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Selecting and breeding rice cultivars that enable strong cadmium (Cd) accumulation in rice straw but low accumulation in brown rice is a promising way to achieve Cd phytoremediation as well as to ensure the food safety of rice. Herein, we isolated a gene OsWNK9 from the quantitative trait locus associated with reducing Cd translocation from rice straw to brown rice and decreasing the Cd concentration in brown rice (BRCdC). Continuous strong expression of OsWNK9 was observed in nodes and internode and was induced after Cd supply. OsWNK9 was localized in the rice cell nucleus and participated in the regulation of Cd transport in yeast. Two independent oswnk9 rice mutants were generated via CRISPR/Cas9 gene-editing and showed significantly higher BRCdC than that of the wild type (WT). The BRCdC of knockout oswnk9 mutants was 0.227â¯mgâ¯kg-1and 0.238â¯mgâ¯kg-1, increased by 14â¯% and 19â¯% compared with that of the WT due to the lower Cd allocation in the basal stem, internode, and node III, which was unrelated to Cd uptake. Interestingly, OsWNK9 could promote iron (Fe) accumulation in rice under Cd-contaminated conditions, suggesting that OsWNK9 is an ideal gene for Cd phytoremediation and Fe biofortification in rice to support safe food production.
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Indoor environments serve as reservoirs for a variety of emerging pollutants (EPs), such as phthalates (PAE), with intricate interactions occurring between these compounds and indoor oxidants alongside dust particles. However, the precise mechanisms governing these interactions and their resulting environmental implications remain unclear. By theoretical simulations, this work uncovers multi-functional compounds and high oxygen molecules as important products arising from the interaction between DEP/DEHP and O3, which are closely linked to SOA formation. Further analysis reveals a strong affinity of DEP/DEHP for mineral dust surfaces, with an adsorption energy of 22.11/30.91 kcal mol-1, consistent with a higher concentration of DEHP on the dust surface. Importantly, mineral particles are found to inhibit every step of the reaction process, albeit resulting in lower product toxicity compared to the parent compounds. Thus, timely removal of dust in an indoor environment may reduce the accumulation and residue of PAEs indoors, and further reduce the combined exposure risk produced by PAEs-dust. This study aims to enhance our understanding of the interaction between PAEs and SOA formation, and to develop a fundamental reaction model at the air-solid surface, thereby shedding light on the microscopic behaviors and pollution mechanisms of phthalates on indoor dust surfaces.
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Tebuconazole (TEB), a prominent chiral triazole fungicide, has been extensively utilized for plant pathogen control globally. Despite experimental evidence of TEB metabolism in mammals, the enantioselectivity in the biotransformation of R- and S-TEB enantiomers by specific CYP450s remains elusive. In this work, integrated in silico simulations were employed to unveil the binding interactions and enantioselective metabolic fate of TEB enantiomers within human CYP1A2, 2B6, 2E1, and 3A4. Molecular dynamics (MD) simulations clearly delineated the binding specificity of R- and S-TEB to the four CYP450s, crucially determining their differences in metabolic activity and enantioselectivity. The primary driving force for robust ligand binding was identified as van der Waals interactions with CYP450s, particularly involving the hydrophobic residues. Mechanistic insights derived from quantum mechanics/molecular mechanics (QM/MM) calculations established C2-methyl hydroxylation as the predominant route of R-/S-TEB metabolism, while C6-hydroxylation and triazol epoxidation were deemed kinetically infeasible pathways. Specifically, the resulting hydroxy-R-TEB metabolite primarily originates from R-TEB biotransformation by 1A2, 2E1 and 3A4, whereas hydroxy-S-TEB is preferentially produced by 2B6. These findings significantly contribute to our comprehension of the binding specificity and enantioselective metabolic fate of chiral TEB by CYP450s, potentially informing further research on human health risk assessment associated with TEB exposure.
