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1.
One-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and high resolution two-dimensional electrophoresis (2-DE) were applied to separate protein molecules in dissolved organic matter (DOM) from oceanic waters. Results were: (1) The 2-DE distinguished a total of 412 protein spots in 10 samples from five water columns over the Pacific, although fewer than 30 proteins were resolved as bands from the identical samples by SDS-PAGE. (2) Major and ubiquitous protein bands (34 and 39 kDa proteins) on the SDS-PAGE gel were resolved into horizontally spread arrays (trains) of spots on the 2-DE gels, indicating that these bands were a mixture of protein species that have the same molecular weight (MW) but different isoelectric points (pIs). (3) Proteins that exhibited such electrophoretic patterns on the 2-DE gels were glycosylated with variable linkages between the sugar and polypeptide chains. (4) N-terminal amino-acid sequencing demonstrated that individual spots within each train of spots had identical N-terminal amino-acid sequences.The N-terminal amino-acid sequences of the 39 and 34 kDa glycoprotein spots in samples collected at different sites were also identical. Protein isoforms with the same amino-acid sequence but different glycosylation profiles, termed glycoforms, were often observed on the 2-DE gel. Thirty-one and 24 spots on the 2-DE gels were glycoforms of two glycoproteins with MWs of 39 and 34 kDa, respectively; they were one protein species. The glycoforms of the 39 kDa protein were identified as a low molecular weight alkaline phosphatase (L-AP) of Pseudomonas aeruginosa PAO1 by a homology search through five amino-acid sequence databases. The present and earlier work indicates that all identified source organisms of dissolved proteins belong to the Pseudomonas group. We propose the hypothesis that proteins associated with membrane vesicles liberated from a minor member of the bacterioplankton assemblage, the marine Pseudomonas group, are one of the important sources of dissolved proteins in oceanic waters. This hypothesis may apply to the source pathway and survival not only of proteins and also to the universally occurring bacterial peptidoglycan and lipopolysaccharide components in DOM.  相似文献   

2.
The molecular distribution of dissolved proteins in seawater from coastal marine environments in Uranouchi Bay, Kochi Prefecture, is first reported in this article. Occurrence of bacteria-derived dissolved proteins and their source bacteria were examined using a probe of the antibody (anti-Omp35La) against a porin outer membrane protein (Omp35La) of the fish pathogenic bacterium Vibrio (Listonella) anguillarum. The electrophoretograms of dissolved proteins from coastal seawater showed a large number of discrete and individual proteins overlapped each other over a wide range of molecular masses indicating active processes in coastal environments in transferring proteins from organisms to the inanimate dissolved protein pool. Among the dissolved proteins, 37 kDa- and 18 kDa-proteins reacted with the Omp35La. In order to isolate the source bacteria of such dissolved proteins, bacteria from seawater and diseased fish were screened by colony Western blotting with anti-Omp35La. The reactive strains were further examined in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blotting to verify the presence of Omp35La homologues among the outer membrane proteins of such strains. Outer membrane proteins reacting with anti-Omp35La were detected in only 4 strains of the 129 strains that were positive in the colony Western blotting. The level of possible source bacteria of 37 kDa- and 18 kDa-dissolved proteins was suggested to be 5–6 orders of magnitude lower than the total bacterial count. The present study leads us to hypothesize that a minor portion of the bacterial assemblage is responsible for the dissolved proteins in the coastal waters.  相似文献   

3.
Microbial degradation and utilization of proteins derived from bacterial detritus were investigated in a microcosm experiment using Pseudomonas aeruginosa detritus as a substrate. To assess the effects of natural marine microbial communities on degradation and utilization of protein derived from P. aeruginosa cells, four microcosms were prepared: natural seawater (containing the natural microbial community) with P. aeruginosa detritus (N+Pa), autoclaved seawater with P. aeruginosa detritus (A+Pa), natural seawater (N) and autoclaved seawater (A) without adding anything as a control. The numbers of total and growing bacterial cells, protease activity, and transition of P. aeruginosa proteins were monitored in the four microcosms. Changes in the numbers of total and growing bacterial cells and protease activities indicated that bacterial detritus significantly stimulated the microbial community in the microcosms. Both the surviving P. aeruginosa in A+Pa and natural microbial community in N+Pa microcosms were able to degrade and utilize P. aeruginosa detritus; however, the community in N+Pa including various microbes maintained high activity longer, indicating that diversity is an important factor in keeping the community active. Even under the very high protease activity in N+Pa, 39-kDa and 48-kDa proteins from P. aeruginosa remained in the microcosm during the entire experiment (150 days). Immunoblotting suggested the 48-kDa protein was an intact molecule of OprP, which had been detected from the dissolved fraction of natural seawater in previous studies. This result suggests that the protein molecules that had been detected from natural seawater actually had a high tolerance to microbial degradation.  相似文献   

