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Genetic diversity of Atlantic Bluefin tuna in the Mediterranean Sea: insights from genome-wide SNPs and microsatellites. 地中海大西洋蓝鳍金枪鱼的遗传多样性:来自全基因组snp和微卫星的见解。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2017-02-16 eCollection Date: 2017-12-01 DOI: 10.1186/s40709-017-0062-2
Aglaia Antoniou, Panagiotis Kasapidis, Georgios Kotoulas, Constantinos C Mylonas, Antonios Magoulas

Background: Elucidating the patterns of the Atlantic Bluefin tuna [ABFT, Thunnus thynnus (Linnaeus, 1758)] population structure constitutes a challenging task of great importance. Most of the unique challenges stem from its biology, as well as the attributes of the marine realm in which it disperses. Accurate information is urgently needed for stock assessment, and the identification of critical features to the persistence and adaptation of populations in order to formulate and adopt effective strategies for ABFT conservation and management. Conclusions of a great number of ABFT genetic studies on the Mediterranean Sea stock structure are rather controversial and not yet conclusive. In this study, ABFT genomic diversity was investigated in the Mediterranean Sea, which is the most important area for the species' reproduction.

Results: Analyzing genome-wide SNPs and microsatellites from ABFT samples collected throughout the Mediterranean Sea did not provide strong evidence of genetic structure, pointing towards the existence of a single panmictic unit. An alternative view would recognize a failure to reject the null hypothesis of a panmictic unit as an effect of the study's sampling design, the type of markers used, and the effectiveness/suitability of analysis methods in respect to the species biological characteristics or any combination of the above.

Conclusions: Unravelling the drivers of ABFT population diversity would require the consideration of important aspects of the species spawning behavior for the determination of the appropriate sampling design. Novel approaches and methods of analysis that will bring together experts in genetics/-omics, ecology and oceanography are deemed necessary. Analyzing ABFT genetic data under the discipline of seascape genetics could provide the analysis framework under which major abiotic and biotic forces controlling ABFT recruitment could be identified, elucidating the complicated population dynamics of the species, while multiple and continuous fisheries monitoring should in all cases be considered as a prerequisite in order to achieve efficient and long-term ABFT conservation.

背景:阐明大西洋蓝鳍金枪鱼[ABFT, Thunnus thynnus (Linnaeus, 1758)]种群结构模式是一项具有挑战性的重要任务。大多数独特的挑战源于它的生物学特性,以及它所分布的海洋领域的特性。为了制定和采取有效的保护和管理策略,迫切需要准确的信息来进行种群评估,并确定种群持久性和适应性的关键特征。大量关于地中海种群结构的ABFT遗传研究的结论颇有争议,尚未得出定论。在本研究中,研究了地中海地区ABFT基因组多样性,这是该物种繁殖最重要的地区。结果:从整个地中海收集的ABFT样本中分析全基因组snp和微卫星并没有提供遗传结构的有力证据,表明存在单一的泛疫单位。另一种观点认为,由于研究的抽样设计、所使用的标记类型以及有关物种生物学特征的分析方法的有效性/适用性或上述任何组合的影响,未能拒绝流感单位的零假设。结论:揭示ABFT种群多样性的驱动因素需要考虑物种产卵行为的重要方面,以确定适当的抽样设计。将遗传学/组学、生态学和海洋学专家聚集在一起的新方法和分析方法被认为是必要的。在海洋遗传学学科下分析ABFT遗传数据可以提供一个分析框架,在此框架下,可以识别控制ABFT招募的主要非生物和生物力量,阐明该物种复杂的种群动态,而在任何情况下,多重和连续的渔业监测都应被视为有效和长期保护ABFT的先决条件。
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引用次数: 1
DNA methylation changes and TE activity induced in tissue cultures of barley (Hordeum vulgare L.). 大麦(Hordeum vulgare L.)组织培养物中诱导的 DNA 甲基化变化和 TE 活性。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-08-08 eCollection Date: 2016-12-01 DOI: 10.1186/s40709-016-0056-5
Renata Orłowska, Joanna Machczyńska, Sylwia Oleszczuk, Janusz Zimny, Piotr Tomasz Bednarek

