RESUMEN
4-Nitrophenol (4-NP), as a toxic and refractory pollutant, has generated significant concern due to its adverse effects. However, the potential toxic effects and mechanism remained unclear. In this study, the reproduction, development, locomotion and reactive oxygen species (ROS) production of Caenorhabditis elegans were investigated to evaluate the 4-NP toxicity. We used metabolomics to assess the potential damage mechanisms. The role of metabolites in mediating the relationship between 4-NP and phenotypes was examined by correlation and mediation analysis. 4-NP (8 ng/L and 8 µg/L) caused significant reduction of brood size, ovulation rate, total germ cells numbers, head thrashes and body bends, and an increase in ROS. However, the oosperm numbers in uterus, body length and body width were decreased in 8 µg/L. Moreover, 36 differential metabolites were enriched in the significant metabolic pathways, including lysine biosynthesis, ß-alanine metabolism, tryptophan metabolism, pentose phosphate pathway, pentose and glucuronate interconversions, amino sugar and nucleotide sugar metabolism, starch and sucrose metabolism, galactose metabolism, propanoate metabolism, glycerolipid metabolism, and estrogen signaling pathway. The mechanism of 4-NP toxicity was that oxidative stress caused by the perturbation of amino acid, which had effects on energy metabolism through disturbing carbohydrate and lipid metabolism, and finally affected the estrogen signaling pathway to exert toxic effects. Moreover, correlation and mediation analysis showed glycerol-3P, glucosamine-6P, glucosamine-1P, UDP-galactose, L-aspartic acid, and uracil were potential markers for the reproduction and glucose-1,6P2 for developmental toxicity. The results provided insight into the pathways involved in the toxic effects caused by 4-NP and developed potential biomarkers to evaluate 4-NP toxicity.
Asunto(s)
Caenorhabditis elegans , Estrógenos , Nitrofenoles , Reproducción , Transducción de Señal , Animales , Caenorhabditis elegans/efectos de los fármacos , Caenorhabditis elegans/fisiología , Reproducción/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Nitrofenoles/toxicidad , Estrógenos/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Estrés Oxidativo/efectos de los fármacosRESUMEN
Bisphenol A (BPA) and p-nitrophenol (PNP) are emerging contaminants of soils due to their wide presence in agricultural and industrial products. Thus, the present study aimed to integrate morpho-physiological, ionic homeostasis, and defense- and antioxidant-related genes in the response of tomato plants to BPA or PNP stress, an area of research that has been scarcely studied. In this work, increasing the levels of BPA and PNP in the soil intensified their drastic effects on the biomass and photosynthetic pigments of tomato plants. Moreover, BPA and PNP induced osmotic stress on tomato plants by reducing soluble sugars and soluble proteins relative to control. The soil contamination with BPA and PNP treatments caused a decline in the levels of macro- and micro-elements in the foliar tissues of tomatoes while simultaneously increasing the contents of non-essential micronutrients. The Fourier transform infrared analysis of the active components in tomato leaves revealed that BPA influenced the presence of certain functional groups, resulting in the absence of some functional groups, while on PNP treatment, there was a shift observed in certain functional groups compared to the control. At the molecular level, BPA and PNP induced an increase in the gene expression of polyphenol oxidase and peroxidase, with the exception of POD gene expression under BPA stress. The expression of the thaumatin-like protein gene increased at the highest level of PNP and a moderate level of BPA without any significant effect of both pollutants on the expression of the tubulin (TUB) gene. The comprehensive analysis of biochemical responses in tomato plants subjected to BPA and PNP stress illustrates valuable insights into the mechanisms underlying tolerance to these pollutants.