4.
Uranium in coastal sediments of Tokyo Bay and Funka Bay   总被引:2,自引:0,他引:2  
The sediment cores from Tokyo Bay and Funka Bay were analyzed for U and its isotopic ratio,234U/238U, after dissolving them in 0.1 M HCl, and 30% H2O2 in 0.05 M HCl. A small fraction of U in the anoxic sediments was dissolved in 0.1M HCl and even the added yield tracer,232U, was lost. The isotopic ratio of H2O2 soluble U in the sediments was equal to that of seawater, suggesting that the H2O2 soluble U in the sediments is authigenic. The 6M HCl solution dissolved part of the lithogenic U besides the authigenic U. The depth profiles of U from the two bays resembled each other. The authigenic U comprised more than half of the total U even at the surface and increased with depth down to 70 cm, showing small maxima at about 20 cm. The concentration of refractory U was nearly constant with depth and similar to that of the pelagic sediments. The highest U concentration, 6 µg g–1 which was about 5 times that of the pelagic sediments, was observed in the layer between 70 and 160 cm depth in Tokyo Bay. The annual sedimentation rates of U in the Tokyo Bay sediments were 2.6 tons at the surface and 7.0 tons at the 70–160 cm depth. The increase in U with depth should be due to the deposition of interstitial U either diffusing downward from the surface indicating the trapping of seawater U, or otherwise diffusing upward from the deeper layer indicating the internal cycling of U within the sediments.  相似文献   

5.
Effects of seawater acidification on hydrolytic enzyme activities   总被引:1,自引:1,他引:0  
We have investigated the effects of seawater acidification on the activities of leucine aminopeptidase (LAPase), β-glucosidase (BGase), phosphatase (P-ase), α-glucosidase (AGase), and lipase (L-ase), which are important promoters of degradation of marine organic matter, including proteins, carbohydrates, organic phosphorus compounds, and lipids. Seawater samples were collected from a eutrophic coastal area, from Tokyo Bay, and from oligotrophic pelagic waters outside the Kuroshio Current. Enzyme activities were measured using fluorogenic substrates added to the seawater samples, which were acidified from pH 8.2 to 5.6 by a chemical buffer. Spontaneous hydrolysis of the substrates was shown to be negligible in heat-inactivated control samples, thus validating our results. LAPase was the most sensitive to acidification; enzyme activity rapidly decreased from pH 8.2 to 7.8, corresponding to a realistic scenario of ocean acidification. L-ase activity also decreased with acidification. Activities of P-ase and BGase were relatively constant across the pH levels examined, suggesting that their activity is not appreciably influenced by acidification. The effect of acidification on P-ase activities differed between the coastal and semipelagic samples, and this was likely due to freshwater influence at the nearshore station. Because of the low activity of AGase in the sample, the effect of acidification on this enzyme could not be examined. The effects of acidification on enzyme activity appear to vary depending on enzyme type and location, but we conclude that acidification will cause changes in the cycling of organic matter in marine ecosystems, in particular to proteinous and lipid substances.  相似文献   

6.
为了建立适于条斑紫菜(Porphyra yezoensis)蛋白质组学分析的的样品提取方法, 作者以条斑紫菜叶状体为材料, 比较了饱和硫酸铵沉淀法、三氯乙酸/丙酮沉淀法和酚提取方法3 种方法对条斑紫菜叶状体总蛋白的提取效果。结果表明, 针对条斑紫菜叶状体细胞中含有大量多糖, 多酚化合物, 色素等干扰物质的特点, 通过酚提取方法能有效去除这些干扰杂质, 得到清晰、稳定、干净的双向电泳图谱和相对较多的蛋白质点数, 为提高其他大型海藻的总可溶性蛋白提取效率提供了重要参考。  相似文献   

