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Adsorption of urinary proteins on the conventionally used urine collection tubes: possible effects on urinary proteome analysis and prevention of the adsorption by polymer coating. 尿蛋白在常规尿收集管上的吸附:对尿蛋白质组分析的可能影响和聚合物涂层对吸附的预防。
Pub Date : 2011-01-01 Epub Date: 2011-09-12 DOI: 10.1155/2011/502845
Iwao Kiyokawa, Kazuyuki Sogawa, Keiko Ise, Fumie Iida, Mamoru Satoh, Toshihide Miura, Ryo Kojima, Katsuhiro Katayama, Fumio Nomura

One possible factor determining recovery of trace amount of protein biomarker candidates during proteome analyses could be adsorption on urine tubes. This issue, however, has not been well addressed so far. Recently, a new technical device of surface coating by poly(2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)) (poly(MPC-co-BMA)) has been developed mainly to prevent the adsorption of plasma proteins. We assessed whether conventionally used urine tubes adsorb trace amount of urinary proteins and, if any, whether the surface coating by poly(MPC-co-BMA) can minimize the adsorption. Proteinuric urine samples were kept in poly(MPC-co-BMA)-coated and noncoated urine tubes for 15 min and possibly adsorbed proteins and/or peptides onto urine tubes were analyzed by SDS-PAGE, 2-DE, and the MALDI-TOF MS. It was found that a number of proteins and/or peptides adsorb on the conventionally used urine tubes and that surface coating by poly(MPC-co-BMA) can minimize the adsorption without any significant effects on routine urinalysis test results. Although it remains to be clarified to what extent the protein adsorption can modify the results of urinary proteome analyses, one has to consider this possible adsorption of urinary proteins when searching for trace amounts of protein biomarkers in urine.

在蛋白质组学分析中,决定痕量蛋白质生物标志物候选物回收率的一个可能因素可能是尿管上的吸附。然而,到目前为止,这个问题还没有得到很好的解决。近年来,为了防止对血浆蛋白的吸附,研制了一种新型的聚(2-甲基丙烯酰氧乙基磷酸胆碱(MPC)-co-n-甲基丙烯酸丁酯(BMA))(聚(MPC-co-BMA))表面涂层技术装置。我们评估了传统的尿管是否能吸附微量的尿蛋白,如果有的话,是否表面涂覆聚乙烯(MPC-co-BMA)可以减少吸附。蛋白尿样品在聚(MPC-co-BMA)包覆和未包覆的尿管中保存15分钟,并通过SDS-PAGE、2-DE和MALDI-TOF质谱分析可能吸附在尿管上的蛋白质和/或肽。结果发现,常规使用的尿管上吸附了许多蛋白质和/或肽,聚(MPC-co-BMA)表面涂层可以减少吸附,而对常规尿液分析结果没有明显影响。尽管蛋白质吸附在多大程度上可以改变尿蛋白质组分析的结果仍有待澄清,但在寻找尿液中痕量蛋白质生物标志物时,必须考虑尿蛋白的这种可能吸附。
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引用次数: 3
Heat shock proteins in association with heat tolerance in grasses. 草耐热性与热休克蛋白的关系。
Pub Date : 2011-01-01 Epub Date: 2011-02-24 DOI: 10.1155/2011/529648
Yan Xu, Chenyang Zhan, Bingru Huang

The grass family Poaceae includes annual species cultivated as major grain crops and perennial species cultivated as forage or turf grasses. Heat stress is a primary factor limiting growth and productivity of cool-season grass species and is becoming a more significant problem in the context of global warming. Plants have developed various mechanisms in heat-stress adaptation, including changes in protein metabolism such as the induction of heat shock proteins (HSPs). This paper summarizes the structure and function of major HSPs, recent research progress on the association of HSPs with grass tolerance to heat stress, and incorporation of HSPs in heat-tolerant grass breeding.