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Antioxidantes , Compuestos de Bencidrilo , Regulación de la Expresión Génica de las Plantas , Nitrofenoles , Fenoles , Solanum lycopersicum , Solanum lycopersicum/genética , Solanum lycopersicum/efectos de los fármacos , Solanum lycopersicum/metabolismo , Fenoles/toxicidad , Compuestos de Bencidrilo/toxicidad , Antioxidantes/metabolismo , Nitrofenoles/toxicidad , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/metabolismo , Hojas de la Planta/genética , Contaminantes del Suelo/toxicidad , Contaminantes del Suelo/efectos adversosRESUMEN
Nitrophenols are environmental pollutants and xenobiotics, the main sources of which are diesel exhaust fumes and pesticides. The biotransformation processes that take place in the liver are defence mechanisms against xenobiotics, such as nitrophenols. Our previous study showed that the chicken ovary is an additional xenobiotic detoxification place and that nitrophenols disrupt steroidogenesis in chicken ovarian follicles. Therefore, the present study aimed to determine the in vivo and in vitro effects of 4-nitrophenol (PNP) and 3-methyl-4-nitrophenol (PNMC) on the expression and activity of phase I (CYP3A) and phase II (COMT) biotransformation enzymes in chicken ovary. In an in vivo study, hens were treated with a vehicle or 10 mg PNP or PNMC/kg b.wt. per day for 6 days. In an in vitro study, prehierarchical white and yellowish follicles, as well as the granulosa and theca layers of the three largest preovulatory follicles (F3, F2 and F1), were isolated and then incubated in a control medium or medium supplemented with PNP (10-6 M) or PNMC (10-6 M) for 24 or 48 h. Both in vivo and in vitro studies showed that nitrophenols exert tissue- and compound-dependent (PNP or PNMC) effects on CYP3A and COMT gene (real-time PCR) protein (Western blot) expression and their activity (colorimetric methods). The inhibitory effect of nitrophenols in vivo on the activity of biotransformation enzymes suggest that the ovary has the capacity to metabolise PNP and PNMC.
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Pollos , Citocromo P-450 CYP3A , Femenino , Animales , Citocromo P-450 CYP3A/genética , Citocromo P-450 CYP3A/metabolismo , Folículo Ovárico/metabolismo , Ovario , Nitrofenoles/toxicidad , Nitrofenoles/metabolismoRESUMEN
Dysfunction of immune regulation plays a crucial role in the pathogenesis of many immune disorders in the body. The underlying mechanism is still not completely understood. Environmental pollution contributes to immune de-regulation. 3-methyl-4-nitrophenol (MNP) is one of the major environmental pollutants. This study aims to investigate the role of MNP in compromising immune regulatory functions in the intestine. A food allergy (FA) mouse model was established using ovalbumin (OVA) as the specific antigen. The activities of regulatory T cells in the mouse intestine were evaluated by flow cytometry and enzyme-linked immunosorbent assay. We found that MNP reduced the CD4+ Foxp3+ Treg frequency, increased Th17 cells, and converted Tregs to Th17 cells in the intestine. MNP induced the expression of IL-6 in regulatory T cells (Tregs). Estrogen receptor (ER) mediated the effects of MNP on promoting IL-6 expression in Tregs. The IL-6 in synergy with transforming growth factor (TGF)-ß to convert Tregs to Th17 cells. The concomitant exposure of MNP and OVA induced FA like response in mice. Modulation of the ER-STAT3-IL-6 signal pathway attenuated mouse FA response. In summary, MNP, an environmental pollutant, acts as an immunoadjuvant for developing FA. By activation of the estrogen receptor, MNP induces Tregs to express IL-6. IL-6 in synergy with TGF-ß converts Tregs to Th17 cells.
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Contaminantes Ambientales , Linfocitos T Reguladores , Ratones , Animales , Nitrofenoles/toxicidad , Nitrofenoles/metabolismo , Contaminantes Ambientales/metabolismo , Receptores de Estrógenos/metabolismo , Interleucina-6/metabolismo , Células Th17 , Ovalbúmina , Factor de Crecimiento Transformador beta/metabolismo , Intestinos , Factores de Transcripción Forkhead/metabolismo , Factores de Transcripción Forkhead/farmacologíaRESUMEN
4-Nitrophenol (PNP) has been extensively used in manufacturing for several decades. Its toxic effects on the male reproductive system have been reported, but the underlying mechanisms remain unclear. In this study, we utilized two testicular somatic cell lines (TM3 and TM4 cells) to explore the possible toxic effects of PNP on the male reproductive system. The activity of the cells after exposure to different doses of PNP (0.01, 0.1, 1, 10 and 100 µM) was evaluated. PNP treatment at 10 µM significantly inhibited cell viability, and 10 µM PNP was thus selected for subsequent experiments. Although PNP (10 µM) inhibited cell proliferation, promoted cell apoptosis, and changed the cell cycle distribution and ultrastructure in both types of cells, these effects were more significant in the TM4 cells. In addition, an Agilent mouse mRNA array was used to identify the gene expression differences between the control and PNP (10 µM) exposed TM3 and TM4 cells. The microarray analysis identified 67 and 1372 differentially expressed genes mainly concentrated in endothelial cell morphogenesis and anatomical structure development in TM3 cells and associated with cardiovascular system development and circulatory system development in TM4 cells. Moreover, a pathway analysis revealed that PNP not only predominately affected meiotic recombination and meiosis in TM3 cells, but also influenced axon guidance and developmental biology in TM4 cells. These results suggest that TM3 and TM4 cells exhibit different responses to PNP, which might mediate different toxic mechanisms.