Background: In vitro plant regeneration via androgenesis or somatic embryogenesis is capable of inducing (epi)mutations that may affect sexual progenies. While epimutations are associated with DNA methylation, mutations could be due to the movement of transposons. The common notion is that both processes are linked. It is being assumed that demethylation activates transposable elements (TEs). Analysis of methylation changes and their relation with TEs activation in tissue cultures requires uniquely derived donor plants (Ds), their regenerants (Rs) and respective progeny (Ps) that would allow discrimination of processes not related to changes introduced via in vitro cultures. Moreover, a set of methods (RP-HPLC, SSAP, and MSTD) is needed to study whether different TEs families are being activated during in vitro tissue culture plant regeneration and whether their activity could be linked to DNA methylation changes or alternative explanations should be considered.

Results: The in vitro tissue culture plant regeneration in barley was responsible for the induction of DNA methylation in regenerants and conservation of the methylation level in the progeny as shown by the RP-HPLC approach. No difference between andro- and embryo-derived Rs and Ps was observed. The SSAP and MSTD approach revealed that Ds and Rs were more polymorphic than Ps. Moreover, Rs individuals exhibited more polymorphisms with the MSTD than SSAP approach. The differences between Ds, Rs and Ps were also evaluated via ANOVA and AMOVA.

Conclusions: Stressful conditions during plant regeneration via in vitro tissue cultures affect regenerants and their sexual progeny leading to an increase in global DNA methylation of Rs and Ps compared to Ds in barley. The increased methylation level noted among regenerants remains unchanged in the Ps as indicated via RP-HPLC data. Marker-based experiments suggest that TEs are activated via in vitro tissue cultures and that, independently of the increased methylation, their activity in Rs is greater than in Ps. Thus, the increased methylation level may not correspond to the stabilization of TEs movement at least at the level of regenerants. The presence of TEs variation among Ds that were genetically and epigenetically uniform may suggest that at least some mobile elements may be active, and they may mask variation related to tissue cultures. Thus, tissue cultures may activate some TEs whereas the others remain intact, or their level of movement is changed. Finally, we suggest that sexual reproduction may be responsible for the stabilization of TEs.

背景:通过雄性或体细胞胚胎发生进行体外植物再生能够诱导(表突变),从而影响有性后代。表突变与 DNA 甲基化有关,而突变则可能是由于转座子的移动。人们普遍认为这两个过程是相互关联的。据推测,去甲基化会激活转座子(TE)。分析组织培养物中的甲基化变化及其与转座元件(TEs)激活的关系需要独特的供体植株(Ds)、其再生植株(Rs)和各自的后代(Ps),这样才能区分与体外培养物引入的变化无关的过程。此外,还需要一套方法(RP-HPLC、SSAP 和 MSTD)来研究体外组织培养植物再生过程中是否有不同的 TEs 家族被激活,以及它们的活性是否与 DNA 甲基化变化有关,还是应该考虑其他解释:结果:RP-HPLC方法显示,大麦的离体组织培养植物再生可诱导再生体的DNA甲基化,并保持后代的甲基化水平。在雄性和胚胎衍生的 Rs 和 Ps 之间没有观察到差异。SSAP 和 MSTD 方法显示,Ds 和 Rs 的多态性高于 Ps。Ds、Rs和Ps之间的差异也通过方差分析和AMOVA进行了评估:结论:通过体外组织培养进行植物再生过程中的应激条件会影响再生体及其有性后代,导致大麦中 Rs 和 Ps 的 DNA 整体甲基化水平高于 Ds。如 RP-HPLC 数据所示,再生体中增加的甲基化水平在 Ps 中保持不变。基于标记的实验表明,TEs 是通过体外组织培养激活的,与甲基化水平的提高无关,Rs 中 TEs 的活性高于 Ps。在遗传和表观遗传一致的 Ds 中存在 TEs 变异,这可能表明至少有一些移动元素是活跃的,它们可能掩盖了与组织培养有关的变异。因此,组织培养可能会激活一些 TEs,而其他 TEs 则保持不变,或者它们的移动水平发生了变化。最后,我们认为有性生殖可能是稳定 TEs 的原因。
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引用次数: 0
Identification and validation of Aeluropus littoralis reference genes for Quantitative Real-Time PCR Normalization. 实时荧光定量PCR归一化的海雀内参基因鉴定与验证。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-07-19 eCollection Date: 2016-12-01 DOI: 10.1186/s40709-016-0053-8
Seyyed Hamidreza Hashemi, Ghorbanali Nematzadeh, Gholamreza Ahmadian, Ahad Yamchi, Markus Kuhlmann

