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Plant NLR receptor proteins and their potential in the development of durable genetic resistance to biotic stresses 植物NLR受体蛋白及其在生物胁迫持久遗传抗性发展中的潜力
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2020.01.002
Amanda Cristina de Araújo, Fernando Campos De Assis Fonseca, Michelle Guitton Cotta, Gabriel Sergio Costa Alves, Robert Neil Gerard Miller

In order to meet global food security demands in the next decades, considerable changes are required for sustainable agriculture in the context of plant disease, with sufficient food production depending on the development of durable genetically disease resistant crops. For this, further advances are required in our understanding of the plant innate immune system and how plants respond to invading pathogenic micro-organisms. Over the past 20 years, considerable research has been conducted into the characterization and cloning of plant nucleotide-binding, leucine-rich repeat (NLR) immune receptors. These intracellular receptors can recognize directly or indirectly pathogen effector proteins, resulting in effector-triggered immunity (ETI). Elucidation, however, of the diversity of NLR resistance gene families and the molecular basis of NLR-driven effector recognition and defense signaling is incomplete. Here, we present a summary of the understanding of NLR structure, function, genomic organization and diversity in plants. Recent advances in target enrichment approaches for NLR characterization and function validation are highlighted in the context of NLR engineering possibilities for accelerated durable genetic resistance to biotic stresses.

为了满足未来几十年的全球粮食安全需求,在植物病害的情况下,需要对可持续农业进行重大变革,粮食生产的充足与否取决于培育耐久的遗传抗病作物。为此,我们需要进一步了解植物先天免疫系统以及植物如何对入侵的致病微生物作出反应。在过去的20年里,对植物核苷酸结合的富含亮氨酸重复序列(NLR)免疫受体的鉴定和克隆进行了大量的研究。这些细胞内受体可以直接或间接识别病原体效应蛋白,从而导致效应触发免疫(ETI)。然而,对NLR抗性基因家族的多样性以及NLR驱动效应物识别和防御信号的分子基础的阐明尚不完整。本文就植物NLR的结构、功能、基因组组织和多样性等方面的研究进展作一综述。在NLR工程可能加速对生物胁迫的持久遗传抗性的背景下,强调了NLR表征和功能验证的靶标富集方法的最新进展。
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引用次数: 26
Differential distribution of polyphenols in plants using multivariate techniques 多酚在植物中的差异分布
Pub Date : 2019-01-01 DOI: 10.1016/J.BIORI.2019.03.001
Vinod Kumar, Anket Sharma, Sukhmeen Kaur Kohli, Shagun Bali, Manik Sharma, Rakesh Kumar, R. Bhardwaj, Ashwani Kumar Thukral
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引用次数: 36
Integrative approach reveals new insights into photosynthetic and redox protection in ex vitro tobacco plantlets acclimatization to increasing light intensity 综合方法揭示了烟草植株离体适应增加光强的光合作用和氧化还原保护的新见解
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2020.04.001
Cinthya F. Vieira , Fabricio E.L. Carvalho , Yugo Lima-Melo , Cristina P.S. Carvalho , Milton C. Lima Neto , Marcio O. Martins , Joaquim A.G. Silveira

Integrative mechanisms involving photosynthetic and antioxidant protection regulated by light intensity during in vitro and ex vitro plantlets acclimatization are poorly understood. Tobacco plantlets grown under in vitro and ex vitro environments were exposed to different light regimes to evaluate the role of photosynthesis and antioxidant protection. In vitro plantlets displayed a narrow photosynthetic capacity to cope with light as revealed by low net CO2 assimilation (PN), decreased actual quantum efficiency of photosystem II (ΦPSII) associated with non-induction of non-photochemical quenching (NPQ). In contrast, acclimated ex vitro plants showed strong stimulation in PN whereas ΦPSII and NPQ remained at high levels. In vitro plantlets exposed to moderate light suffered strong oxidative stress associated with increased activities of superoxide dismutases, catalases and ascorbate peroxidases, revealing an ineffective antioxidant system. In contrast, ex vitro plants presented lower oxidative damage in parallel to unchanged enzymatic activities, indicating an efficient antioxidant steady state. The levels of reduced ascorbate (ASC) and glutathione (GSH) were also increased only in ex vitro plants in response to excess light. An integrative study based on correlation networks and principal component analyses (PCA) corroborate that the two plant groups indeed displayed contrasting acclimation processes. In conclusion, during in vitro to ex vitro transition, tobacco plantlets exposed to increasing light display physiological adjustments involving photosynthesis and improvement of the enzymatic and non-enzymatic antioxidant systems. These findings highlight the importance of integrative approaches to understand ex vitro acclimatization to environmental stimuli.