Asunto(s)
Células Intersticiales del Testículo/efectos de los fármacos , Nitrofenoles/toxicidad , Células de Sertoli/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Orientación del Axón/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Regulación de la Expresión Génica/efectos de los fármacos , Células Intersticiales del Testículo/metabolismo , Masculino , Meiosis/efectos de los fármacos , Ratones , Nitrofenoles/administración & dosificación , Reproducción/efectos de los fármacos , Células de Sertoli/metabolismo , Testículo/citología , Testículo/efectos de los fármacosRESUMEN
Para-nitrophenol (PNP) is often detected in industrial wastewater that is discharged into municipal wastewater treatment plants. Intermittent discharge of PNP into municipal treatment facilities puts their biological process at risk of inhibition, and the risk is especially great for nitrification. In this work, nitrifying biomass was acclimated to PNP. The acclimated biomass retained most of its ammonium-removal activity when it was exposed to PNP at up to 100 mg/L, while the normal (unacclimated) biomass had nearly complete inhibition. PNP was effectively biodegraded by the acclimated biomass, but the normal biomass had minimal PNP biodegradation. After PNP disappeared, the acclimated biomass recovered its ability for NH4+-N removals within one to two days, but the normal biomass did not fully recovery even after seven days. The acclimated biomass had superior ability to sustain nitrification due to its ability to biodegrade PNP and its selection of nitrifying bacteria more resistant to PNP. The PNP-acclimated community was enriched in genera that could have been active in the biodegradation of PNP, such as Chloroflexi. Although the abundance of well-known nitrifiers, Nitrosomonas and Nitrospira, decreased, Nitrosospira and other genera within the Proetobacteria phylum increased, presumably because they were more resistant to PNP.
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Nitrificación , Aguas del Alcantarillado , Biomasa , Reactores Biológicos , Nitrofenoles/toxicidad , NitrosomonasRESUMEN
The toxic unit and additive index approaches were used to understand how 2 pesticides, 3-trifluoromethyl-4-nitrophenol (TFM) and 2,5-dichloro-4-nitrosalicylanilide (niclosamide; Nic), interact in mixtures. Our first objective was to determine whether the interaction was strictly additive or greater than additive at doses comparable to those used to control invasive sea lamprey (Petromyzon marinus) in the Laurentian Great Lakes, and our second was to compare the utility of the toxic unit and additive index models for determining how TFM and Nic interacted. Typically, TFM is mixed with Nic (1-2%, w/v) to increase its potency and reduce TFM use. However, there is little information on how the 2 chemicals interact. Using a well-studied, resident nontarget fish, the rainbow trout (Oncorhynchus mykiss), we conducted toxicity tests with TFM, Nic, and TFM:Nic (100:1, w/v; TFM/1% Nic) mixtures over 12 h to determine if the interaction was strictly additive, less than additive (antagonistic), or greater than additive (synergistic). The toxic unit and additive index approaches indicated synergistic interactions at environmentally relevant concentrations, suggesting that both are valid approaches for predicting how TFM and Nic interact. The toxic unit approach was simpler to conceptualize and to calculate, and we recommend that it be used when describing how TFM and Nic, and other similar organic compounds, interact with each other in aquatic ecosystems. Environ Toxicol Chem 2021;40:1419-1430. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.
