抗杀虫双小菜蛾和抗杀螟丹小菜娥酯酶同工酶的研究

用聚丙烯酰胺凝胶电泳法对抗杀虫双、抗杀螟丹的两个小菜蛾品系的酯酶同工酶研究结果表明：小菜蛾对杀虫双和杀螟丹的抗性与特异性酯酶同工酶的形成及部分非特异性酯酶同工酶活性的变化有关;杀虫双和杀螟丹在选育小菜蛾抗性的过程中对其酯酶同工酶的影响有很大的相似性。     

抗杀虫双小菜蛾和抗杀螟丹小菜蛾酯酶同工酶的研究

用聚丙烯酰胺凝胶电泳法对抗杀虫双、抗杀螟丹的两个小菜蛾品系的酯酶同工酶研究结果表明：小菜蛾对杀虫双和杀螟丹的抗性与特异酯酶同工酶的形成及部分非特异性酯酶同工酶活性的变化有关;杀虫和杀螟丹在选育小菜蛾抗性的过程中过程中对其酯酶同工酶的影响有很大的相似性。     

中国东部假俭草种质资源多样性初步研究Ⅳ--同工酶分析

利用聚丙烯酰胺垂直凝胶电泳技术,对中国东部地区16份代表性假俭草种质资源两种同工酶进行了测定。结果表明,种源间过氧化物酶和酯酶同工酶酶谱在酶带数目、相对迁移率上均不尽相同,呈现出较丰富的多样性。     

中国东部假俭草种质资源多样性初步研究IV——同工酶分析

利用聚丙烯酰胺垂直凝胶电泳技术,对中国东部地区16份代表性假俭草种质资源两种同工酶进行了测定。结果表明,种源间过氧化物酶和酯酶同工酶酶谱在酶带数目、相对迁移率上均不尽相同,呈现出较丰富的多样性。     

高粱属A-PAGE 图谱的建立及在种子鉴定中的应用

以高粱、苏丹草等5种高粱属植物的种子为材料,利用酸性聚丙烯酰胺凝胶电泳(A-PAGE)检测其醇溶蛋白.同时研究了丙酮处理、样品上样量和凝胶浓度对电泳的影响.结果表明:样品经丙酮处理后,在上样量为32μg、凝胶质量分数为15%时电泳可以得到清晰的A-PAGE图谱,据此可以鉴定5种高粱属植物.     

高粱属植物对土壤镉吸收及亚细胞的分配

通过盆栽模拟试验研究了3种高粱属植物(甜高粱、高丹草和苏丹草)对土壤cd的富集效应及其亚细胞的分配.结果表明:cd在3种高粱属植物不同部位的富集量随着cd处理浓度的增加而增加,生育期内3种高粱属植物根系中Cd的富集量大于茎鞘和叶片中的富集量;高丹草对Cd的富集量高,甜高梁次之,苏丹草最低;Cd在高粱属植物叶片、茎鞘和根系中各亚细胞组分中的分布相似表现为细胞壁>可溶性部分>细胞核、叶绿体组分>线粒体.     

意蜂和中蜂四种同工酶的研究

用聚丙烯酰胺凝胶等电聚焦电泳分析了意蜂和中蜂的酯酶(Est)、异柠檬酸脱氢酶(Idh)、苹果酸酶(Me)和苹果酸脱氢酶(Mdh)同工酶.两个蜂种的四种同工酶谱有不同程度的差别、意蜂酯酶Ⅳ和苹果酸脱氢酶Ⅲ是多态性的;中蜂的四种同工酶没有多态现象.     

叶甲科三亚科十三种叶甲酯酶同工酶的研究(鞘翅目,叶甲总科)

应用聚丙烯酰胺凝胶电泳方法分析研究了叶甲科3亚科13种昆虫的酯酶同工酶.结果显示,其聚类结果与传统分类结果基本相一致,说明以酯酶同工酶作为研究手段来进行叶甲类昆虫亚科以下阶元的分类是可行的,同时也说明了它们酯酶同工酶酶谱的差异和其分类地位是一致的;但跳甲亚科和叶甲亚科先聚为一类,再与萤叶甲亚科聚为一类,与前人的研究有差异,作者认为:酯酶同工酶的编码基因可能是快进化单位,在解决亚科以下阶元的系统关系时是很好的分子标记,而对于研究叶甲科、亚科间的系统关系,就不一定很合适.     

