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Molecular Characterization and Benefits of the Intestinal Ecosystem 肠道生态系统的分子特征和益处
Pub Date : 2006-07-01 DOI: 10.12938/BIFIDUS.25.67
M. Blaut
The microbial community resident in the human gastrointestinal tract has a major impact on host physiology. Manipulating this complex ecosystem by dietary intervention requires knowledge of the parameters that influence its composition and the activity. More recent developments have taken advantage of culture-independent molecular methods for bacterial identification on the basis of the highly discriminatory sequence database of 16S rRNA. This culture-independent approach was applied to asses the fecal microbiota of human subjects from different European countries in relation to nutrition. In one of two human studies, a positive correlation was observed between the intake of fruits and vegetables and the proportion of bacteria belonging to the Clostridium leptum group. In the second study, the relative proportion of bifidobacteria was much higher in the Italian study group than in any other study group. This finding correlated positively with a consumption of water-soluble fiber. Intestinal bacteria also play a role in the activation and inactivation of plant-derived polyphenolic substances such as lignans and flavonoids. To assess the influence of bacterial metabolism on the bioavailability of flavonoids in the human intestinal tract, two model organisms were used to characterize the transformation of flavones in comparison to that of flavonols. Lignans require activation by intestinal bacteria to exert estrogenic and antioxidant activities. Several bacterial species involved in lignan activation were isolated and the catalytic steps involved were characterized. The results indicate that the activation of dietary lignans involves phylogenetically diverse bacteria, most of which are common members of the dominant human intestinal microbiota.
居住在人体胃肠道的微生物群落对宿主生理有重要影响。通过饮食干预来操纵这个复杂的生态系统需要了解影响其组成和活动的参数。最近的发展利用了不依赖培养的分子方法,在16S rRNA高度区分序列数据库的基础上进行细菌鉴定。这种与培养无关的方法被应用于评估来自不同欧洲国家的人类受试者的粪便微生物群与营养的关系。在两项人体研究中的一项中,人们观察到水果和蔬菜的摄入量与瘦梭菌群细菌的比例呈正相关。在第二项研究中,意大利研究组中双歧杆菌的相对比例远高于其他任何研究组。这一发现与摄入水溶性纤维呈正相关。肠道细菌也在植物源性多酚物质(如木脂素和类黄酮)的激活和失活中发挥作用。为了评估细菌代谢对黄酮类化合物在人体肠道内生物利用度的影响,我们使用了两种模式生物来表征黄酮类化合物与黄酮醇的转化。木脂素需要肠道细菌的激活才能发挥雌激素和抗氧化活性。分离了几种参与木脂素活化的细菌,并对其催化步骤进行了表征。结果表明,膳食木脂素的激活涉及系统发育多样性的细菌,其中大多数是人类肠道优势菌群的共同成员。
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引用次数: 2
Clinical Potential of Lactococcus lactis Mediated Delivery of Human Interleukin-10 and Trefoil Factors 乳酸乳球菌介导的人白细胞介素-10和三叶因子输送的临床潜力
Pub Date : 2006-07-01 DOI: 10.12938/BIFIDUS.25.81
E. Remaut, H. Braat, K. Vandenbroucke, P. Rottiers, L. Steidler, L. Steidler
Systemic treatment with infliximab, a monoclonal antibody that binds TNFa is among the most potent therapies for Crohn's disease. Despite initial successes with intravenous addition of IL-10, this treatment was abandoned in later trials. Both treatments suffered a serious drawback from the fact that they had to be administered systemically and thus caused potentially serious side effects. We used two mouse models for IBD to evaluate the efficacy of L. lactis mediated topical delivery of IL-10. In the Dextran Sulphate Sodium (DSS)-induced chronic colitis intragastric administration of mIL-10-secreting L. lactis led to a 50% reduction in inflammation. Also, daily administration of the engineered strain prevented the onset of colitis, normally associated with the IL-10 knockout genotype. In both models, the effect was strictly dependent on delivery of live bacteria. We engineered a strain exhibiting biological containment. To this end, the thymidilate synthase (thy A) gene of L. lactis was replaced with a synthetic human IL-10 gene. ThyA-deficient bacteria are suicidal in the absence of thymine or thymidine and therefore cannot accumulate in the environment. A limited clinical trail in Crohn's patients under physical containment proved that the treatment was safe, the biological containment strategy was effective and the results obtained suggested a clinical effect. In contrast to oral administration of purified protein, intragastric administration of Trefoil factor-secreting L. lactis was very effective in prevention and healing of acute DSS colitis. In addition this approach was successful in improving established chronic colitis in IL-10 knockout mice.