Asunto(s)
Oncorhynchus mykiss , Animales , Ecosistema , Niclosamida , Nitrofenoles/toxicidadRESUMEN
Nitroaromatics belong to the group of toxic components of aerosol particles and atmospheric hydrometeors that enter the atmosphere through biomass burning and fuel combustion. In the present work, we report on the cytotoxic effects of a 2-, 3- and 4-nitrophenol mixture on a model eukaryotic-like cell membrane and compared it with in vitro cellular models BEAS-2B (immortalized bronchial epithelial cells) and A549 (cancerous alveolar epithelial cells). A selected model biomembrane comprised of DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) was studied. The electrochemical-based method, combined with atomic force microscopy (AFM) and phase-contrast microscopy imaging, allowed to get insights into the mechanism of cellular function disruption caused by airborne nitrophenols. The efficacy of the method is supported by the data obtained from in vitro experiments performed on cell models. The nitrophenol mixture exhibited cytotoxic effects at concentrations above 100 µg mL-1, as demonstrated by phase-contrast microscopy in real lung cell lines. Electrochemical impedance spectroscopy (EIS) revealed the formation of membrane defects at a nitrophenol concentration of 200 µg mL-1. AFM imaging confirmed the model membrane disintegration and phospholipids rearrangement in the presence of nitrophenols. These observations indicate that particle-bound nitrophenols induce substantial changes in cell membranes and make them more permeable to aerosol, resulting in major cellular damage in the lungs when inhaled. The study provides initial evidence of cellular membrane damage induced by three important nitrated phenols present in the environment.
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Células Eucariotas , Membrana Dobles de Lípidos , Membrana Celular , Nitrofenoles/toxicidad , Fosfatidilcolinas , FosfolípidosRESUMEN
3-Trifluoromethyl-4-nitrophenol (TFM) has been used for more than 60 yr to control the invasive parasitic sea lamprey (Petromyzon marinus) in the Great Lakes Basin (USA/Canada). In the early 1990s, researchers reported that TFM induced vitellogenin in fish and that TFM was an agonist for the rainbow trout estrogen receptor. To support continued registration of TFM for sea lamprey control, regulatory agencies required further testing to evaluate potential endocrine disruption effects. Fathead minnow (Pimephales promelas) were exposed to TFM at measured concentrations of 0.0659, 0.181, 0.594, 1.79, and 5.11 mg active ingredient (a.i.)/L for 21 d. No-observable- and lowest-observable-effect concentrations (NOEC and LOEC, respectively) were determined to be 1.79 mg/L or greater for each endpoint. Male survival in the highest treatment group was reduced relative to the controls. Percentage of egg fertility was reduced in the highest treatment group, resulting in an estimated NOEC of 1.79 mg/L. Whereas no effect on the gonadosomatic index (GSI) was observed for males, female GSI was increased in the 5.11-mg/L treatment. Vitellogenin production was not altered relative to the controls for all TFM treatment groups. However, female testosterone was elevated in the 5.11-mg/L treatment. The results suggest that prolonged exposure to TFM at concentrations exceeding 1.79 mg/L has the potential to disrupt endocrine function. Biologically relevant effects were found at the highest exposure concentration following a 21-d exposure. However, the duration of exposure in our study is not consistent with typical treatment durations (12 h) for sea lamprey control. Environ Toxicol Chem 2020;39:1599-1607. © 2020 SETAC.
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Bioensayo/métodos , Cyprinidae/fisiología , Disruptores Endocrinos/toxicidad , Nitrofenoles/toxicidad , Reproducción/efectos de los fármacos , Animales , Cyprinidae/sangre , Estradiol/sangre , Femenino , Larva/efectos de los fármacos , Masculino , Oncorhynchus mykiss/sangre , Oncorhynchus mykiss/fisiología , Petromyzon/fisiología , Control de Calidad , Testosterona/sangre , Vitelogeninas/metabolismoRESUMEN
The main source of nitroaromatic compounds in nature is anthropogenic activities. In the present report, a facile and green method was employed to fabricate highly efficient catalyst to turn toxic nitrophenols to more safe aminophenols. The Fe3O4/Ag@Ca-Al LDH hybrid (FAL hybrid) was synthesized by the Fe3O4 and Ag nanoparticles supported on layer double hydroxide (LDH) and the bionanocomposite (BNC) of this hybrid and starch was prepared via an in situ growth method. The FAL hybrid and BNC were characterized by different techniques. The reduction reaction of 4-nitrophenol in the presence of these catalysts indicated that both of them have high catalytic activity due to their large numbers of hydroxyl groups as capping agent. The comparison rate kinetics study between 4-nitrophenol and 2-nitrophenol showed that the FALS-BNC has higher potential in the case of the para derivative. Also, catalysts were recovered without difficulty and reused after completion of the reduction reaction.