禾本科包括作为主要粮食作物种植的一年生物种和作为饲料或草坪草种植的多年生物种。热应激是限制冷季草生长和生产力的主要因素,在全球变暖的背景下,热应激正成为一个越来越重要的问题。植物对热胁迫的适应机制多种多样,包括诱导热休克蛋白(HSPs)等蛋白质代谢的变化。本文综述了主要热休克蛋白的结构和功能、热休克蛋白与牧草耐热性的关系以及热休克蛋白在耐热草育种中的应用研究进展。
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引用次数: 97
Proteomics-based disease biomarkers. 基于蛋白质组学的疾病生物标志物。
Pub Date : 2011-01-01 Epub Date: 2011-10-30 DOI: 10.1155/2011/894618
David E Misek, Tadashi Kondo, Mark W Duncan
Sequencing of the human genome has greatly impacted the proteomics-based analysis of disease by providing a framework for understanding the proteome of diseased cells, tissues, and biological fluids. Consequently, there is a growing interest in applying proteomics technologies to define protein pathways involved in various diseases, to identify new biomarkers that correlate with diseases, ideally in their early stages, and to accelerate the development of new therapeutic targets. However, disease-related proteomics applications require that we improve our ability to separate and characterize the components of complex protein mixtures in such a way as to boost both throughput and sensitivity. In response to these demands, the proteomics technologies have been improved markedly over recent years. Today, proteomics, in all its various forms, is proving to be invaluable to our understanding of the biochemistry of health and disease and will likely play a central role in the evolution of personalized medicine. In this special issue, we include reports of novel research findings together with several reviews that highlight advances in key areas. The first two papers of this special issue focus on lung cancer. The first paper, by H. C. Gong et al., addresses the profiling of receptor tyrosine kinase pathway activation and the role of key genetic mutations in human lung tumor cell lines and human lung tumors. The authors defined molecular pathways which may assist in development of targeted lung tumor therapies. Within the second paper, Q. Zhang et al. used proteomic profiling to delineate expression and subcellular localization of multiple forms of aldehyde dehydrogenase in lung adenocarcinoma cell lines. The next two papers focus on pancreatic cancer. The third paper, by R. S. Kwon and D. M. Simeone, reviews the use of protein-based biomarkers for the diagnosis of cystic tumors of the pancreas. The fourth paper, by M. Abulaizi et al., utilizes a three-step proteomic protocol (immunodepletion of abundant serum proteins, followed by fractionation by RP-HPLC and further separation by 2D-PAGE) to discover candidate early detection biomarkers of pancreatic cancer. The next two papers focus on breast cancer, with the fifth paper, by D. E. Misek and E. H. Kim, reviewing the development of protein biomarkers for the early detection of breast cancer. The sixth paper, by J. He et al., addresses LC-MS/MS identification of protein biosignatures in breast tumors, as protein-based markers that correctly classify tumor subtypes and predict therapeutic response would be of great clinical utility in guiding patient treatment. The next two papers are both by M. S. Sabel et al., and focus on melanoma. The seventh paper reviews the use of proteomics for the discovery of new prognostic and predictive biomarkers. The eighth paper explores the clinical utility of serum autoantibodies that were detected in melanoma patients. The investigators profiled serum antibodies a
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引用次数: 2
The Role of Proteomics in the Diagnosis and Treatment of Women's Cancers: Current Trends in Technology and Future Opportunities. 蛋白质组学在女性癌症的诊断和治疗中的作用:目前的技术趋势和未来的机会。
Pub Date : 2011-01-01 DOI: 10.1155/2011/373584
Eun-Kyoung Yim Breuer, Mandi M Murph

Technological and scientific innovations over the last decade have greatly contributed to improved diagnostics, predictive models, and prognosis among cancers affecting women. In fact, an explosion of information in these areas has almost assured future generations that outcomes in cancer will continue to improve. Herein we discuss the current status of breast, cervical, and ovarian cancers as it relates to screening, disease diagnosis, and treatment options. Among the differences in these cancers, it is striking that breast cancer has multiple predictive tests based upon tumor biomarkers and sophisticated, individualized options for prescription therapeutics while ovarian cancer lacks these tools. In addition, cervical cancer leads the way in innovative, cancer-preventative vaccines and multiple screening options to prevent disease progression. For each of these malignancies, emerging proteomic technologies based upon mass spectrometry, stable isotope labeling with amino acids, high-throughput ELISA, tissue or protein microarray techniques, and click chemistry in the pursuit of activity-based profiling can pioneer the next generation of discovery. We will discuss six of the latest techniques to understand proteomics in cancer and highlight research utilizing these techniques with the goal of improvement in the management of women's cancers.