用英夫利昔单抗(一种结合TNFa的单克隆抗体)进行全身治疗是治疗克罗恩病最有效的方法之一。尽管静脉注射IL-10取得了初步成功,但在后来的试验中,这种治疗被放弃了。这两种治疗方法都有一个严重的缺点,即它们必须全身使用,因此可能会产生严重的副作用。我们使用两种IBD小鼠模型来评估乳杆菌介导的局部递送IL-10的效果。在葡聚糖硫酸钠(DSS)诱导的慢性结肠炎中,胃内给予分泌mil -10的乳酸乳杆菌可使炎症减少50%。此外,每天给药工程菌株防止结肠炎的发作,通常与IL-10敲除基因型有关。在这两种模型中,效果都严格依赖于活细菌的输送。我们设计了一种具有生物控制能力的菌株。为此,用人工合成的人IL-10基因代替乳杆菌胸腺嘧啶合成酶(thymidate synthase, thy A)基因。胸腺嘧啶缺乏的细菌在缺乏胸腺嘧啶或胸腺嘧啶的情况下是自杀的,因此不能在环境中积累。对克罗恩病患者进行物理隔离的有限临床试验证明,该治疗是安全的,生物隔离策略是有效的,所获得的结果表明临床效果。与口服纯化蛋白相比,灌胃分泌三叶因子乳杆菌对急性DSS结肠炎的预防和愈合非常有效。此外,这种方法成功地改善了IL-10基因敲除小鼠的慢性结肠炎。
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引用次数: 2
Pathways for the Recognition of the Intestinal Microbiota 肠道微生物群的识别途径
Pub Date : 2006-04-01 DOI: 10.12938/BIFIDUS.25.33
H. Reinecker
The specific role of most of these newly discovered DC subsets in antigen sampling and presentation is unknown and it remains therefore unclear whether they work synergistically, as alternatives or have distinct functions in the recognition of the intestinal microbiota. The focus of effort in this field will be to understand how these diverse DC subsets cooperate in regulating the complex homeostasis and host defense in the different intestinal immune compartments. The intestinal lamina propria contains a DC network that serves as a newly discovered gateway for the uptake and transport of the intestinal microbiota. Specialized vesicular structures at the end of transepithelial dendrites serve as 'luminal sensors' for the mucosal immune system. Characterization of the surface components of these luminal sensors may aid in developing strategies to prevent bacterial and viral pathogen entry. We propose that the lamina propria and M cell-dependent antigen sampling systems are associated with specific DC subsets. The discovery of functionally defined subsets of dendritic cells associated with distinct mechanisms responsible for the uptake of antigens across the intestinal barrier opens the path for strategies for targeting them specifically in the development of vaccines or treatment approaches for inflammatory bowel diseases. For these exciting possibilities to progress into practical applications, it needs to be established whether distinct DCs subsets are associated with multiple pathways or if their function is linked to specific innate or adaptive immune responses. Defining DC function in the intestine will be pivotal in finally progressing our understanding on how the mucosal immune system makes the distinction between commensal microbiota, pathogens and self antigens.