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Catálisis , Tecnología Química Verde/métodos , Nanopartículas de Magnetita/química , Nanocompuestos/química , Nitrofenoles/química , Aminofenoles/química , Contaminantes Ambientales/química , Sustancias Peligrosas/química , Cinética , Nitrofenoles/toxicidad , Oxidación-Reducción , Plata/química , Almidón/químicaRESUMEN
Although health hazards of 4-nitrophenol (PNP) exposure have been reported, the adverse effects of PNP exposure on cancer biological features are still unknown. We investigated the effects of administration of PNP in T24 human bladder cancer cells. The results showed that PNP exposure promoted cellular proliferation, migration and invasion, inhibited adhesion and apoptosis in vitro. Using quantitative real-time PCR, we found that (1) the mRNA expression levels of cell-cycle regulators PCNA, cyclin D1 and COX-2 were increased in PNP-treated cells compared to controls, however, that of pro-apoptotic gene Bax was decreased; (2) the expression level of EMT-associated gene E-cadherin was decreased in PNP-treated cells, whereas those of N-cadherin, vimentin, snail, and slug were increased; (3) the expression levels of cancer-promoting genes HIF-1, IL-1ß, VEGFα and K-Ras were enhanced, but those of tumor suppressors p53, PTEN and BRCA were decreased. There was a positive association between PNP exposure times and the promotion effects. Finally, we found that the expression level of PPARγ (γ1 isoform) was increased in PNP-treated T24 cells. GW9662, a specific PPARγ antagonist, attenuated PNP-induced cell migration and invasion. These findings indicate that PNP exposure may promote bladder cancer growth and progression involving PPARγ signaling. PPARγ is a potential target for development of novel intervention study on environment pollution. Environ. Mol. Mutagen. 61:316-328, 2020. © 2019 Wiley Periodicals, Inc.
Asunto(s)
Carcinógenos Ambientales/toxicidad , Proliferación Celular/efectos de los fármacos , Disruptores Endocrinos/toxicidad , Transición Epitelial-Mesenquimal/efectos de los fármacos , Nitrofenoles/toxicidad , Neoplasias de la Vejiga Urinaria/inducido químicamente , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Progresión de la Enfermedad , Humanos , Invasividad Neoplásica/patología , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
p-Nitrophenol (PNP) is one type of environmental pollutant, which is difficult to degrade and soluble in water. To investigate the effects of PNP on economically important marine fish species, we subjected Larimichthys crocea juvenile to five different concentrations of PNP for 96â¯h, and the semi-lethal concentration (LC50) was 6.218â¯mg/L. Then we collected the liver, kidney, and gill tissues to determine enzyme activity and gene expression levels, and analyzed histological changes. In histological analysis, the gills showed curling of lamella, epithelial lifting and hyperplasia; the parenchymal structure of hepatocytes was significantly damaged, with severe vacuolation and loss of original structure. The renal cells were damaged too, with congestion and renal tubular necrosis. Catalase and superoxide dismutase both showed an up- and down-tendency with the rise of concentration in the three tissues, and GSH-px had similar trend in the kidney, which decreased at 8â¯mg/L in the liver but showed no significant differences in the gills. Malondialdehyde of three tissues was increased with an increase in PNP concentration. The expression of four detoxification (cyp450, gst, gpx, hsp70) and one immune-related (mhc II) genes was induced at low PNP concentrations but inhibited at high PNP concentrations in the kidney. In liver, cyp450, hsp70 and mhc II showed similar trend but gst and gpx didn't increase at low PNP concentrations. Our results indicate that the fish possesses the ability to detoxify PNP; however, at high concentrations, PNP still causes serious damage to them. Our data not only help in understanding the ability of L. crocea to detoxify PNP but also should serve as a basis for the study of toxic effects of nitrobenzenes on marine fish.
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Branquias/metabolismo , Riñón/metabolismo , Hígado/metabolismo , Nitrofenoles/toxicidad , Perciformes/metabolismo , Contaminantes Químicos del Agua/toxicidad , Animales , Catalasa/metabolismo , Proteínas de Peces/metabolismo , Inactivación Metabólica , Malondialdehído/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
The toxicity of an organophosphorus (OP) insecticide, methyl parathion (MP), and its hydrolysis product, p-nitrophenol (PNP), to the native Australian cladoceran species, Daphnia carinata, was assessed. Both MP and PNP were stable in cladoceran water during the test period. D. carinata was sensitive to both MP and PNP; however, the parent compound was more toxic than its metabolite. This is the first study that demonstrated the acute toxicity of MP and PNP towards an Australian daphnid species. The present investigation emphasizes the need for including the native taxa as non-target test organisms while evaluating the toxicity of environmental pollutants.
