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Detoxification of ochratoxin A by Lysobacter sp. CW239 and characteristics of a novel degrading gene carboxypeptidase cp4
2020
Wei, Wei | Qian, Yingying | Wu, Yanbo | Chen, Ying | Peng, Cheng | Luo, Mingzhong | Xu, Junfeng | Zhou, Yu
Ochratoxin A (OTA) is a potent mycotoxin that frequently contaminates agro-products and threatens food safety. A highly efficient OTA degrading strain Lysobacter sp. CW239 was isolated, and the OTA degradation characteristics were investigated. A novel OTA degrading gene carboxypeptidase cp4 was successfully cloned and characterized from CW239. The heterologous recombinant was constructed by gene cp4 and expression vector pET-32a⁽⁺⁾ and overexpressed by E. coli BL21 CodonPlus™ (DE3). The recombinant protein rCP4 was purified, and the OTA-degrading activity was evaluated. Although OTA was efficiently degraded by CW239 (24-h degradation ratio of 86.2%), the 24-h OTA degradation ratio for rCP4 was only 36.8% at fairly high concentration (0.25 mg/mL) protein. The degraded product was obtained by immune affinity column (IAC) and determined by mass spectrometry (MS), and the degraded product was the less toxic ochratoxin α (OTα). Based on the serial investigations of this study, OTA might be simultaneously co-degraded by CP4 and another unknown degrading agent in that degrading strain.
Show more [+] Less [-]Accumulation of free and covalently bound microcystins in tissues of Lymnaea stagnalis (Gastropoda) following toxic cyanobacteria or dissolved microcystin-LR exposure
2010
Lance, Emilie | Neffling, Milla-Riina | Gérard, Claudia | Meriluoto, Jussi | Bormans, Myriam
Accumulation of free microcystins (MCs) in freshwater gastropods has been demonstrated but accumulation of MCs covalently bound to tissues has never been considered so far. Here, we follow the accumulation of total (free and bound) MCs in Lymnaea stagnalis exposed to i) dissolved MC-LR (33 and 100 μg L−1) and ii) Planktothrix agardhii suspensions producing 5 and 33 μg MC-LR equivalents L−1 over a 5-week period, and after a 3-week depuration period. Snails exposed to dissolved MC-LR accumulated up to 0.26 μg total MCs g−1 dry weight (DW), with no detection of bound MCs. Snails exposed to MCs producing P. agardhii accumulated up to 69.9 μg total MCs g−1 DW, of which from 17.7 to 66.7% were bound. After depuration, up to 15.3 μg g−1 DW of bound MCs were detected in snails previously exposed to toxic cyanobacteria, representing a potential source of MCs transfer through the food web. The study concerns accumulation and elimination of both free and bound microcystins (MCs) in tissues of a gastropod exposed to MCs producing cyanobacteria or dissolved MC-LR.
Show more [+] Less [-]Sulforaphane prevents chromium-induced lung injury in rats via activation of the Akt/GSK-3β/Fyn pathway
2020
Lv, Yueying | Jiang, Huijie | Li, Siyu | Han, Bing | Liu, Yan | Yang, Daqian | Li, Jiayi | Yang, Qingyue | Wu, Pengfei | Zhang, Zhigang
Chromium (Cr) is an internationally recognized carcinogenic hazard that causes serious pulmonary toxicity. However, Cr-induced pulmonary toxicity lacks effective treatment to date. Sulforaphane (SFN), a well-known organosulfur compound, has gained increasing attention because of its unique biological function. This study investigates if SFN could decrease K₂Cr₂O₇-induced pulmonary toxicity and a potential mechanism involved using a rat 35-day Cr-induced pulmonary toxicity model and the mouse alveolar type II epithelial cell line (MLE-12). The results showed that SFN prevented Cr-induced oxidative stress, histopathological lesions, inflammation, apoptosis, and changes in protein kinase B (Akt) and glycogen synthase kinase 3 beta (GSK-3β) levels in vivo and in vitro. However, SFN can not play the protective effect against K₂Cr₂O₇-induced cell injury after treating by an Akt-specific inhibitor (MK-2206 2HCl) in MLE-12 cells. Furthermore, SFN increased the expression of nuclear factor-E2-related factor-2 (Nrf2) phase II detoxification enzymes. Collectively, this study demonstrates that SFN prevents K₂Cr₂O₇-induced lung toxicity in rats through enhancing Nrf2-mediated exogenous antioxidant defenses via activation of the Akt/GSK-3β/Fyn signaling pathway. SFN may be a novel natural substance to cure Cr-induced lung toxicity.