柑桔近缘植物酯酶同工酶的研究

本文用聚丙烯酰胺凝胶电泳测定了柑桔近缘植物14个种群的种子及幼苗的酯酶同工酶,根据酶谱及扫描图的异同,分析了彼此的亲缘关系,试验结果表明,柑桔近缘植物种属间的酯酶同工酶的酶带数目,酶活性,迁移率及酶谱扫描均有不同程度的差异,同一品种不同发育时期的同工酶也具有不同表现形式,特别是柑桔种子的酯酶同工酶谱一般较稳定,可以作为柑桔亲缘关系的生化遗传指标。     

柑桔近缘植物酯酶同工酶的研究

本文用聚丙烯酰胺凝胶电泳测定了柑桔近缘植物14个种群的种子及幼苗的酯酶同工酶,根据酶谱及扫描图的异同,分析了彼此的亲缘关系,试验结果表明,柑桔近缘植物种属间的酯酶同工酶的酶带数目,酶活性,迁移率及酶谱扫描均有不同程度的差异,同一品种不同发育时期的同工酶也具有不同表现形式,特别是柑桔种子的酯酶同工酶谱一般较稳定,可以作为柑桔亲缘关系的生化遗传指标。     

档案馆库房中微生物的研究

通过对安徽省不同地区档案馆库房中微生物的研究,结果表明：被污染的档案馆库房中的微生物主要优势菌为霉菌,细菌为其次,同时也有一定数量放线菌。不同管理层次档案馆库房中的微生物数量相差较大。     

Proteomic analysis of formalin-fixed, paraffin-embedded lung neuroendocrine tumor samples from hospital archives

Hospital tissue repositories host an invaluable supply of diseased samples with matched retrospective clinical information. In this work, a recently optimized method for extracting full-length proteins from formalin-fixed, paraffin-embedded (FFPE) tissues was evaluated on lung neuroendocrine tumor (LNET) samples collected from hospital repositories. LNETs comprise a heterogeneous spectrum of diseases, for which subtype-specific diagnostic markers are lacking. Six archival samples diagnosed as typical carcinoid (TC) or small cell lung carcinoma (SCLC) were subjected to a full-length protein extraction followed by a GeLC-MS/MS analysis, enabling the identification of over 300 distinct proteins per tumor subtype. All identified proteins were categorized through DAVID software, revealing a differential distribution of functional classes, such as those involved in RNA processing, response to oxidative stress and ion homeostasis. Moreover, using spectral counting for protein abundance estimation and beta-binomial test as statistical filter, a list of 28 differentially expressed proteins was generated and submitted to pathway analysis by means of Ingenuity Pathway Analysis software. Differential expression of chromogranin-A (more expressed in TCs) and stathmin (more expressed in SCLCs) was consistently confirmed by immunohistochemistry. Therefore, FFPE hospital archival samples can be successfully subjected to proteomic investigations aimed to biomarker discovery following a GeLC-MS/MS label-free approach.     

Public proteomic MS repositories and pipelines: available tools and biological applications

As proteomic MS has increased in throughput, so has the demand to catalogue the increasing number of peptides and proteins observed by the community using this technique. As in other 'omics' fields, this brings obvious scientific benefits such as sharing of results and prevention of unnecessary repetition, but also provides technical insights, such as the ability to compare proteome coverage between different laboratories, or between different proteomic platforms. Journals are also moving towards mandating that proteomics data be submitted to public repositories upon publication. In response to these demands, several web-based repositories have been established to store protein and peptide identifications derived from MS data, and a similar number of peptide identification software pipelines have emerged to deliver identifications to these repositories. This paper reviews the latest developments in public domain peptide and protein identification databases and describes the analysis pipelines that feed them. Recent applications of the tools to pertinent biological problems are examined, and through comparing and contrasting the capabilities of each system, the issues facing research users of web-based repositories are explored. Future developments and mechanisms to enhance system functionality and user-interfacing opportunities are also suggested.     

Application of 2-D DIGE to formalin-fixed, paraffin-embedded tissues

The ability to investigate the proteome of formalin-fixed, paraffin-embedded (FFPE) tissues can be considered a major recent achievement in the field of clinical proteomics. However, gel-based approaches to the investigation of FFPE tissue proteomes have lagged behind, mainly because of insufficient quality of full-length protein extracts. Here, the 2-D DIGE technology was investigated for applicability to FFPE proteins, for internal reproducibility among replicate FFPE extracts, and for comparability between FFPE and fresh-frozen tissue profiles. The 2-D DIGE patterns obtained upon labeling and electrophoresis of replicate FFPE tissue extracts were highly reproducible, with satisfactory resolution and complexity. Moreover, the implementation of DIGE enabled to highlight and characterize the consistent differences found in the FFPE profiles compared with fresh-frozen profiles, represented by an acidic shift, directly correlated to the protein pI value, and by a reduction in spot signal intensity, directly correlated to molecular weight and percentage of lysine residues. Being constantly and reproducibly present in all FFPE tissue extract replicates at similar extents, these modifications do not appear to hinder the comparative analysis of FFPE tissue extracts by 2-D DIGE, opening the way to its application for the differential proteomic investigation of archival tissue repositories.     