大多数这些新发现的DC亚群在抗原采样和递呈中的具体作用尚不清楚,因此仍不清楚它们是否协同工作,作为替代品或在识别肠道微生物群中具有不同的功能。该领域的工作重点将是了解这些不同的DC亚群如何在不同的肠道免疫室中协同调节复杂的稳态和宿主防御。肠固有层包含一个DC网络,作为肠道微生物群摄取和运输的新发现的门户。上皮树突末端的特化囊泡结构充当粘膜免疫系统的“管腔传感器”。表征这些管腔传感器的表面成分可能有助于制定防止细菌和病毒病原体进入的策略。我们认为固有层和M细胞依赖抗原取样系统与特定的DC亚群有关。功能明确的树突状细胞亚群的发现与负责通过肠道屏障摄取抗原的不同机制相关,为在开发针对炎症性肠病的疫苗或治疗方法中特异性靶向它们的策略开辟了道路。为了使这些令人兴奋的可能性进入实际应用,需要确定不同的dc亚群是否与多种途径相关,或者它们的功能是否与特定的先天或适应性免疫反应相关。确定DC在肠道中的功能将是最终推进我们对粘膜免疫系统如何区分共生微生物群、病原体和自身抗原的理解的关键。
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引用次数: 0
The Effects of a Probiotic Milk Product Containing Lactobacillus casei Strain Shirota on the Defecation Frequency and the Intestinal Microflora of Sub-optimal Health State Volunteers: A Randomized Placebo-controlled Cross-over Study 含干酪乳杆菌Shirota菌的益生菌乳制品对亚理想健康状态志愿者排便频率和肠道菌群的影响:一项随机安慰剂对照交叉研究
Pub Date : 2006-04-01 DOI: 10.12938/BIFIDUS.25.39
Kazumasa Matsumoto, T. Takada, K. Shimizu, Y. Kado, K. Kawakami, I. Makino, Y. Yamaoka, Koichi Hirano, A. Nishimura, O. Kajimoto, K. Nomoto
Forty healthy individuals with a low defecation frequency were selected, and the effects of intake of a fermented milk beverage that contains Lactobacillus casei strain Shirota (LcS) at 4 x 10 9 bacteria/bottle for 2 weeks (1 bottle/day) on the defecation frequency and intestinal microflora were evaluated by the placebo-controlled double-blind cross-over scheme. Defecation frequency both in times per week and days per week significantly increased in the LcS beverage period compared with the frequency before the beginning of intake. The differences were more notable in those with a stronger tendency to constipation (frequency of defecation before intake ≤ 4.0 times/week, n=21), and the frequency of defecation in the LcS beverage period was significantly higher than in the placebo period. The stool smell and feeling of completion of voiding improved significantly in the LcS beverage period compared with the placebo period, and in those with a stronger tendency to constipation, the stools were significantly softened compared with the state before intake. The number of bifidobacteria and their percentage in the total number of fecal bacteria in the LcS beverage period increased significantly compared with the levels before intake and were significantly higher than the values in the placebo period. No marked change due to the intake of the LcS beverage was observed in the other components of the microflora, the organic acid contents, stool pH, water content, or contents of putrefactive metabolites. These results suggest that intake of the probiotic fermented milk beverage conditions the intestines by improving the state of bowel movements and stool quality and increasing the fecal population level of bifidobacteria.
选择排便频率较低的健康个体40例,采用安慰剂对照双盲交叉试验方案,以4 × 10 9个细菌/瓶(LcS)的剂量,连续2周(1瓶/天)饮用含干酪乳杆菌(Lactobacillus Shirota)的发酵乳饮料,评价其对排便频率和肠道菌群的影响。与开始摄入LcS饮料之前相比,LcS饮料期间每周排便次数和每周排便天数均显著增加。便秘倾向较强者(摄入前排便次数≤4.0次/周,n=21)差异更显著,且LcS饮料期排便次数显著高于安慰剂期。LcS饮料期大便气味和排便完成感较安慰剂期明显改善,便秘倾向较强者大便较摄入前明显软化。饮用LcS饮料期间,双歧杆菌的数量及其占粪便细菌总数的百分比与摄入前相比显著增加,且显著高于安慰剂期。摄入LcS饮料后,在微生物菌群的其他成分、有机酸含量、粪便pH值、水分含量或腐烂代谢物含量方面没有观察到明显的变化。综上所述,摄入益生菌发酵乳饮料可以改善肠道蠕动状态和大便质量,增加粪便中双歧杆菌的数量,从而改善肠道状况。
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引用次数: 80
Human Evolution, Nutritional Ecology and Prebiotics in Ancient Diet 人类进化、营养生态学与古代饮食中的益生元
Pub Date : 2006-01-01 DOI: 10.12938/BIFIDUS.25.1
J. Leach, G. Gibson, J. Loo
Modern studies of prebiotic non digestible carbohydrates continue to expand and demonstrate their colonic and systemic benefits. However, virtually nothing is known of their use among ancient populations. In this paper we discuss evidence for prebiotic use in the archaeological record from select areas of the world. It is suggested that members of our genus Homo would have had sufficient ecological opportunity to include prebiotic-bearing plants in diet as early as ~ 2 million years ago, but that significant dietary intake would not have taken place until the advent of technological advances that characterized the Upper Paleolithic of ~40,000 years ago. Throughout human evolution, hominid populations that diversified their diet to include prebiotic-bearing plants would have had a selective advantage over competitors.
