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1.
The susceptibility to trypsin of conglutin gamma, a lupin seed glycoprotein affected by this enzyme only when in a non-native conformation, was used to study the effect of Zn(2+) and other metal ions on the structural dynamics of the protein. When acid-treated trypsin-susceptible conglutin gamma was incubated at neutral pH in the presence of Zn(2+), it became resistant to tryptic attack, contrary to the protein treated in the absence of Zn(2+). The time course of this refolding event has been quantitatively evaluated by SDS-PAGE. Amino acid sequencing of the major polypeptide fragments, produced by trypsin before completion of the refolding process, indicated that only a few cleavable bonds were accessible to the enzyme. This suggested that the presence of metal ions affected the pathway of degradation of the protein, by inducing its folding. Among the other metal ions tested, Ni(2+) also promoted the adoption of a trypsin-resistant conformation of conglutin gamma, whereas Mn(2+) and Ca(2+) had only much lower effects. The relevance of these findings for a deeper understanding of the in vivo degradation of plant food proteins and how it is affected by metal ions are discussed.  相似文献   

2.
The presence of ethylenediamine-N-(o-hydroxyphenylacetic)-N'-(p-hydroxyphenylacetic) acid (o,p-EDDHA) as the second largest component in commercial EDDHA iron chelates has recently been demonstrated. Here is reported the speciation of o,p-EDDHA by the application of a novel methodology through the determination of the complexing capacity, protonation, and Ca(2+), Mg(2+), Cu(2+), and Fe(3+) stability constants. The pM values and species distribution in solution, hydroponic, and soil conditions were obtained. Due to the para position of one phenol group in o,p-EDDHA, the protonation constants and Ca and Mg stability constants have different values from those of o,o-EDDHA and p,p-EDDHA regioisomers. o,p-EDDHA/Fe(3+) stability constants are higher than those of EDTA/Fe(3+) but lower than those of o,o-EDDHA/Fe(3+). The sequence obtained for pFe is o,o-EDDHA/Fe(3+) >/= o,p-EDDHA/Fe(3+) > EDTA/Fe(3+). o,p-EDDHA/Fe(3+) can be used as an iron chelate in hydroponic conditions. Also, it can be used in soils with limited Cu availability.  相似文献   

3.
The β-carotene bleaching assay, a common method for evaluating antioxidant activity, has been widely criticized due to its low reproducibility, problematic quantification, complex reagent preparation, and interference of different factors (temperature, pH, solvents, and metals). In this work we have examined the effects of these factors and developed a highly reproducible procedure for microplate assay, evaluated the critical points of the method, and proposed a kinetic model for quantifying both antioxidant and prooxidant activities. The application of these tools produced very consistent results, which provide robust and meaningful criteria to compare in detail the characteristics of several well-known commercial antioxidants, as well as several predictable prooxidants, and can be easily applied to natural extracts, food samples, and many other type of compounds. As an example, we have tested a set of commercial antioxidants and some typical lipophilic prooxidants. The activity of the tested antioxidants decreased in the following order: ethoxyquin ? α-tocopherol > butylhydroxyanisole > butylhydroxytoluene ? propyl gallate. On the other hand, hemoglobin and Fe(2+), Fe(3+), Co(2+), and Cu(2+) showed a strong prooxidant effect, and the activity was null in Cd(2+), Ni(2+), and Sr(2+), slightly antioxidant in Mg(2+), and strongly antioxidant in Zn(2+) and Mn(2+).  相似文献   

