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GroEL, a chaperone protein responsible for peptide and denatured protein folding, undergoes substantial conformational changes driven by ATP binding and hydrolysis during folding. Utilizing these conformational changes, we demonstrated the GroEL-mediated regioselective photocyclodimerization of 2-anthracenecarboxylic acid (AC) using ATP hydrolysis as an external stimulus. We designed and prepared an optimal GroEL mutant to employ in a docking simulation that has been actively used in recent years. Based on the large difference in the motif of hydrogen bonds between AC and GroEL mutant compared with the wild-type, we predicted that GroEL MEL , in which the 307‒309th amino acid residues were mutated to Ala, could alter the orientation of bound AC in GroEL. The GroEL MEL -mediated photocyclodimerization of AC can be used for regioselective inversion upon ATP addition to a moderate extent., (Copyright © 2024 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The persistence and ubiquity of polycyclic aromatic hydrocarbons (PAHs) in the environment necessitate effective remediation strategies. Hence, this study investigated the potential of purified Laccases, TlFLU1L and TpFLU12L, from two indigenous fungi Trichoderma lixii FLU1 (TlFLU1) and Talaromyces pinophilus FLU12 (TpFLU12), respectively for the oxidation and detoxification of anthracene. Anthracene was degraded with v max values of 3.51 ± 0.06 mg/L/h and 3.44 ± 0.06 mg/L/h, and K m values of 173.2 ± 0.06 mg/L and 73.3 ± 0.07 mg/L by TlFLU1L and TpFLU12L, respectively. The addition of a mediator compound 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) to the reaction system significantly increased the degradation of anthracene, with up to a 2.9-fold increase in v max value and up to threefold decrease in K m values of TlFLU1L and TpFLU12L. The GC-MS analysis of the metabolites suggests that anthracene degradation follows one new pathway unique to the ABTS system-hydroxylation and carboxylation of C-1 and C-2 position of anthracene to form 3-hydroxy-2-naphthoic acid, before undergoing dioxygenation and side chain removal to form chromone which was later converted into benzoic acid and CO 2 . This pathway contrasts with the common dioxygenation route observed in the free Laccase system, which is observed in the second degradation pathways. Furthermore, toxicity tests using V. parahaemolyticus and HT-22 cells, respectively, demonstrated the non-toxic nature of Laccase-ABTS-mediated metabolites. Intriguingly, analysis of the expression level of Alzheimer's related genes in HT-22 cells exposed to degradation products revealed no induction of neurotoxicity unlike untreated cells. These findings propose a paradigm shift for bioremediation by highlighting the Laccase-ABTS system as a promising green technology due to its efficiency with the discovery of a potentially less harmful degradation pathway, and the production of non-toxic metabolites., (© 2024. The Author(s).)

Laccases (EC 1.10.3.2) are multicopper oxidases with the capability to oxidize diverse phenolic and non-phenolic substrates. While the molecular mechanism of their activity towards phenolic substrates is well-established, their reactivity towards non-phenolic substrates, such as polycyclic aromatic hydrocarbons (PAHs), remains unclear. To elucidate the oxidation mechanism of PAHs, particularly the activation mechanism of the sp 2 aromatic C-H bond, we conducted a density functional theory investigation on the oxidation of two PAHs (anthracene and benzo[ a ]pyrene) using an extensive model of the T1 copper catalytic site of the fungal laccase from Trametes versicolor .

Background: The search for new bioactive natural compounds with anticancer activity is still of great importance. Even though their potential for diagnostics and treatment of cancer has already been proved, the availability is still limited. Hypericin, a naphthodianthrone isolated essentially from plant source Hypericum perforatum L. along with other related anthraquinones and bisanthraquinones belongs to this group of compounds. Although it has been proven that hypericin is synthesized by the polyketide pathway in plants, none of the candidate genes coding for key enzymes has been experimentally validated yet. Despite the rare occurrence of anthraquinones in plants, their presence in microorganisms, including endophytic fungi, is quite common. Unlike plants, several biosynthetic genes grouped into clusters (BGCs) in fungal endophytes have already been characterized., Results: The aim of this work was to predict, identify and characterize the anthraquinone BGCs in de novo assembled and functionally annotated genomes of selected endophytic fungal isolates (Fusarium oxysporum, Plectosphaerella cucumerina, Scedosporium apiospermum, Diaporthe eres, Canariomyces subthermophilus) obtained from different tissues of Hypericum spp. The number of predicted type I polyketide synthase (PKS) BGCs in the studied genomes varied. The non-reducing type I PKS lacking thioesterase domain and adjacent discrete gene encoding protein with product release function were identified only in the genomes of C. subthermophilus and D. eres. A candidate bisanthraquinone BGC was predicted in C. subthermophilus genome and comprised genes coding the enzymes that catalyze formation of the basic anthraquinone skeleton (PKS, metallo-beta-lactamase, decarboxylase, anthrone oxygenase), putative dimerization enzyme (cytochrome P450 monooxygenase), other tailoring enzymes (oxidoreductase, dehydrogenase/reductase), and non-catalytic proteins (fungal transcription factor, transporter protein)., Conclusions: The results provide an insight into genetic background of anthraquinone biosynthesis in Hypericum-borne endophytes. The predicted bisanthraquinone gene cluster represents a basis for functional validation of the candidate biosynthetic genes in a simple eukaryotic system as a prospective biotechnological alternative for production of hypericin and related bioactive anthraquinones., (© 2024. The Author(s).)

The anthracene biodegradation potential of Serratia quinivorans HP5 was studied under a controlled laboratory environment. The green TiO 2 nanoparticles (NPs) synthesized from Paenibacillus sp. HD1PAH was used to accelerate the biodegradation process. The synergistic application of TiO 2 NPs and S. quinivorans HP5 resulted in a reduction of anthracene concentration by 1.2 folds in liquid-medium and 1.5 folds in contaminated soil. Gas-chromatography and mass-spectrometric investigation showed the production of four anthracene derivatives, namely 1,2-anthracene dihydrodiol, 6,7-benzocoumarin, anthrone, and 9,10-anthraquinoneat the termination of experimental periods. Furthermore, bacterial biomass increased by 23.3 folds in the presence of TiO 2 NPs, and overall soil enzyme activities were enhanced by 4.2 folds in the treated samples. In addition, there was a negative correlation observed between the biomass of S. quinivorans HP5 and the concentrations of anthracene, suggesting the involvement of bacterium in anthracene biodegradation processes. The degradation pathway of anthracene revealed its transformation into the less toxic compound 9,10-anthraquinone. Overall, this study elucidates a novel biodegradation pathway for anthracene and highlights the potential of nano-assisted bacterial remediation as a promising approach for environmental cleanup., (© 2024 Wiley‐VCH GmbH.)

The environment in India is contaminated with polycyclic aromatic hydrocarbons (PAHs) due to the occurrence of large anthropogenic activities, i.e., fuel combustion, mineral roasting, and biomass burning. Hence, 13 toxic PAHs were detected: phenanthrene, anthracene, fluoranthene, pyrene, and benz(a) anthracene, ben-zo; (b) fluoranthene, benzo(k) fluoranthene, benzo(a) pyrene, benzo(ghi)perylene, dibenz (ah) anthracene, indeno1,2,3-(cd) pyrene, coronene and coronene in the environment (i.e., ambient particulate matter, road dust, sludge, and sewage) of the most industrialized area. Pollutants such as heavy metals and polycyclic aromatic hydrocarbons co-contaminate the soil and pose a significant hazard to the ecosystem because these pollutants are harmful to both humans and the environment. Phytoremediation is an economical plant-based natural approach for soil clean-up that has no negative impact on ecosystems. The aim of this study was to investigate the effects of pyrene (500 mg kg -1 ), Zn (150 mg kg -1 ), Pb (150 mg kg -1 ), and Cd (150 mg kg -1 ) alone and in combination on the phytoextraction efficiency of Medicago sativa growing in contaminated soil. Plant biomass, biochemical activities, translocation factors, accumulation of heavy metals, and pyrene removal were determined. After 60 days of planting, compared with those of the control plants, the growth parameters, biomass, and chlorophyll content of the M. sativa plants were significantly lower, and the reactive oxygen species activity, such as proline and polyphenol content and metallothionein protein content, was markedly greater in the pyrene and heavy metal-polluted soils. Furthermore, the combined toxicity of pyrene and all three metals on M. sativa growth and biochemical parameters was significantly greater than that of pyrene, Zn, Pb, or Cd alone, indicating the synergistic effect of pyrene and heavy metals on cytotoxicity. Pyrene stress increased Cd accumulation in M. sativa. After pyrene exposure alone or in combination with Zn-pyrene, a greater pyrene removal rate (85.5-81.44%) was observed than that in Pb-pyrene, Cd-pyrene, and Zn-Pb-Cd-pyrene polluted soils (62.78-71.27%), indicating that zinc can enhance the removal of pyrene from contaminated soil. The resulting hypotheses demonstrated that Medicago sativa can be used as a promising phytoremediation agent for co-contaminated soil., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Poly aromatic hydrocarbons (PAHs) are considered as hazardous compounds which causes serious threat to the environment dua to their more carcinogenic and mutagenic impacts. In this study, Pseudomonas aeruginosa PP4 strain and synthesized iron nanoparticles were used to evaluate the biodegradation efficiency (BE %) of residual anthracene. The BE (%) of mixed degradation system (Anthracene + PP4+ FeNPs) was obtained about 67 %. The FTIR spectra result revealed the presence of functional groups (C-H, -CH 3 , CC, =C-H) in the residual anthracene. The FESEM and TEM techniques were used to determine the surface analysis of the synthesized FeNPs and the average size was observed by TEM around 5-50 nm. The crystalline nature of the synthesized iron nanoparticles was confirmed by the observed different respective peaks of XRD pattern. The various functional constituents (OH, C-H, amide I, CH 3 ) were identified in the synthesized iron nanoparticles by FTIR spectrum. In conclusion, this integrated nano-bioremediation approach could be an promising and effective way for many environmental fields like cleanup of hydrocarbon rich environment., 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 © 2023 Elsevier Inc. All rights reserved.)