对益生元不可消化碳水化合物的现代研究不断扩大,并证明了它们对结肠和全身的益处。然而,几乎没有人知道它们在古代人群中的使用情况。在本文中,我们讨论了来自世界上一些地区的考古记录中关于益生元使用的证据。研究表明,早在约200万年前,我们属的人就有足够的生态机会在饮食中加入含有益生元的植物,但直到约4万年前旧石器时代晚期的技术进步出现,才会出现大量的饮食摄入。在整个人类进化过程中,将饮食多样化,包括含有益生元的植物的原始人种群将比竞争对手具有选择优势。
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引用次数: 20
Effects of Wheat Bran and Brown Rice Cereals on the Intestinal Environment and Skin Conditions 麦麸和糙米谷物对肠道环境和皮肤状况的影响
Pub Date : 2006-01-01 DOI: 10.12938/BIFIDUS.25.9
K. Hirayama, Ryutarou Arai, K. Itoh, Rumi Ide, Shimako Mutoh
Ingestion of brown rice cereal has been found to improve skin conditions, while the effects of wheat bran cereal were limited (Ide et al., J. Integr. Study Diet. Habits, 2005, in press). The effects of these breakfast cereals on the composition of intestinal flora and the intestinal environment were investigated. Two different types of breakfast cereals, wheat bran cereal that is particularly rich in dietary fiber and brown rice cereal fortified with vitamins and minerals, were consumed twice a day by young female volunteers. The control group ingested their usual diet. Composition of fecal flora, fecal moisture, fecal pH, fecal enzymic activities and concentrations of intestinal putrefactive products and short chain fatty acids as well as skin conditions were analyzed. The populations of Enterobacteriaceae and Streptococcaceae significantly decreased after two weeks of wheat bran cereal consumption. Moisture of feces was significantly decreased and the properties of feces and defecation frequency were improved by wheat bran cereal. Activity of β-glucosidase increased significantly and the concentrations of putrefactive products decreased slightly. The effects of brown rice cereal on composition of intestinal flora and intestinal environment were not obvious. No direct correlation between the effects on the intestinal environment and skin conditions was found. The results suggest that the two different types of breakfast cereals used in the present study have different impacts on the intestinal environment and skin conditions.
研究发现,摄入糙米谷物可以改善皮肤状况,而麦麸谷物的效果有限(Ide et al., J. Integr.)。研究饮食。《习惯》,2005年出版)。研究了这些早餐谷物对肠道菌群组成和肠道环境的影响。年轻的女性志愿者每天吃两次不同类型的早餐谷物,一种是富含膳食纤维的麦麸谷物,另一种是富含维生素和矿物质的糙米谷物。对照组摄食正常饮食。分析粪便菌群组成、粪便水分、粪便pH、粪便酶活性、肠道腐烂产物和短链脂肪酸浓度以及皮肤状况。食用麦麸谷物两周后,肠杆菌科和链球菌科的数量显著减少。麦麸谷物显著降低了粪便水分,改善了粪便性质和排便频率。β-葡萄糖苷酶活性显著升高,腐烂产物浓度略有下降。糙米谷物对肠道菌群组成和肠道环境的影响不明显。对肠道环境的影响和皮肤状况之间没有直接的联系。结果表明,本研究中使用的两种不同类型的早餐谷物对肠道环境和皮肤状况有不同的影响。
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引用次数: 0
Stimulation of Cytokine Gene Expression in a Murine Macrophage Cell Line by Lactic Acid Bacteria Isolated from Bhutanese Dried Yak Cheese 不丹干牦牛奶酪乳酸菌对小鼠巨噬细胞细胞因子基因表达的刺激作用
Pub Date : 2005-10-01 DOI: 10.12938/BIFIDUS.24.125
Y. Ohashi, K. Nagai, M. Tokunaga, K. Ushida, Tomomi Yamada
Lactic acid bacteria (LAB) isolated from dried yak cheese from the Bhutan Himalayas were examined for their ability to stimulate the expression of cytokine mRNA on the murine macrophage. The expression of tumor necrosis factor-a, interleukin (IL) 6 or IL-12 p40 gene was stimulated by several strains. The dried yak cheese, Chugo, contains LAB that possess immuno-modulating activity.