7.
不同保存方法对厚壳贻贝样品蛋白提取效果的比较   总被引:1,自引:0,他引:1  
为了解厚壳贻贝(Mytilus coruscus)稚贝样品在冷冻保存过程中的蛋白变化情况,作者采用直接冷冻法、添加海水冷冻法、蛋白裂解液冷冻法等分别对保存1周至6个月的厚壳贻贝样品蛋白提取的不同效果进行了比较。SDS-PAGE胶结果显示,3种冷冻保存方法均出现4个有规律条带(a、b、c、d),其相对分子量依次约为55、46、38和26kDa。SDS-PAGE蛋白条带灰度值的检测结果表明,1个月内蛋白裂解液法保存的样品最佳;直接冷冻法和添加海水冷冻法的研究发现保存过程中的蛋白均出现不同程度的降解现象,因而可能不适宜用于该种稚贝样品的保存。  相似文献   

8.
好氧不产氧光合(AAP)细菌因其具有利用溶解性有机物及光能的能力,在海洋碳循环及能量流动中发挥着重要的作用。该类细菌广泛分布在海洋环境中,其在不同生境中的多样性已被调查。但到目前为止,人们对于高纬度地区好氧不产氧光合细菌的认识还较为缺乏。有鉴于此,本研究基于编码光反应复合物上一个色素结合蛋白亚基的pufM基因,对北极王湾及南极乔治王岛近岸水体中夏季好氧不产氧光合细菌的多样性进行了检测。针对2个王湾站位和2个南极麦克斯维尔湾站位构建了4个pufM基因克隆文库,获得674个阳性克隆子。北极克隆子全部由α-变形细菌组成,而南极克隆子则包括α-变形细菌及β-变形细菌。来源于类似红细菌科的pufM基因在所有样品中皆占据优势。此外,与一株滴状亚硫酸盐杆菌中质粒编码的pufM基因存在亲缘关系的序列,在南、北极样品中均占优势。结果表明海洋环境中的pufM基因存在跨极甚至是环球分布。与此同时,南、北极序列之间的差异,也表明了极地地方种的存在。这些结果显示,作为好氧不产氧光合细菌的红细菌科在两极的近岸水体中具有重要的地位。  相似文献   

9.
To examine the possibility that outer membrane proteins (OMP) of Synechococcus sp. remain in seawater, we investigated the stability of OMPs in vitro and in situ. Some fractions prepared from Synechococcus sp. CSIRO-94 were treated with trypsin and proteinase K. Four tightly bound OMPs were separated from Synechococcus. We designated the two major OMPs of 52 kDa and 48 kDa as Omp52Sy and Omp48Sy, respectively. Degradation of the OMP in natural seawater was monitored in microcosms to which intact Synechococcus cells and outer membrane (OM) were added. Omp52Sy and Omp48Sy were the most stable against trypsin and proteinase K among the OMPs when they were embedded in the OM. However, in the microcosm experiment using intact cells, Omp52Sy and Omp48Sy were detected in the particulate fraction only during the first 4 days, after which they could not longer be detected. Omp52Sy and Omp48Sy were the most stable proteins among the Synechococcus OMPs in vitro, but they might be degraded in situ. This indicates that stability of Synechococcus porin differs depending on complex formation with other membrane molecules, which might cause different preservation of microbial membrane proteins in the dissolved protein pool in the ocean. This study suggests that Gram negative bacterial OM with thin peptidoglycan forms a lipid bilayer that proptects OMP, but Synechococcus OM with thick peptidoglycan cannot form a lipid bilayer. The incomplete bilayer might not be able to protect from protease attack in the natural environment.  相似文献   