The northeastern Pacific (NEP) Ocean spans the coast of British Columbia (Canada) and is impacted by anthropogenic activities including oil pipeline developments, maritime fossil fuel tanker traffic, industrial chemical effluents, agricultural and urban emissions in tandem with stormwater and wastewater discharges, and forest wildfires. Such events may expose surrounding marine environments to toxic polycyclic aromatic hydrocarbons (PAHs) and impact critical habitats of threatened killer whales (Orcinus orca). We analyzed skeletal muscle and liver samples from stranded Bigg's killer whales and endangered Southern Resident killer whales (SRKWs) for PAH contamination using LRMS. C3-phenanthrenes/anthracenes (mean: 632 ng/g lw), C4-dibenzothiophenes (mean: 334 ng/g lw), and C4-phenanthrenes/anthracenes (mean: 248 ng/g lw) presented the highest concentrations across all tissue samples. Diagnostic ratios indicated petrogenic-sourced contamination for SRKWs and pyrogenic-sourced burdens for Bigg's killer whales; differences between ecotypes may be attributed to habitat range, prey selection, and metabolism. A mother-fetus skeletal muscle pair provided evidence of PAH maternal transfer; low molecular weight compounds C3-fluorenes, dibenzothiophene, and naphthalene showed efficient and preferential exposure to the fetus. This indicates in-utero exposure of PAH-contamination to the fetus. Our results show that hydrocarbon-related anthropogenic activities are negatively impacting these top predators; preliminary data found here can be used to improve oil spill and other PAH pollution management and regulation efforts, and inform policy to conserve killer whale habitats in the NEP., (© 2023. The Author(s).)

Biosurfactants are surface active molecules generated by various microorganisms, including bacteria, actinobacteria, algae, and fungi. In this study, bacterial strains are isolated from soil contaminated with used motor oil and examined for potential biosurfactant production. A minimum salt medium (MSM), with crude oil as the only carbon source, is used to isolate potential biosurfactant-producing bacterial strains. About 23 strains are isolated, and all are subjected to the primary screening methods for biosurfactant production. Based on the emulsification index, oil displacement, and drop collapse screening methods, two isolates with potential biosurfactant-producing ability are selected for further studies. The synthesis of biosurfactants, crude oil and anthracene biodegradation is carried out with strains DTS1 and DTS2. Both strains show significant outcomes in crude oil degradation. In addition, both strains can utilize anthracene as the sole carbon source. During the degradation course, changes in the growth conditions are continuously monitored by measuring turbidity and pH. In this degradation study, the biosurfactant production aptitude of the isolated strains plays an essential role in increasing the bioavailability of hydrophobic hydrocarbons. These strains are identified down to the molecular level by employing 16S rRNA gene sequencing, and the acquired sequences are submitted to get the accession numbers. These prospective strains can be utilized to remediate hydrocarbon-contaminated environments., 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 © 2023 Elsevier Ltd. All rights reserved.)

Mammary cancer incidence varies greatly across species and underlying mechanisms remain elusive. We previously showed that mammosphere-derived epithelial cells from species with low mammary cancer incidence, such as horses, respond to carcinogen 7, 12-Dimethylbenz(a)anthracene-induced DNA damage by undergoing apoptosis, a postulated anti-cancer mechanism. Additionally, we found that miR-214-3p expression in mammosphere-derived epithelial cells is lower in mammary cancer-resistant as compared to mammary cancer-susceptible species. Here we show that increasing miR-214 expression and decreasing expression of its target gene nuclear factor kappa B subunit 1 in mammosphere-derived epithelial cells from horses abolishes 7,12-Dimethylbenz(a)anthracene-induced apoptosis. A direct interaction of miR-214-3p with another target gene, unc-5 netrin receptor A, is also demonstrated. We propose that relatively low levels of miR-214 in mammosphere-derived epithelial cells from mammals with low mammary cancer incidence, allow for constitutive gene nuclear factor kappa B subunit 1 expression and apoptosis in response to 7, 12-Dimethylbenz(a)anthracene. Better understanding of the mechanisms regulating cellular responses to carcinogens improves our overall understanding of mammary cancer resistance mechanisms., (© 2023. Springer Nature Limited.)

The size of microplastics (MPs) plays an important role in combined toxic effects including synergistic or antagonistic effects. However, the influence of the size of MPs on the combined toxicity of contaminants remains unclear. In this study, we employed a zebrafish model to investigate the effects of MP size on the combined toxicity of benz[a]anthracene (BaA), a representative polyaromatic hydrocarbon, using three different sizes of polystyrene MPs (PSMPs) (0.2, 1.0, and 10 μm). Treatment of all groups did not result in any mortality of the zebrafish larvae. However, small-sized PSMPs (0.2 μm) enhanced the toxic effect of BaA in larvae such as cardiac defect and disruption of vessel formation. Medium-sized PSMPs (1.0 μm) were boundary in terms of the combined toxic effect; however, large-sized PSMPs (10 μm) alleviated the cardiotoxicity of BaA, including cardiac defect, ROS levels, and cell death. The combined effects showed a correlation with the body burden of MPs and BaA in larvae according to particle size (in the order of 0.2 μm > 1.0 μm > 10 μm). The synergistic effects occurred likely because the small PSMPs facilitated the body burden of BaA, induced excessive ROS by Ahr-mediated activity, and caused cell death in the heart, resulting in increased heart defects in the larvae. In contrast, large PSMPs abated the combined toxic effect through decreased body burden, whereas medium PSMPs form a boundary in combined effects. Therefore, the combined toxic effects of MPs are dependent on their size, which plays an important role in the transport and accumulation of environmental pollutants., 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 © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The present study evaluated the aryl hydrocarbon receptor (AhR), estrogen receptor-α (ER-α), and retinoic acid receptor (RAR) mediated activities of nine 4- and 5-ring unsubstituted and monomethylated polycyclic aromatic hydrocarbons (PAHs) using a series of Chemical-Activated LUciferase gene eXpression (CALUX) assays. The potential role of these aforementioned receptors in relation to the developmental toxicity of these PAHs was further assessed in the zebrafish embryotoxicity test (ZET). The results show that all nine tested PAHs were AhR agonists, benz[a]anthracene (BaA) and 8-methyl-benz[a]anthracene (8-MeBaA) were ER-α agonists, and none of the tested PAHs induced ER-α antagonistic or RAR (ant)agonistic activities. In the AhR CALUX assay, all the methylated PAHs showed higher potency (lower EC50) in activating the AhR than their respective unsubstituted PAHs, implying that the addition of a methyl substituent on the aromatic ring of PAHs could enhance their AhR-mediated activities. Co-exposure of zebrafish embryos with each individual PAH and an AhR antagonist (CH223191) counteracted the observed developmental retardations and embryo lethality to a certain extent, except for 8-methyl-benzo[a]pyrene (8-MeBaP). Co-exposure of zebrafish embryos with either of the two estrogenic PAHs (i.e., BaA and 8-MeBaA) and an ER-α antagonist (fulvestrant) neutralized embryo lethality induced by 50 μM BaA and the developmental retardations induced by 15 μM 8-MeBaA. Altogether, our findings suggest that the observed developmental retardations in zebrafish embryos by the PAH tested may partially be AhR- and/or ER-α-mediated, whereas the RAR seems not to be relevant for the PAH-induced developmental toxicity in the ZET., (© 2022 The Authors. Journal of Applied Toxicology published by John Wiley & Sons Ltd.)