在离体和离体植株驯化过程中,光强调控的光合作用和抗氧化保护的综合机制尚不清楚。在离体和离体环境下培养烟草植株,研究不同光照条件下烟草植株的光合作用和抗氧化保护作用。体外植株表现出较窄的光合能力来应对光,这主要表现在低净CO2同化(PN)、光系统II的实际量子效率降低(ΦPSII)和未诱导非光化学猝灭(NPQ)。驯化的离体植株PN受到强烈刺激,而ΦPSII和NPQ仍保持较高水平。暴露在中等光照下的试管苗遭受强烈的氧化应激,其超氧化物歧化酶、过氧化氢酶和抗坏血酸过氧化物酶的活性增加,表明抗氧化系统无效。相比之下,离体植株在保持酶活性不变的同时表现出较低的氧化损伤,表明其处于有效的抗氧化稳定状态。抗坏血酸(ASC)和谷胱甘肽(GSH)的还原水平也仅在离体植物中因过度光照而增加。基于相关网络和主成分分析(PCA)的综合研究证实,两个植物类群确实表现出不同的驯化过程。综上所述,在离体向离体过渡的过程中,暴露于增强光下的烟草植株表现出包括光合作用在内的生理调节以及酶和非酶抗氧化系统的改善。这些发现强调了综合方法对了解体外环境刺激适应的重要性。
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引用次数: 5
Differential distribution of polyphenols in plants using multivariate techniques 多酚在植物中的差异分布
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2019.03.001
Vinod Kumar , Anket Sharma , Sukhmeen Kaur Kohli , Shagun Bali , Manik Sharma , Rakesh Kumar , Renu Bhardwaj , Ashwani Kumar Thukral

Plants form a substantive portion of human diet that contains a plethora of structurally diverse polyphenols. These polyphenols extend both nutritional as well as disease preventive benefits to the consumer. Many ancient medicinal systems like Ayurveda reiterate the consumption of particular polyphenol rich plant in specific medical condition. Therefore, it is necessary to create a database of the contents of different polyphenols in various plants. This study compiles a bibliographic review of polyphenol distribution in different plants along with their statistical analysis like Pearson's correlation matrix, cluster analysis, principal component and factor analysis, and non-metric multidimensional scaling. It was observed that caffeic acid and quercetin were the most abundant polyphenols in the plants. A significant positive correlation of umbelliferone was found with kaempferol and epicatechin. Species having close proximities on the basis of polyphenols were Parthenium hysterophorus, Rumex dentatus, Achyranthus aspera, Chenopodium ambrosoides, Cannabis sativa, Rhododendron arboreum, Alternanthera philoxeroides, Debregeasia longifolia and C. album. Factor analysis showed four underlying factors for polyphenols. Factor-1 had maximum loadings on epicatechin, umbelliferone and kaempferol. Gallic acid and catechin had maximum loadings on factor-2. Factor-3 had maximum loadings on chlorogenic acid and quercetin, and factor-4 had maximum loading on coumaric acid.

