 1.5 to 200 μgL(-1), and the detection limit was 0.51 μgL(-1) (2.01 × 10(-9)M) after 2-min accumulation. Finally, this method was used in traditional Chinese medicines, and the results consisted with the values that obtained by high-performance liquid chromatography., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Characterization of polyphenolic compounds in the stems of P. multiflorum was conducted using HPLC, high resolution LC-MS and LC-MSn. Proanthocyanidins in particular were isolated in 4.8% yield using solvent extraction followed by Sephadex LH-20 fractionation. HPLC analysis using a diol column revealed oligomers (from dimer to nonamer) as minor components, with (epi)catechin monomeric units predominating, and oligomers with higher degree of polymerization being dominant. Thiolysis treatment of the proanthocyanidins using mercaptoacetic acid produced thioether derivatives of (epi)catechin as the major product and a mean value of the degree of polymerization of 32.6 was estimated from the ratio of terminal and extension units of the (epi)catechin. The isolated proanthocyanidins were shown to strongly inhibit α-amylase with an acarbose equivalence (AE) value of 1,954.7 µmol AE/g and inhibit α-glucosidase with an AE value of 211.1 µmol AE/g.

Cassia fistula is well known for its laxative and antifungal properties due to anthraquinone compounds in the pods. This study quantitatively analyzed rhein in the C. fistula pod pulp decoction extracts kept under various storage conditions using HPLC. The antifungal activity of the extracts and their hydrolyzed mixture was also evaluated against dermatophytes. The contents of rhein in all stored decoction extracts remained more than 95% (95.69-100.66%) of the initial amount (0.0823 ± 0.001% w/w). There was no significant change of the extracts kept in glass vials and in aluminum foil bags. The decoction extract of C. fistula pod pulp and its hydrolyzed mixture containing anthraquinone aglycones were tested against clinical strains of dermatophytes by broth microdilution technique. The results revealed good chemical and antifungal stabilities against dermatophytes of C. fistula pod pulp decoction extracts stored under various accelerated and real time storage conditions.

Hollow fiber liquid phase microextraction (HFLPME) coupled with high performance liquid chromatography (HPLC) has been developed to analyze the emodin and its metabolites in plasma and urine samples. The abilities of the absorption and metabolism for the active components in traditional Chinese medicines between the male and female rats were compared, and the biological metabolism and transmutation of the analyte were detailed discussed. Emodin and its metabolites in plasma and urine samples were extracted into n-octanol (acceptor) in hollow fiber. The acceptor phase was dried and dissolved by 50 microL methanol and then analyzed by HPLC. Under the optimal conditions, the linearities of the analytes were all very good in biological samples (r > 0.9960), the detection limits of the analytes were within the ranges of 0.1-3.0 microg/L. The enrichment factors were 12.2 to 26.3. The relative standard deviations for intra-day and inter-day precision were lower than 11.0%. The average recoveries of the analytes in plasma and urine samples were all in the range of 97.9% to 103%. HFLPME-HPLC can eliminate interference from complex biological samples, improve the sensitivity and reduce the detection limit, thus this method is suitable for the determination of trace compounds in complex sample.

Background: Fumaria parviflora Linn. (Fumariaceae), is a small branched annual herb found in many parts of the world including Saudi Arabia and Pakistan. This study was designed to provide pharmacological basis for the medicinal use of Fumaria parviflora in gut motility disorders., Methods: The in-vivo prokinetic and laxative assays were conducted in mice. Isolated intestinal preparations (ileum and jejunum) from different animal species (mouse, guinea-pig and rabbit) were separately suspended in tissue baths containing Tyrode's solution bubbled with carbogen and maintained at 37°C. The spasmogenic responses were recorded using isotonic transducers coupled with PowerLab data acquisition system., Results: The aqueous-methanol extract of Fumaria parviflora (Fp.Cr), which tested positive for the presence of alkaloids, saponins, tannins and anthraquinones showed partially atropine-sensitive prokinetic and laxative activities in the in-vivo in mice at 30 and 100 mg/kg. In the in-vitro studies, Fp.Cr (0.01-1 mg/ml) caused a concentration-dependent atropine-sensitive stimulatory effect both in mouse tissues (jejunum and ileum), and rabbit jejunum but had no effect in rabbit ileum. In guinea-pig tissues (ileum and jejunum), the crude extract showed a concentration-dependent stimulatory effect with higher efficacy in ileum and the effect was partially blocked by atropine, indicating the involvement of more than one types of gut-stimulant components (atropine-sensitive and insensitive). This could be a plausible reason for the greater efficacy of Fp.Cr in gut preparations of guinea-pig than in rabbit or mouse., Conclusions: This study shows the prokinetic, laxative and spasmodic effects of the plant extract partially mediated through cholinergic pathways with species and tissue-selectivity, and provides a sound rationale for the medicinal use of Fumaria parviflora in gut motility disorders such as, indigestion and constipation. This study also suggests using different species to know better picture of pharmacological profile of the test material.