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1.
A hydrophilic form of acetylcholinesterase (AChE) was purified from N-methyl carbamate susceptible (SA) and highly N-methyl carbamate-resistant (N3D) strains of the green rice leafhopper (GRLH), Nephotettix cincticeps Uhler. Both of purified AChE from SA and N3D strains displayed the highest activities toward acetylthiocholine (ATCh) at pH 8.5. In the SA strain, the optimum concentrations for ATCh, propionylthiocholine (PTCh), and butyrylthiocholine (BTCh) were about 1 × 10−3, 2.5 × 10−3, and 1 × 10−3 M, respectively. However, in the N3D strain, substrate inhibition was not identified for ATCh, PTCh, and BTCh to 1 × 10−2 M. The Km value in the SA strain was 51.1, 39.1, and 41.6 μM and that in the N3D strain was 91.8, 88.1, and 85.2 μM for ATCh, PTCh, and BTCh, respectively. The Km value in the N3D strain indicated about 1.80-, 2.25-, and 2.05-fold lower affinity than that of the SA strain for ATCh, PTCh, and BTCh, respectively. The Vmax value in the SA strain was 70.2, 30.5, and 4.6 U/mg protein and that in the N3D strain was 123.0, 27.0, and 14.5 U/mg protein for ATCh, PTCh, and BTCh, respectively. The Vmax value in the N3D strain was 1.75- and 3.15-fold higher for ATCh and BTCh than that in the N3D strain. However, it was 1.13-fold lower for PTCh. The increased activity of AChE in the N3D strain is due to the qualitatively modified enzyme with a higher catalytic efficiency. The bimolecular rate constant (ki) for propoxur was 27.1 × 104 and 0.51 × 104 M−1 min−1 in the SA and N3D strain and that for monocrotophos was 0.031 × 104 and 2.0 × 104 M−1 min−1 in the SA and N3D strain. AChE from the N3D strain was 53-fold less sensitive than SA strain to inhibition by propoxur. In contrast, AChE from the N3D strain was 65-fold more sensitive to inhibition by monocrotophos than AChE from the SA strain. This indicated negatively correlated cross-insensitivity of AChE to propoxur and monocrotophos.  相似文献   

2.
The oriental tobacco worm, Helicoverpa assulta Guenée, is one of the most destructive pests of tobacco and peppers in China. We determined the susceptibility of H. assulta reared on an artificial diet, chili pepper and tobacco to four insecticides (fenvalerate, phoxim, methomyl, indoxacarb) under laboratory conditions associated with the activities of acetylcholinesterase (AChE), carboxylesterase (CarE) and glutathione S-transferase (GST) in its larvae. H. assulta larvae that were fed with chili pepper were more susceptible to fenvalerate, indoxacarb, and phoxim than those that were fed with tobacco and the artificial diet, but not to methomyl. The larvae that were fed with chili pepper were 3.65-, 2.49-, 1.92- and 2.44-fold more susceptible to fenvalerate, phoxim, methomyl, and indoxacarb than those fed with tobacco, respectively. The AChE activities of H. assulta larvae that were fed with chili pepper and tobacco were 2.12 and 1.07 μmol mg−1 15 min−1, respectively, almost 2-fold difference. The CarE activity of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 4.12, 7.40 and 7.12 μmol mg−1 30 min−1, respectively. Similarly, the GST activities of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 52.02, 79.37 and 80.02 μmol mg−1 min−1, respectively. H. assulta larvae that were fed with chili pepper were more resistance to the tested insecticides. The low activities of AChE and the high activities of CarE and GST lead to H. assulta become more susceptible to the tested insecticides.  相似文献   

3.
The toxicity of fenvalerate to the prawn Penaeus monodon was evaluated using biomarkers of stress. In a preliminary bioassay test, P. monodon was exposed to a series of fenvalerate concentrations, which showed 4, 6.5 and 8.5 μg L−1 to be sublethal, median lethal and lethal, respectively. Sublethal effect of fenvalerate was further evaluated in hepatopancreas, muscle and gills of prawns with reference to oxidative stress biomarkers. Significant induction of lipid peroxidation and glutathione-S-transferase activity was found in hepatopancreas, muscle and gills of prawns exposed to fenvalerate when compared to control (P < 0.001, P < 0.05 and P < 0.05). On the contrary, the activities of Superoxide dismutase, catalase, glutathione peroxidase, vitamin C, vitamin E and glutathione were found to be reduced in the experimental group of prawns when compared to control. The results suggest that the animals were under oxidative stress when exposed to sublethal concentration of fenvalerate.  相似文献   