Background: The use of stably expressed genes as normalizers has crucial role in accurate and reliable expression analysis estimated by quantitative real-time polymerase chain reaction (qPCR). Recent studies have shown that, the expression levels of common housekeeping genes are varying in different tissues and experimental conditions. The genomic DNA contamination in RNA samples is another important factor that also influence the interpretation of the data obtained from qPCR. It is estimated that the gDNA contamination in gene expression analysis lead to an overestimation of the RNA transcript level. The aim of this study was to validate the most stably expressed reference genes in two different tissues of Aeluropus littoralis-halophyte grass at salt stress and recovery condition. Also, a qPCR-based approach for monitoring contamination with gDNA was conducted.

Results: Ten candidate reference genes participating in different biological processes were analyzed in four groups of samples including root and leaf tissues, salt stress and recovery condition. To determine the most stably expressed reference genes, three statistical methods (geNorm, NormFinder and BestKeeper) were applied. According to results obtained, ten candidate reference genes were ranked based on the stability of their expression. Here, our results show that a set of four housekeeping genes (HKGs) e.g. RPS3, EF1A, GTF and RPS12 could be used as general reference genes for the all selected conditions and tissues. Also, four set of reference genes were proposed for each tissue and condition including: RPS3, EF1A and UBQ for salt stress and root samples; RPS3, EF1A, UBQ as well as GAPDH for recovery condition; U2SURP and GTF for leaf samples. Additionally, for assessing DNA contamination in RNA samples, a set of unique primers were designed based on the conserved region of ribosomal DNA (rDNA). The universality, specificity and sensitivity of these primer pairs were also evaluated in Poaceae.

Conclusions: Overall, the sets of reference genes proposed in this study are ideal normalizers for qPCR analysis in A. littoralis transcriptome. The novel reference gene e.g. RPS3 that applied this study had higher expression stability than commonly used housekeeping genes. The application of rDNA-based primers in qPCR analysis was addressed.

背景:使用稳定表达的基因作为正常化因子在定量实时聚合酶链反应(qPCR)准确可靠的表达分析中起着至关重要的作用。最近的研究表明,在不同的组织和实验条件下,常见管家基因的表达水平是不同的。RNA样品中的基因组DNA污染也是影响qPCR数据解释的另一个重要因素。据估计,基因表达分析中的dna污染导致了对RNA转录水平的高估。本研究的目的是验证盐胁迫和恢复条件下滨海海茅两种不同组织中最稳定表达的内参基因。此外,还进行了基于qpcr的dna污染监测方法。结果:在根和叶组织、盐胁迫和恢复条件4组样品中,分析了10个参与不同生物过程的候选内参基因。为了确定最稳定表达的内参基因,采用了三种统计方法(geNorm、NormFinder和BestKeeper)。根据得到的结果,根据10个候选内参基因的表达稳定性进行排序。本研究结果表明,RPS3、EF1A、GTF和RPS12等4个内参基因可作为所有选择条件和组织的一般内参基因。同时,针对不同的组织和条件提出了4组内参基因,分别为:盐胁迫和根系样品的RPS3、EF1A和UBQ;RPS3、EF1A、UBQ、GAPDH为恢复状态;叶片样品的U2SURP和GTF。此外,为了评估RNA样品中的DNA污染,基于核糖体DNA (rDNA)的保守区域设计了一套独特的引物。并对这些引物对在禾科植物中的普遍性、特异性和敏感性进行了评价。结论:总的来说,本研究中提出的内参基因集是对滨荆转录组进行qPCR分析的理想归一化基因。应用于本研究的新型内参基因如RPS3比常用的内参基因具有更高的表达稳定性。介绍了rdna引物在qPCR分析中的应用。
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引用次数: 15
Hominin interbreeding and the evolution of human variation. 古人类杂交与人类变异的进化。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-07-16 eCollection Date: 2016-12-01 DOI: 10.1186/s40709-016-0054-7
Kwang Hyun Ko