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The newly discovered ClO⢠and BrO⢠contribute to pollutant degradation in advanced oxidation processes, while acrylamide (AM) and acrylonitrile (ACN) are always the focus of scientists concerned due to their continuous production and highly toxic effects. Moreover, various particles with a graphene-like structure are the companions of AM/ACN in dry/wet sedimentation or aqueous phase existence, which play an important role in heterogeneous oxidation. Thus, this work focuses on the reaction mechanism and environmental effect of AM/ACN with ClOâ¢/BrOâ¢/HO⢠in the water environment under the influence of graphene (GP). The results show that although the reactivity sequence of AM and ACN takes the order of with HO⢠> with BrO⢠> with ClOâ¢, the easiest channel always occurs at the same C-position of the two reactants. The reaction rate constants (k) of AM with three radicals are 2 times larger than that with ACN, and amide groups have a better ability to activate CC bonds than cyanide groups. The existence of GP can accelerate the target reaction, and the k increased by 9-13 orders of magnitude. The toxicity assessment results show that the toxic effect of most products is lower than that of parent compounds, but the environmental risk of products from ClOâ¢/BrOâ¢-adducts is higher than those from HOâ¢-adducts. The oxidative degradation process based on ClO⢠and BrO⢠deserves special attention, and the catalytic effect of GP and its derivatives on the oxidation process is non-negligible.
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Acrilamida , Acrilonitrila , Grafite , Oxirredução , Acrilonitrila/química , Acrilamida/química , Grafite/química , Poluentes Químicos da Água/química , Modelos Teóricos , Radical Hidroxila/químicaRESUMO
Over the past decade, Pickering emulsions (PEs) stabilized by protein particles have been the focus of researches. The characteristics of protein particles at the oil-water interface are crucial for stabilizing PEs. The unique adsorption behaviors of protein particles and various modification methods enable oil-water interface to exhibit controllable regulation strategies. However, from the perspective of the interface, studies on the regulation of PEs by the adsorption behaviors of protein particles at oil-water interface are limited. Therefore, this review provides an in-depth study on oil-water interfacial adsorption of protein particles and their regulation on PEs. Specifically, the formation of interfacial layer and effects of their interfacial characteristics on PEs stabilized by protein particles are elaborated. Particularly, complicated behaviors, including adsorption, arrangement and deformation of protein particles at the oil-water interface are the premise of affecting the formation of interfacial layer. Moreover, the particle size, surface charge, shape and wettability greatly affect interfacial adsorption behaviors of protein particles. Importantly, stabilities of protein particles-based PEs also depend on properties of interfacial layers, including interfacial layer thickness and interfacial rheology. This review provides useful insights for the development of PEs stabilized by protein particles based on interfacial design.
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Emulsões , Óleos , Proteínas , Água , Emulsões/química , Adsorção , Água/química , Óleos/química , Proteínas/química , Tamanho da Partícula , Propriedades de Superfície , Reologia , MolhabilidadeRESUMO
In situ immobilization is a widely used measure for passivating Cd-contaminated soils. Amendments need to be continuously applied to achieve stable remediation effects. However, few studies have evaluated the impact of consecutive application of amendments on soil health and the microecological environment. A field experiment was conducted in a Cd-contaminated paddy (available Cd concentration 0.40 mg kg-1) on the Chengdu Plain to investigate the changes in soil Cd availability and response characteristics of soil bacterial communities after consecutive application of rice straw biochar (SW), fly ash (FM) and marble powder (YH) amendments from 2018 to 2020. Compared with control treatment without amendments (CK), soil pH increased by 0.6, 0.5 and 1.5 under SW, FM and YH amendments, respectively, and the soil available Cd concentration decreased by 10.71%, 21.42% and 25.00%, respectively. The Cd concentration in rice grain was less than 0.2 mg kg-1 under YH amendment, which was within the Chinese Contaminant Limit in Food of National Food Safety Standards (GB2762-2022) in the second and third years. The three amendments had different effects on the transformation of Cd fractions in soil, which may be relevant to the specific bacterial communities shaped under different treatments. The proportion of Fe-Mn oxide-bound fraction Cd (OX-Cd) increased by 11% under YH treatment, which may be due to the promotion of Fe(III) and Cd binding by some enriched iron-oxidizing bacteria, such as Lysobacter, uncultured_Pelobacter sp. and Sulfurifusis. Candidatus_Tenderia and Sideroxydans were enriched under SW and FM amendments, respectively, and were likely beneficial for reducing Cd availability in soil through Cd immobilization. These results revealed the significance of the bacterial community in soil Cd immobilization after consecutive application of amendments and highlighted the potential of applying YH amendment to ensure the safe production of rice in Cd-contaminated soil.