4.
In vitro inhibition of electric eel acetylcholinesterase (AChE) by single and simultaneous exposure to organophosphorus insecticides diazinon and chlorpyrifos, and their transformation products, formed due to photoinduced degradation, was investigated. Increasing concentrations of diazinon, chlorpyrifos and their oxidation products, diazoxon and chlorpyrifos-oxon, inhibited AChE in a concentration-dependent manner. IC50 (20 min) values, obtained from the inhibition curves, were (in mol/l): (5.1 ± 0.3) × 10−8, (4.3 ± 0.2) × 10−6 and (3.0 ± 0.1) × 10−8 for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively, while maximal diazinon concentration was lower than its IC50 (20 min). Calculated KI values, in mol/l, of 7.9 × 10−7, 9.6 × 10−6 and 4.3 × 10−7 were obtained for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively. However, 2-isopropyl-4-methyl-6-pyrimidinol (IMP) and 3,5,6-trichloro-2-pyridinol, diazinon and chlorpyrifos hydrolysis products, did not noticeably affect the enzyme activity at all investigated concentrations. Additive inhibition effect was achieved for lower concentrations of the inhibitors (diazinon/diazoxon ?1 × 10−4/1 × 10−8 mol/l i.e., chlorpyrifos/chlorpyrifos-oxon ?2 × 10−6/3 × 10−8 mol/l), while an antagonistic effect was obtained for all higher concentrations of the organophosphates. Inhibitory power of 1 × 10−4 mol/l diazinon irradiated samples can be attributed mostly to the formation of diazoxon, while the presence of non-inhibiting photodegradation product IMP did not affect diazinon and diazoxon inhibitory efficiencies.  相似文献   

5.
The mechanism of resistance to quinclorac was investigated in a smooth crabgrass biotype [Digitaria ischaemum (Schreb. ex Schweig) Schreb. ex Muhl] from Tulare County, California. Quinclorac (8.96 kg a.i. ha−1) had no effect (P = 0.18) on the resistant (R) biotype, but reduced fresh weight of a susceptible (S) biotype by 93%. After treatment with 4.48 kg a.i. quinclorac ha−1, the S biotype produced about three times more ethylene than the R biotype and accumulated cyanide in tissues. Similar amounts of endogenous cyanide resulting from treatment with KCN reproduced quinclorac phytotoxicity. Pre-treatment with the ACC synthase inhibitor AVG reduced quinclorac phytotoxicity by 37% and ethylene production by 89%. These data suggest a target site-based mechanism of resistance involving stimulation of ACC synthesis and accumulation of cyanide. Also, the R biotype had four times more β-cyanoalanine synthase activity than the S biotype, suggesting a higher ability to detoxify cyanide.  相似文献   

6.
Topical laboratory selection of tobacco budworm larvae, Heliothis virescens, with technical spinosad for multiple generations resulted in larvae 1068-fold resistant to topical applications of the insecticide and 316.6-fold resistant to insecticide treated diet as compared to the parental strain. The penetration of 2′-O-methyl[14C]spinosyn A across the cuticle of the susceptible (parental) and selected (resistant) tobacco budworms increased with time 3-12 h after application. A trend of reduced penetration in the resistant strain was found but the differences were not statistically significant. 2′-O-methyl[14C]spinosyn A when injected into the hemocoel was not metabolized 96 h after treatment in both the susceptible and resistant strain, suggesting that a change in metabolism was not the mechanism of resistance. Electrophysiological studies indicated that dose-dependent spinosyn A-induced currents occurred in neurons from spinosyn resistant and susceptible (adult) tobacco budworms. At both 10 and 100 nM spinosyn A, however, the amplitude of these currents in the resistant insects was significantly smaller than the amplitude of currents observed from neurons from susceptible tobacco budworm adults. This suggests that neurons from resistant insects have decreased sensitivity to spinosyn A. However, the reduced inward currents in the resistant strain may or may not be related to the mode of action of the spinosyns. No statistically significant cross-resistance was noted for the spinosad resistant tobacco budworms for topical applications of permethrin (Pounce®), profenofos (Curacron®), emamectin benzoate (Denim®), or indoxacarb (Steward®). A statistically significant reduction in susceptibility to acetamiprid (Mospilan®) in artificial diet as determined from a resistance ratio of 0.482 was found.  相似文献   