Anthropogenic activities have been increasing Polycyclic Aromatic Hydrocarbons (PAHs) release, promoting an urgent need for decontamination methods. Therefore, anthracene biodegradation by endophytic, extremophilic, and entomophilic fungi was studied. Moreover, a salting-out extraction methodology with the renewable solvent ethanol and the innocuous salt K 2 HPO 4  was employed. Nine of the ten employed strains biodegraded anthracene in liquid medium (19-56% biodegradation) after 14 days at 30 °C, 130 rpm, and 100 mg L -1 . The most efficient strain Didymellaceae sp. LaBioMMi 155, an entomophilic strain, was employed for optimized biodegradation, aiming at a better understanding of how factors like pollutant initial concentration, pH, and temperature affected this process. Biodegradation reached 90 ± 11% at 22 °C, pH 9.0, and 50 mg L -1 . Futhermore, 8 different PAHs were biodegraded and metabolites were identified. Then, experiments with anthracene in soil ex situ were performed and bioaugmentation with Didymellaceae sp. LaBioMMi 155 presented better results than natural attenuation by the native microbiome and biostimulation by the addition of liquid nutrient medium into soil. Therefore, an expanded knowledge about PAHs biodegradation processes was achieved with emphasis to the action of Didymellaceae sp. LaBioMMi 155, which can be further employed for in situ biodegradation (after strain security test), or for enzyme identification and isolation aiming at oxygenases with optimal activity under alkaline conditions., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Gap junctional intercellular communication (GJIC) is composed of connexin (Cx) and plays an important role in maintaining intracellular homeostasis. Loss of GJIC is involved in the early stages of cancer pathways of non-genotoxic carcinogens; however, the effect of genotoxic carcinogens, including polycyclic aromatic hydrocarbons (PAHs), on GJIC function remains unclear. Therefore, we determined whether and how a representative PAH 7,12-dimethylbenz[a]anthracene (DMBA) suppresses GJIC in WB-F344 cells. First, DMBA significantly inhibited GJIC and dose-dependently reduced Cx43 protein and mRNA expression. In contrast, Cx43 promoter activity was upregulated after DMBA treatment via the induction of specificity protein 1 and hepatocyte nuclear factor 3β, indicating that the promoter-independent loss of Cx43 mRNA can be associated with the inhibition of mRNA stability, which was verified by actinomycin D assay. In addition to a decrease in mRNA stability involved in human antigen R, we also observed DMBA-induced acceleration of Cx43 protein degradation, which was closely related to the loss of GJIC through Cx43 phosphorylation via MAPK activation. In conclusion, the genotoxic carcinogen DMBA suppresses GJIC by inhibiting post-transcriptional and post-translational processing of Cx43. Our findings suggest that the GJIC assay is an efficient short-term screening test for predicting the carcinogenic potential of genotoxic carcinogens., 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 © 2023 Elsevier Ltd. All rights reserved.)

The ability to degrade aromatic hydrocarbons, including (i) benzene, toluene, o- xylene, naphthalene, anthracene, phenanthrene, benzo[a]anthracene, and benzo[a]pyrene; (ii) polar substituted derivatives of benzene, including phenol and aniline; (iii) N-heterocyclic compounds, including pyridine; 2-, 3-, and 4-picolines; 2- and 6-lutidine; 2- and 4-hydroxypyridines; (iv) derivatives of aromatic acids, including coumarin, of 133 Rhodococcus strains from the Regional Specialized Collection of Alkanotrophic Microorganisms was demonstrated. The minimal inhibitory concentrations of these aromatic compounds for Rhodococcus varied in a wide range from 0.2 up to 50.0 mM. o- Xylene and polycyclic aromatic hydrocarbons (PAHs) were the less-toxic and preferred aromatic growth substrates. Rhodococcus bacteria introduced into the PAH-contaminated model soil resulted in a 43% removal of PAHs at an initial concentration 1 g/kg within 213 days, which was three times higher than that in the control soil. As a result of the analysis of biodegradation genes, metabolic pathways for aromatic hydrocarbons, phenol, and nitrogen-containing aromatic compounds in Rhodococcus , proceeding through the formation of catechol as a key metabolite with its following ortho-cleavage or via the hydrogenation of aromatic rings, were verified.

Various mutagenic, carcinogenic pollutants such as Polycyclic Aromatic Hydrocarbons (PAHs) are released into the environment posing a negative effect on plant metabolism. All the pollutants that are emitted into the atmosphere, ultimately find their way into the plant. Soil salinity stress is one of the major determinants of crop productivity. Different plants respond differently to different abiotic stress present alone or in combination. One such combination of abiotic stress is PAHs and salinity stress. The present research aims to study the effect of the application of NaCl and Anthracene alone and in various combinations on two chickpea genotypes GPF2 and PDG4. A 21 days laboratory experiment was conducted in petriplates and growth pouches. Different concentrations of NaCl and Anthracene were given to two chickpea genotypes viz. GPF2 and PDG4, alone as well as in combinations to study morphological, physiological and antioxidant responses. Results obtained were further analyzed by using various statistical measures such as Principle Component Analysis and Two-way ANOVA. Results indicated that under the dual presence of NaCl and Anthracene, GPF2 exhibited higher activities of antioxidant enzymes and was shown to have a negative correlation with plant height and chlorophyll content. Based on the results of the present investigation, it was concluded that GPF2 was a better performing chickpea genotype towards the combined presence of Anthracene and NaCl as compared to PDG4., 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 © 2022 Elsevier Ltd. All rights reserved.)

Environmentally persistent free radicals (EPFRs) as intermediate products exist widely in the PAHs-contaminated soils, but toxicity assessment associated with EPFRs for terrestrial invertebrates remains unclear. Using the model organism Eisenia fetida, we compared the adverse effects among anthracene (ANT), anthraquinone (ANQ), and EPFRs induced by ANT transformation on clay surfaces. Our results showed that EPFRs-exposed earthworms experienced histopathological damage, which was more severe than ANT and ANQ-exposed earthworms. The source of EPFRs damage was associated with the obvious dysbiosis of reactive oxygen species in earthworms. Specifically, EPFRs trigged more severe antioxidant responses and oxidative damages (e.g., membrane lipid and DNA injury) in comparison with ANT and ANQ exposure, as evidenced by the values of integrated biomarker response (IBR) following the order of EPFRs (14.5) > ANT (12.8) > ANQ (10.9). Moreover, high-throughput sequencing found that EPFRs induced dramatic changes in the composition and structure of earthworm gut microbiota, which may involve immune and metabolism dysfunction, in turn aggravated EPFRs toxicity. Overall, the obtained information highlights the more severe injury of EPFRs to terrestrial organisms, deserving more attentions for the assessment of potential risks associated with radical intermediates in PAHs-contaminated soils., 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 © 2022 Elsevier B.V. All rights reserved.)

The impacts of crude oil contamination on soil microbial populations were explored in seven different polluted areas near oil and gas drilling sites and refineries of Assam, India. Using high-throughput sequencing techniques, the functional genes and metabolic pathways involved in the bioconversion of crude oil contaminants by the indigenous microbial community were explored. Total petroleum hydrocarbon (TPH) concentrations in soil samples ranged from 1109.47 to 75,725.33 mg/kg, while total polyaromatic hydrocarbon (PAH) concentrations ranged from 0.780 to 560.05 mg/kg. Pyrene, benzo[a]anthracene, naphthalene, phenanthrene, and anthracene had greater quantities than the maximum permitted limits, suggesting a greater ecological risk, in comparison to other polyaromatic hydrocarbons. According to the metagenomic data analysis, the bacterial phyla Proteobacteria, Actinobacteria, Acidobacteria, and Bacteroides were the most prevalent among all polluted areas. The most prominent hydrocarbon degraders in the contaminated sites included Burkholderia, Mycobacterium, Polaromonas, and Pseudomonas. However, the kinds of pollutants and their concentrations did not correlate with the abundances of respective degrading genes for all polluted locations, as some of the sites with little to low PAH contamination had significant abundances of corresponding functional genes for degradation. Thus, the findings of this study imply that the microbiome of hydrocarbon-contaminated areas, which are biologically involved in the degradation process, has various genes, operons and catabolic pathways that are independent of the presence of a specific kind of contaminant., 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 © 2022 Elsevier Inc. All rights reserved.)