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Unveiling the role of heteroatom compounds in heavy oil viscosity is pivotal for finding targeted viscosity reduction methods to improve oil recovery. This research investigates the impact of heteroatoms in asphaltene molecules by utilizing quantum chemical calculations and molecular dynamics simulations to analyze their electrostatic potential characteristics, pairwise interactions, and dynamic behavior within realistic reservoirs. Heteroatom compounds can influence the molecular-level properties of asphaltenes and thus impact the macroscopic behavior of heavy oils. Research results suggest that the presence of ketone and aromatic rings in asphaltene molecules leads to the unrestricted movement of pi electrons due to their collective electronegativity. Two distinct configurations of asphaltene dimers, face-to-face, and edge-to-face, were observed. Intermolecular interactions were predominantly governed by van der Waals forces, highlighting their significant role in stabilizing asphaltene aggregates. The distribution of asphaltene molecules in the oil phase can be summarized as the "rebar-cement" theory. In the heteroatom-free system, the face-to-face peaks in the radial distribution function exhibit significantly reduced magnitudes compared to those in the heteroatom-containing system. This emphasizes the pivotal function of heteroatoms in connecting molecular components to form a more compact asphaltene structure, which may result in a higher viscosity of heavy oil. These findings give insight into the significance of heteroatoms in bridging molecular components and shaping the intricate structure of asphaltene and advance our understanding of heavy oil viscosity properties.
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Phosphorus (P)-hyperaccumulators for phytoextraction from P-polluted areas generally show rapid growth and accumulate large amounts of P without any toxicity symptom, which depends on a range of physiological processes and gene expression patterns that have never been explored. We investigated growth, leaf element concentrations, P fractions, photosynthetic traits, and leaf metabolome and transcriptome response in amphibious P-hyperaccumulators, Polygonum hydropiper and P. lapathifolium, to high-P exposure (5 mmol L-1), with 0.05 mmol L-1 as the control. Under high-P exposure, both species demonstrated good growth, allocating more P to metabolite P and inorganic P (Pi) accompanied by high potassium and calcium. The expression of a cluster of unigenes associated with photosynthesis was maintained or increased in P. lapathifolium, explaining the increase in net photosynthetic rate and the rapid growth under high-P exposure. Metabolites of trehalose metabolism, including trehalose 6-phosphate and trehalose, were sharply increased in both species by the high-P exposure, in line with the enhanced expression of associated unigenes, indicating that trehalose metabolic pathway was closely related to high-P tolerance. These findings elucidated the physiological and molecular responses involved in the photosynthesis and trehalose metabolism in P-hyperaccumulators to high-P exposure, and provides potential regulatory pathways to improve the P-phytoextraction capability.