7.
Effects of deltamethrin on voltage-sensitive calcium channels (VSCC) from rat brain (Cav2.2) expressed in Xenopus oocytes were assessed electrophysiologically. Deltamethrin reduced peak current of wild-type Cav2.2 in a stereospecific and concentration-dependent manner with an EC50 of 1 × 10−9 M. Phosphorylation of threonine 422 enhances voltage-sensitive calcium current, increases the probability that Cav2.2 will open under depolarizing conditions and antagonizes the inhibition of the channel by the betagamma subunit of heterotrimeric G-protein (Gβγ). Site-directed mutagenesis of threonine 422 to glutamic acid (T422E) results in a channel that acts as if it were permanently phosphorylated. Deltamethrin (10−7 M) significantly enhanced peak current via the T422E channel (1.5-fold) compared to the nontreated control and the increase was significantly greater than for either the wild-type (T422) or T422A (permanently unphosphorylated mutant) channels. The effect of deltamethrin on T422E Cav2.2 was stereospecific and concentration-dependent with an EC50 of 9.8 × 10−11 M. Thus, Cav2.2 is modified by deltamethrin but the resulting perturbation is dependent upon the phosphorylation state of threonine 422.  相似文献   

8.
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9.
Trehalase, with the target to control insects, nematodes and fungi, is of increasing interest and has been investigated extensively in recent years. Validamycin compounds, as competitive trehalase inhibitors and lead compounds with broad applications have attracted substantial attention as well. In this study, the characterizations of termites trehalase were investigated and the inhibitory effects of validamycin compounds on the termites trehalase were studied as well. Results showed that the termites trehalase is presumably belonging to the acid trehalase with optimal pH of 3.3 and optimal temperature of 37 °C. It was investigated that the concentrations of validoxylamine A (VAA), validoxylamine B (VBB), validamycin A (VA) and validamycin B (VB) required for 50% inhibition IC50 of termites trehalase were calculated to be 14.73 mg l−1, 20.80 mg l−1, 3.17 × 103 mg l−1and 2.24 × 103 mg l−1, respectively. The inhibition kinetic constant Ki values for the above validamycin compounds were 3.2 × 10−6 mol l−1, 1.03 × 10−5 mol l−1, 4.02 × 10−4 mol l−1and 2.69 × 10−4 mol l−1, respectively. Validoxylamine A appeared to be the most potential termites trehalase inhibitor among the four compounds.  相似文献   

10.
The present study was conducted to determine the 96 h-LC50 of benomyl to the Nile tilapia, Oreochromis niloticus and to investigate the biochemical or hematological indices of blood and the alterations in the antioxidant enzymes of this fish in response to sublethal concentrations of benomyl. Fish weighing 71.61 ± 12.05 g were used in this study; they were subjected to fasting for 4 weeks before treatment. An aqueous solution of benomyl (0, 0.5, 1, 2, 4, 8, and 16 mg L−1) was administered for 96 h to determine the LC50. The 96 h-LC50 value of benomyl was 4.39 (3.23-5.60) mg L−1 in the present study. For 5 weeks, the aqueous solution of benomyl (0, 100, 200, and 400 μg L−1) was administered to investigate its effect on the hematological parameters and antioxidant enzymes. The predominant hematological findings in fish exposed to benomyl were as follows: no significant change in the Hb (g dL−1) level, MCV (μm3), MCH (pg) and MCHC (%) as compared to the control. Benomyl exposure led to greater increases in the GPT, GOT (Karmen-unit), LDH (Wroblewski unit), total cholesterol, Fe, and Ca (mg dL−1) values, whereas the levels of ALP (KA unit), total protein, triglyceride, albumin, and Mg (mg dL−1) did not increase. Benomyl increased the in vivo HSI (%), GST (nmol min−1 mg protein−1), and SOD (U mg protein−1) values in the fish livers in the test group, unlike those in the control group for 5 weeks. At concentrations higher than 100 μg L−1, benomyl affected the GST and SOD levels of Nile tilapia in a dose- and time-dependent manner. The present findings suggest that the in vivo hepatotoxicity associated with benomyl may, in part, result from the hematological index, and antioxidants may provide limited protection against benomyl toxicity.  相似文献   