Mitochondrial Eve confirms the "out of Africa" theory, but the evidence also supports interbreeding between Homo sapiens and other hominins: Neanderthals, Denisovans, and Homo heidelbergensis. This article explains how interbreeding between early H. sapiens and archaic hominins occurred. The availability of edible insects in East Asia aided the spread of the unaggressive, highly cooperative Neanderthals, who interbred with H. sapiens in Asia, resulting in a higher admixture of Neanderthal DNA in East Asian populations. Geographical variation in degree of interbreeding between H. sapiens and Neanderthals likely contributed to neurological and behavioral differences in modern humans. Similarly, people with Denisovan genetic admixture were better able to dwell in mountainous regions, allowing their genetic legacy to cross the Himalayas and persist in Southeast Asian and Oceanian H. sapiens. In the Sub-Saharan region, unaffected by Denisovan or Neanderthal interbreeding, H. sapiens interbred with H. heidelbergensis, because high humidity militated against fire-making and allowed the survival of these non-fire-making hominins.

线粒体夏娃证实了“走出非洲”的理论,但证据也支持智人与其他人类之间的杂交:尼安德特人、丹尼索瓦人和海德堡人。这篇文章解释了早期智人与古人类的杂交是如何发生的。东亚地区可食用昆虫的出现,帮助了尼安德特人的传播,他们与亚洲的智人杂交,导致了东亚人群中尼安德特人DNA的高度混合。智人与尼安德特人杂交程度的地理差异可能导致现代人神经和行为上的差异。同样,拥有丹尼索瓦人遗传混合基因的人更能在山区生活,使他们的遗传遗产能够跨越喜马拉雅山脉,在东南亚和大洋洲的智人中留存下来。在撒哈拉以南地区,没有受到丹尼索瓦人或尼安德特人杂交的影响,智人与海德堡人杂交,因为高湿度不利于生火,使这些不生火的古人类得以生存。
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引用次数: 14
Ganges: special at its origin. 恒河:它的起源很特别。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-07-15 eCollection Date: 2016-12-01 DOI: 10.1186/s40709-016-0055-6
Krishna Khairnar
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引用次数: 14
Prediction of protein-protein interaction sites by means of ensemble learning and weighted feature descriptor. 基于集成学习和加权特征描述子的蛋白质相互作用位点预测。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-07-04 eCollection Date: 2016-05-01 DOI: 10.1186/s40709-016-0046-7
Xiuquan Du, Shiwei Sun, Changlin Hu, Xinrui Li, Junfeng Xia

Background: Reliable prediction of protein-protein interaction sites is an important goal in the field of bioinformatics. Many computational methods have been explored for the large-scale prediction of protein-protein interaction sites based on various data types, including protein sequence, structural and genomic data. Although much progress has been achieved in recent years, the problem has not yet been satisfactorily solved.

Results: In this work, we presented an efficient approach that uses ensemble learning algorithm with weighted feature descriptor (EL-WFD) to predict protein-protein interaction sites. Moreover, weighted feature descriptor was designed to describe the distance influence of neighboring residues on interaction sites. The results on two dataset (Hetero and Homo), show that the proposed method yields a satisfactory accuracy with 83.8 % recall and 96.3 % precision on the Hetero dataset and 84.2 % recall and 96.3 % precision on the Homo dataset, respectively. In both datasets, our method tend to obtain high Mathews correlation coefficient compared with state-of-the-art technique random forest method.

Conclusions: The experimental results show that the EL-WFD method is quite effective in predicting protein-protein interaction sites. The novel weighted feature descriptor was proved to be promising in discovering interaction sites. Overall, the proposed method can be considered as a new powerful tool for predicting protein-protein interaction sites with excellence performance.