4.
Antioxidative ability of lactic acid bacteria   总被引：24，自引：0，他引：24  
Nineteen strains of lactic acid bacteria were investigated for antioxidative activity. These includedLactobacillus acidophilus B, E, N1, 4356, LA-1, and Farr; Lactobacillus bulgaricus 12 278, 448, 449, Lb, 1006, and 11 842; Streptococcus thermophilus 821, MC, 573, 3641, and 19 987; and Bifidobacterium longum B6 and 15 708. Intracellular cell-free extract of all strains demonstrated antioxidative activity with inhibition rates of ascorbate autoxidation in the range of 7-12%. Antioxidative mechanisms including metal ion chelating ability, scavenge of reactive oxygen species, enzyme inhibition, and reducing activity of intracellular cell-free extract of lactic acid bacteria were studied. S. thermophilus 821 had the highest metal ion chelating ability for Fe(2+), and B. longum 15 708 showed the highest Cu(2+) chelating ability among the 19 strains tested. All strains demonstrated reactive oxygen species scavenging ability. L. acidophilus E showed the highest hydroxyl radical scavenging ability, and B. longum B6 had the best hydrogen peroxide scavenging ability. Reducing activity was also found in all strains. Most of the strains tested demonstrated excellent reducing activity. B. longum B6 showed the highest reducing activity among the 19 strains tested. In enzyme inhibition, superoxide dismutase activity was not found in these 19 strains, and the activity of superoxide dismutase was not induced when metal ion Mn(2+), Fe(2+), or Cu(2+)Zn(2+) was present.  相似文献   

5.
The hyphomycete Chalara (syn. Thielaviopsis) paradoxa produces endoglucanase activity during the late trophophase. The low molecular mass (35 kDa) endoglucanase purified from cultured broths works optimally at 37 degrees C and pH 5.0. The enzyme inactivates at pH below 3.0 and also at temperatures of 50 degrees C or higher, but it is stable at lower temperatures, including refrigeration temperature and freezing. The enzyme is inhibited by detergents, by EDTA, and by the divalent cations Hg(2+) and Ag(2+). It is also inhibited to some extent by 10 mM Zn(2+), Fe(2+), and Mg(2+), but it is stimulated by Mn(2+). Enzyme activity is not affected by reducing agents. In the presence of low concentrations of water miscible organic solvents (20%) endoglucanase activity is inhibited by 7% (for methanol) to 50% (for acetonitrile), and it is totally inhibited at higher solvent concentrations (50%). Enzyme activity is not affected by the water immiscible solvent ethyl acetate. Carboxymethylcellulose is the preferred substrate (K(m(app)) = 8.3 g/L; V(max(app)) = 1.1 microM/min). Hydrolysis of crystalline cellulosic substrates is very limited, but it is greatly enhanced by phosphoric acid swelling. The purified enzyme shows no activity toward disaccharides or aryl-glucosides. Its activity is inhibited by cellobiose.  相似文献   

6.
Xylan-rich hemicelluloses-based hydrogel was developed as a novel porous bioadsorbent by graft co-polymerization of acrylic acid (AA) and xylan-rich hemicelluloses for adsorption of heavy metal ions (Pd(2+), Cd(2+), and Zn(2+)) from aqueous solutions. The chemical structure, the interaction between the hydrogel and metal ions, and the porous structure of xylan-rich hemicelluloses-g-AA hydrogel were revealed by Fourier transform infrared spectroscopy and scanning electron microscopy. The effects of AA and cross-linker dosage, pH value, contacting time, and initial concentration of metal ion on the adsorption capacity were studied. The adsorption equilibrium time was about 60 min from the adsorption kinetics study. The maximum adsorption capacities of Pd(2+), Cd(2+), and Zn(2+) were 859, 495, and 274 mg/g, respectively. Furthermore, xylan-rich hemicelluloses-g-AA hydrogel also exhibited highly efficient regeneration and metal ion recovery efficiency and can be reused without noticeable loss of adsorption capacity for Pd(2+), Cd(2+), and Zn(2+) after quite a number of repeated adsorption/desorption cycles.  相似文献   

7.
Aluminum (Al) and chromium (Cr) stresses often occur simultaneously in agricultural soils, and pose a great damage to crop growth, yield formation and product safety. In the current study, the influence of combined Al and Cr stresses on plant biomass, metal and nutrient contents was determined in comparison with that of Al or Cr stress alone. A hydroponic experiment was conducted to investigate the effect of pH, Al and Cr in the medium solution on the uptake of mineral elements as well as Al and Cr in the two barley genotypes differing in Al tolerance. Aluminum sensitive genotype Shang 70-119 had significantly higher Cr and Al contents in plants than Al-tolerant genotype Gebeina. Barley roots had much higher Al and Cr contents than above-ground plant parts. Chromium contents were much higher in the solution with pH 4.0 than in that with pH 6.5. Aluminum stress reduced phosphorus (P), calcium (Ca), magnesium (Mg), sulfur (S), copper (Cu), manganese (Mn), zinc (Zn) and boron (B) contents in roots and restrained potassium (K) and iron (Fe) from being translocated into shoots and leaves. Chromium stress resulted in reduced P, K, Mg, S, Fe, Zn and Mn contents in roots at pH 6.5 and P, K, Ca, Mg, S, Zn and Mn contents at pH 4.0. Translocation of all nutrients from roots to upper parts of plants was inhibited except Ca in pH 6.5 with Cr addition. Lower contents of all nutrients were observed at pH 4.0 as compared to pH 6.5. Combined stress of Cr and Al, on the whole, caused further reduction in mineral content in all plant parts of the two barley genotypes as compared to Al or Cr stress alone. Moreover, the reduction was more pronounced in Al sensitive genotype Shang 70-119.  相似文献   