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Regulação da Expressão Gênica de Plantas , Fósforo , Fotossíntese , Folhas de Planta , Polygonum , Fósforo/metabolismo , Polygonum/metabolismo , Polygonum/genética , Polygonum/efeitos dos fármacos , Polygonum/fisiologia , Folhas de Planta/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Trealose/metabolismo , Metaboloma , TranscriptomaRESUMO
Dihalogenated nitrophenols (2,6-DHNPs), an emerging group of aromatic disinfection byproducts (DBPs) detected in drinking water, have limited available information regarding their persistence and toxicological risks. The present study found that 2,6-DHNPs are resistant to major drinking water treatment processes (sedimentation and filtration) and households methods (boiling, filtration, microwave irradiation, and ultrasonic cleaning). To further assess their health risks, we conducted a series of toxicology studies using zebrafish embryos as the model organism. Our findings reveal that these emerging 2,6-DHNPs showed lethal toxicity 248 times greater than that of the regulated DBP, dichloroacetic acid. Specifically, at sublethal concentrations, exposure to 2,6-DHNPs generated reactive oxygen species (ROS), caused apoptosis, inhibited cardiac looping, and induced cardiac failure in zebrafish. Remarkably, the use of a ROS scavenger, N-acetyl-l-cysteine, considerably mitigated these adverse effects, emphasizing the essential role of ROS in 2,6-DHNP-induced cardiotoxicity. Our findings highlight the cardiotoxic potential of 2,6-DHNPs in drinking water even at low concentrations of 19 µg/L and the beneficial effect of N-acetyl-l-cysteine in alleviating the 2,6-DHNP-induced cardiotoxicity. This study underscores the urgent need for increased scrutiny of these emerging compounds in public health discussions.
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In the face of ongoing water pollution challenges, the intricate interplay between dissolved organic matter and disinfectants like chlorine gives rise to potentially harmful disinfection byproducts (DBPs) during water treatment. The exploration of DBP formation originating from amino acids (AA) is a critical focus of global research. Aromatic DBPs, in particular, have garnered considerable attention due to their markedly higher toxicity compared to their aliphatic counterparts. This work seeks to advance the understanding of DBP formation by investigating chlorination disinfection and kinetics using tyrosine (Tyr), phenylalanine (Phe), and tryptophan (Trp) as precursors. Via rigorous experiments, a total of 15 distinct DBPs with accurate molecular structures were successfully identified. The chlorination of all three AAs yielded highly toxic chlorophenylacetonitriles (CPANs), and the disinfectant dosage and pH value of the reaction system potentially influence chlorination kinetics. Notably, Phe exhibited the highest degradation rate compared to Tyr and Trp, at both the CAA:CHOCl ratio of within 1:2 and a wide pH range (6.0 to 9.0). Additionally, a neutral pH environment triggered the maximal reaction rates of the three AAs, while an acidic condition may reduce their reactivity. Overall, this study aims to augment the DBP database and foster a deeper comprehension of the DBP formation and relevant kinetics underlying the chlorination of aromatic AAs.
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Aminoácidos Aromáticos , Desinfecção , Halogenação , Purificação da Água , Cinética , Aminoácidos Aromáticos/química , Purificação da Água/métodos , Desinfetantes/química , Poluentes Químicos da Água/química , Concentração de Íons de HidrogênioRESUMO
In this study, a novel coordination polymer {Co2(Oaobtc)(bpe)(H2O)4]}n (1) was synthesized under hydrothermal conditions using a hybrid ligand synthesis method, where H4Oobtc represents 2,3,3'-tricarboxylate azobenzene, and bpe represents 1,2-bis(4-pyridyl)ethylene. The obtained CP1 was characterized by elemental analysis (EA), powder X-ray diffraction (PXRD), and thermal gravimetric analysis (TGA). Fluorescence testing confirmed the excellent photoluminescent performance of compound 1, indicating its potential as a cyan-emitting fluorescent material. Hyaluronic acid (HA) and carboxymethyl chitosan (CMCS) are natural polysaccharides known for their biocompatibility. HA/CMCS hydrogels were synthesized using a chemical synthesis method, featuring a three-dimensional network structure with interconnected pores, and an average pore size of 314.75 ± 11.25 µm. The characterization of the taxotere-loaded hydrogel was performed using infrared spectroscopy, confirming the effective encapsulation of the drug within the hydrogel. Utilizing taxotere as a model drug, a novel taxotere-loaded metal gel was synthesized, and its anticancer efficacy was evaluated. Furthermore, the influence of different pH levels on drug release rate was investigated. Finally, the encapsulation and release of taxotere in the hydrogel were studied using UV-visible spectroscopy.