背景:蛋白质相互作用位点的可靠预测是生物信息学领域的一个重要目标。基于各种数据类型,包括蛋白质序列、结构和基因组数据,已经探索了许多计算方法来大规模预测蛋白质-蛋白质相互作用位点。尽管近年来取得了很大进展,但这一问题尚未得到圆满解决。结果:在这项工作中,我们提出了一种有效的方法,使用加权特征描述符(EL-WFD)的集成学习算法来预测蛋白质-蛋白质相互作用位点。此外,设计了加权特征描述符来描述相邻残基对相互作用位点的距离影响。在两个数据集(Hetero和Homo)上的结果表明,该方法在Hetero数据集上的查全率和查准率分别为83.8%和96.3%,在Homo数据集上的查全率和查准率分别为84.2%和96.3%。在这两个数据集上,与目前最先进的随机森林方法相比,我们的方法倾向于获得较高的马修斯相关系数。结论:实验结果表明,EL-WFD方法在预测蛋白-蛋白相互作用位点方面是非常有效的。结果表明,该加权特征描述符在发现交互点方面具有较好的应用前景。总之,该方法是预测蛋白质-蛋白质相互作用位点的一种强有力的新工具,具有优良的性能。
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引用次数: 7
An approach to semantic query expansion system based on Hepatitis ontology. 一种基于肝炎本体的语义查询扩展系统方法。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-07-04 eCollection Date: 2016-05-01 DOI: 10.1186/s40709-016-0044-9
Chen Yunzhi, Lu Huijuan, Linda Shapiro, Ravensara S Travillian, Li Lanjuan

Background: Ontology development, as an increasingly practical vehicle applied in various fields, plays a significant role in knowledge management. This paper, focusing on constructing and querying a hepatitis ontology, aims to provide a framework for ontology-based medical services. The paper is devoted to the algorithm of query expansion for the hepatitis ontology, including synonym expansion, hypernym/hyponym expansion and expansion of similar words. It applies semantic similarity calculation to judge the similarity of retrieval terms.

Results: The paper proposes a new prototype system. The accuracy of query expansion is improved in both precision@40 and AP@40, which indicates that query expansion improves the accuracy of the query after using the method proposed in this paper.

Conclusions: The paper has adopted semantic similarity computing to improve retrieval performance. Experiments show that search precision of query expansion is higher based on domain concept relationship.

背景:本体开发作为一种越来越实用的工具应用于各个领域,在知识管理中发挥着重要的作用。本文主要研究肝炎本体的构建和查询,旨在为基于本体的医疗服务提供一个框架。本文研究了肝炎本体的查询扩展算法,包括同义词扩展、上下词扩展和相似词扩展。它采用语义相似度计算来判断检索词的相似度。结果:本文提出了一种新的原型系统。在precision@40和AP@40中都提高了查询扩展的准确性,说明使用本文提出的方法后,查询扩展提高了查询的准确性。结论:本文采用语义相似度计算提高了检索性能。实验表明,基于领域概念关系的查询扩展具有较高的搜索精度。
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引用次数: 26
PredPhos: an ensemble framework for structure-based prediction of phosphorylation sites. PredPhos:一个基于结构预测磷酸化位点的集成框架。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-07-04 eCollection Date: 2016-05-01 DOI: 10.1186/s40709-016-0042-y
Yong Gao, Weilin Hao, Jing Gu, Diwei Liu, Chao Fan, Zhigang Chen, Lei Deng

Background: Post-translational modifications (PTMs) occur on almost all proteins and often strongly affect the functions of modified proteins. Phosphorylation is a crucial PTM mechanism with important regulatory functions in biological systems. Identifying the potential phosphorylation sites of a target protein may increase our understanding of the molecular processes in which it takes part.

Results: In this paper, we propose PredPhos, a computational method that can accurately predict both kinase-specific and non-kinase-specific phosphorylation sites by using optimally selected properties. The optimal combination of features was selected from a set of 153 novel structural neighborhood properties by a two-step feature selection method consisting of a random forest algorithm and a sequential backward elimination method. To overcome the imbalanced problem, we adopt an ensemble method, which combines bootstrap resampling technique, support vector machine-based fusion classifiers and majority voting strategy. We evaluate the proposed method using both tenfold cross validation and independent test. Results show that our method achieves a significant improvement on the prediction performance for both kinase-specific and non-kinase-specific phosphorylation sites.