Show more [+] Less [-]Detoxification and repair process of ozone injury: From O3 uptake to gene expression adjustment
2009
Castagna, A. | Ranieri, A.
Plants react to O3 threat by setting up a variety of defensive strategies involving the co-ordinated modulation of stress perception, signalling and metabolic responses. Although stomata largely controls O3 uptake, differences in O3 tolerance cannot always be ascribed to changes in stomatal conductance but cell protective and repair processes should be taken into account. O3-driven ROS production in the apoplast induces a secondary, active, self-propagating generation of ROS, whose levels must be finely tuned, by many enzymatic and non-enzymatic antioxidant systems, to induce gene activation without determining uncontrolled cell death. Additional signalling molecules, as ethylene, jasmonic and salicylic acid are also crucial to determine the spreading and the containment of leaf lesions. The main recent results obtained on O3 sensing, signal transduction, ROS formation and detoxification mechanisms are here discussed. A dissection of the complex network of interacting mechanisms which determine the cell fate under ozone stress.
Show more [+] Less [-]Pteris vittata - Revisited: Uptake of As and its speciation, impact of P, role of phytochelatins and S
2009
Vetterlein, Doris | Wesenberg, Dirk | Nathan, Petra | Bräutigam, Anja | Schierhorn, Angelika | Mattusch, Jürgen | Jahn, Reinhold
Pteris vittata is known to hyperaccumulate As but the mechanism is poorly understood. We found an increase of As concentration with increasing soil solution As concentrations, but P application had no impact, although plant P concentrations responded to different rates of P supply. As in fronds was dominantly (82-89%) present in the form of AsIII. In roots we detected 45% as AsIII which is higher than reported in previous studies and supports substantial As-reduction to take place in roots. We detected PC2/3GS-AsIII, PC2-GS-AsIII and (PC2)2-AsIII in increasing amounts with application of As. The total amount of PC was in the range reported previously and far too small to assign a significant role in As detoxification to PCs. The close correlation between S and As in fronds and the lack of data on sulphur uptake and metabolism indicates the need for a detailed investigation on sulphur nutritional status and As metabolism in P. vittata. As-PC complexes were detected in increasing amounts with increasing As availability, but total amounts were small and do not explain the close correlation between S and As in fronds.
Show more [+] Less [-]Ca2+-dependent plant response to Pb2+ is regulated by LCT1
2007
Wojas, S. | Ruszczyńska, A. | Bulska, E. | Wojciechowski, M. | Antosiewicz, D.M.
Tobacco plants transformed with TaLCT1 were cultured on Knop's medium with modified calcium concentrations (0.01-3 mM) in the presence of Pb2+, and in soil contaminated by lead. A 4-5 μM Pb2+ administered in the presence of 1 mM Ca2+ inhibited the root growth of transgenic plants to much lesser degree than of control plants, whereas in the presence of 3 mM Ca2+ no differences were found between the studied lines. The reduction of Pb2+ toxicity in the presence of 1 mM Ca2+ was not accompanied by a change in the lead tissue concentration. However, when Ca2+ level in the medium was lowered to 0.01 mM, several fold higher root/shoot Pb ratio in transgenic plants was observed, twofold increase in the total amount of metal accumulated, and lower concentration of Pb in the xylem sap. Results suggest the involvement of TaLCT1 in the regulation of Ca-dependent Pb-detoxification, and under conditions of low calcium in lead uptake and distribution. Ca2+-dependent Pb2+ detoxification and uptake was regulated by TaLCT1.