11.
Elevated oxidative detoxification is a major mechanism responsible for pyrethroid resistance in Helicoverpa armigera from Asia. Constitutive overexpression of CYP9A12 and CYP9A14 was associated with pyrethroid resistance in the YGF strain of H. armigera. CYP9A12 and CYP9A14 were functionally expressed in the W(R) strain of yeast (Saccharomyces cerevisiae) transformed with a plasmid shuttle vector pYES2. The cell lysates prepared from yeast transformed with CYP9A12 and CYP9A14, respectively, exhibited considerable O-demethylation activities against two model substrates p-nitroanisole (0.59 and 0.42 nmol p-nitrophenol min−1 mg protein−1) and methoxyresorufin (2.98 and 5.41 pmol resorufin min−1 mg protein−1), and clearance activity against the pyrethroid esfenvalerate (8.18 and 4.29 pmol esfenvalerate min−1 mg protein−1). These results provide important evidence on the role of CYP9A12 and CYP9A14 in conferring pyrethroid resistance in H. armigera, and also demonstrate that the yeast expression system can provide necessary redox environment for insect P450s to metabolize xenobiotics.  相似文献   

12.
Evolution of resistance by pests is the greatest threat to the continuous success of theBacillus thuringiensis (Bt) toxins used in conventional sprays or in transgenic plants. The most common mechanism of insect resistance to Bt is reduced binding of toxins to target sites in the brush border membrane of the larval mid-gut. In this paper, binding experiments were performed with three 125I-Cry1A toxins and the brush border membrane vesicles from Cry1Ac resistant or susceptible strains of Helicoverpa armigera. The homologous competition test showed that there was no significant difference in Cry1Ac-binding affinity, but the concentration of Cry1Ac-binding sites dramatically decreased in the resistant strain (Rt decreased from 5.87 ± 1.40 to 2.23 ± 0.80). The heterologous competition test showed that there were three Cry1Ac-binding sites in the susceptible strain. Among them, site 1 bound with all three Cry1A toxins, site 2 bound with both Cry1Ab and Cry1Ac, and site 3 only bound with Cry1Ac. In the Cry1Ac resistant strain, the binding capability of site 1 with Cry1Ab decreased and site 2 did not bind with Cry1Ac. It is suggested that the absence of one binding site is responsible for H. armigera resistance to Cry1Ac. This result also showed that the resistance fitted the “mode 1” pattern of Bt resistance described previously.  相似文献   

13.
Organophosphorus pesticides (OPs) are of environmental significance due to their high toxicity to animals. Binding to plasma proteins may effective influence the toxicological properties of xenobiotics. In an attempt to evaluate the affinity of phenthoate (PTA) to bovine serum albumin (BSA) and inhibitory ability of bound PTA to acetylcholinesterase (AChE), we investigated the interactions between phenthoate (PTA) and bovine serum albumin (BSA) using tryptophan fluorescence quenching and subsequent inhibition on AChE activity by PTA. The results showed that PTA caused the fluorescence quenching of BSA because of the formation of a PTA-BSA complex. Quenching constants (Ksv), determined using the Sterns-Volmer equation to provide a measure of the binding affinity between PTA and BSA at 303, 306, 310 and 313 K were (3.4295 ± 0.0763) × 10−4, (3.2446 ± 0.0635) × 10−4, (3.0434 ± 0.0856) × 10−4 and (2.8262 ± 0.0569) × 10−4 M−1, respectively. The thermodynamic parameters, ΔH and ΔS were −25.04 kJ mol−1 and 168.94 J mol−1 K−1, respectively, which indicated that the electrostatic interactions played a major role in PTA-BSA association. The presence of BSA consistently reduced the inhibitory ability of PTA on AChE, with the relative activity being increased from 46.98 to 61.71% for the concentration range of BSA between 0 and 4.0 g L−1.  相似文献   