8.
研究了酵母膏、葡萄糖、蛋白胨、土壤浸液及Ca2 、Mg2 、Fe2 、Fe3 、Al3 、Mn2 、Co2 、Zn2 、Li 、Cu 、Cu2 、Ba2 、Ni2 等 13种金属离子对甲基对硫磷降解菌DLL E4降解对硝基苯酚的影响。结果表明 :适量添加酵母膏、葡萄糖和蛋白胨都能有效提高菌株对对硝基苯酚的降解 ,土壤浸液没有影响 ;金属离子中 ,除Li 和 0 1mmolL-1Fe3 外均对DLL E4降解对硝基苯酚的性状有一定的影响 ,其中Ca2 、Mg2 、Mn2 影响不大 ,Fe3 、Fe2 、Al3 、Ba2 、Zn2 高浓度时影响较大 ,Co2 、Cu2 、Cu 、Ni2 对DLL E4降解对硝基苯酚有较大的影响  相似文献   

9.
《Journal of plant nutrition》2013,36(10-11):2253-2265
Abstract

T2 tobacco lines overexpressing soybean ferritin in the plastids (+TPs) or apoplasm (AFs) under the regulation of CaMV 35S promoter were grown on MS nutrient solution. After 1 month growth, statuses of six major divalent‐metals (Ca, Cu, Fe, Mg, Mn, and Zn) were measured in leaves and roots. Both +TPs and AFs showed enhanced growth (max. 1.7×) in leaves than the control line. The Fe contents in leaves of +TPs and AFs were significantly larger (1.9–2.8×) than that of the control line. The other metal contents in leaves of +TPs and AFs were almost the same as or less than those of the control line. In contrast to the result of leaves, the growth enhancement in roots was not clear in +TPs, but in AFs. Also, some of the non‐ferrous metal contents in roots of +TPs and AFs were dramatically increased compared with those of the control line (Mn, 1.9–10.4×; Zn, 1.6–2.3×), whereas the differences in content of Fe, Cu, Ca, and Mg were insignificant. These results demonstrated that the ferritin overexpression in apoplasm was as effective for inducing Fe accumulation as that in plastid. Under the normal metal‐balanced condition, even if the activation of Fe uptake related enzymes leads to the accumulation of non‐specific accumulation of divalent metal ions in roots, an Fe loading/unloading system and/or an internal translocator in xylem and phloem might specifically deliver Fe to the upper part of plants.  相似文献   

10.
A novel extracellular tripeptidyl peptidase (TPP) was homogenously purified from the culture supernatant of Rhizopus oligosporus by sequential fast protein liquid chromatography. The purified enzyme was a 136.5 kDa dimer composed of identical subunits. The effects of inhibitors and metal ions indicated that TPP is a metallo- and serine protease. TPP was activated by divalent cations, such as Co(2+) and Mn(2+), and completely inhibited by Cu(2+). Enzyme activity was optimal at pH 7.0 and 45 °C with a specific activity of 281.9 units/mg for the substrate Ala-Ala-Phe-pNA. The purified enzyme catalyzed cleavage of various synthetic tripeptides but not when proline occupied the P1 position. Purified TPP cleaved the pentapeptide Ala-Ala-Phe-Tyr-Tyr and tripeptide Ala-Ala-Phe, confirming the TPP activity of the enzyme.  相似文献   