研究了从不丹喜马拉雅地区牦牛干奶酪中分离的乳酸菌(LAB)刺激小鼠巨噬细胞细胞因子mRNA表达的能力。几种菌株均能刺激肿瘤坏死因子-a、白细胞介素(IL) 6或IL- 12p40基因的表达。Chugo干牦牛奶酪含有具有免疫调节作用的乳酸菌。
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引用次数: 0
Comparison of Growth Promoting Effects on Bifidobacterium spp. by Bovine Lactoferrin Hydrolysates 牛乳铁蛋白水解物对双歧杆菌促生长作用的比较
Pub Date : 2005-10-01 DOI: 10.12938/BIFIDUS.24.119
Woan-sub Kim, M. Rahman, H. Kumura, K. Shimazaki
The effect of bovine lactoferrin hydrolysates on the growth of four species of bifidobacteria, B. bifidum, B. longum, B. breve and B. infantis was investigated. It was observed that the growth of 3 species of bifidobacteria (B. bifidum, B. breve and B. infantis) was stimulated by bovine lactoferrin hydrolysates. These results suggest the possibility that lactoferrin, digested in the intestine, acts as a bifidogenic factor for the growth of bifidobacteria.
研究了牛乳铁蛋白水解物对两歧双歧杆菌、长双歧杆菌、短双歧杆菌和婴儿双歧杆菌4种双歧杆菌生长的影响。结果表明,牛乳铁蛋白水解物可促进3种双歧杆菌(双歧双歧杆菌、短双歧杆菌和婴儿双歧杆菌)的生长。这些结果表明,在肠道中消化的乳铁蛋白可能是双歧杆菌生长的双歧因子。
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引用次数: 11
Apoptosis induced by Short-chain Fatty Acids Modulates Immunoresponses : Role of Cell-to-cell Communication in Inhibiting Butyric Acid-induced T Cell Apoptosis 短链脂肪酸诱导的细胞凋亡调节免疫反应:细胞间通讯在抑制丁酸诱导的T细胞凋亡中的作用
Pub Date : 2005-07-01 DOI: 10.12938/BIFIDUS.24.91
K. Ochiai, T. Kurita-Ochiai
Butyric acid present in the culture filtrates of Porphyromonas gingivalis, Prevotella loescheii and Fusobacterium nucleatum induced cytotoxicity and apoptosis in murine thymocytes, splenic T cells, and human Jurkat T cells. A pronounced accumulation of ROS occurred during butyric acid-induced apoptosis. Butyric acid induced apoptosis via the mitochondrial apoptotic pathway, e.g. cytochrome c, AIF, and Smac, and by the ceramide pathway. Up-regulation of JNK and p38, and down-regulation of ERK occurred immediately after butyric acid treatment. In microarray analysis, butyric acid treatment resulted in increased expression of the proapoptotic gene, whereas the expression of anti-apoptotic mediators was decreased. These data suggest that butyric acid is an apoptosis-inducing agent in most lymphoreticular cells. In contrast, epithelial cells and fibroblasts were insensitive to butyric acid. Fibroblasts from healthy gingival tissue rescued butyric acid-induced T cell apoptosis via proinflammatory cytokines such as IL-6 and IL-11, which were produced by the fibroblasts stimulated by butyric acid. Furthermore, the T cell apoptosis was also down-regulated by T cell adhesion to gingival fibroblasts through interaction with CD44, VLA-2, and VLA-5 expressed on T cells stimulated with butyric acid. Also, gingival fibroblasts from periodontal patients were highly susceptible to apoptosis induced by butyric acid when compared to healthy gingival fibroblasts. In conclusion, since short-chain fatty acids produced by periodontopathic bacteria induce apoptosis in immunological cells and the fibroblasts from periodontal patients, the results strongly suggested that they are concerned with the progress of periodontal disease.