Asunto(s)
Daphnia/efectos de los fármacos , Insecticidas/toxicidad , Metil Paratión/toxicidad , Nitrofenoles/toxicidad , Animales , Australia , Relación Dosis-Respuesta a Droga , Pruebas de Toxicidad AgudaRESUMEN
The invasion of the Laurentian Great Lakes of North America by sea lampreys (Petromyzon marinus) in the early 20th century contributed to the depletion of commercial, recreational and culturally important fish populations, devastating the economies of communities that relied on the fishery. Sea lamprey populations were subsequently controlled using an aggressive integrated pest-management program which employed barriers and traps to prevent sea lamprey from migrating to their spawning grounds and the use of the piscicides (lampricides) 3-trifluoromethyl-4-nitrophenol (TFM) and niclosamide to eliminate larval sea lampreys from their nursery streams. Although sea lampreys have not been eradicated from the Great Lakes, populations have been suppressed to less than 10% of their peak numbers in the mid-1900s. The ongoing use of lampricides provides the foundation for sea lamprey control in the Great Lakes, one of the most successful invasive species control programs in the world. Yet, significant gaps remain in our understanding of how lampricides are taken-up and handled by sea lampreys, how lampricides exert their toxic effects, and how they adversely affect non-target invertebrate and vertebrates species. In this review we examine what has been learned about the uptake, handling and elimination, and the mode of TFM and niclosamide toxicity in lampreys and in non-target animals, particularly in the last 10 years. It is now clear that the mode of TFM toxicity is the same in non-target fishes and lampreys, in which TFM interferes with oxidative phosphorylation by the mitochondria leading to decreased ATP production. Vulnerability to TFM is related to abiotic factors such as water pH and alkalinity, which we propose changes the relative amounts of the bioavailable un-ionized form of TFM in the gill microenvironment. Niclosamide, which is also a molluscicide used to control snails in areas prone to schistosomiasis infections of humans, also likely works by uncoupling oxidative phosphorylation, but less is known about other aspects of its toxicology. The effects of TFM include reductions in energy stores, particularly glycogen and high energy phosphagens. However, non-target fishes readily recover from sub-lethal TFM exposure as demonstrated by the rapid restoration of energy stores and clearance of TFM. Although both TFM and niclosamide are non-persistent in the environment and critical for sea lamprey control, increasing public and institutional concerns about pesticides in the environment makes it imperative to explore other means of sea lamprey control. Accordingly, we also address possible "next-generation" strategies of sea lamprey control including genetic tools such as RNA interference and CRISPR-Cas9 to impair critical physiological processes (e.g. reproduction, digestion, metamorphosis) in lamprey, and the use of green chemistry to develop more environmentally benign chemical methods of sea lamprey control.
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Especies Introducidas , Niclosamida/toxicidad , Nitrofenoles/toxicidad , Plaguicidas/toxicidad , Petromyzon/crecimiento & desarrollo , Contaminantes Químicos del Agua/toxicidad , Animales , Humanos , Lagos/química , Larva/efectos de los fármacos , América del Norte , Fosforilación OxidativaRESUMEN
The hepatic cytochrome p450 enzymes 1 A, 2A19 and 2E1 is very important for the elimination of skatole from the body of pigs. Impaired skatole metabolism, results in skatole accumulation, which give rise to off flavor of the meat. Several metabolites of skatole has been identified, however the role of these metabolites in the inhibition of the skatole metabolizing enzymes are not documented. Using microsomes from pigs and fish, we determined the ability of several skatole metabolites to inhibit CYP1 A, CYP2A19 and CYP2E1 dependent activity. Our results show that 2-aminoacetophenone is an inhibitor of porcine CYP2A19 and CYP2E1 activity, but not the piscine orthologues. In conclusion, there is species specific differences in the inhibition of CYP1 A and CYP2A19 dependent metabolism of probe substrates. This is relevant to the evaluation of different model systems and to the reduction of off flavor of meat.