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Rhamnolipids (RLs) have emerged as one of the most promising classes of biosurfactants. The ratio of mono-RL to di-RL plays a significant role in determining its performance. Therefore, strains whose production of mono-RL and di-RL are manuplable, have advantage on applications in various scenarios. In this study, we developed a rhlC deletion mutant strain in Pseudomonas aeruginosa PAO1, which produced primarily mono-RL. Subsequently, we generated two complemented strains by integrating the arabinose-induced PBAD-rhlC gene, either directly into the chromosomes or expressing it on plasmids. Our results indicate that the ratio of mono-RL to di-RL synthesized by the complemented strain gradually decreased as the concentration of arabinose (the inducer) increased. Consequently, there was a decrease in emulsification ability and an increase in surface tension and critical micelle concentration (CMC) of the corresponding rhamnolipids. The complemented strains without inducer can produce a small amount of di-rhamnolipids, which enhanced the surfactant properties. Notably, the rhamnolipids induced by 0.10% arabinose exhibited the most potent antibacterial effect.
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Arabinose , Glicolipídeos , Glicolipídeos/farmacologia , Antibacterianos/farmacologia , Tensoativos/farmacologia , Pseudomonas aeruginosa/genéticaRESUMO
The escalating focus on the environmental occurrence and toxicology of emerging pollutants underscores the imperative need for a profound exploration of their metabolic transformations mediated by human CYP450 enzymes. Such investigations have the potential to unravel the intricate metabolite profiles, substantially altering the toxicological outcomes. In this study, we integrated the computational simulations with in vitro metabolism experiments to investigate the metabolic activity and mechanism of an emerging pollutant, 1,3,5-tris(2,3-dibromopropyl)-1,3,5-triazinane-2,4,6-trione (TDBP-TAZTO), catalyzed by human CYP450s. The results highlight the important contributions of CYP2E1, 3A4 and 2C9 to the biotransformation of TDBP-TAZTO, leading to the identification of four distinct metabolites. The effective binding conformations governing biotransformation reactions of TDBP-TAZTO within active CYP450s are unveiled. Structural instability of primary hydroxyTDBP-TAZTO products suggests three potential outcomes: (1) generation of an alcohol metabolite through successive debromination and reduction reactions, (2) formation of a dihydroxylated metabolite through secondary hydroxylation by CYP450, and (3) production of an N-dealkylated metabolite via decomposition and isomerization reactions in the aqueous environment. The formation of a desaturated debrominated metabolite may arise from H-abstraction and barrier-free Br release during the primary oxidation, potentially competing with the generation of hydroxyTDBP-TAZTO. These findings provide detailed mechanistic insight into TDBP-TAZTO biotransformation by CYP450s, which can enrich our understanding of the metabolic fate and associated health risk of this chemical.
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Poluentes Ambientais , Retardadores de Chama , Humanos , Retardadores de Chama/metabolismo , Triazinas/análise , Sistema Enzimático do Citocromo P-450/metabolismo , Biotransformação , OxirreduçãoRESUMO
3-bromine carbazole (3-BCZ) represents a group of emerging aromatic disinfection byproducts (DBP) detected in drinking water; however, limited information is available regarding its potential cardiotoxicity. To assess its impacts, zebrafish embryos were exposed to 0, 0.06, 0.14, 0.29, 0.58, 1.44 or 2.88 mg/L of 3-BCZ for 120 h post fertilization (hpf). Our results revealed that ≥1.44 mg/L 3-BCZ exposure induced a higher incidence of heart malformation and an elevated pericardial area in zebrafish larvae; it also decreased the number of cardiac muscle cells and thins the walls of the ventricle and atrium while increasing cardiac output and impeding cardiac looping. Furthermore, 3-BCZ exposure also exhibited significant effects on the transcriptional levels of genes related to both cardiac development (nkx2.5, vmhc, gata4, tbx5, tbx2b, bmp4, bmp10, and bmp2b) and cardiac function (cacna1ab, cacna1da, atp2a1l, atp1b2b, atp1a3b, and tnnc1a). Notably, N-acetyl-L-cysteine, a reactive oxygen species scavenger, may alleviate the failure of cardiac looping induced by 3-BCZ but not the associated cardiac dysfunction or malformation; conversely, the aryl hydrocarbon receptor agonist CH131229 can completely eliminate the cardiotoxicity caused by 3-BCZ. This study provides new evidence for potential risks associated with ingesting 3-BCZ as well as revealing underlying mechanisms responsible for its cardiotoxic effects on zebrafish embryos.