植物是人类饮食的重要组成部分,其中含有大量结构多样的多酚。这些多酚对消费者既具有营养作用,又具有预防疾病的作用。许多古老的医疗系统,如阿育吠陀,重申在特定的医疗条件下食用富含多酚的植物。因此,有必要建立各种植物中不同多酚含量的数据库。本文对多酚在不同植物中的分布进行了文献综述,并对其进行了Pearson相关矩阵、聚类分析、主成分与因子分析、非度量多维标度等统计分析。结果表明,咖啡酸和槲皮素是该植物中含量最多的多酚类物质。伞花素与山奈酚、表儿茶素呈显著正相关。多酚含量相近的植物有Parthenium hysterophorus、Rumex dentatus、Achyranthus aspera、Chenopodium ambrosoides、Cannabis sativa、Rhododendron arboreum、Alternanthera philoxeroides、Debregeasia longifolia和C. album。因子分析显示了多酚的四个潜在因素。因子1对表儿茶素、伞花素和山奈酚的负荷最大。没食子酸和儿茶素对因子-2的负荷最大。因子-3对绿原酸和槲皮素的负荷最大,因子-4对香豆酸的负荷最大。
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引用次数: 35
Absorption of polycyclic aromatic hydrocarbons onto depolymerized lignocellulosic wastes by Streptomyces viridosporus T7A 毒孢链霉菌T7A对解聚木质纤维素废弃物多环芳烃的吸附研究
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2019.04.002
Ana Caroline de Oliveira , Ana Aguilar-Galvez , David Campos , Hervé Rogez

Preparations containing mainly lignified cell walls are effective adsorbents of hydrophobic carcinogens in vitro. Therefore, this innovative study aimed to structurally modify lignocellulosic materials in an enzymatic and environmentally friendly manner to improve the efficiency of sequestering three polycyclic aromatic hydrocarbons (PAHs) by adsorption. Submerged fermentations were performed to produce lignin peroxidase (LiP) in medium supplemented with three different lignocellulosic wastes (0.5% (w/v); açai seed, sugarcane bagasse, and seed coat of Brazil nut), followed by posterior hydrolysis of these three residues and subsequent adsorption to generate hydrolysate wastes. Among the three wastes, the açai seed was the most favourable carbon source for LiP production because a high enzyme activity peak was quickly achieved. Sugarcane bagasse residue was most readily hydrolysed (82.44%), and it had the highest increase in adsorption of the three PAHs tested dissolved in olive oil, from 15.67% of benzo[a]pyrene adsorbed before treatment to 39.45% after treatment. The depolymerisation of wastes may have increased binding sites for olive oil favouring the adsorption of PAHs on hydrolysed residues.

以木质化细胞壁为主的制剂是疏水致癌物的有效体外吸附剂。因此,本创新研究旨在通过酶和环境友好的方式对木质纤维素材料进行结构修饰,以提高吸附三种多环芳烃(PAHs)的效率。在培养基中添加三种不同的木质纤维素废弃物(0.5% (w/v);甘蔗种子、甘蔗渣和巴西坚果种皮),然后对这三种残留物进行后水解,随后吸附产生水解产物废物。在这三种废弃物中,阿帕拉泰种子是最有利的碳源,因为它的酶活性峰值很快就能达到。蔗渣最易水解(82.44%),对橄榄油中溶解的三种多环芳烃的吸附量增幅最大,对苯并[a]芘的吸附量由处理前的15.67%增至处理后的39.45%。废物的解聚合可能增加了橄榄油的结合位点,有利于在水解残基上吸附多环芳烃。
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引用次数: 3
Microbial fuel cells potential of marine actinobacteria Actinoalloteichus sp. MHA15 from the Havelock island of the Andamans, India 来自印度安达曼群岛Havelock岛的海洋放线菌Actinoalloteichus sp. MHA15的微生物燃料电池潜力
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2019.01.003
Rajagopal Gobalakrishnan , Rameshbabu Bhuvaneswari