Conclusions: The experimental results demonstrate that the proposed method is quite effective in predicting phosphorylation sites. Promising results are derived from the new structural neighborhood properties, the novel way of feature selection, as well as the ensemble method.

翻译后修饰(Post-translational modification, PTMs)发生在几乎所有的蛋白质上,并且常常强烈地影响被修饰蛋白质的功能。磷酸化是一个重要的PTM机制,在生物系统中具有重要的调控功能。确定目标蛋白的潜在磷酸化位点可能会增加我们对其参与的分子过程的理解。结果:在本文中,我们提出了PredPhos,这是一种计算方法,可以通过最优选择的性质准确预测激酶特异性和非激酶特异性磷酸化位点。采用随机森林算法和顺序反向消去法两步特征选择方法,从153个新结构邻域属性中选择最优特征组合。为了克服不平衡问题,我们采用了一种集成方法,该方法结合了自举重采样技术、基于支持向量机的融合分类器和多数投票策略。我们使用十倍交叉验证和独立检验来评估所提出的方法。结果表明,我们的方法对激酶特异性和非激酶特异性磷酸化位点的预测性能都有显著提高。结论:实验结果表明,该方法对磷酸化位点的预测是非常有效的。新的结构邻域特性、新的特征选择方法以及集成方法都取得了令人满意的结果。
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引用次数: 8
Iterative ensemble feature selection for multiclass classification of imbalanced microarray data. 不平衡微阵列数据多类分类的迭代集成特征选择。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-07-04 eCollection Date: 2016-05-01 DOI: 10.1186/s40709-016-0045-8
Junshan Yang, Jiarui Zhou, Zexuan Zhu, Xiaoliang Ma, Zhen Ji

Background: Microarray technology allows biologists to monitor expression levels of thousands of genes among various tumor tissues. Identifying relevant genes for sample classification of various tumor types is beneficial to clinical studies. One of the most widely used classification strategies for multiclass classification data is the One-Versus-All (OVA) schema that divides the original problem into multiple binary classification of one class against the rest. Nevertheless, multiclass microarray data tend to suffer from imbalanced class distribution between majority and minority classes, which inevitably deteriorates the performance of the OVA classification.

Results: In this study, we propose a novel iterative ensemble feature selection (IEFS) framework for multiclass classification of imbalanced microarray data. In particular, filter feature selection and balanced sampling are performed iteratively and alternatively to boost the performance of each binary classification in the OVA schema. The proposed framework is tested and compared with other representative state-of-the-art filter feature selection methods using six benchmark multiclass microarray data sets. The experimental results show that IEFS framework provides superior or comparable performance to the other methods in terms of both classification accuracy and area under receiver operating characteristic curve. The more number of classes the data have, the better performance of IEFS framework achieves.

Conclusions: Balanced sampling and feature selection together work well in improving the performance of multiclass classification of imbalanced microarray data. The IEFS framework is readily applicable to other biological data analysis tasks facing the same problem.

背景:微阵列技术允许生物学家监测各种肿瘤组织中数千种基因的表达水平。识别相关基因对各种肿瘤类型进行样本分类,有利于临床研究。对于多类分类数据,最广泛使用的分类策略之一是One- versus - all (OVA)模式,它将原始问题划分为一个类对其他类的多个二元分类。然而,多类微阵列数据往往存在多数类和少数类之间的类分布不平衡的问题,这不可避免地会影响OVA分类的性能。结果:在本研究中,我们提出了一种新的迭代集成特征选择(IEFS)框架,用于不平衡微阵列数据的多类分类。特别是,过滤器特征选择和平衡采样是迭代地或交替地执行的,以提高OVA模式中每个二元分类的性能。使用六个基准多类微阵列数据集对所提出的框架进行了测试,并与其他具有代表性的最先进的滤波器特征选择方法进行了比较。实验结果表明,IEFS框架在分类精度和接收者工作特征曲线下面积方面均优于或与其他方法相当。类的数量越多,IEFS框架的性能越好。结论:平衡采样和特征选择相结合可以很好地提高对不平衡微阵列数据的多类分类性能。IEFS框架很容易适用于面临同样问题的其他生物数据分析任务。
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引用次数: 30
Targeted CYP2E1 quantification and its correlation to currently acceptable clinical biochemical indices. 靶向CYP2E1定量及其与目前可接受的临床生化指标的相关性。
IF 3.3 3区 生物学 Q1 Agricultural and Biological Sciences Pub Date : 2016-06-30 eCollection Date: 2016-12-01 DOI: 10.1186/s40709-016-0052-9
Christina Gertrude Yap, Anuar Zaini, Iekhsan Othman