牙龈卟啉单胞菌、罗氏普氏菌和核梭杆菌培养滤液中的丁酸可诱导小鼠胸腺细胞、脾脏T细胞和人Jurkat T细胞的细胞毒性和凋亡。在丁酸诱导的细胞凋亡过程中,ROS显著积累。丁酸通过细胞色素c、AIF和Smac等线粒体凋亡途径和神经酰胺途径诱导细胞凋亡。丁酸处理后,JNK和p38上调,ERK下调。在微阵列分析中,丁酸处理导致促凋亡基因的表达增加,而抗凋亡介质的表达减少。这些数据表明,在大多数淋巴网状细胞中,丁酸是一种凋亡诱导剂。相反,上皮细胞和成纤维细胞对丁酸不敏感。健康牙龈组织成纤维细胞在丁酸刺激下产生IL-6和IL-11等促炎细胞因子,挽救了丁酸诱导的T细胞凋亡。此外,丁酸刺激T细胞表达的CD44、VLA-2和VLA-5与牙龈成纤维细胞的黏附作用也下调了T细胞的凋亡。此外,与健康牙龈成纤维细胞相比,牙周病患者的牙龈成纤维细胞对丁酸诱导的细胞凋亡高度敏感。综上所述,由于牙周病细菌产生的短链脂肪酸可诱导牙周患者免疫细胞和成纤维细胞凋亡,结果强烈提示它们与牙周病的进展有关。
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引用次数: 1
Aerobic and Anaerobic Biotransformation of Bile Acids by Escherichia coli (II): No Conversion of α-Muricholic Acid by 7α-Hydroxysteroid Dehydrogenase of E. coli 大肠杆菌胆汁酸的好氧和厌氧生物转化(II):大肠杆菌7α-羟基类固醇脱氢酶不转化α-胆酸
Pub Date : 2005-04-01 DOI: 10.12938/BIFIDUS.24.41
Y. Ogura, N. Yamaga, Takao Suzuki, K. Kimura, Y. Kido, Kazuo Yamada, K. Uchida
The effect of Escherichia coli K-12 on the transformation of hyocholic acid, α-muricholic acid, β-muricholic acid and 3α,7β,12α-trihydroxy-5β-cholanoic acid was examined in aerobic and anaerobic cultures for 4 days and the activities were compared with those on cholic acid and chenodeoxycholic acid. Hyocholic acid was dehydrogenated, but to a lesser extent than cholic acid or chenodeoxycholic acid. However, α-muricholic acid, which has a 7α-hydroxyl group, was not dehydrogenated even after 4 days. This was confirmed by experiments using a cell-free crude enzyme preparation and by gas chromatographic analysis of bile acids in the medium culture. β-Muricholic acid and 3α,7β,12α-trihydroxy-5β-cholanoic acid were not changed by Esherichia coli K-12.
在好氧和厌氧培养条件下,研究了大肠杆菌K-12对牛胆酸、α-胆酸、β-胆酸和3α,7β,12α-三羟基-5β-胆酸转化的影响,并与胆酸和鹅去氧胆酸的活性进行了比较。犬胆酸脱氢,但脱氢程度低于胆酸或鹅去氧胆酸。而含有7α-羟基的α-苦胆酸在4天后仍未脱氢。用无细胞粗酶制备的实验和培养液中胆汁酸的气相色谱分析证实了这一点。大肠杆菌K-12对β-胆酸和3α,7β,12α-三羟基-5β-胆酸没有影响。
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引用次数: 0
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