In the recent past, use of fossil fuel is on the rise and has triggered global energy crisis. So, renewable bioenergy is viewed as one of the means to tackle this problem. In this context, interest in microbial fuel cells (MFCs) is increasing and gaining popularity due to their ability to convert the organic wastes into renewable energy. Present investigation is on the bioelectricity production by a marine actinobacterium isolated from the Havelock island of the Andamans. Actinobacterial colonies were isolated from the sediment samples, using Kuster's agar. 19 morphologically distinct strains were subjected to cellulase enzyme screening. Among them, higher cellulose degradation capacity was found in the strain MHA15. This potential strain was selected and identified as a species close to Actinoalloteichus cyanogriseus. The strain was subjected to bioelectricity generation using sugarcane bagasse as substrate and was evaluated in a dual chambered microbial fuel cell. In the MFC, initial voltage output started at 160 mV and it gradually increased, reaching a maximum of 257 mV at the 3rd hour in the actinobacterial incubated bagasse solution. From the MFC analyzed sludge bagasse, microbial diversity was searched using different media and only actinobacterial colonies were observed. Conventional taxonomic characters of the isolates were identical to the potential actinobacterial strain MHA15 which produced bioelectricity from bagasse. Marine actinobacteria, with their unique nature, differ very much in many aspects from their terrestrial counterparts and are known to produce diverse spectra of novel and useful substances and excellent bioactivity. Results of the present study have ascertained that the marine actinobacterial strain Actinoalloteichus sp. MHA15 is capable of generating bioelectricity and there is much scope for utilizing such marine actinobacteria for large scale production of bioelectricity, after further in-depth studies.

近年来,化石燃料的使用呈上升趋势,并引发了全球能源危机。因此,可再生生物能源被视为解决这一问题的手段之一。在这种背景下,由于微生物燃料电池能够将有机废物转化为可再生能源,人们对其的兴趣越来越大,越来越受欢迎。目前的研究是关于从安达曼群岛的哈夫洛克岛分离的一种海洋放线菌的生物电生产。利用库斯特琼脂从沉积物样品中分离放线菌菌落。对19株形态不同的菌株进行纤维素酶筛选。其中,菌株MHA15的纤维素降解能力较高。筛选出该潜在菌株,并鉴定为与cyanogriseus Actinoalloteichus接近。该菌株以甘蔗渣为底物进行生物发电,并在双室微生物燃料电池中进行了评价。在MFC中,初始电压输出从160 mV开始逐渐增加,在放线菌培养的甘蔗渣溶液中,在第3小时达到257 mV的最大值。从MFC分析的污泥甘蔗渣中,用不同的培养基搜索微生物多样性,只观察到放线菌菌落。该菌株的常规分类特征与利用甘蔗渣发电的潜在放线菌菌株MHA15相同。海洋放线菌具有独特的性质,在许多方面与陆地放线菌有很大的不同,可以产生多种光谱的新型有用物质和优异的生物活性。本研究结果确定了海洋放线菌菌株Actinoalloteichus sp. MHA15具有产生生物电的能力,在进一步深入研究后,利用这种海洋放线菌大规模生产生物电的空间很大。
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引用次数: 4
Effects of Avicennia marina extracts on Labeo rohita (Ham) challenged with Pseudomonas fluorescens 海棠提取物对荧光假单胞菌侵染的红唇海棠的影响
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2018.10.002
G. Dinesh Kumar , M. Karthik , R. Rajakumar , K. Baskar

The medicinal plant Avicennia marina was evaluated for their immunostimulatory activity on Pseudomonas fluorescens infested fish, Labeo rohita. The fish was dosed intraperitoneally at 10, 20 and 30 ppm concentrations of ethanolic leaves extract of A. marina and control. After 10, 20 and 30 days of treatments, the immunological, hematological and serum protein level of fish was assessed in control and treatments. All the concentration of plant leaves extract significantly enhanced the agglutination, hematological parameters and total serum protein on 30th days after treatment. The highest agglutination activity was observed in the group treated with 30 ppm concentration of A. marina on 20 days. The WBC, RBC and hemoglobin content was increased with increasing concentration of the treatments. The results, clearly indicates that A. marina leaves extract will be used as immunostimulatory agent to aquaculture for mass production of healthy fish.