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Proteínas de Peixe-Zebra , Peixe-Zebra , Animais , Proteínas de Peixe-Zebra/genética , Coração , Bromo/farmacologia , Cardiotoxicidade , Receptores de Hidrocarboneto Arílico/genética , Larva , Desinfecção , Embrião não MamíferoRESUMO
Effectively identifying persistent organic pollutants (POPs) with extensive organic chemical datasets poses a formidable challenge but is of utmost importance. Leveraging machine learning techniques can enhance this process, but previous models often demanded advanced programming skills and high-end computing resources. In this study, we harnessed the simplicity of PyCaret, a Python-based package, to construct machine-learning models for POP screening based on 2D molecular descriptors. We compared the performance of these models against a deep convolutional neural network (DCNN) model. Utilising minimal Python code, we generated several models that exhibited superior or comparable performance to the DCNN. The most outstanding performer, the Light Gradient Boosting Machine (LGBM), achieved an accuracy of 96.20 %, an AUC of 97.70 %, and an F1 score of 82.58 %. This model outshone the DCNN model. Furthermore, it excelled in identifying POPs within the REACH PBT and compiled industrial chemical lists. Our findings highlight the accessibility and simplicity of PyCaret, requiring only a few lines of code, rendering it suitable for non-computing professionals in environmental sciences. The ability of low code machine learning tools (e.g. PyCaret) to facilitate model comparison and interpretation holds promise, encouraging prompt assessment and management of chemical substances.
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The unrestricted utilization of antibiotics poses a critical challenge to global public health and safety. Levofloxacin (LEV) and sulfaphenazole (SPN), widely employed broad-spectrum antimicrobials, are frequently detected at the terminal stage of water treatment, raising concerns regarding their potential conversion into detrimental disinfection byproducts (DBPs). However, current knowledge is deficient in identifying the potential DBPs and elucidating the precise transformation pathways and influencing factors during the chloramine disinfection process of these two antibiotics. This study conducts a comprehensive analysis of reaction pathways, encompassing piperazine ring opening/oxidation, Cl-substitution, OH-substitution, desulfurization, and S-N bond cleavage, during chloramine disinfection. Twelve new DBPs were identified in this study, exhibiting stability and persistence even after 24 h of disinfection. Additionally, an examination of DBP generation under varying disinfectant concentrations and pH values revealed peak levels at a molar ratio of 25 for LEV and SPN to chloramine, with LEV contributing 11.5% and SPN 23.8% to the relative abundance of DBPs. Remarkably, this research underscores a substantial increase in DBP formation within the molar ratio range of 1:1 to 1:10 compared to 1:10 to 1:25. Furthermore, a pronounced elevation in DBP generation was observed in the pH range of 7 to 8. These findings present critical insights into the impact of the disinfection process on these antibiotics, emphasizing the innovation and significance of this research in assessing associated health risks.