Polycyclic aromatic hydrocarbons (PAHs) are key organic pollutants in the environment that pose threats to the ecosystem and human health. The degradation of high molecular weight (HMW) PAHs by enriched bacterial consortia has been previously studied, while the involved metabolisms and microbial communities are still unclear and warrant further investigations. In this study, five bacterial consortia capable of utilizing different PAHs (naphthalene, anthracene, and pyrene) as the sole carbon and energy sources were enriched from PAH-contaminated soil samples. Among the five consortia, consortium TC exhibited the highest pyrene degradation efficiency (91%) after 19 d of incubation. The degradation efficiency was further enhanced up to 99% by supplementing yeast extract. Besides, consortium TC showed tolerances to high concentrations of pyrene (up to 1000 mg/L) and different heavy metal stresses (including Zn 2+ , Cd 2+ , and Pb 2+ ). The dominant genus in consortium TC, GS, and PL showing relatively higher degradation efficiency for anthracene and pyrene was Pseudomonas , whereas consortium PG and GD were predominated by genus Achromobacter and class Enterobacteriaceae, respectively. Consortium TC, as a highly efficient HMW PAH-degrading consortium, could be applied for synergistic biodegradation of HMW PAHs and in situ bioremediation of the sites contaminated with both PAHs and heavy metals.

Polycyclic aromatic hydrocarbons (PAHs) are structurally diverse organic chemicals that can have adverse effects on the health of fishes through activation of aryl hydrocarbon receptor 2 (AhR2). They are ubiquitous in the environment, but alkyl PAHs are more abundant in some environmental matrices. However, relatively little is known regarding the effects of alkylation on the toxicity of PAHs to fishes in vivo and how this relates to potency for activation of AhR2 in vitro. Therefore, the objectives of the present study were to determine the toxicity of benz[a]anthracene and three alkylated homologs representing various alkylation positions to early life stages of zebrafish (Danio rerio) and to assess the potency of each for activation of the zebrafish AhR2 in a standardized in vitro AhR transactivation assay. Exposure of embryos to each of the PAHs caused a dose-dependent increase in mortality and malformations characteristic of AhR2 activation. Each alkyl homolog had in vivo toxicities and in vitro AhR2 activation potencies different from those of the parent PAH in a position-dependent manner. However, there was no statistically significant linear relationship between responses measured in these assays. The results suggest a need for further investigation into the effect of alkylation on the toxicity of PAHs to fishes and greater consideration of the contribution of alkylated homologs in ecological risk assessments. Environ Toxicol Chem 2022;41:1993-2002. © 2022 SETAC., (© 2022 SETAC.)

We investigated the use of POXA1b laccase from Pleurotus ostreatus for the oxidation of anthracene into anthraquinone. We show that different pathways can occur depending on the nature of the redox mediator combined to laccase, leading to different structural isomers. The laccase combined with 2,2'-azine-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) leads to the formation of 1,4-anthraquinone and/or 1,2-anthraquinone. The unprecedented role of carbon nanotubes (CNTs) as redox mediators for oxidation of anthracene into 9,10-anthraquinone is shown and corroborated by density-functional theory (DFT) calculations. Owing to the efficient adsorption of anthraquinones at CNT electrodes, anthracene can be detected with low limit-of-detection using either laccase in solution, CNT-supported laccase or laccase immobilized at magnetic beads exploiting the adhesive property of a chimeric hydrophobin-laccase.

Medicinal phytochemicals, such as artemisinin and taxol, have impacted the world, and hypericin might do so if its availability issue could be addressed. Hypericin is the hallmark component of Saint John's wort (Hypericum perforatum L.), an approved depression alleviator documented in the US, European, and British pharmacopoeias with its additional effectiveness against diverse cancers and viruses. However, the academia-to-industry transition of hypericin remain hampered by its low in planta abundance, unfeasible bulk chemical synthesis, and unclear biosynthetic mechanism. Here, we present a strategy consisting of the hypericin-structure-centered modification and reorganization of microbial biosynthetic steps in the repurposed cells that have been tamed to enable the designed consecutive reactions to afford hypericin (43.1 mg L -1 ), without acquiring its biosynthetic knowledge in native plants. The study provides a synthetic biology route to hypericin and establishes a platform for biosustainable access to medicinal phytochemicals., (© 2021 Wiley-VCH GmbH.)

The standalone metallo-β-lactamase-type thioesterase (MβL-TE), belongs to the group V nonreducing polyketide synthase agene cluster, catalyzes the rate-limiting step of product releasing. Our work first investigated on the orthologous MβL-TEs from different origins to determine which nonconserved amino acid residues are important to the hydrolysis efficiency. A series of chimeric MβL-TEs were constructed by fragment swapping and site-directed mutagenesis, in vivo enzymatic assay showed that two nonconserved residues A19 and E75 (numbering in HyTE) were critical to the catalytic performance. Protein structure modeling suggested that these two residues are located in different areas of HyTE. A19 is on the entrance to the active sites, whereas E75 resides in the linker between the two β strands which hold the metal-binding sites. Combining with computational simulations and comparative enzymatic assay, different screening criteria were set up for selecting the variants on the two noncatalytic and nonconserved key residues to improve the catalytic activity. The rational design on A19 and E75 gave five candidates in total, two (A19F and E75Q) of which were thus found significantly improved the enzymatic performance of HyTE. The double-point mutant was constructed to further improve the activity, which was increased by 28.4-fold on product accumulation comparing to the wild-type HyTE. This study provides a novel approach for engineering on nonconserved residues to optimize enzymatic performance., (© 2021 Wiley Periodicals LLC.)

Every petroleum-processing plant produces sewage sludge containing several types of polycyclic aromatic hydrocarbons (PAHs). The degradation of PAHs via physical, biological, and chemical methods is not yet efficient. Among biological methods, the use of marine sponge symbiont bacteria is considered an alternative and promising approach in the degradation of and reduction in PAHs. This study aimed to explore the potential performance of a consortium of sponge symbiont bacteria in degrading anthracene and pyrene. Three bacterial species ( Bacillus pumilus strain GLB197, Pseudomonas stutzeri strain SLG510A3-8, and Acinetobacter calcoaceticus strain SLCDA 976) were mixed to form the consortium. The interaction between the bacterial consortium suspension and PAH components was measured at 5 day intervals for 25 days. The biodegradation performance of bacteria on PAH samples was determined on the basis of five biodegradation parameters. The analysis results showed a decrease in the concentration of anthracene (21.89%) and pyrene (7.71%), equivalent to a ratio of 3:1, followed by a decrease in the abundance of anthracene (60.30%) and pyrene (27.52%), equivalent to a ratio of 2:1. The level of pyrene degradation was lower than that of the anthracene due to fact that pyrene is more toxic and has a more stable molecular structure, which hinders its metabolism by bacterial cells. The products from the biodegradation of the two PAHs are alcohols, aldehydes, carboxylic acids, and a small proportion of aromatic hydrocarbon components.

Hypericin (Hy) is a hydrophobic photosensitizer used in photodynamic therapy for cancer therapeutic. In this study, Hy-loaded oil-in-water (O/W) nanoemulsions (NEs) were produced by the ultrasonication method combing different biocompatible oils and surfactants to enhance Hy aqueous solubility and bioavailability. Experimental parameters were optimized by the characterization of droplet size, zeta potential, and physicochemical properties. In vitro studies based on the release profile, cytotoxicity, cell morphology, and Hy intracellular accumulation were assayed. Hy at 100 mg L -1 was incorporated into the low viscosity (~0.005 Pa s) NEs with spherical droplets averaging 20-40 nm in size and polydispersity index <0.02. Hy release from the NE was significantly higher (4-fold) than its suspension (p < 0.001). The NEs demonstrated good physical stability during storage at 5 °C for at least six months. The Hy-loaded NEs exhibited an IC50 value 6-fold lower than Hy suspension during PDT against breast cancer cell lines (MCF-7). Cell microscopy imaging confirmed the increased cytotoxic effects of Hy-loaded NEs, showing damaged and apoptotic cells. Confocal laser scanning microscopy evidenced greater Hy delivery through NE into MCF-7 cells followed by improved intracellular ROS generation. Our results suggest that the Hy-loaded NEs can improve hypericin efficacy and assist Hy-PDT's preclinical development as a cancer treatment., (Copyright © 2021 Elsevier B.V. All rights reserved.)