14.
Multiwalled carbon nanotubes-polymeric methyl red film modified electrode (MWNT-PMRE) was made. The electrochemical behavior of carbendazim on modified electrode was studied with Cyclic Voltammetry, Linear Sweep Voltammetry, Stable Polarization Method and Chronocoulometry. The results indicated that the electrical oxidation of carbendazim on MWNT-PMRE in H2SO4 supporting electrolyte with concentration of 0.6 mol/L was irreversible and was mainly controlled by diffusion. Some parameters of the electrochemical process were evaluated. The impacts of experiment conditions on the electrochemical behavior of carbendazim were studied. A good linearity relationship between peak current and concentration of carbendazim in the range of 2.0 × 10−7-1.0 × 10−5 mol/L was found, of which the equation was Ip(A) = −1.149 × 10−5 − 2.301c (mol/L), the correlative coefficient R = −0.9953 and detection limit was 9.0 × 10−9 mol/L. The recovery was between 90.3% and 94.7%.  相似文献   

15.
The in vitro inhibition potency of some organophosphates (OPs) and carbamates (CAs) which are widely used to control plant-parasitic nematodes on acetylcholinesterase (AChE) of Meloidogyne javanica, Heterodera avenae and Tylenchulus semipenetrans, the major pathogens responsible for the damage of a wide range of crops in Al-Qassim region, Saudi Arabia was examined. AChE of H. avenae activity was 1.58- and 1.51-fold greater than that of T. semipenetrans or M. javanica, respectively. The order of inhibition potency of the tested compounds against T. semipenetrans AChE was: carbofuran > paraoxon > oxamyl > fenamiphos > phorate-sulfoxide > aldicarb, where the corresponding concentrations that inhibited 50% of the nematode AChE activity (I50) were 5 × 10−8, 7 × 10−7, 7.5 × 10−7, 2 × 10−6, 2 × 10−4 and 2 × 10−3 M, respectively. Paraoxon, fenamiphos and carbofuran exhibited high inhibition potency against M. javanica AChE where the I50 values were below 1 nM. Phorate-sulfoxide and aldicarb were potent inhibitors of M. javanica AChE with I50 values of 3.8 and 8 nM, respectively, while oxamyl exhibited low inhibition potency with I50 of 15 nM. Fenamiphos and paraoxon showed the highest I50 values of <100 μM against H. avenae followed by oxamyl (I50 < 1 mM), whereas paraoxon, carbofuran and aldicarb showed low potency with I50 values >1 mM. All the tested compounds exhibited high inhibition potency to AChE of M. javanica than T. semipenetrans or H. avenae. Except phorate-sulfoxide in M. javanica the inhibition pattern and implied mechanism for all the tested compounds for the three nematodes is suggested to be a linear mixed type (a combination of competitive and non-completive type).  相似文献   

16.
Recovery study was performed at regular intervals to establish the time course of 50% and 100% recovery in neurotransmitter enzyme (acetylcholinesterase, AChE, EC 3.1.1.7) and locomotor behaviour response of mosquito fish, Gambusia affinis exposed to lethal concentration (20.49 mg L−1) of an organophosphorous pesticide, monocrotophos (MCP) for 96 h. In vitro AChE activity studies indicated that MCP could cause 50% inhibition (I50) at 10.2 × 10−5 M. A positive correlation was observed between brain AChE activity and swimming speed during the recovery study. Also, the recovery response of the antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) as well as lipid peroxidation (LPO) as biomarkers of oxidative stress were assessed in viscera of G. affinis. The results showed that the MCP besides its inhibitory effect on target enzyme AChE activity and induction in antioxidant enzyme activities as a characteristic of oxidative stress, which can be used as biomarkers in the pesticide contaminated aquatic streams.  相似文献   