11.
An experiment was conducted to find out how humic substances affected nutrient uptake of plants. The test plants, oregano, thyme, and basil, were grown in nutrient film technique at two pH levels (4.5 and 6.5), in two substrates (peat and perlite), and at three levels of humic substance that was a peat extract (control, low, and high concentration). Nutrient uptake of potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) were determined by elemental contents in aerial parts of the plant and its weight. Humic substance had no effect on K, Ca, and Mg uptake but lowered the uptake of Fe, Mn, Zn, and Cu for the three test plants, more pronounced with perlite than peat and more at low pH than at high pH. The lowering of the uptake might be caused by complexation of Fe, Mn, Zn, and Cu by the humic substance and the lower availability of these metals in complexed form than as a cation or as EDTA‐chelate in the case of Fe. It is not clear why the effect of the humic substance on micro‐element uptake is larger at low than at high pH. The complexation is expected to be stronger at pH 6.5 than at pH 4.5. At low pH, the high concentration of humic substance caused a low fresh weight of the shoots, perhaps caused by a toxicity of the humic substance at low pH. This was less pronounced at high pH.  相似文献   

12.
Evidence clearly shows that cationic micronutrients in spray solutions reduce the herbicidal effectiveness of glyphosate for weed control due to the formation of metal-glyphosate complexes. The formation of these glyphosate-metal complexes in plant tissue may also impair micronutrient nutrition of nontarget plants when exposed to glyphosate drift or glyphosate residues in soil. In the present study, the effects of simulated glyphosate drift on plant growth and uptake, translocation, and accumulation (tissue concentration) of iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) were investigated in sunflower (Helianthus annuus L.) plants grown in nutrient solution under controlled environmental conditions. Glyphosate was sprayed on plant shoots at different rates between 1.25 and 6.0% of the recommended dosage (i.e., 0.39 and 1.89 mM glyphosate isopropylamine salt). Glyphosate applications significantly decreased root and shoot dry matter production and chlorophyll concentrations of young leaves and shoot tips. The basal parts of the youngest leaves and shoot tips were severely chlorotic. These effects became apparent within 48 h after the glyphosate spray. Glyphosate also caused substantial decreases in leaf concentration of Fe and Mn while the concentration of Zn and Cu was less affected. In short-term uptake experiments with radiolabeled Fe (59Fe), Mn (54Mn), and Zn (65Zn), root uptake of 59Fe and 54Mn was significantly reduced in 12 and 24 h after application of 6% of the recommended dosage of glyphosate, respectively. Glyphosate resulted in almost complete inhibition of root-to-shoot translocation of 59Fe within 12 h and 54Mn within 24 h after application. These results suggest that glyphosate residues or drift may result in severe impairments in Fe and Mn nutrition of nontarget plants, possibly due to the formation of poorly soluble glyphosate-metal complexes in plant tissues and/or rhizosphere interactions.  相似文献   

13.
转大麦烟酰胺合成酶基因提高水稻逆境胁迫耐受性的研究   总被引：1，自引：0，他引：1  
维持铁等金属离子的动态平衡是保持植物细胞功能正常的先决条件。烟酰胺合成酶基因在单子叶禾本科的缺铁胁迫应答反应中起着关键作用,它的催化产物烟酰胺(NA)是铁及其他二价金属离子在体内吸收和转运的重要载体,且能与Fe2+结合形成Fe2+-NA复合物,从而使植物在酸性土壤中免受铁毒害。利用农杆菌介导法,将大麦烟酰胺合成酶基因NASHOR1转入水稻台北309中,经PCR及PCR-Southern杂交检测,确定目的基因已经整合到水稻基因组中。铁、锰、铜和锌含量的检测结果显示:与非转基因对照植株相比,转基因植株的金属元素含量都明显提高,铜、锌、锰和铁元素含量分别增加了15%、80%、31%和44%,但铁、锰和锌元素增量在株系间差异较大。在干旱胁迫下,转基因植株的超氧化物歧化酶(SOD)活性和脯氨酸含量都高于非转基因对照植株的,暗示大麦烟酰胺合酶基因在一定程度上提高了水稻的耐旱性。  相似文献   