10.
In Tokyo Bay the concentrations of dissolved gaseous mercury (DGM) in the surface seawater and total gaseous mercury (TGM) over the sea were measured during December 2003, October 2004 and January 2005. Based on these data, the evasional fluxes of mercury from the sea surface were estimated using a gas exchange model. In addition, an automatic wet and dry deposition sampler was used to measure the wet and dry depositional fluxes of mercury from December 2003 to November 2004 at three locations in and near Tokyo Bay. The results indicate that the average DGM and TGM levels of seven locations are 52 ± 26 ng m−3 and 1.9 ± 0.6 ng m−3, respectively, which shows that the surface seawater in Tokyo Bay is supersaturated with gaseous mercury, leading to an average mercury evasional flux of 140 ± 120 ng m−2d−1. On the other hand, the annual average wet and dry depositional fluxes of mercury at three locations were 19 ± 3 μg m−2yr−1 and 20 ± 9 μg m−2yr−1, respectively. These depositional fluxes correspond to the daily average total depositional flux of 110 ± 20 ng m−2d−1. Thus, it is suggested that in Tokyo Bay, the evasional fluxes of mercury are comparable to the depositional fluxes.  相似文献   

11.
夏季外海水入侵对大亚湾浮游植物群落结构的影响   总被引:1,自引:0,他引:1  
杨熙  谭烨辉 《海洋科学》2019,43(7):96-105
夏季大亚湾存在由粤东沿岸上升流所引起的外海水入侵现象,且入侵强度存在年际差异,作者利用大亚湾2004~2017年历年夏季航次调查数据,将弱入侵年份与强入侵年份进行对比分析,以探讨外海水入侵对大亚湾浮游植物群落结构的影响。结果显示,当外海水入侵由弱变强时,湾内水体理化特征发现显著变化,水体由高温低盐转变为低温高盐,N、P等营养盐含量出现下降。海水理化性质的改变导致了浮游植物群落结构的变化,硅藻、甲藻种类数以及浮游植物Shannon-wiener指数均出现升高;浮游植物总丰度和硅藻丰度下降,甲藻丰度变化不明显;常见浮游植物种类伪菱形藻属(Pseudo-nitzschiasp.)、角毛藻属(Chaetocerossp.)和叉角藻(Ceratiumfurca)丰度出现下降,而中肋骨条藻(Skeletonemacostatum)和菱形海线藻(Thalassionemanitzschioides)丰度出现升高;优势种由单一硅藻种类向硅藻和甲藻共为优势转变。此外,外海水入侵还会通过改变海水理化因子的空间分布以及湾内上层水体流向来影响浮游植物群落结构的空间分布。  相似文献   

12.
提取中国明对虾的血浆蛋白用SDS-PAGE进行分离.将电泳条带从上到下平均切成4份,分别胶内酶解并利用液相色谱.串联质谱系统进行质谱分析.利用SEQUEST软件对NCBInr蛋白数据库进行检索,鉴定了66个中国明对虾血浆蛋白质.运用生物信息学手段,对鉴定的蛋白进行功能分类,包括转录、细胞结构和骨架、信号转导、蛋白质合成与处理、蛋白质折叠、分拣和降解、免疫功能、生长和发育、能量代谢、细胞周期、碳水化合物代谢、钙离子平衡、抗氧化剂和氨基酸和脂类代谢等.本研究丰富了中国对虾血浆蛋白质表达谱,可为进一步研究对虾免疫机制提供有力的支持.  相似文献   

13.
Dissolved organic matter (DOM) composition and dynamics in temperate shallow coastal bays are not well described although these bays may be important as local sources of organic carbon to ocean waters and are often sites of economically-important fisheries and aquaculture. In this study surface water samples were collected on a monthly to bi-monthly basis over two years from a mid-Atlantic coastal bay (Chincoteague Bay, Virginia and Maryland, USA). Dissolved organic carbon (DOC) concentrations and light absorbance characteristics were measured on sterile-filtered water, and high-molecular weight (> 1 kDa) dissolved OM (DOM) was isolated to determine stable isotope composition and molecular-level characteristics. Our time series encompassed both a drought year (2002) and a year of above-average rainfall (2003). During the dry year, one of our sites developed a very intense bloom of the brown tide organism Aureococcus anophagefferens while during the wet year there were brown tide bloom events at both of our sampling sites. During early spring of the wet year, there were higher concentrations of > 1 kDa DOC; this fraction represented a larger proportion of overall DOC and appeared considerably more allochthonous. Based upon colored dissolved organic matter (CDOM) and high-molecular weight DOM analyses, the development of extensive phytoplankton blooms during our sampling period significantly altered the quality of the DOM. Throughout both years Chincoteague Bay had high DOC concentrations relative to values reported for the coastal ocean. This observation, in conjunction with the observed effects of phytoplankton blooms on DOM composition, indicates that Chincoteague Bay may be a significant local source of “recently-fixed” organic carbon to shelf waters. Estimating inputs of DOC from Chincoteague Bay to the Mid-Atlantic Bight suggests that shallow productive bays should be considered in studies of organic carbon on continental shelves.  相似文献   