本研究评价了药用植物龙葵对荧光假单胞菌侵染鱼的免疫刺激活性。分别以10、20和30 ppm浓度的黄颡鱼叶乙醇提取物和对照品腹腔注射。处理10、20和30 d后,分别测定对照组和处理组鱼的免疫学、血液学和血清蛋白水平。各浓度植物叶提取物均能显著提高大鼠第30天的凝集率、血液学指标和血清总蛋白。在30 ppm浓度的水芹处理20天后,凝集活性最高。白细胞、红细胞和血红蛋白含量随处理浓度的增加而增加。结果表明,沙蚕叶提取物可作为免疫刺激剂用于水产养殖,实现健康鱼的批量生产。
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引用次数: 5
Effects of Avicennia marina extracts on Labeo rohita (Ham) challenged with Pseudomonas fluorescens 海棠提取物对荧光假单胞菌侵染的红唇海棠的影响
Pub Date : 2019-01-01 DOI: 10.1016/J.BIORI.2018.10.002
G. D. Kumar, M. Karthik, R. Rajakumar, K. Baskar
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引用次数: 5
Endogenous soybean peptide overexpression: an alternative to protect plants against root-knot nematodes 内源性大豆肽过表达:保护植物免受根结线虫侵害的一种替代方法
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2019.12.002
Ciliana Rechenmacher , Beatriz Wiebke-Strohm , Luisa Abruzzi de Oliveira-Busatto , Ricardo Luis Mayer Weber , Mariana Cristina Moraes Corso , Valéria Stefania Lopes-Caitar , Suellen Mika Hishinuma Silva , Waldir Pereira Dias , Francismar Correa Marcelino-Guimarães , Celia Regina Carlini , Maria Helena Bodanese-Zanettini

Nematodes are pathogens of many important crops, including soybean. The main species found in Brazil are root-knot (Meloidogyne spp.), cyst (Heterodera glycines), root lesion (Pratylenchus brachyurus) and reniform (Rotylenculus reniformis) nematodes. Ureases are traditionally known for catalyzing the hydrolysis of urea to ammonia and carbon dioxide. Besides the main function, they present other independent biological roles, including toxic activities against insects, specially Coleoptera and Hemiptera, and fungi. In previous work, the DNA sequence encoding an insecticidal peptide - named Jaburetox - was identified in a Canavalia ensiformes urease gene. The recombinant Jaburetox exhibited toxicity against insects. Subsequently, the DNA sequence corresponding to Jaburetox was identified as part of the soybean Eu4 urease gene, with the resulting peptide named Soyuretox. In the present study, explants of soybean were transformed with Agrobacterium rhizogenes and ‘composite’ plants produced consisting of wild-type shoots and transgenic hairy roots overexpressing Soyuretox. Thereafter soybean plants overexpressing Soyuretox were obtained through bombardment transformation. Due to similarity between nematode and insect digestion mechanisms, we challenged composite and whole-transgenic plants with the nematode Meloidogyne javanica. Hairy roots overexpressing Soyuretox exhibited a significant reduction (48 %; p < 0.05) in the average reproductive factor when compared with empty-vector transformed hairy roots. Transgenic plants overexpressing Soyuretox also exhibited significant reduction (37.5 %; p < 0.05) in reproductive factor when compared with non-transformed plants. This study demonstrates the potential of Soyuretox in confering resistance against nematodes, representing a new alternative control method for nematodes in economically important crops.