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Levofloxacino , Purificação da Água , Levofloxacino/farmacologia , Sulfafenazol , Cloraminas/farmacologia , Desinfecção , Antibacterianos/farmacologiaRESUMO
Chemotherapy resistance is the major cause of treatment failure in osteosarcoma, the most common primary bone malignancy, and sensitizing therapeutic strategy is required to improve the clinical outcome. In this study, we discovered that navitoclax, a selective inhibitor of Bcl-2/Bcl-xL, effectively combats chemoresistance in osteosarcoma. Our research revealed that Bcl-2, but not Bcl-xL, is upregulated in osteosarcoma cells that are resistant to doxorubicin. However, venetoclax, a specific inhibitor of Bcl-2, did not exhibit activity against doxorubicin-resistant cells. Further analysis showed that depleting either Bcl-2 or Bcl-xL alone was insufficient to overcome doxorubicin resistance. Only by depleting both Bcl-2 and Bcl-xL significantly reduce the viability of doxorubicin-resistant cells. Similarly, navitoclax not only decreased the viability of doxorubicin-resistant cells but also acted synergistically with doxorubicin in cells sensitive to the drug. To confirm the ability of navitoclax to overcome doxorubicin resistance, we conducted experiments using multiple mouse models of osteosarcoma, both doxorubicin-sensitive and doxorubicin-resistant. The results provided confirmation that navitoclax is effective in overcoming doxorubicin resistance. Our findings demonstrate that simultaneous inhibition of Bcl-2 and Bcl-xL could serve as a novel strategy to sensitize chemoresistant osteosarcoma cells. Moreover, our study presents preclinical evidence supporting the potential of a navitoclax and doxorubicin combination therapy for the treatment of osteosarcoma, paving the way for future clinical investigations.
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Compostos de Anilina , Neoplasias Ósseas , Osteossarcoma , Sulfonamidas , Animais , Camundongos , Proteína bcl-X/farmacologia , Proteína bcl-X/uso terapêutico , Apoptose , Linhagem Celular Tumoral , Proteínas Proto-Oncogênicas c-bcl-2/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/uso terapêutico , Doxorrubicina/farmacologia , Doxorrubicina/uso terapêutico , Osteossarcoma/tratamento farmacológico , Osteossarcoma/patologia , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/patologia , Resistencia a Medicamentos AntineoplásicosRESUMO
The root-associated microbiome assembly substantially promotes (hyper)accumulator plant growth and metal accumulation and is influenced by multiple factors, especially host species and environmental stress. Athyrium wardii (Hook.) is a phytostabilizer that grows in lead (Pb)-zinc (Zn) mine tailings and shows high root Pb accumulation. However, there remains little information on the assembly of the root-associated microbiome of A. wardii and its role in phytostabilization. A field study investigated the structural and functional variation in the root-associated bacterial microbiome of Athyrium wardii (Hook.) exposed to different levels of contamination in Pb-Zn mine tailings. The root compartment dominated the variation in the root-associated bacterial microbiome but the levels of contaminants showed less impact. Bacterial co-occurrence was enhanced in the rhizosphere soil and rhizoplane but tended to be much simpler in the endosphere in terms of network complexity and connectivity. This indicates that the microbial community assembly of A. wardii was non-random and shaped by root selective effects. Proteobacteria, Chloroflexi, Actinobacteria, Cyanobacteria, and Acidobacteriota were generally the dominant bacterial phyla. The genera Crossiella and Bradyrhizobium were enriched in the rhizosphere and cyanobacterial genera were enriched in the endosphere, demonstrating substantial advantages to plant survival and adaptation in the harsh mine environment. Functional categories involved in amino acid and carbohydrate metabolism were abundant in the rhizosphere soil, thus contributing to metal solubility and bioavailability in the rhizosphere. Membrane transporters, especially ATP-binding cassette transporters, were enriched in the endosphere, indicating a potential role in metal tolerance and transportation in A. wardii. The study shows substantial variation in the structure and function of microbiomes colonizing different compartments, with the rhizosphere and endophytic microbiota potentially involved in plant metal tolerance and accumulation during phytostabilization.