Show more [+] Less [-]Regioselective hydroxylation of carbendazim by mammalian cytochrome P450: A combined experimental and computational study
2022
Lv, Xia | Li, Jing-Xin | Wang, Jia-Yue | Tian, Xiang-Ge | Feng, Lei | Sun, Cheng-Peng | Ning, Jing | Wang, Chao | Zhao, Wen-Yu | Li, Ya-Chen | Ma, Xiao-Chi
Carbendazim (CBZ), a broad-spectrum pesticide frequently detected in fruits and vegetables, could trigger potential toxic risks to mammals. To facilitate the assessment of health risks, this study aimed to characterize the cytochrome P450 (CYPs)-mediated metabolism profiles of CBZ by a combined experimental and computational study. Our results demonstrated that CYPs-mediated region-selective hydroxylation was a major metabolism pathway for CBZ in liver microsomes from various species including rat, mouse, minipig, dog, rabbit, guinea pig, monkey, cow and human, and the metabolite was biosynthesized and well-characterized as 6-OH-CBZ. CYP1A displayed a predominant role in the region-selective hydroxylation of CBZ that could attenuate its toxicity through converting it into a less toxic metabolite. Meanwhile, five other common pesticides including chlorpyrifos-methyl, prochloraz, chlorfenapyr, chlorpyrifos, and chlorothalonil could significantly inhibit the region-selective hydroxylation of CBZ, and consequently remarkably increased CBZ exposure in vivo. Furthermore, computational study clarified the important contribution of the key amino acid residues Ser122, and Asp313 in CYP1A1, as well as Asp320 in CYP1A2 to the hydroxylation of CBZ through hydrogen bonds. These results would provide some useful information for the metabolic profiles of CBZ by mammalian CYPs, and shed new insights into CYP1A-mediated metabolic detoxification of CBZ and its health risk assessment.
Show more [+] Less [-]Bioengineered magnetic graphene oxide microcomposites for bioremediation of chromium in ex situ - A novel strategy for aggrandized recovery by electromagnetic gadgetry
2022
Aravind, Manikka Kubendran | Kappen, Jincymol | Narayanamoorthi, Eswaran | Sanjaykumar, Ashokkumar | Varalakshmi, Perumal | Arockiadoss, Thevasahayam | John, Swamidoss Abraham | Ashokkumar, Balasubramaniem
Novel magnetic microcomposites consisting of graphene oxide and iron oxide was synthesized to immobilize metabolically versatile Paracoccus sp. MKU1 and Leucobacter sp. AA7 and tested for the simultaneous adsorption and enhanced biological detoxification of hexavalent chromium (Cr(VI)) from tannery wastewater. This study reports highest chromium adsorption of 272.6 mg/g and 179.3 mg/g with complete reduction of Cr(VI) to Cr(III) by the microcomposites of AA7 and MKU1 from wastewater in a bioreactor (10 L) at large-scale for first time in ex situ. Furthermore, both the microcomposites displayed an enhanced detoxification of tannery wastewater by reducing various physicochemical conditions such as ammonia, nitrate, TDS, fluoride, CaCO₃, Ca, Mg, NO₃ and SO₂ under the permissible limits. Use of electromagnetic device for magnetic microcomposites recovery from bioreactor yielded a maximum of 88% and 80.6% recovery for AA7 and MKU1, respectively. The rate of chromium recuperation achieved following desorption from the microcomposites of AA7 and MKU1 was 90.71% and 93.97%, respectively. Thus, the multifarious benefits including adsorption, metabolic detoxification, recovery, and recuperation by single functional microcomposites seems to be an intriguing and profitable approach for practicing in real-time operations to effectively remove heavy metals from the contaminated wastewater for environmental protection.