线虫是包括大豆在内的许多重要作物的病原体。在巴西发现的主要种类是根结线虫(Meloidogyne spp.)、囊肿线虫(Heterodera glycines)、根瘤线虫(Pratylenchus brachyurus)和肾形线虫(Rotylenculus reniformis)。脲酶传统上以催化尿素水解成氨和二氧化碳而闻名。除主要功能外,它们还具有其他独立的生物学作用,包括对昆虫(特别是鞘翅目和半翅目昆虫)和真菌的毒性作用。在之前的工作中,编码杀虫肽的DNA序列-命名为Jaburetox -在Canavalia ensiformes脲酶基因中被鉴定出来。重组Jaburetox对昆虫具有一定的毒性。随后,Jaburetox对应的DNA序列被鉴定为大豆Eu4脲酶基因的一部分,由此产生的肽被命名为Soyuretox。在本研究中,用根状农杆菌转化大豆外植体,产生由野生型芽和表达Soyuretox的转基因毛状根组成的“复合”植株。然后通过轰击转化获得过表达Soyuretox的大豆植株。由于线虫和昆虫消化机制的相似性,我们用javanica Meloidogyne线虫挑战复合和全转基因植物。过表达Soyuretox的毛状根表现出显著的减少(48%;p & lt;与空载体转化毛状根相比,毛状根的平均繁殖因子降低了0.05)。过表达Soyuretox的转基因植物也表现出显著降低(37.5%;p & lt;与未转化植株相比,生殖因子差异显著(0.05)。本研究证明了Soyuretox具有抗线虫的潜力,为重要经济作物的线虫防治提供了一种新的替代方法。
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引用次数: 4
Brazilian red propolis extracts: study of chemical composition by ESI-MS/MS (ESI+) and cytotoxic profiles against colon cancer cell lines 巴西红蜂胶提取物:ESI-MS/MS (ESI+)化学成分及对结肠癌细胞系的细胞毒性研究
Pub Date : 2019-01-01 DOI: 10.1016/j.biori.2019.02.001
Denis Amilton dos Santos , Fernanda Mosena Munari , Caroline Olivieri da Silva Frozza , Sidnei Moura , Thiago Barcellos , João Antonio Pêgas Henriques , Mariana Roesch-Ely

Red propolis is a natural resin mixture produced by honeybees and presents a source of active compounds with a variety of biological activities. In this study, we describe the chemical characterization and potential antitumor activity of total extract of Brazilian red propolis and its fractions. Fractions were obtained through column chromatography revealing 14 different compounds in all samples, which were determined and distinguished of other isobar molecules by fragmentation pathways by ESI-MS/MS in positive mode. Some molecules as cis-asarone or trans-isoelemicin were identified and distinguish from elemicin compound and vestitol or isovestitol were also distinguished from neovestitol by fragmention pathway. Other important compounds as liquiritigenin was differentiated from isoliquiritigenin and formononetin from dalbergin.

MTT viability assay showed different toxicity in cell lines after exposition to total extract and fractions. Fractions 05 and 06 had more selectivity against HT-29 and HCT-116 cancer cells, respectively, in relation to normal cells. IC50 (ranging of 72.45 ± 6.57 to 73.58 ± 1.00 μg/mL) in cancer cells were lower than reported in total extracts of propolis. May-Grunwald/Giemsa staining revealed cellular morphological changes after exposition to higher concentrations of red propolis extracts. Fractionation techniques can contribute to reduce chemical diversity verified in propolis mixtures, generating fractions with improved biological activity and contributing to the development of new strategies for discovery of natural compounds against cancer.

红蜂胶是一种由蜜蜂生产的天然树脂混合物,是具有多种生物活性的活性化合物的来源。在本研究中,我们描述了巴西红蜂胶总提取物及其组分的化学性质和潜在的抗肿瘤活性。通过柱层析得到14种不同的化合物,并在阳性模式下通过ESI-MS/MS裂解途径与其他等压线分子进行鉴定。通过片段途径将顺式细辛烷和反式异茴香素等分子与茴香素化合物区分开来,并将异茴香醇和新茴香醇区分开来。从黄豆素中分离出了异黄豆素和刺芒柄花素等重要化合物。MTT活性试验表明,总提取物和部分提取物对细胞系的毒性不同。与正常细胞相比,组分05和组分06分别对HT-29和HCT-116癌细胞具有更高的选择性。蜂胶总提取物的IC50值(72.45±6.57 ~ 73.58±1.00 μg/mL)低于文献报道。May-Grunwald/Giemsa染色显示暴露于高浓度红色蜂胶提取物后的细胞形态学变化。分离技术有助于减少蜂胶混合物中已证实的化学多样性,产生具有更高生物活性的组分,并有助于开发新的策略来发现抗癌的天然化合物。
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引用次数: 19
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