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Microbiota , Traqueófitas , Chumbo/toxicidade , Chumbo/metabolismo , Plantas , Bactérias , Zinco/toxicidade , Zinco/metabolismo , Solo/química , Rizosfera , Raízes de Plantas/metabolismo , Microbiologia do SoloRESUMO
BACKGROUND: Extrahepatic cholangiocarcinoma sarcoma is extremely rare in clinical practice. These cells consist of both epithelial and mesenchymal cells. Patient-derived cell lines that maintain tumor characteristics are valuable tools for studying the molecular mechanisms associated with carcinosarcoma. However, cholangiocarcinoma sarcoma cell lines are not available in cell banks. AIM: To establish and characterize a new extrahepatic cholangiocarcinoma sarcoma cell line, namely CBC2T-2. METHODS: We conducted a short tandem repeat (STR) test to confirm the identity of the CBC2T-2 cell line. Furthermore, we assessed the migratory and invasive properties of the cells and performed clonogenicity assay to evaluate the ability of individual cells to form colonies. The tumorigenic potential of CBC2T-2 cells was tested in vivo using non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. The cells were injected subcutaneously and tumor formation was observed. In addition, immunohistochemical analysis was carried out to examine the expression of epithelial marker CK19 and mesenchymal marker vimentin in both CBC2T-2 cells and xenografts. The CBC2T-2 cell line was used to screen the potential therapeutic effects of various clinical agents in patients with cholangiocarcinoma sarcoma. Lastly, whole-exome sequencing was performed to identify genetic alterations and screen for somatic mutations in the CBC2T-2 cell line. RESULTS: The STR test showed that there was no cross-contamination and the results were identical to those of the original tissue. The cells showed round or oval-shaped epithelioid cells and mesenchymal cells with spindle-shaped or elongated morphology. The cells exhibited a high proliferation ratio with a doubling time of 47.11 h. This cell line has migratory, invasive, and clonogenic abilities. The chromosomes in the CBC2T-2 cells were polyploidy, with numbers ranging from 69 to 79. The subcutaneous tumorigenic assay confirmed the in vivo tumorigenic ability of CBC2T-2 cells in NOD/SCID mice. CBC2T-2 cells and xenografts were positive for both the epithelial marker, CK19, and the mesenchymal marker, vimentin. These results suggest that CBC2T-2 cells may have both epithelial and mesenchymal characteristics. The cells were also used to screen clinical agents in patients with cholangiocarcinoma sarcoma, and a combination of paclitaxel and gemcitabine was found to be the most effective treatment option. CONCLUSION: We established the first human cholangiocarcinoma sarcoma cell line, CBC2T-2, with stable biogenetic traits. This cell line, as a research model, has a high clinical value and would facilitate the understanding of the pathogenesis of cholangiocarcinoma sarcoma.
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Neoplasias dos Ductos Biliares , Colangiocarcinoma , Sarcoma , Camundongos , Animais , Humanos , Vimentina , Linhagem Celular Tumoral , Camundongos SCID , Camundongos Endogâmicos NOD , Sarcoma/genética , Sarcoma/patologia , Colangiocarcinoma/genética , Colangiocarcinoma/patologia , Neoplasias dos Ductos Biliares/patologia , Ductos Biliares Intra-Hepáticos/patologiaRESUMO
Biofilms are complex microbial communities embedded in extracellular matrix. Pathogens within the biofilm become more resistant to the antibiotics than planktonic counterparts. Novel strategies are required to encounter biofilms. Exopolysaccharides are one of the major components of biofilm matrix and play a vital role in biofilm architecture. In previous studies, a glycosyl hydrolase, PslGPA, from Pseudomonas aeruginosa was found to be able to inhibit biofilm formation by disintegrating exopolysaccharide in biofilms. Here, we investigate the potential spectrum of PslG homologous protein with anti-biofilm activity. One glycosyl hydrolase from Pseudomonas fluorescens, PslGPF, exhibits anti-biofilm activities and the key catalytic residues of PslGPF are conserved with those of PslGPA. PslGPF at concentrations as low as 50 nM efficiently inhibits the biofilm formation of P. aeruginosa and disassemble its preformed biofilm. Furthermore, PslGPF exhibits anti-biofilm activity on a series of Pseudomonads, including P. fluorescens, Pseudomonas stutzeri and Pseudomonas syringae pv. phaseolicola. PslGPF stays active under various temperatures. Our findings suggest that P. fluorescens glycosyl hydrolase PslGPF has potential to be a broad spectrum inhibitor on biofilm formation of a wide range of Pseudomonads.