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Inhibidores del Citocromo P-450 CYP2E1/toxicidad , Hígado/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Escatol/toxicidad , Acetofenonas/toxicidad , Animales , Cumarinas/toxicidad , Citocromo P-450 CYP1A1/antagonistas & inhibidores , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP2E1/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Peces , Hígado/metabolismo , Masculino , Microsomas Hepáticos/efectos de los fármacos , Nitrofenoles/toxicidad , Oxazinas/toxicidad , Carne Roja/análisis , Alimentos Marinos/análisis , Especificidad de la Especie , PorcinosRESUMEN
PURPOSE: To evaluate if the common field lampricide 3-trifluoromethyl-4-nitrophenol (TFM) that is intended to eradicate the invasive species sea lampreys in the Great Lakes has the potential to sensitize radiation responses in cells from non-targeted native fish MATERIALS AND METHODS: The TFM toxicity was assessed acutely and chronically with the clonogenic fish cell line eelB. The acute toxicity (24-h exposure) was determined by the fluorescent cell viability probe Alamar Blue. The chronic toxicity was determined either by Alamar Blue (7-d exposure) or the clonogenic survival assay (14-d exposure). Pre- and post-exposure of fish cells to environmentally relevant TFM concentrations following gamma irradiation were performed. Clonogenic survival was determined to assess the damage level of radiation-induced reproductive cell death. RESULTS: The chronic toxicity tests were more sensitive than the acute toxicity tests. The 14-d EC50 using the clonogenic survival endpoint was 2.09⯱â¯0.28⯵g/mL and was statistically similar to the 7-d EC50 (1.85⯱â¯0.07⯵g/mL) based on the Alamar Blue-based cytotoxicity endpoint. Post-exposure of cells to environmentally relevant TFM concentrations following irradiation did not have any effect as compared to the irradiation alone group. In contrast, pre-exposure of cells to TFM following irradiation had a negative additive effect when the total radiation dose was 2â¯Gy, but not 0.1 or 0.5â¯Gy. CONCLUSION: Our results suggest that the common field lampricide TFM is a potential radiation sensitizer in cells from non-targeted native fish. This could be a health problem of concern for non-targeted native fish if a large accidental radioactive release occurs.
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Nitrofenoles/toxicidad , Fármacos Sensibilizantes a Radiaciones/toxicidad , Animales , Supervivencia Celular , Peces , Petromyzon/fisiologíaRESUMEN
CYP2E1 is a mammalian cytochrome P450 enzyme, which oxidizes a structurally diverse class of endogenous and exogenous (xenobiotic) compounds. Best studied is the role of CYP2E1 in phase I metabolism of xenobiotics including alcohol. CYP2E1 metabolizes ethanol and is active in generating reactive oxygen species (ROS) and subsequent oxidative stress in the hepatic tissues. Several studies have shown and discussed the importance of CYP2E1 in the hepatotoxic actions of alcohol. However, the vast majority assessed the CYP2E1 activity only in isolated microsomes. Here, we aimed to develop and optimize a fast and easy method to assess alcohol-induced CYP2E1 activity in hepatocytes in vitro applying oxidation of para-nitrophenol to para-nitrocatechol as specific substrate probe. Using hepatoma cells with and without stable CYP2E1 expression and primary human hepatocytes, we established specific methodology to assess CYP2E1 catalytic activity and its induction by ethanol in a small number of cells and in a very short time.
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Citocromo P-450 CYP2E1/metabolismo , Hepatocitos/efectos de los fármacos , Nitrofenoles/toxicidad , Bioensayo , Relación Dosis-Respuesta a Droga , Etanol/farmacología , Células Hep G2 , Hepatocitos/enzimología , Humanos , Nitrofenoles/metabolismo , Oxidación-Reducción , Cultivo Primario de CélulasRESUMEN
The pesticide 3-trifluoromethyl-4-nitrophenol (TFM) is used to control sea lamprey populations in the Great Lakes and Lake Champlain. Little is known about the effects of this pesticide on gene expression in eukaryotic species. This study used microarray analysis to determine the effects of short term, high dose TFM exposure on gene expression in Saccharomyces cerevisiae. Yeast grown in standard glucose-containing media showed significant variation in gene expression in pathways related to cytoplasmic translation with the majority of these genes being downregulated. These findings were supported by the analysis of a similar but glucose-free experiment revealing that these cytoplasmic translation genes, mostly ribosomal subunit expressing genes, were similarly downregulated. The repression of the ribosomal subunit genes suggests that TFM exposure, regardless of glucose availability, evokes features of the environmental stress response in yeast.