Using next-generation sequencing for high resolution multiplex analysis of copy number variation from nanogram quantities of DNA from formalin-fixed paraffin-embedded specimens

The use of next-generation sequencing technologies to produce genomic copy number data has recently been described. Most approaches, however, reply on optimal starting DNA, and are therefore unsuitable for the analysis of formalin-fixed paraffin-embedded (FFPE) samples, which largely precludes the analysis of many tumour series. We have sought to challenge the limits of this technique with regards to quality and quantity of starting material and the depth of sequencing required. We confirm that the technique can be used to interrogate DNA from cell lines, fresh frozen material and FFPE samples to assess copy number variation. We show that as little as 5 ng of DNA is needed to generate a copy number karyogram, and follow this up with data from a series of FFPE biopsies and surgical samples. We have used various levels of sample multiplexing to demonstrate the adjustable resolution of the methodology, depending on the number of samples and available resources. We also demonstrate reproducibility by use of replicate samples and comparison with microarray-based comparative genomic hybridization (aCGH) and digital PCR. This technique can be valuable in both the analysis of routine diagnostic samples and in examining large repositories of fixed archival material.     

A comparative evaluation of technical solutions for long-term data repositories in integrative biodiversity research

The current study investigates existing infrastructure, its technical solutions and implemented standards for data repositories related to integrative biodiversity research. The storage and reuse of complex biodiversity data in central databases are becoming increasingly important, particularly in attempts to cope with the impacts of environmental change on biodiversity and ecosystems. From the data side, the main challenge of biodiversity repositories is to deal with the highly interdisciplinary and heterogeneous character of standardized and unstandardized data and metadata covering information from genes to ecosystems. Furthermore, the technical improvements in data acquisition techniques produce ever larger data volumes, which represent a challenge for database structure and proper data exchange.The current study is based on comprehensive in-depth interviews and an online survey addressing IT specialists involved in database development and operation. The results show that metadata are already well established, but that non-meta data still is largely unstandardized across various scientific communities. For example, only a third of all repositories in our investigation use internationally unified semantic standard checklists for taxonomy. The study also showed that database developers are mostly occupied with the implementation of state of the art technology and solving operational problems, leaving no time to implement user's requirements. One of the main reasons for this dissatisfying situation is the undersized and unreliable funding situation of most repositories, as reflected by the marginally small number of permanent IT staff members. We conclude that a sustainable data management system that fosters the future use and reuse of these valuable data resources requires the development of fewer, but more permanent data repositories using commonly accepted standards for their long-term data. This can only be accomplished through the consolidation of hitherto widely scattered small and non-permanent repositories.     

Centralized mouse repositories

Because the mouse is used so widely for biomedical research and the number of mouse models being generated is increasing rapidly, centralized repositories are essential if the valuable mouse strains and models that have been developed are to be securely preserved and fully exploited. Ensuring the ongoing availability of these mouse strains preserves the investment made in creating and characterizing them and creates a global resource of enormous value. The establishment of centralized mouse repositories around the world for distributing and archiving these resources has provided critical access to and preservation of these strains. This article describes the common and specialized activities provided by major mouse repositories around the world.     

The Pig PeptideAtlas: A resource for systems biology in animal production and biomedicine

Biological research of Sus scrofa, the domestic pig, is of immediate relevance for food production sciences, and for developing pig as a model organism for human biomedical research. Publicly available data repositories play a fundamental role for all biological sciences, and protein data repositories are in particular essential for the successful development of new proteomic methods. Cumulative proteome data repositories, including the PeptideAtlas, provide the means for targeted proteomics, system‐wide observations, and cross‐species observational studies, but pigs have so far been underrepresented in existing repositories. We here present a significantly improved build of the Pig PeptideAtlas, which includes pig proteome data from 25 tissues and three body fluid types mapped to 7139 canonical proteins. The content of the Pig PeptideAtlas reflects actively ongoing research within the veterinary proteomics domain, and this article demonstrates how the expression of isoform‐unique peptides can be observed across distinct tissues and body fluids. The Pig PeptideAtlas is a unique resource for use in animal proteome research, particularly biomarker discovery and for preliminary design of SRM assays, which are equally important for progress in research that supports farm animal production and veterinary health, as for developing pig models with relevance to human health research.     

Flow cytometric DNA ploidy analysis of soft tissue sarcomas. A comparative study of preoperative fine needle aspirates and postoperative fresh tissues and archival material

Flow cytometric (FCM) DNA ploidy measurements on frozen fresh samples of soft tissue sarcomas were compared with the corresponding analyses on preoperative fine needle aspirates and postoperative formalin-fixed archival tissues from the same tumors. A concordance in ploidy status (diploid versus non-diploid) was obtained for 63% of the fresh tissue-fine needle aspiration (FNA) sample comparisons and for 85% of the fresh tissue-archival material comparisons. The majority of discordances in the fresh tissue-FNA sample comparisons could be explained by FNA sampling errors. In the remaining discordant cases (3 of 27 FNA sample comparisons and 6 of 40 archival material comparisons), sampling errors could not explain the differences in ploidy status. The discordant cases were evenly distributed among the different sampling methods. Method reproducibility was not responsible for the differences in ploidy determinations; tumor heterogeneity may be an explanation for the discrepancies. This study showed that archival soft tissue sarcoma samples are as well suited for DNA ploidy analysis as are fresh frozen tissues.