α -glucosidase is a major enzyme that is involved in starch digestion and type 2 diabetes mellitus. In this study, the inhibition of hypericin by α-glucosidase and its mechanism were firstly investigated using enzyme kinetics analysis, real-time interaction analysis between hypericin and α -glucosidase by surface plasmon resonance (SPR), and molecular docking simulation. The results showed that hypericin was a high potential reversible and competitive α-glucosidase inhibitor, with a maximum half inhibitory concentration (IC 50 ) of 4.66 ± 0.27 mg/L. The binding affinities of hypericin with α -glucosidase were assessed using an SPR detection system, which indicated that these were strong and fast, with balances dissociation constant (KD) values of 6.56 × 10 -5 M and exhibited a slow dissociation reaction. Analysis by molecular docking further revealed that hydrophobic forces are generated by interactions between hypericin and amino acid residues Arg-315 and Tyr-316. In addition, hydrogen bonding occurred between hypericin and α -glucosidase amino acid residues Lys-156, Ser-157, Gly-160, Ser-240, His-280, Asp-242, and Asp-307. The structure and micro-environment of α-glucosidase enzymes were altered, which led to a decrease in α-glucosidase activity. This research identified that hypericin, an anthracene ketone compound, could be a novel α-glucosidase inhibitor and further applied to the development of potential anti-diabetic drugs.

The present study reports the performance of the pigment hypericin (HYP)-loaded poloxamer-based mucoadhesive in situ gelling liquid crystalline precursor system (LCPS) for the treatment of vulvovaginal candidiasis (VVC) in mice. LCPS composed of 40% of ethoxylated and propoxylated cetyl alcohol, 30% of oleic acid and cholesterol (7:1), 30% of a dispersion of 16% poloxamer 407 and 0.05% of HYP (HYP-LCPS) was prepared and characterized by polarized light microscopy (PLM), small-angle X-ray scattering (SAXS) and ex vivo permeation and retention studies across vaginal porcine mucosa were performed. In addition, the antifungal properties of the HYP-LCPS were evaluated in a murine in vivo model; for this, infected C57BL female mice groups were treated with both HYP in solution and HYP-LCPS, and after 6 days colony forming unit (CFU)/ml count was performed. PLM and SAXS confirmed that HYP-LCPS is a microemulsion situated in boundary transition region confirming its action as an LCPS. When in contact with simulated vaginal fluid, HYP-LCPS became rigid and exhibited maltase crosses and bragg peaks characteristics of lamellar phase. Ex vivo permeation and retention studies showed that HYP-LCPS provides a localized treatment on the superficial layers of porcine vaginal mucosa. HYP-LCPS induced a significant reduction in the number of CFU/ml in the mice; thus this formulation indicated it is as effective as a commercial dosage form. It was concluded that LCPS maintains the biological activity of HYP and provides an adequate drug delivery system for this lipophilic molecule at the vaginal mucosa, being a promising option in cases of VVC., (© The Author(s) 2021. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

HepG2 cells continue to be a valuable tool in early drug discovery and pharmaceutical development. In the current study we develop a 3D in vitro liver model, using HepG2/C3A cells that is predictive of human genotoxic exposure. HepG2/C3A cells cultured for 7-days in agarose-coated microplates formed spheroids which were uniform in shape and had well defined outer perimeters and no evidence of a hypoxic core. Quantitative real-time-PCR analysis showed statistically significant transcriptional upregulation of xenobiotic metabolising genes (CYP1A1, CYP1A2, UG1A1, UGT1A3, UGT1A6, EPHX, NAT2) and genes linked to liver function (ALB, CAR) in 3D cultures. In response to three model pro-genotoxicants: benzo[a]pyrene, amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-aminoanthracene (2-AA), we observed further transcriptional upregulation of xenobiotic metabolising genes (CYP1A1, CYP1A2, NAT1/2, SULT1A2, UGT1A1, UGT1A3) compared to untreated spheroids. Consistent with this, spheroids were more sensitive than 2D monolayers to compound induced single- and double- stranded DNA-damage as assessed by the comet assay and γH2AX phosphorylation respectively. In contrast, levels of DNA-damage induced by the direct acting mutagen 4-nitroquinoline N-oxide (4NQO) was the same in spheroids and monolayers. In support of the enhanced genotoxic response in spheroids we also observed transcriptional upregulation of genes relating to DNA-damage and cellular stress response (e.g. GADD45A and CDKN1A) in spheroids. In conclusion, HepG2/C3A 3D spheroids are a sensitive model for in vitro genotoxicity assessment with potential applications in early stage drug development., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

An unusual, new anthracene natural product named filamentos A ( 1 ), two new anthraquinone derivatives named filamentos B ( 2 ) and C ( 3 ), together with sixteen known compounds ( 4 - 19 ), were isolated from the roots of the tree Prismatomeris filamentosa . Their structures were identified by analysis of mass and spectroscopic data (IR, 1D and 2D NMR). Twelve metabolites ( 1 - 6 , 9 , 12 - 14 , 16 - 17 ) showed moderate antibacterial activities against a wide range of Gram-positive and Gram-negative bacteria such as B . subtilis , B . cereus , S . aureus , E . coli , P . aeruginosa and S . sonnei .

The use of nanocarriers for intracellular transport of actives has been extensively studied in recent years and represents a central area of nanomedicine. The main novelty of this paper lies on the use of nanogels formed by a low-molecular-weight gelator ( 1 ). Here, non-polymeric, molecular nanogels are successfully used for intracellular transport of two photodynamic therapy (PDT) agents, Rose Bengal (RB) and hypericin (HYP). The two photosensitizers (PSs) exhibit different drawbacks for their use in clinical applications. HYP is poorly water-soluble, while the cellular uptake of RB is hindered due to its dianionic character at physiological pH values. Additionally, both PSs tend to aggregate precluding an effective PDT. Despite the different nature of these PSs, nanogels from gelator 1 provide, in both cases, an efficient intracellular transport into human colon adenocarcinoma cells (HT-29) and a notably improved PDT efficiency, as assessed by confocal laser scanning microscopy and flow cytometry. Furthermore, no significant dark toxicity of the nanogels is observed, supporting the biocompatibility of the delivery system. The developed nanogels are highly reproducible due to their non-polymeric nature, and their synthesis is easily scaled up. The results presented here thus confirm the potential of molecular nanogels as valuable nanocarriers, capable of entrapping both hydrophobic and hydrophilic actives, for PDT of cancer.

The study explored the polycyclic aromatic hydrocarbon tolerance of indigenous biosurfactant producing microorganisms. Three bacterial species were isolated from crude oil contaminated sites of Haldia, West Bengal. The three species were screened for biosurfactant production and identified by 16S rRNA sequencing as Brevundimonas sp. IITISM 11, Pseudomonas sp. IITISM 19 and Pseudomonas sp. IITISM 24. The strains showed emulsification activities of 51%, 57% and 63%, respectively. The purified biosurfactants were characterised using FT-IR, GC-MS and NMR spectroscopy and found to have structural similarities to glycolipopeptides, cyclic lipopeptides and glycolipids. The biosurfactants produced were found to be stable under a wide range of temperature (0-100 °C), pH (4-12) and salinity (up to 20% NaCl). Moreover, the strains displayed tolerance to high concentrations (275 mg/L) of anthracene and fluorene and showed a good amount of cell surface hydrophobicity with different hydrocarbons. The study reports the production and characterisation of biosurfactant by Brevundimonas sp. for the first time. Additionally, the kinetic parameters of the bacterial strains grown on up to 300 mg/L concentration of anthracene and fluorene, ranged between 0.0131 and 0.0156 µ max (h -1 ), while the K s (mg/L) ranged between 59.28 and 102.66 for Monod's Model. For Haldane-Andrew's model, µ max (h -1 ) varied between 0.0168 and 0.0198. The inhibition constant was highest for Pseudomonas sp. IITISM 19 on anthracene and Brevundimonas sp. IITISM 11 on fluorene. The findings of the study suggest that indigenous biosurfactant producing strains have tolerance to high PAH concentrations and can be exploited for bioremediation purposes., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The prediction of host-guest binding affinities with computational modelling is still a challenging task. In the 7th statistical assessment of the modeling of proteins and ligands (SAMPL) challenge, a new host named TrimerTrip was synthesized and the thermodynamic parameters of 16 structurally diverse guests binding to the host were characterized. In the TrimerTrip-guest challenge, only structures of the host and the guests are provided, which indicates that the predictions of both the binding poses and the binding affinities are under assessment. In this work, starting from the binding poses obtained from our previous enhanced sampling simulations in the configurational space, we perform extensive alchemical and end-point free energy calculations to calculate the host-guest binding affinities retrospectively. The alchemical predictions with two widely accepted charge schemes (i.e. AM1-BCC and RESP) are in good agreement with the experimental reference, while the end-point estimates perform poorly in reproducing the experimental binding affinities. Aside from the absolute value of the binding affinity, the rank of binding free energies is also crucial in drug design. Surprisingly, the end-point MM/PBSA method seems very powerful in reproducing the experimental rank of binding affinities. Although the length of our simulations is long and the intermediate spacing is dense, the convergence behavior is not very good, which may arise from the flexibility of the host molecule. Enhanced sampling techniques in the configurational space may be required to obtain fully converged sampling. Further, as the length of sampling in alchemical free energy calculations already achieves several hundred ns, performing direct simulations of the binding/unbinding event in the physical space could be more useful and insightful. More details about the binding pathway and mechanism could be obtained in this way. The nonequilibrium method could also be a nice choice if one insists to use the alchemical method, as the intermediate sampling is avoided to some extent.