过去十年的技术和科学创新极大地促进了影响妇女的癌症的诊断、预测模型和预后。事实上,这些领域信息的爆炸式增长几乎保证了未来几代人对癌症的治疗效果将继续改善。在这里,我们讨论乳腺癌、宫颈癌和卵巢癌的现状,因为它与筛查、疾病诊断和治疗方案有关。在这些癌症的差异中,令人惊讶的是,乳腺癌有多种基于肿瘤生物标志物的预测测试和复杂的、个性化的处方治疗选择,而卵巢癌缺乏这些工具。此外,宫颈癌在预防癌症的创新疫苗和预防疾病进展的多种筛查选择方面处于领先地位。对于每一种恶性肿瘤,基于质谱分析、氨基酸稳定同位素标记、高通量ELISA、组织或蛋白质微阵列技术以及追求基于活性分析的点击化学的新兴蛋白质组学技术可以引领下一代的发现。我们将讨论六种最新的技术来理解癌症中的蛋白质组学,并重点介绍利用这些技术改善女性癌症管理的研究。
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引用次数: 45
Urine glycoprotein profile reveals novel markers for chronic kidney disease. 尿液糖蛋白图谱揭示了慢性肾脏病的新标记。
Pub Date : 2011-01-01 Epub Date: 2011-10-10 DOI: 10.1155/2011/214715
Anuradha Vivekanandan-Giri, Jessica L Slocum, Carolyn L Buller, Venkatesha Basrur, Wenjun Ju, Rodica Pop-Busui, David M Lubman, Matthias Kretzler, Subramaniam Pennathur

Chronic kidney disease (CKD) is a significant public health problem, and progression to end-stage renal disease leads to dramatic increases in morbidity and mortality. The mechanisms underlying progression of disease are poorly defined, and current noninvasive markers incompletely correlate with disease progression. Therefore, there is a great need for discovering novel markers for CKD. We utilized a glycoproteomic profiling approach to test the hypothesis that the urinary glycoproteome profile from subjects with CKD would be distinct from healthy controls. N-linked glycoproteins were isolated and enriched from the urine of healthy controls and subjects with CKD. This strategy identified several differentially expressed proteins in CKD, including a diverse array of proteins with endopeptidase inhibitor activity, protein binding functions, and acute-phase/immune-stress response activity supporting the proposal that inflammation may play a central role in CKD. Additionally, several of these proteins have been previously linked to kidney disease implicating a mechanistic role in disease pathogenesis. Collectively, our observations suggest that the human urinary glycoproteome may serve as a discovery source for novel mechanism-based biomarkers of CKD.

慢性肾脏病(CKD)是一个重大的公共卫生问题,进展到终末期肾病会导致发病率和死亡率急剧上升。疾病进展的机制尚不明确,目前的非侵入性标记物与疾病进展的相关性也不完全。因此,亟需发现治疗慢性肾功能衰竭的新型标志物。我们利用糖蛋白组剖析方法来验证一个假设,即慢性肾功能衰竭患者的尿液糖蛋白组剖析将有别于健康对照组。我们从健康对照组和 CKD 患者的尿液中分离并富集了 N-连接的糖蛋白。这一策略确定了 CKD 中多种不同表达的蛋白质,包括具有内肽酶抑制剂活性、蛋白质结合功能和急性期/免疫应激反应活性的多种蛋白质,支持了炎症可能在 CKD 中起核心作用的观点。此外,这些蛋白质中有几个以前曾与肾脏疾病相关,这暗示了它们在疾病发病机制中的作用。总之,我们的观察结果表明,人类尿液糖蛋白组可作为基于机制的新型 CKD 生物标记物的发现源。
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引用次数: 0
Protein biomarkers for the early detection of breast cancer. 早期检测乳腺癌的蛋白质生物标志物。
Pub Date : 2011-01-01 Epub Date: 2011-08-11 DOI: 10.1155/2011/343582
David E Misek, Evelyn H Kim

Advances in breast cancer control will be greatly aided by early detection so as to diagnose and treat breast cancer in its preinvasive state prior to metastasis. For breast cancer, the second leading cause of cancer-related death among women in the United States, early detection does allow for increased treatment options, including surgical resection, with a corresponding better patient response. Unfortunately, however, many patients' tumors are diagnosed following metastasis, thus making it more difficult to successfully treat the malignancy. There are, at present, no existing validated plasma/serum biomarkers for breast cancer. Only a few biomarkers (such as HER-2/neu, estrogen receptor, and progesterone receptor) have utility for diagnosis and prognosis. Thus, there is a great need for new biomarkers for breast cancer. This paper will focus on the identification of new serum protein biomarkers with utility for the early detection of breast cancer.