Background: The Cytochrome P450 enzymes are commonly known for their major role in metabolism. Besides its metabolic role, CYP2E1 gene expression has been associated with the onset of diabetic nephropathy. CYP2E1 protein elevation has also been reported to be responsible for the production of reactive oxygen species. The aims of this study were (i) to optimize and validate a targeted proteomic approach for quantitating CYP2E1 and validating it as a suitable clinical test, (ii) to investigate the concurrency between ESI-LCMS-MS quantitated circulating CYP2E1 and gold standard indices in the context of outpatient point-of-care clinical settings involving various groups of diabetic patients and (iii) to investigate the concurrency profile of circulating CYP2E1 protein, CYP2E1 gene expression and reactive oxygen species (ROS). This is a cross sectional study involving three groups of subjects (n = 166): control, pre-diabetes, and diabetes. We optimized a targeted proteomic approach for absolute quantification of CYP2E1. "YPEIEEK" and "GTVVVPTLYDNQEFPDPEK" were the representative peptides of CYP2E1 for our analytical method. Deuterated forms of "YPEIEEK" and "GTVVVPTLYDNQEFPDPEK" were used as internal standards. Lymphocytes were isolated from whole blood, microsomes were prepared, followed by in-solution digestion for production of tryptic peptides. Amounts of "YPEIEEK" and "GTVVVPTLYDNQEFPDPEK" from patients' samples were calculated from a calibration curve.

Results: "YPEIEEK" is a unique and reliable representative peptide for CYP2E1 quantification. "GTVVVPTLYDNQEFPDPEK" showed poor reproducibility and sensitivity. Incremental amounts of CYP2E1 protein in the peripheral circulation clearly showed concurrency with CYP2E1 gene expression and ROS levels in our study population. Elevations of CYP2E1 were observed even when gold standard clinical indicator for glycemic control (HbA1c) was within normal reference limits. Quantitated amounts of CYP2E1 protein in the pre-diabetes and diabetes groups showed significant difference relative to control group (p < 0.001). No significant differences were observed between the medians of pre-diabetes and diabetes groups (p = 0.870).

Conclusions: CYP2E1 protein in peripheral blood can be reliably quantitated by the validated targeted proteomic approach method. Quantifiable amounts of CYP2E1 preceded abnormal HbA1C levels which indicates quantitation of CYP2E1 could be useful as an additional tool for early indication of diabetic risks and it complications.