A small library of derivatives carrying a polycyclic scaffold recently identified by us as a new privileged structure in medicinal chemistry was designed and synthesized, aiming at obtaining potent MDR reverting agents also endowed with antitumor properties. In particular, as a follow-up of our previous studies, attention was focused on the role of the spacer connecting the polycyclic core with a properly selected nitrogen-containing group. A relevant increase in reverting potency was observed, going from the previously employed but-2-ynyl- to a pent-3-ynylamino moiety, as in compounds 3d and 3e, while the introduction of a triazole ring proved to differently impact on the activity of the compounds. The docking results supported the data obtained by biological tests, showing, for the most active compounds, the ability to establish specific bonds with P-glycoprotein. Moreover, a multifaceted anticancer profile and dual in vitro activity was observed for all compounds, showing both revertant and antitumor effects on leukemic cells. In this respect, 3c emerged as a "triple-target" agent, endowed with a relevant reverting potency, a considerable antiproliferative activity and a collateral sensitivity profile., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Fluorescent nanoparticles based on 9,10-distyrylanthracene (DSA) derivatives (4,4'-((1E,1'E)-anthracene-9,10-diylbis(ethene-2,1-diyl))bis(N,N-dimethylaniline) (NDSA) and 4,4'-((1E,1'E)-anthracene-9,10-diylbis(ethene-2,1-diyl))dibenzonitrile (CNDSA)) were prepared using an ultrasound aided nanoprecipitation method. The morphologies of the fluorescent nanoparticles could be controlled by adjusting the external ultrasonication time. NDSA or CNDSA could form spherical nanodots (NDSA NDs, CNDSA NDs) in a THF-H2O mixture with an 80% or 70% water fraction when the ultrasonication time was 30 s. When the ultrasonication time was prolonged to 10 min, NDSA and CNDSA could assemble into nanorods (NDSA NRs, CNDSA NRs). Meanwhile, the sizes of NDSA NRs and CNDSA NRs could be controlled by adjusting the water content in the mixture. As the water fraction was increased from 60% to 80%, the sizes of NDSA and CNDSA nanorods or nanodots reduced from 238.4 nm to 140.3 nm, and 482 nm to 198.4 nm, respectively. When the water fraction was up to 90%, irregular morphologies of NDSA and CNDSA could be observed. The nanoparticles exhibited intense fluorescence emission, good anti-photobleaching properties, as well as excellent stability and biocompatibility. In vitro cell imaging experiments indicated that the nanorods prepared by this simple method had the potential to be used for efficient and noninvasive long-term bioimaging.

Sirtuin isoform 2 (SIRT2) is an enzyme that catalyzes the removal of acyl groups from lysine residues. SIRT2's catalytic domain has a hydrophobic tunnel where its substrate acyl groups bind. Here, we report that the fluorescent probe 1-aminoanthracene (AMA) binds within SIRT2's hydrophobic tunnel in a substrate-dependent manner. AMA's interaction with SIRT2 was characterized by its enhanced fluorescence upon protein binding (>10-fold). AMA interacted weakly with SIRT2 alone in solution ( K d = 37 μM). However, when SIRT2 was equilibrated with a decanoylated peptide substrate, AMA's affinity for SIRT2 was enhanced ∼10-fold ( K d = 4 μM). The peptide's decanoyl chain and AMA co-occupied SIRT2's hydrophobic tunnel when bound to the protein. In contrast, binding of AMA to SIRT2 was competitive with a myristoylated substrate whose longer acyl chain occluded the entire tunnel. AMA competitively inhibited SIRT2 demyristoylase activity with an IC 50 of 21 μM, which was significantly more potent than its inhibition of other deacylase activities. Finally, binding and structural analysis suggests that the AMA binding site in SIRT2's hydrophobic tunnel was structurally stabilized when SIRT2 interacted with a decanoylated or 4-oxononanoylated substrate, but AMA's binding site was less stable when SIRT2 was bound to an acetylated substrate. Our use of AMA to explore changes in SIRT2's hydrophobic tunnel that are induced by interactions with specific acylated substrates has implications for developing ligands that modulate SIRT2's substrate specificity.

Benz(a)anthracene (BaA) is a polycyclic aromatic hydrocarbons (PAHs), that belongs to a group of carcinogenic and mutagenic persistent organic pollutants found in a variety of ecological habitats. In this study, the efficient biodegradation of BaA by a green alga Chlamydomonas reinhardtii (C. reinhardtii) CC-503 was investigated. The results showed that the growth of C. reinhardtii was hardly affected with an initial concentration of 10 mg/L, but was inhibited significantly under higher concentrations of BaA (>30 mg/L) (p < 0.05). We demonstrated that the relatively high concentration of 10 mg/L BaA was degraded completely in 11 days, which indicated that C. reinhardtii had an efficient degradation system. During the degradation, the intermediate metabolites were determined to be isomeric phenanthrene or anthracene, 2,6-diisopropylnaphthalene, 1,3-diisopropylnaphthalene, 1,7-diisopropylnaphthalene, and cyclohexanol. The enzymes involved in the degradation included the homogentisate 1,2-dioxygenase (HGD), the carboxymethylenebutenolidase, the ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and the ubiquinol oxidase. The respective genes encoding these proteins were significantly up-regulated ranging from 3.17 fold to 13.03 fold and the activity of enzymes, such as HGD and Rubisco, was significantly induced up to 4.53 and 1.46 fold (p < 0.05), during the BaA metabolism. This efficient degradation ability suggests that the green alga C. reinhardtii CC-503 may be a sustainable candidate for PAHs remediation., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A new class of non-ionic amphiphiles have been synthesised using a combination of polyethylene glycol (PEG) and oligoglycerol dendrons as hydrophilic units and an alkoxy aryl moiety as hydrophobic unit. The resulting amphiphiles were found to aggregate in aqueous medium. Their aggregation behaviour was studied using dynamic light scattering (DLS), fluorescence spectroscopy, and cryogenic electron microscopy (cryo-TEM). The inner hydrophobic core of these aggregates in aqueous medium is capable of encapsulating lipophilic guest molecules. The encapsulation behaviour was studied using Nile red as a hydrophobic dye as well as Curcumin and Dexamethasone as hydrophobic drug candidates. Furthermore, for biological evaluation, cytotoxicity and cellular uptake was studied using different cancer cell lines. The biomedical application of synthesised amphiphiles was further investigated for dermal drug delivery on excised human skin using Nile red encapsulated in the nanocarrier. The release profile of drug/dye encapsulated amphiphiles was studied under physiochemical conditions in the presence of immobilized lipase Novozym 435., 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 B.V. All rights reserved.)