14.
Bidimensional and monodimensional polyacrylamide gel electrophoresis were used to study protein expression from zooplankton collected in thirteen stations of Toulon Bay (NW Mediterranean). In this ecosystem, Little Bay showed higher trace metal concentrations (13.5–23.8 nM for Cu, 0.73–1.24 nM for Pb, 27.8–58.7 nM for Zn) than Large Bay (Cu 2.2–15.6 nM; Pb 0.19–0.78 nM; Zn 9.0–38.8 nM). Trace metals positively correlated (p < 0.05) with expression of four zooplankton proteins (MW in kDa/pI: 25.0/5.6; 48.8/4.1; 38.2/4.4; 38.3/5.8) and with biomass of Oithona nana, predominant copepod in Little Bay. Sequencing by LC–MS/MS putatively provided zooplankton identity of these proteins: they were cytoskeleton actin, except one protein that was the chaperone calreticulin. We suggest that actin and calreticulin could be regarded as zooplankton markers of metal stress and be involved in a possible tolerance of O. nana to contamination, contributing to its development in a marine perturbed ecosystem.  相似文献   

15.
《Marine Chemistry》2007,103(1-2):1-14
We succeeded in determining the Ce isotopic composition (138Ce/142Ce) in seawater with an error of 2σm = 0.3–0.7 of ε unit. In this study, 1000–3000 L of seawater samples were passed through MnO2 fibers to concentrate Ce and Nd for precise measurement of their isotope ratios. Four surface seawater samples of the northwestern Pacific and a coastal sample in Tokyo Bay were analyzed. Most Ce isotope ratios in the surface water showed positive εCe values (+ 0.8 to + 1.4) in the northwestern Pacific Ocean. These values indicate that Ce in the surface water originates from the continental crust preferentially over mantle-derived materials. We examined binary mixing model between the continental crust and mid-ocean ridge basalt. However the model could not explain both isotopic compositions and concentrations, which implies that the atmospheric input was a possible pathway for Ce into the ocean. A negative εCe value was observed in Tokyo Bay, suggesting mantle-derived sources.  相似文献   

16.
A risk assessment of Tributyltin (TBT) in Tokyo Bay was conducted using the Margin of Exposure (MOE) method at the species level using the Japanese short-neck clam, Ruditapes philippinarum. The assessment endpoint was defined to protect R. philippinarum in Tokyo Bay from TBT (growth effects). A No Observed Effect Concentration (NOEC) for this species with respect to growth reduction induced by TBT was estimated from experimental results published in the scientific literature. Sources of TBT in this study were assumed to be commercial vessels in harbors and navigation routes. Concentrations of TBT in Tokyo Bay were estimated using a three-dimensional hydrodynamic model, an ecosystem model and a chemical fate model. MOEs for this species were estimated for the years 1990, 2000, and 2007. Estimated MOEs for R. philippinarum for 1990, 2000, and 2007 were approximately 1–3, 10, and 100, respectively, indicating a declining temporal trend in the probability of adverse growth effects.A simplified software package called RAMTB was developed by incorporating the chemical fate model and the databases of seasonal flow fields and distributions of organic substances (phytoplankton and detritus) in Tokyo Bay, simulated by the hydrodynamic and ecological model, respectively.  相似文献   