背景:细胞色素P450酶因其在代谢中的主要作用而广为人知。除了代谢作用外,CYP2E1基因表达与糖尿病肾病的发病有关。据报道,CYP2E1蛋白的升高也与活性氧的产生有关。本研究的目的是(i)优化和验证一种靶向蛋白质组学方法,用于定量CYP2E1,并将其作为一种合适的临床试验,(ii)调查ESI-LCMS-MS定量循环CYP2E1和金标准指标在门诊护理点临床环境下的并发性,涉及不同组的糖尿病患者,(iii)调查循环CYP2E1蛋白的并发性。CYP2E1基因表达与活性氧(ROS)。这是一项横断面研究,涉及三组受试者(n = 166):对照组、糖尿病前期和糖尿病。我们优化了一种靶向蛋白质组学方法,用于CYP2E1的绝对定量。“YPEIEEK”和“GTVVVPTLYDNQEFPDPEK”是我们分析方法中CYP2E1的代表性肽段。“YPEIEEK”和“GTVVVPTLYDNQEFPDPEK”的氘化形式被用作内标。从全血中分离淋巴细胞,制备微粒体,然后在溶液中消化产生色氨酸肽。根据校准曲线计算患者样品中的“YPEIEEK”和“GTVVVPTLYDNQEFPDPEK”的量。结果:“YPEIEEK”是一种独特、可靠的CYP2E1定量代表肽。“GTVVVPTLYDNQEFPDPEK”重现性和灵敏度较差。在我们的研究人群中,外周循环中CYP2E1蛋白的增量量明显与CYP2E1基因表达和ROS水平同步。即使血糖控制的金标准临床指标(HbA1c)在正常参考范围内,也观察到CYP2E1升高。糖尿病前期组和糖尿病组CYP2E1蛋白定量与对照组相比有显著差异(p)。结论:经验证的靶向蛋白质组学方法可以可靠地定量外周血CYP2E1蛋白。可量化的CYP2E1量先于异常HbA1C水平,这表明CYP2E1的定量可以作为糖尿病风险及其并发症早期指示的额外工具。
{"title":"Targeted CYP2E1 quantification and its correlation to currently acceptable clinical biochemical indices.","authors":"Christina Gertrude Yap,&nbsp;Anuar Zaini,&nbsp;Iekhsan Othman","doi":"10.1186/s40709-016-0052-9","DOIUrl":"https://doi.org/10.1186/s40709-016-0052-9","url":null,"abstract":"<p><strong>Background: </strong>The Cytochrome P450 enzymes are commonly known for their major role in metabolism. Besides its metabolic role, CYP2E1 gene expression has been associated with the onset of diabetic nephropathy. CYP2E1 protein elevation has also been reported to be responsible for the production of reactive oxygen species. The aims of this study were (i) to optimize and validate a targeted proteomic approach for quantitating CYP2E1 and validating it as a suitable clinical test, (ii) to investigate the concurrency between ESI-LCMS-MS quantitated circulating CYP2E1 and gold standard indices in the context of outpatient point-of-care clinical settings involving various groups of diabetic patients and (iii) to investigate the concurrency profile of circulating CYP2E1 protein, CYP2E1 gene expression and reactive oxygen species (ROS). This is a cross sectional study involving three groups of subjects (n = 166): control, pre-diabetes, and diabetes. We optimized a targeted proteomic approach for absolute quantification of CYP2E1. \"YPEIEEK\" and \"GTVVVPTLYDNQEFPDPEK\" were the representative peptides of CYP2E1 for our analytical method. Deuterated forms of \"YPEIEEK\" and \"GTVVVPTLYDNQEFPDPEK\" were used as internal standards. Lymphocytes were isolated from whole blood, microsomes were prepared, followed by in-solution digestion for production of tryptic peptides. Amounts of \"YPEIEEK\" and \"GTVVVPTLYDNQEFPDPEK\" from patients' samples were calculated from a calibration curve.</p><p><strong>Results: </strong>\"YPEIEEK\" is a unique and reliable representative peptide for CYP2E1 quantification. \"GTVVVPTLYDNQEFPDPEK\" showed poor reproducibility and sensitivity. Incremental amounts of CYP2E1 protein in the peripheral circulation clearly showed concurrency with CYP2E1 gene expression and ROS levels in our study population. Elevations of CYP2E1 were observed even when gold standard clinical indicator for glycemic control (HbA1c) was within normal reference limits. Quantitated amounts of CYP2E1 protein in the pre-diabetes and diabetes groups showed significant difference relative to control group (p < 0.001). No significant differences were observed between the medians of pre-diabetes and diabetes groups (p = 0.870).</p><p><strong>Conclusions: </strong>CYP2E1 protein in peripheral blood can be reliably quantitated by the validated targeted proteomic approach method. Quantifiable amounts of CYP2E1 preceded abnormal HbA1C levels which indicates quantitation of CYP2E1 could be useful as an additional tool for early indication of diabetic risks and it complications.</p>","PeriodicalId":50251,"journal":{"name":"Journal of Biological Research-Thessaloniki","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2016-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s40709-016-0052-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34699118","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 3
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Journal of Biological Research-Thessaloniki
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