The biological active compound rishirilide B is produced by Streptomyces bottropensis . The cosmid cos4 contains the complete rishirilide B biosynthesis gene cluster. Its heterologous expression in the host Streptomyces albus J1074 led to the production of rishirilide B as a major compound and to small amounts of rishirilide A, rishirilide D and lupinacidin A. In order to gain more insights into the biosynthesis, gene inactivation experiments and gene expression experiments were carried out. This study lays the focus on the functional elucidation of the genes involved in the early biosynthetic pathway. A total of eight genes were deleted and six gene cassettes were generated. Rishirilide production was not strongly affected by mutations in rslO2, rslO6 and rslH . The deletion of rslK4 and rslO3 led to the formation of polyketides with novel structures. These results indicated that RslK4 and RslO3 are involved in the generation or selection of the starter unit for rishirilide biosynthesis. In the rslO10 mutant strain, two novel compounds were detected, which were also produced by a strain containing solely the genes rslK1 , rslK2 , rslK3 , rslK4 , and rslA . rslO1 and rslO4 mutants predominately produce galvaquinones. Therefore, the ketoreductase RslO10 is involved in an early step of rishirilide biosynthesis and the oxygenases RslO1 and RslO4 are most probably acting on an anthracene moiety. This study led to the functional elucidation of several genes of the rishirilide pathway, including rslK4 , which is involved in selecting the unusual starter unit for polyketide synthesis.

The structural diversity of type II polyketides is largely generated by tailoring enzymes. In rishirilide biosynthesis by Streptomyces bottropensis , 13 C-labeling studies previously implied extraordinary carbon backbone and side-chain rearrangements. In this work, we employ gene deletion experiments and in vitro enzyme studies to identify key biosynthetic intermediates and expose intricate redox tailoring steps for the formation of rishirilides A, B, and D and lupinacidin A. First, the flavin-dependent RslO5 reductively ring-opens the epoxide moiety of an advanced polycyclic intermediate to form an alcohol. Flavin monooxygenase RslO9 then oxidatively rearranges the carbon backbone, presumably via lactone-forming Baeyer-Villiger oxidation and subsequent intramolecular aldol condensation. While RslO9 can further convert the rearranged intermediate to rishirilide D and lupinacidin A, an additional ketoreductase RslO8 is required for formation of the main products rishirilide A and rishirilide B. This work provides insight into the structural diversification of aromatic polyketide natural products via unusual redox tailoring reactions that appear to defy biosynthetic logic.

Smoothened (SMO) antagonist Vismodegib effectively inhibits the Hedgehog pathway in proliferating cancer cells. In early stage of treatment, Vismodegib exhibited promising outcomes to regress the tumors cells, but ultimately relapsed due to the drug resistive mutations in SMO mostly occurring before (primary mutations G497W) or after (acquired mutations D473H/Y) anti-SMO therapy. This study investigates the unprecedented insights of structural and functional mechanism hindering the binding of Vismodegib with sensitive and resistant mutant variants of SMO (SMO Mut ). Along with the basic dynamic understanding of Vismodegib-SMO complexes, network propagation theory based on heat diffusion principles is first time applied here to identify the modules of residues influenced by the individual mutations. The allosteric modulation by GLY497 residue in Vismodegib bound SMO wild-type (SMO WT ) conformation depicts the interconnections of intermediate residues of SMO with the atom of Vismodegib and identify two important motifs (E-X-P-L) and (Q-A-N-V-T-I-G) mediating this allosteric regulation. In this study a novel computational framework based on the heat diffusion principle is also developed, which identify significant residues of allosteric site causing drug resistivity in SMO Mut . This framework could also be useful for assessing the potential allosteric sites of different other proteins. Moreover, previously reported novel inhibitor "ZINC12368305," which is proven to make an energetically favorable complex with SMO WT is chosen as a control sample to assess the impact of receptor mutation on its binding and subsequently identify the important factors that govern binding disparity between Vismodegib and ZINC12368305 bound SMO WT/Mut conformations., (© 2019 Wiley Periodicals, Inc.)

Halophytic plants growing in harsh desert environments are rich reservoirs of unique endophytic microorganisms. Here, healthy fresh plants of the families Tamaricaceae and Amarantaceae at three saline locations in Iran were investigated for their bioactive endophytic fungi. Among a vast number of isolates, eight isolates were identified as Humicola fuscoatra (Sordariomycetes, Pezizomycotina, Ascomycota) by microscopy and representative DNA sequences of the 5.8S rDNA (ITS) and partial β-tubulin (TUB2). Those isolates were halotolerant, and highly bioactive, so that their intra- and extra-cellular metabolites possessed in vitro antifungal, antibacterial and antiproliferative activities, against a number of fungal and bacterial plant pathogens including the fungi Arthrobotrys conoides, Pyrenophora graminea, Pyricularia grisea and the bacteria Agrobacterium tumefaciens, Pseudomonas syringae and Xanthomonas oryzae. Chemical analyses of metabolites from the endophytes using HNMR, CNMR, NOESY, COSY, HMBC, HSQC, DEPT, TOCSY and EI MASS techniques identified 3,8-dihydroxy-1-methyl-9,10-anthracenedione (aloesaponarin II; an anthraquinone derivative), 1,8,9-anthracenetriol structure (chrysarobin; an anthranol derivative) and 2,4-di-tert-butylthiophenol in fungal extracts. To the best of our knowledge, this is the first report of endophytic association of halotolerant H. fuscoatra isolates with Tamaricaceae and Amarantaceae, and their bioactivity against plant pathogens. Also, the capability of chrysarobin and aloesaponarin II production is new to the fungal kingdom. These findings may find application in agriculture, pharmacology, and biotechnology.

Polycyclic aromatic hydrocarbons (PAHs) are universal organic pollutants in the agro ecosystems in China, therefore, it is important to understand the uptake and accumulation of PAHs in crops growing on PAHs contaminated soils for human health risk assessments. Water management is a common practice to maintain high grain yields during wheat production. However, the effects of soil water content on the accumulation and translocation of PAHs in wheat are still not clear. The main objectives of the present study were to investigate the effects of soil water content on the accumulation of three selected PAHs (Σ 3 PAHs, phenanthrene, anthracene and pyrene) in wheat during whole plant growth stage and on translocation or remobilization of Σ 3 PAHs from vegetative tissues to wheat grains. Winter wheat (Triticum aestivum cv. Xiaoyan22) were grown on Σ 3 PAHs spiked soils maintaining 80%, 60% or 40% water-holding capacity during the whole plant growth stage. Plant samplings were performed at jointing, anthesis or maturity stage, respectively. The present study showed that grain yield and biomass of the crop increased with soil water content increasing. Transpiration rate of wheat leaf under 80% and 60% water-holding capacity treatments was significantly (p < 0.05) higher than that under 40% water-holding capacity treatment at both anthesis and filling stage. Soil water content and plant growth stage had significant (p < 0.0001) effects on concentrations of phenanthrene, anthracene and pyrene in winter wheat. When exposed to 0, 15, 60, and 150 mg kg -1 Σ 3 PAHs in soils, Σ 3 PAHs concentrations in the grains under 60% water-holding capacity treatment were 46.6%, 69.9%, 89.5% and 81.7% of those under 80% water-holding capacity treatment, respectively. The highest concentrations of Σ 3 PAHs in the crop were recorded at anthesis stage. The distribution of PAHs in different tissues of wheat varied among different soil water treatments and plant growth stages. The present study indicated that optimizing soil water content during winter wheat production could apparently reduce concentrations of Σ 3 PAHs in grains via influence root uptake of Σ 3 PAHs and translocation of Σ 3 PAHs from stem or leaf into grain, suggesting the potential of water management to cope with PAHs contamination in crops growing on PAHs contaminated soils., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Polycyclic aromatic hydrocarbons (PAHs) are primarily produced during the incomplete combustion of organic matter. PAHs are suspected endocrine disruptors and possible carcinogenic materials. The major sources of human exposure to PAHs are inhaled fumes and food. The aim of this study was to provide an alternative drying method to mitigate PAH formation in dried red peppers. We prepared dried red pepper samples using air-drying and heat pump-assisted drying methods, and measured the concentrations of four PAHs (PAH4), benzo[a] anthracene (B[a]A), chrysene (CHR), benzo[b]fluoranthene (B[b]F), and benzo[a]pyrene (B[a]P), in the resulting pepper samples. The PAH concentrations ranged from 3.61 to 18.0 µg/kg and from 2.22 to 8.35 µg/kg in the air-dried and heat pump-dried pepper samples, respectively. Overall, the results have shown that dried peppers contain PAH4, that the drying conditions for these contaminants should be optimized for mitigating the PAH formation in dried red peppers., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