17.
Dissolved and particulate concentrations of silver in Tokyo Bay estuarine waters and Japanese rivers were determined in this study. The dissolved silver concentrations in the surface water of Tokyo Bay range from 5.9 to 15.1 pmol kg−1, which is comparable to those in the surface water of the Japan Sea, but two or three times higher than those in the surface water of the open ocean. However, elevated concentrations of dissolved silver are not found in Tokyo Bay compared with those in other highly urbanized estuaries, such as San Francisco Bay (20∼243 pmol kg−1). In the Tokyo Bay estuary, silver typically exhibits non-conservative mixing behavior, which is a common feature in the other estuaries reported previously. Dissolved silver concentrations decrease with salinity from the rivers to the mouth of Tokyo Bay. Silver is efficiently scavenged by suspended particulates, as evidenced by the high conditional distribution coefficients for silver throughout the estuary (log Kd > 5.0 ± 0.6). The silver fluxes into Tokyo Bay via inflowing rivers and atmospheric deposition were estimated as 83 kg y−1 and 15 kg y−1, respectively. A simple budget calculation shows that the silver supplied from rivers and atmosphere must be rapidly scavenged within the Tokyo Bay estuary.  相似文献   

18.
自2007年以来,浒苔绿潮已经连续15年在南黄海暴发。浒苔(Ulva prolifera)作为主要肇事藻种,在暴发过程中向海水释放大量的溶解有机碳(DOC)。然而,这些藻源DOC能否长期保存在海洋中,主要取决于它们的生物可利用性,目前关于此方面的研究甚少。本研究在浒苔绿潮大规模暴发时期(2019年6月),分别在浒苔暴发海区和无浒苔海区各选择3个站位富集表层海水,在实验室进行长期(300 d)的DOC降解实验。结果发现,在60 d内,不同站位富集海水中的DOC浓度随着微生物的利用快速下降,微生物丰度也在第60天达到峰值,表明这些被消耗的DOC是生物可利用性高的活性DOC(LDOC)。60 d后,剩余的DOC可抵抗微生物的降解,在60~300 d内保持稳定,表明这些DOC是具有强稳定性的惰性DOC(RDOC)。最终发现,浒苔暴发海水的RDOC占富集DOC的46%,明显高于无浒苔海水的(36%)。并且,荧光溶解有机物(FDOM)中活性的类蛋白组分随着微生物的利用被快速消耗,惰性的类腐殖质组分逐渐积累,暗示了在降解过程中LDOC逐渐向RDOC转化。可见,浒苔绿潮暴发除了在短时间内增加海水中的D...  相似文献   

19.
采集胶州湾表层和底层海水样品,分析了Cu、Cr、Cd、Pb、Ni、Co等痕量金属在海水中的空间分布特征及其在不同分子量溶解有机质中的分配特征,并探讨了痕量金属?溶解有机质分配机理及浮游生物活动与盐度等环境因素对该分配过程的影响。结果表明,胶州湾海水中痕量金属呈近岸浓度较高的分布特征,在湾东北部出现高值区,Cd和Pb还分别在湾口与湾中部出现高值区。胶州湾海水中痕量金属平均有70.1%分配于低分子量(<1 kDa)组分中,其中Cu和Cd低分子量组分所占平均比例分别达79.0%与77.6%,Cr、Ni和Co稍低,分别为71.5%、67.3%及66.9%,Pb则仅为58.2%。海水中的溶解有机碳也以低分子量组分为主,所占比例平均达73.1%,且光谱特征显示低分子量溶解有机质中类腐殖质含量更高,含有丰富的羧基和羟基,金属配合能力较高,导致痕量金属多分配于低分子量溶解有机质中。高分子量溶解有机质(>1 kDa)所占比例与叶绿素a浓度呈显著正相关,表明浮游植物初级生产通过释放高分子量溶解有机质影响海水痕量金属?溶解有机质的分配过程。胶州湾湾顶盐度较低海域痕量金属高分子量组分略高,可能是生物活动及陆源输入(产生更多高分子量溶解有机质)与盐度(低盐有利于高分子量有机质的稳定性)共同作用的结果。  相似文献   

20.
We performed a comprehensive metaproteomic analysis of the dissolved organic matter (DOM) in Japanese coastal waters using liquid chromatography–tandem mass spectrometry and demonstrated that these proteomes were characterized by proteins with various functions, including metabolic enzymes, membranes, and photosynthetic proteins. The protein sources included cyanobacteria, heterotrophic bacteria, and eukaryotic phytoplankton. Most of the components were similar among samples and also similar to pelagic components. We also observed differences in the compositions of the microbial communities of origin among the different dissolved protein samples and differences in the relative abundance of specific dissolved protein types (e.g., cytoskeletal proteins), possibly indicating potential dynamics in the coastal DOM pool.  相似文献   

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