早期发现有助于乳腺癌控制的进步,从而在乳腺癌转移前的侵袭前状态进行诊断和治疗。乳腺癌是美国女性癌症相关死亡的第二大原因,早期发现确实可以增加治疗选择,包括手术切除,患者的反应也相应更好。然而,不幸的是,许多患者的肿瘤是在转移后被诊断出来的,这使得成功治疗恶性肿瘤变得更加困难。目前,还没有有效的乳腺癌血浆/血清生物标志物。只有少数生物标志物(如HER-2/neu、雌激素受体和孕激素受体)对诊断和预后有实用价值。因此,迫切需要新的乳腺癌生物标志物。本文将着重于鉴定新的血清蛋白生物标志物,用于乳腺癌的早期检测。
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引用次数: 80
Comprehensive proteomic profiling of aldehyde dehydrogenases in lung adenocarcinoma cell lines. 肺腺癌细胞系醛脱氢酶的综合蛋白质组学分析。
Pub Date : 2011-01-01 Epub Date: 2011-10-29 DOI: 10.1155/2011/145010
Qing Zhang, Ayumu Taguchi, Mark Schliekelman, Chee-Hong Wong, Alice Chin, Rork Kuick, David E Misek, Samir Hanash

We have explored the potential of proteomic profiling to contribute to the delineation of the range of expression and subcellular localization of aldehyde dehydrogenases (ALDHs) in lung adenocarcinoma. In-depth quantitative proteomics was applied to 40 lung adenocarcinoma cell lines resulting in the identification of the known members of the ALDH family. Substantial heterogeneity in the level and occurrence of ALDHs in total lysates and on the cell surface and in their release into the culture media was observed based on mass spectrometry counts. A distinct pattern of expression of ALDHs was observed in cells exhibiting epithelial features relative to cells exhibiting mesenchymal features. Strikingly elevated levels of ALDH1A1 were observed in two cell lines. We also report on the occurrence of an immune response to ALDH1A1 in lung cancer.

我们已经探索了蛋白质组学分析的潜力,有助于描绘肺腺癌中醛脱氢酶(ALDHs)的表达范围和亚细胞定位。将深度定量蛋白质组学应用于40个肺腺癌细胞系,鉴定出已知的ALDH家族成员。基于质谱计数,观察到总裂解物和细胞表面以及释放到培养基中的ALDHs的水平和发生率存在很大的异质性。ALDHs在上皮细胞和间充质细胞中的表达模式不同。在两种细胞系中观察到显著升高的ALDH1A1水平。我们还报道了ALDH1A1在肺癌中发生的免疫反应。
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引用次数: 15
Signatures of drug sensitivity in nonsmall cell lung cancer. 非小细胞肺癌的药物敏感性特征。
Pub Date : 2011-01-01 Epub Date: 2011-08-07 DOI: 10.1155/2011/215496
Hua C Gong, Sean Wang, Gary Mayer, Guoan Chen, Glen Leesman, Sharat Singh, David G Beer

We profiled receptor tyrosine kinase pathway activation and key gene mutations in eight human lung tumor cell lines and 50 human lung tumor tissue samples to define molecular pathways. A panel of eight kinase inhibitors was used to determine whether blocking pathway activation affected the tumor cell growth. The HER1 pathway in HER1 mutant cell lines HCC827 and H1975 were found to be highly activated and sensitive to HER1 inhibition. H1993 is a c-MET amplified cell line showing c-MET and HER1 pathway activation and responsiveness to c-MET inhibitor treatment. IGF-1R pathway activated H358 and A549 cells are sensitive to IGF-1R inhibition. The downstream PI3K inhibitor, BEZ-235, effectively inhibited tumor cell growth in most of the cell lines tested, except the H1993 and H1650 cells, while the MEK inhibitor PD-325901 was effective in blocking the growth of KRAS mutated cell line H1734 but not H358, A549 and H460. Hierarchical clustering of primary tumor samples with the corresponding tumor cell lines based on their pathway signatures revealed similar profiles for HER1, c-MET and IGF-1R pathway activation and predict potential treatment options for the primary tumors based on the tumor cell lines response to the panel of kinase inhibitors.