Asunto(s)
Nitrofenoles/toxicidad , Plaguicidas/toxicidad , Saccharomyces cerevisiae/efectos de los fármacos , Saccharomyces cerevisiae/fisiología , Estrés Fisiológico , Perfilación de la Expresión Génica , Análisis por Micromatrices , Biosíntesis de Proteínas/efectos de los fármacos , Ribosomas/efectos de los fármacos , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crecimiento & desarrolloRESUMEN
The piscicide 3-trifluoromethyl-4-nitrophenol (TFM) has been used to control invasive sea lamprey (Petromyzon marinus) populations in the Great Lakes for almost 60 years. Applied to rivers and streams containing larval lampreys, TFM seldom harms non-target fishes, but the effects of sub-lethal treatments on fish physiology are not well understood. We examined the effects of 9 h exposure to TFM on the stress axis and liver metabolic capacity of rainbow trout (Oncorhynchus mykiss) using in vivo and in vitro approaches. The fish that had been acutely exposed to TFM in vivo had increased plasma cortisol levels at 12 h post-treatment, but TFM exposure did not interfere with in vitro cortisol production in head kidney preparations. Subjecting trout to an acute handling stressor 12 h post-TFM exposure resulted in a relative attenuation of the plasma cortisol and glucose response compared to pre-stress levels. We conclude that routine TFM treatments can lead to elevations of plasma cortisol following exposure, plus a relative dampening of the stress response in rainbow trout, with high cortisol levels lasting at least 12 h post-treatment. Since the ability of the fish to produce cortisol and the liver metabolic capacity were not compromised following TFM exposure, it is likely that their ability to cope with other stressors is not altered in the long-term.
Asunto(s)
Riñón Cefálico/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Nitrofenoles/toxicidad , Oncorhynchus mykiss/fisiología , Estrés Fisiológico , Animales , Glucosa/metabolismo , Hidrocortisona/sangre , Hidrocortisona/metabolismo , Lagos , Larva/efectos de los fármacos , Plaguicidas/toxicidad , Petromyzon , Ríos , Factores de TiempoRESUMEN
Lampricides are currently being applied to streams and rivers to control the population of sea lamprey, an invasive species, in the Great Lakes. The most commonly used lampricide agent used in the field is 3-trifluoromethyl-4-nitrophenol (TFM), which targets larval sea lamprey in lamprey-infested rivers and streams. The specificity of TFM is due to the relative inability of sea lamprey to detoxify the agent relative to non-target fishes. There is increasing concern, however, about non-target effects on fishes, particularly threatened populations of juvenile lake sturgeon (LS; Acipenser fulvescens). There is therefore a need to develop models to better define lake sturgeon's response to TFM. Here we report the establishment of five LS cell lines derived from the liver, gill, skin and intestinal tract of juvenile LS and some of their cellular characteristics. All LS cell lines grew well at 25⯰C in Leibovitz's (L)-â¯15 medium supplemented with 10% FBS. All cell lines demonstrated high senescence-associated ß-galactosidase activity and varying levels of Periodic acid Schiff-positive polysaccharides, indicating substantial production of glycoproteins and mucosubstances by the cells. Comparative toxicity of TFM in the five LS cell lines was assessed by two fluorescent cell viability dyes, Alamar Blue and CFDA-AM, in conditions with and without serum and at 24 or 72â¯h exposure. Deduced EC50 values were compared between the cell lines and to the reported in vivo LC50s. Tissues sensitive to the effects of TFM in vivo correlated with cell lines from the same tissues being most sensitive to TFM in vitro. EC50 values for the LSliver-e cells was significantly lower than the EC50 for the rainbow trout (RBT) liver cells RTL-W1, reaffirming the in vivo observation that LS was generally more TFM-sensitive than rainbow trout. Our data suggests that whole-fish sensitivity of LS to TFM is likely attributable to sensitivity at the cellular level. Thus, LS cell lines, as well as those of RBT, can be used to screen and evaluate the toxicity of the next generation of lampricides on non-target fish such as lake sturgeon.