17.
The effects of lethal and sublethal concentrations of chlorpyrifos and endosulfan on oxygen consumption and ammonia excretion rate of the crab Trichodactylus borellianus were evaluated. Oxygen consumption and energy expenditure had significant effect in relation to exposure times. Regarding endosulfan, a significant difference in consumption among times of exposure was registered in 625 μg L−1. Moreover, at the highest concentration, energy expenditure rate was observed stabilized during 1-3 h. A significant increase in ammonia excretion was evidenced in 150 and 300 μg L−1 of chlorpyrifos. The O:N ratio showed a decrease in chlorpyrifos and in 2500 μg L−1 of endosulfan. This indicated a shift towards protein primary metabolism. An increment in the O:N ratio was observed in the lower endosulfan solutions. The relation oxygen:nitrogen showed a shift towards lipid and carbohydrate primary metabolism. This work indicated the complexity of the metabolism in the freshwater crab affected by xenobiotic elements.  相似文献   

18.
Molecular mechanisms of monocrotophos resistance in the two-spotted spider mite (TSSM), Tetranychus urticae Koch, were investigated. A monocrotophos-resistant strain (AD) showed ca. 3568- and 47.6-fold resistance compared to a susceptible strain (UD) and a moderately resistant strain (PyriF), respectively. No significant differences in detoxification enzyme activities, except for the cytochrome P450 monooxygenase activity, were found among the three strains. The sensitivity of acetylcholinesterase (AChE) to monocrotophos, however, was 90.6- and 41.9-fold less in AD strain compared to the UD and PyriF strains, respectively, indicating that AChE insensitivity mechanism plays a major role in monocrotophos resistance. When AChE gene (Tuace) sequences were compared, three point mutations (G228S, A391T and F439W) were identified in Tuace from the AD strain that likely contribute to the AChE insensitivity as predicted by structure analysis. Frequencies of the three mutations in field populations were predicted by quantitative sequencing (QS). Correlation analysis between the mutation frequency and actual resistance levels (LC50) of nine field populations suggested that the G228S mutation plays a more crucial role in resistance (r2 = 0.712) compared to the F439W mutation (r2 = 0.419). When correlated together, however, the correlation coefficient was substantially enhanced (r2 = 0.865), indicating that both the F439W and G228S mutations may work synergistically. The A391T mutation was homogeneously present in all field populations examined, suggesting that it may confer a basal level of resistance.  相似文献   

19.
The silphinenes are tricyclic sesquiterpenes that have antifeedant and toxic effects in insects and structural similarity to the known GABA antagonist, picrotoxinin. In murine synaptoneurosomes, silphinenes block GABA-stimulated influx of 36Cl− with EC50s in the range of 10-30 μM. In insects, silphinenes were tested in neurophysiological recordings of central neurons from third instar Drosophila melanogaster larvae. Silphinenes reversed the blockage of neuronal firing induced by GABA, but had little effect below 100 μM. The structure-activity profile observed in the murine chloride flux assay was also observed in the larval neurophysiological assay, indicating little selectivity for the silphinenes. A reference silphinene was equally active on nerve preparations from the rdl strain of D. melanogaster, which is resistant to channel-blocking antagonists via an altered GABA receptor. This latter finding suggests that silphinenes interact with the insect GABA receptor in a manner somewhat different from PTX, and that rdl resistance in the field may have little effect on silphinene efficacy.  相似文献   

20.
Experiment was carried out to determine the median lethal concentration (LC50) of fenvalerate to Labeo rohita fingerlings. After determining the LC50 value of fenvalerate, a sub-lethal concentration (1/3rd of LC50) of fenvalerate was exposed for 15 days. Significant alterations in SOD (P < 0.05) activity of liver and gill was observed due to fenvalerate. Catalase activity in gills of fishes was also affected significantly (P < 0.05). WBC, NBT and Hct values were reduced significantly in fenvalerate exposed fishes as compared to control group, whereas blood glucose level showed higher values in fenvalerate exposed group. Serum total protein and albumin were also reduced significantly as a result of fenvalerate exposure. Significant increase in the serum GOT, serum GPT, creatinine, triglyceride and serum ACP was noticed after 15 days of fenvalerate exposure. Results indicated that short term exposure of fenvalerate can induce biochemical and haematological alterations causing stress to L. rohita fingerlings.  相似文献   

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