我们分析了8个人肺肿瘤细胞系和50个人肺肿瘤组织样本中受体酪氨酸激酶途径的激活和关键基因突变,以确定分子途径。一组8种激酶抑制剂被用来确定阻断通路激活是否影响肿瘤细胞生长。在HER1突变细胞系HCC827和H1975中发现HER1通路高度激活,并且对HER1抑制敏感。H1993是一种c-MET扩增细胞系,显示c-MET和HER1途径的激活以及对c-MET抑制剂治疗的反应性。IGF-1R通路激活的H358和A549细胞对IGF-1R抑制敏感。下游PI3K抑制剂BEZ-235在除H1993和H1650细胞外的大多数细胞系中都能有效抑制肿瘤细胞的生长,而MEK抑制剂PD-325901能有效抑制KRAS突变细胞系H1734的生长,但对H358、A549和H460细胞无抑制作用。基于相应肿瘤细胞系的通路特征对原发肿瘤样本进行分层聚类,揭示了HER1、c-MET和IGF-1R通路激活的相似特征,并根据肿瘤细胞系对激酶抑制剂的反应预测了原发肿瘤的潜在治疗方案。
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引用次数: 24
The application of a three-step proteome analysis for identification of new biomarkers of pancreatic cancer. 三步蛋白质组分析在胰腺癌新生物标志物鉴定中的应用。
Pub Date : 2011-01-01 Epub Date: 2011-10-17 DOI: 10.1155/2011/628787
Mayinuer Abulaizi, Takeshi Tomonaga, Mamoru Satoh, Kazuyuki Sogawa, Kazuyuki Matsushita, Yoshio Kodera, Jurat Obul, Shigetsugu Takano, Hideyuki Yoshitomi, Masaru Miyazaki, Fumio Nomura

We searched for novel tumor markers of pancreatic cancer by three-step serum proteome analysis. Twelve serum abundant proteins were depleted using immunoaffinity columns followed by fractionation by reverse-phase high-performance liquid chromatography. Proteins in each fraction were separated by two-dimensional gel electrophoresis. Then the gel was stained by Coomassie Brilliant Blue. Protein spots in which the expression levels were significantly different between cancer and normal control were identified by LC-MS/MS. One hundred and two spots were upregulated, and 84 spots were downregulated in serum samples obtained from patients with pancreatic cancers, and 58 proteins were identified by mass spectrometry. These candidate proteins were validated using western blot analysis and enzyme-linked immunosorbent assay (ELISA). As a result of these validation process, we could confirm that the serum levels of apolipoprotein A-IV, vitamin D-binding protein, plasma retinol-binding protein 4, and tetranectin were significantly decreased in patients with pancreatic cancer.

我们通过三步血清蛋白质组分析寻找新的胰腺癌肿瘤标志物。采用免疫亲和柱分离12种血清富集蛋白,反相高效液相色谱分离。通过双向凝胶电泳分离各组分中的蛋白质。然后用考马斯亮蓝染色。采用LC-MS/MS技术鉴定癌组织与正常对照组表达水平有显著差异的蛋白点。在胰腺癌患者的血清样本中,有102个位点表达上调,84个位点表达下调,并通过质谱法鉴定出58种蛋白。这些候选蛋白通过western blot分析和酶联免疫吸附试验(ELISA)进行验证。通过这些验证过程,我们可以证实胰腺癌患者血清载脂蛋白a - iv、维生素d结合蛋白、血浆视黄醇结合蛋白4和四联素水平显著降低。
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引用次数: 18
Proteomics in pancreatic cancer research. 胰腺癌研究中的蛋白质组学。
Pub Date : 2011-01-01 Epub Date: 2011-08-14 DOI: 10.1155/2011/365350
Ruihui Geng, Zhaoshen Li, Shude Li, Jun Gao

Pancreatic cancer is a highly aggressive malignancy with a poor prognosis and deeply affects the life of people. Therefore, the earlier diagnosis and better treatments are urgently needed. In recent years, the proteomic technologies are well established and growing rapidly and have been widely applied in clinical applications, especially in pancreatic cancer research. In this paper, we attempt to discuss the development of current proteomic technologies and the application of proteomics to the field of pancreatic cancer research. This will explore the potential perspective in revealing pathogenesis, making the diagnosis earlier and treatment.

胰腺癌是一种高度侵袭性的恶性肿瘤,预后差,严重影响人们的生活。因此,迫切需要早期诊断和更好的治疗。近年来,蛋白质组学技术发展迅速,已广泛应用于临床,特别是胰腺癌的研究。本文就蛋白质组学技术的发展现状及在胰腺癌研究中的应用作一综述。这将为揭示发病机制、早期诊断和治疗提供潜在的视角。
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引用次数: 30
期刊
International journal of proteomics
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