14.
The application of nuclear magnetic resonance (NMR) for the quality control of fertilizers based on Fe(3+), Mn(2+), and Cu(2+) chelates and complexes is precluded by the strong paramagnetism of metals. Recently, a method based on the use of ferrocyanide has been described to remove iron from commercial iron chelates based on the o,o-EDDHA [ethylenediamine-N,N'bis(2-hydroxyphenylacetic)acid] chelating agent for their analysis and quantification by NMR. The present work extended that procedure to other paramagnetic ions, manganese and copper, and other chelating, EDTA (ethylenediaminetetraacetic acid), IDHA [N-(1,2-dicarboxyethyl)-d,l-aspartic acid], and complexing agents, gluconate and heptagluconate. Results showed that the removal of the paramagnetic ions was complete, allowing us to obtain (1)H NMR spectra characterized by narrow peaks. The quantification of the ligands by NMR and high-performance liquid chromatography showed that their complete recovery was granted. The NMR analysis enabled detection and quantification of unknown impurities without the need of pure compounds as internal standards.  相似文献   

15.
Ferulic acid esterase (EC 3.1.1.73) cleaves the feruloyl groups substituted at the 5'-OH group of arabinosyl residues of arabinoxylans and is known to modulate their functional properties. In this study, ferulic acid esterase from 96 h finger millet malt was purified to apparent homogeneity by three-step purification with a recovery of 3% and a fold purification of 22. The substrate p-nitrophenylferulate (PNPF) was synthesized and used to assay this enzyme spectrophotometrically. The products liberated from ragi and wheat water-soluble polysaccharides by the action of purified ragi ferulic acid esterase were identified by ESI-MS. The pH and temperature optima of the enzyme were found to be 6.0 and 45 degrees C, respectively. The pH and temperature stabilities of the enzyme were found to be in the range of 5.5-9.0 and 30 degrees C, respectively. The activation energy of the enzymatic reaction was found to be 4.08 kJ mol(-1). The apparent K m and V max of the purified ferulic acid esterase for PNPF were 0.053 microM and 0.085 unit mL(-1), respectively. The enzyme is a monomer with a molecular mass of 16.5 kDa. Metal ions such as Ni(2+), Zn(2+), Co(2+), and Cu(2+) and oxalic and citric acids enhanced the enzyme activity. The enzyme was completely inhibited by Fe(3+). Group specific reagents such as p-chloromercuric benzoate and iodoacetamide inhibited the enzyme, indicating the possible presence of cysteine residues in the active site pocket.  相似文献   

16.
Glyphosate is largely used to control weeds in wetland soils of Brazil. We investigated changes in the chemistry of soluble manganese (Mn) and iron (Fe) in these soils as affected by glyphosate dosage. Triplicate samples of the A horizon of wetland soils with different organic-matter contents were incubated with deionized water (1:2) for 1, 3, and 30 days under flooding. Three different glyphosate doses (0, 0.048, and 0.096 g L?1 m?2) were spiked on the flooded water at the beginning of the incubation periods. After incubation, pH was measured and samples of the supernatant were collected for determination of Mn/Fe concentrations by atomic absorption. Glyphosate application impacted Mn but had no effect on pH and Fe. Soluble Mn concentrations decreased as glyphosate dosage increased for the high organic-matter soil after 3 days of incubation. It indicated that glyphosate application can change the chemistry of soil metals. The intensity of these changes depends on the glyphosate dosage, evolved metal, incubation time, and soil properties.  相似文献   

17.
Weekly or half-monthly bulk rainwater samples were collected over 3 yr (1993–1995) at Porspoder, an Atlantic station near Brest, France. They were analyzed for 0.08 M HNO3-leachable trace metals Cd, Cu, Fe, Mn, Ni, Pb and Zn by direct electrothermal atomic absorption spectrometry. Major ions and nitrogen species (NO3- and NH4+) in precipitation were determined by ion chromatography. In coastal precitation, a high correlation was observed for major ions having a seawater origin (Na+, CL- and Mg2+. Zn, Cd, Cu, Ni and Pb were well correlated indicating their anthropogenic origin. In the case of nitrogen species and nssSO42- a high correlation with Zn was observed, taken as an anthropogenic pollution tracer. Major ions and nitrogen species were determined according to wind sector direction showing clearly the impact of the pollution at this station attributed to continental air masses. A relative constancy of pH values in all wind directions was observed. Due to an important oceanic influence, trace metal concentrations and annual deposition densities were significantly lower at Porspoder than at North Sea coastal stations. In the case of nitrogen species, the relatively higher concentration levels probably indicated a significant local agricultural contribution.  相似文献   