Show more [+] Less [-]Identification of epigenetic mechanisms in paddy crop associated with lowering environmentally related cadmium risks to food safety
2020
Feng, Sheng Jun | Liu, Xue Song | Ma, Li Ya | khan, Irfan ullah | Rono, Justice Kipkoir | Yang, Zhi Min
Cadmium (Cd) is a toxic metal that contributes to human diseases such as pediatric cancer and cardiovascular dysfunction. Epigenetic modification caused by Cd exposure is the major factor in etiology of environmentally-relevant diseases. However, the underlying epigenetic mechanism for Cd uptake and accumulation in food crops, particularly those growing in Cd-contaminated environments, is largely unknown. This study investigated uncharacterized regulatory mechanisms and biological functions of global DNA hypomethylation at CG sites that are associated with gene expression for Cd detoxification and accumulation in the food crop rice. Mutation of the CG maintenance enzyme OsMET1 confers rice tolerance to Cd exposure. Genome-wide analysis of OsMET1 loss of function mutant Osmet1 and its wild type shows numerous loci differentially methylated and upregulated genes for Cd detoxification, transport and accumulation. We functionally identified a new locus for a putative cadmium tolerance factor (here termed as OsCTF) and demonstrated that Cd-induced DNA demethylation is the drive of OsCTF expression. The 3′-UTR of OsCTF is the primary site of DNA and histone (H3K9me2) demethylation, which is associated with higher levels of OsCTF transcripts detected in the Osmet1 and Ossdg714 mutant lines. Mutation of OsCTF in rice led to hypersensitivity to Cd and the Osctf line accumulated more Cd, whereas transfer of OsCTF back to the Osctf mutant completely restored the normal phenotype. Our work unveiled an important epigenetic mechanism and will help develop breeding crops that contribute to food security and better human health.
Show more [+] Less [-]Nuclear receptor AHR-mediated xenobiotic detoxification pathway involves in atrazine-induced nephrotoxicity in quail (Coturnix C. coturnix)
2019
Zhang, Cong | Li, Huixin | Qin, Lei | Ge, Jing | Qi, Zhang | Talukder, Milton | Li, Yan-Hua | Li, Jin-Long
Atrazine (ATR), one of the most widely used pesticides in agricultural production, are gradually concerned due to potential ecosystem and health risks. Further, the induction of ATR nephrotoxicity and detoxification response is still unknown. To evaluate ATR-induced nephrotoxicity, quails were treated with 0, 50, 250 or 500 mg/kg ATR by gavage administration for 45 days. Histopathology indicated that ATR exposure caused renal tubular epithelial cell swelling and endoplasmic reticulum degeneration, suggesting that ATR exposure causes renal impairment even renal diseases. Notably, ATR interfered cytochrome P450 system (CYP450s) homeostasis by enhancing contents or activities of CYP450s (total CYP450, Cyt b5, AH, APND, NCR and ERND) and the expression of CYP450 isoforms (CYP1A, CYP1B, CYP2C and CYP3A). ATR triggered phase II detoxifying reaction, reflected by the elevated GSH level, GST activity and the up-regulation of GST isoforms (GSTa, GSTa3 and GSTt1) and GSH synthetase (GCLC). Moreover, ABC transporters were activated to expel ATR from the body by increasing expression of MRP1 and P-GP gene. Accompanying these alterations, the nuclear receptors (AHR, CAR and PXR) were activated by ATR in a dose-dependent manner. Analysis results of present study demonstrated that the induction of phase II detoxifying enzyme system and ABC transporters could be modulated by nuclear receptors response and CYP450s disturbance in low-dose ATR-treated quail. In conclusion, all data suggested that nuclear receptors AHR-mediated detoxification pathway was involved in ATR-induced nephrotoxicity. These results provided new evidence about the nephrotoxic effects of ATR on the response of biotransformation and detoxification system.
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