There is a growing interest in materials that can dynamically change their properties in the presence of cells to study mechanobiology. Herein, we exploit the 365 nm light mediated [4+4] photodimerization of anthracene groups to develop cytocompatible PEG-based hydrogels with tailorable initial moduli that can be further stiffened. A hydrogel formulation that can stiffen from 10 to 50 kPa, corresponding to the stiffness of a healthy and fibrotic heart, respectively, was prepared. This system was used to monitor the stiffness-dependent localization of NFAT, a downstream target of intracellular calcium signaling using a reporter in live cardiac fibroblasts (CFbs). NFAT translocates to the nucleus of CFbs on stiffening hydrogels within 6 h, whereas it remains cytoplasmic when the CFbs are cultured on either 10 or 50 kPa static hydrogels. This finding demonstrates how dynamic changes in the mechanical properties of a material can reveal the kinetics of mechanoresponsive cell signaling pathways that may otherwise be missed in cells cultured on static substrates., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Fluorophores are used for sensing biologically relevant ions, toxic metals or pathogenic markers. However, the mode of entry of such fluorophores into the cell greatly depends on their size, shape, surface charge, functional groups, and hydrophobicity. In particular, the influence of hydrophobicity on the intracellular uptake of fluorophores is poorly investigated. Self-assembly is a recent strategy to tune the intracellular uptake of fluorophores, facilitating increased intracellular sensing and fluorescence. Herein, self-assembly of three novel poly(aryl ether) dendron derivatives that contain rhodamine units was used to investigate the effect of hydrophobicity on the intracellular uptake of self-assembled fluorophores. The results suggest that monomer hydrophobicity plays an important role in the uptake. The dendron-based fluorophores, which upon self-assembly, formed stable spherical aggregates ranging from 300 to 500 nm. The rhodamine-based dendrons could selectively sense Hg 2+ ions in the presence of other competing metal cations. Intracellular imaging of the dendron-based fluorophores displayed bright red fluorescence in human embryonic kidney cells. The rate of intracellular uptake of the three dendron-based fluorophores was analyzed by flow cytometry. The results establish the importance of the hydrophilic-lipophilic balance of the self-assembled amphiphiles for tuning the intracellular uptake., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The anthraquinones endocrocin and emodin are synthesized by a special class of type I NR-PKSs and a discrete MβL-TE. In this work, we first reconstituted a biosynthetic pathway of endocrocin and emodin in S. cerevisiae by combining enzymes from different sources. We functionally characterized a TE-less NR-PKS (SlACAS) and a MβL-TE (SlTE) from S. lycopersici as well as four orthologous MβL-TEs. SlACAS was coexpressed with different MβL-TEs in S. cerevisiae. SlACAS generated the highest amount of endocrocin when coupled with HyTE, the yield was 115.6% higher than that with the native SlTE. To accumulate more emodin, seven decarboxylases with high homology to HyDC were identified and introduced into the biosynthetic pathway. Among these orthologs, AfDC exhibited the highest catalytic activity and the conversion rate reached 98.6%. A double-point mutant acetyl-CoA carboxylase, ACC1 S659A, S1157A , was further introduced to increase the production of malonyl-CoA as a precursor of these anthraquinones. The production of endocrocin (233.6 ± 20.3 mg/L) and emodin (253.2 ± 21.7 mg/L) then dramatically increased. We also optimized the carbon source in the medium and conducted fed-batch fermentation with the engineered strains. The titers of endocrocin and emodin obtained were 661.2 ± 50.5 mg/L and 528.4 ± 62.7 mg/L, respectively, which are higher than previously reported. In this work, by screening a small library of orthologous biosynthetic bricks, an efficient biosynthetic pathway of endocrocin and emodin was first created in S. cerevisiae. This study provides a novel metabolic engineering approach for optimization of the production of desired molecules., (Copyright © 2019 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)

Introduction: The demand to develop efficient and reliable analytical methods for the quality control of nutraceuticals is on the rise, together with an increase in the legal requirements for safe and consistent levels of its active principles., Objective: To establish a reliable model for the quality control of widely used Senna preparations used as laxatives and assess its phyto-equivalency., Methods: A comparative metabolomics approach via NMR and MS analyses was employed for the comprehensive measurement of metabolites and analyzed using chemometrics., Results: Under optimized conditions, 30 metabolites were simultaneously identified and quantified including anthraquinones, bianthrones, acetophenones, flavonoid conjugates, naphthalenes, phenolics, and fatty acids. Principal component analysis (PCA) was used to define relative metabolite differences among Senna preparations. Furthermore, quantitative 1 H NMR (qHNMR) was employed to assess absolute metabolites levels in preparations. Results revealed that 6-hydroxy musizin or tinnevellin were correlated with active metabolites levels, suggesting the use of either of these naphthalene glycosides as markers for official Senna drugs authentication., Conclusion: This study provides the first comparative metabolomics approach utilizing NMR and UPLC-MS to reveal for secondary metabolite compositional differences in Senna preparations that could readily be applied as a reliable quality control model for its analysis.

A urea derivative L1 exhibits Aggregation-Induced Emission (AIE) activity in an acetonitrile-water mixed solvent. The aggregation phenomenon has been corroborated by microscopy and light scattering studies. The ligand (L1) also displays a selective turn-on fluorescence response towards human serum albumin (HSA) in 100% aqueous medium over various other comparable proteins (even bovine serum albumin (BSA)) and enzymes. The weakly emissive probe L1 showed a substantial increase in emission intensity upon binding with HSA through electrostatic interactions. The good linear relationship between the fluorescence enhancement (I/I0 - 1) and the concentration of HSA provided the scope to attain an impressive detection limit as low as 5 μg mL-1. A drug displacement experiment and molecular docking study were employed to ascertain the likely protein (HSA)-ligand binding interactions.

Laccases produced by Leucoagaricus gongylophorus act in lignocellulose degradation and detoxification processes. Therefore, the use of L. gongylophorus laccase (Lac1Lg) was proposed in this work for degradation of anthracene and others polycyclic aromatic hydrocarbons without the use of mediators. Degradation reactions were performed in buffer aqueous solution with 10 ppm of anthracene and other PAHs, Tween-20 in 0.25% v/v and a laccase preparation of 50 U. The optimum condition (pH 6.0 and 30 °C) was determined by response surface methodology with an excellent coefficient of determination (R 2 ) of 0.97 and an adjusted coefficient of determination (R 2 adj ) of 0.93. In addition, the employment of the mediator ABTS decreased the anthracene biodegradation from 44 ± 1% to 30 ± 1%. This optimum pH of 6.0 suggests that the reaction occurs by a hydrogen atom transfer mechanism. Additionally, in 24 h Lac1Lg biodegraded 72 ± 1% anthracene, 40 ± 3% fluorene and 25 ± 3% phenanthrene. The yellow laccase from L. gongylophorus biodegraded anthracene and produced anthrone and anthraquinone, which are interesting compounds for industrial applications. Moreover, this enzyme also biodegraded the PAHs phenanthrene and fluorene justifying the study of Lac1Lg for bioremediation of these compounds in the environment.

Lichens are generally known as self-sufficient, symbiotic life-forms between fungi and algae/cyanobacteria, and they also provide shelter for a wide range of beneficial bacteria. Currently, bacterial-derived biodegradable polyhydroxyalkanoate (PHA) is grabbing the attention of many researchers as a promising alternative to non-degradable plastics. This study was conducted to develop a new method of PHA production using unexplored lichen-associated bacteria, which can simultaneously degrade two ubiquitous industrial toxins, anthracene and naphthalene. Here, 49 lichen-associated bacteria were isolated and tested for PHA synthesis. During the GC-MS analysis, a potential strain of EL19 was found to be a 3-hydroxyhexanoate (3-HHx) accumulator and identified as Pseudomonas sp. based on the 16S rRNA sequencing. GC analysis revealed that EL19 was capable of accumulating 30.62% and 19.63% of 3-HHx from naphthalene and anthracene, respectively, resulting in significant degradation of 98% and 96% of naphthalene and anthracene, respectively, within seven days. Moreover, the highly expressed phaC gene verified the genetic basis of PHA mcl production under nitrogen starvation conditions. Thus, this study strongly supports the hypothesis that lichen-associated bacteria can detoxify naphthalene and anthracene, store energy for extreme conditions, and probably help the associated lichen to live in extreme conditions. So far, this is the first investigation of lichen-associated bacteria that might utilize harmful toxins as feasible supplements and convert anthracene and naphthalene into eco-friendly 3-HHx. Implementation of the developed method would reduce the production cost of PHA mcl while removing harmful waste products from the environment.