18.
The enrichment factor, multivariate analysis and metal speciation studies were used to identify degree, source and dispersal of metal contamination in Khli Ti watershed, Thailand. Topsoil samples were collected throughout the watershed, analyzed for total metal concentration. Sequential extraction was also carried out to determine geochemical phases of metals which were identified as exchangeable and bound to carbonates, Fe–Mn oxides, organic matter and residuals. Soil characteristics including pH, total organic carbon, redox potential, cation exchange capacity and texture were also analyzed. Principal component analysis yielded three metal groups which explained 83% of the variance. The concentrations of metals which were derived from lithogenic origin, such as Co, Cr, Fe, Ni and V were in natural background levels and were mostly bound to the residual phase. The remaining elements (i.e. Ba, Cd, Cu, Pb, Sb and Zn) were associated with the contamination from previous activities of the Pb-ore concentrator and Zn–Pb mining. Anthropogenic contamination mainly increased Pb and Zn bound to Fe–Mn oxides at the expense of residual fraction. Even though low exchangeable Pb contents in Khli Ti soils indicated low availability to plants, Pb bound to Fe–Mn oxides fraction might increase its mobility under reducing conditions.  相似文献   

19.
Quantitative parameters for the interactions between phytate (Phy) and Al(3+), Fe(3+), and Cr(3+) were determined potentiometrically in NaNO(3) aqueous solutions at I = 0.10 mol L(-1) and T = 298.15 K. Different complex species were found in a wide pH range. The various species are partially protonated, depending on the pH in which they are formed, and are indicated with the general formula MH(q)Phy (with 0 ≤ q ≤ 6). In all cases, the stability of the FeH(q)Phy species is several log K units higher than that of the analogous AlH(q)Phy and CrH(q)Phy species. For example, for the MH(2)Phy species, the stability trend is log K(2) = 15.81, 20.61, and 16.70 for Al(3+), Fe(3+), and Cr(3+), respectively. The sequestering ability of phytate toward the considered metal cations was evaluated by calculating the pL(0.5) values (i.e., the total ligand concentration necessary to bind 50% of the cation present in trace in solution) at different pH values. In general, phytate results in a quite good sequestering agent toward all three cations in the whole investigated pH range, but the order of pL(0.5) depends on it. For example, at pH 5.0 it is pL(0.5) = 5.33, 5.44, and 5.75 for Fe(3+), Cr(3+), and Al(3+), respectively (Fe(3+) < Cr(3+) < Al(3+)); at pH 7.4 it is pL(0.5) = 9.94, 9.23, and 8.71 (Al(3+) < Cr(3+) < Fe(3+)), whereas at pH 9.0 it is pL(0.5) = 10.42, 10.87, and 8.34 (Al(3+) < Fe(3+) < Cr(3+)). All of the pL(0.5) values, and therefore the sequestering ability, regularly increase with increasing pH, and the dependence of pL(0.5) on pH was modeled using some empirical equations.  相似文献   

20.
Barley seedlings were grown in hydroponic culture in the presence of toxic concentrations of cadmium (Cd), molybdenum (Mo), nickel (Ni), and zinc (Zn) and analyzed for element composition [boron (B), calcium (Ca), Cd, iron (Fe), potassium (K), magnesium (Mg), manganese (Mn), molybdenum (Mo), nickel (Ni), phosphorus (P), and Zn]. In a first survey, heavy metal concentrations were selected which resulted in a similar inhibition of root growth. Toxic concentrations of Cd, Mo, Ni, and Zn revealed both similar and distinct effects on specific leaf and root element contents. Examples for such responses were decreasing contents in root Mn and Mg at elevated levels of all heavy metals, including Mo, in the medium. In contrast, changes in root contents of B were specific for the applied type of heavy metal stress. The heavy metal dependent changes in B, Ca, Mg, and Mn contents were studied in more detail. In some cases, severe heavy metal toxicity caused excessive accumulation or depletion of nutrient elements that may be deleterious to the plants in addition to other primary damages caused by the heavy metal ions.  相似文献   

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