Prospects for incorporation of epigenetic biomarkers in human health and environmental risk assessment of chemicals

Epigenetic mechanisms have gained relevance in human health and environmental studies, due to their pivotal role in disease, gene × environment interactions and adaptation to environmental change and/or contamination. Epigenetic mechanisms are highly responsive to external stimuli and a wide range of chemicals has been shown to determine specific epigenetic patterns in several organisms. Furthermore, the mitotic/meiotic inheritance of such epigenetic marks as well as the resulting changes in gene expression and cell/organismal phenotypes has now been demonstrated. Therefore, epigenetic signatures are interesting candidates for linking environmental exposures to disease as well as informing on past exposures to stressors. Accordingly, epigenetic biomarkers could be useful tools in both prospective and retrospective risk assessment but epigenetic endpoints are currently not yet incorporated into risk assessments. Achieving a better understanding on this apparent impasse, as well as identifying routes to promote the application of epigenetic biomarkers within environmental risk assessment frameworks are the objectives of this review. We first compile evidence from human health studies supporting the use of epigenetic exposure‐associated changes as reliable biomarkers of exposure. Then, specifically focusing on environmental science, we examine the potential and challenges of developing epigenetic biomarkers for environmental fields, and discuss useful organisms and appropriate sequencing techniques to foster their development in this context. Finally, we discuss the practical incorporation of epigenetic biomarkers in the environmental risk assessment of chemicals, highlighting critical data gaps and making key recommendations for future research within a regulatory context.     

Biomarker/bioindicator response profiles of organisms can help differentiate between sources of anthropogenic stressors in aquatic ecosystems

Aquatic ecosystems can be chronically stressed by multiple environmental factors which originate from a variety of point and non-point sources. In addition, these stressors may vary both spatially and temporally, and, combined with synergestic and cumulative interactions of these stressors, complicate the interpretation and evaluation of stress responses in organisms. To help identify and differentiate between sources of anthropogenic stressors in aquatic systems, a diagnostic approach based on exposure-response profiles in sentinel organisms was developed from the known effects of various anthropogenic activities on biological systems. To generate these exposure-effects profiles, biomarkers of exposure were plotted against bioindicators of corresponding effects for several major anthropogenic activities including petrochemical, pulp and paper, domestic sewage, mining operations, land-development, and agricultural activities. Biomarkers of exposure to environmental stressors varied widely depending on the type of anthropogenic activity involved. Bioindicator effects, however, including histopathological lesions, bioenergetic status, growth, reproductive impairment, and community-level endpoints were similar among several of the major anthropogenic activities because responses at these higher levels are less specific to stressors than are biomarkers. This approach appears useful for helping to identify and diagnose sources of stress in environments impacted by multiple stressors. By identifying the types and sources of environmental stressors impacting key components of biological systems, aquatic ecosystems can be more effectively protected, regulated, and managed to help improve and maintain environmental quality and ecosystem fitness.     

Toxicogenomic approach for assessing toxicant-related disease

The problems of identifying environmental factors involved in the etiology of human disease and performing safety and risk assessments of drugs and chemicals have long been formidable issues. Three principal components for predicting potential human health risks are: (1) the diverse structure and properties of thousands of chemicals and other stressors in the environment; (2) the time and dose parameters that define the relationship between exposure and disease; and (3) the genetic diversity of organisms used as surrogates to determine adverse chemical effects. The global techniques evolving from successful genomics efforts are providing new exciting tools with which to address these intractable problems of environmental health and toxicology. In order to exploit the scientific opportunities, the National Institute of Environmental Health Sciences has created the National Center for Toxicogenomics (NCT). The primary mission of the NCT is to use gene expression technology, proteomics and metabolite profiling to create a reference knowledge base that will allow scientists to understand mechanisms of toxicity and to be able to predict the potential toxicity of new chemical entities and drugs. A principal scientific objective underpinning the use of microarray analysis of chemical exposures is to demonstrate the utility of signature profiling of the action of drugs or chemicals and to utilize microarray methodologies to determine biomarkers of exposure and potential adverse effects. The initial approach of the NCT is to utilize proof-of-principle experiments in an effort to "phenotypically anchor" the altered patterns of gene expression to conventional parameters of toxicity and to define dose and time relationships in which the expression of such signature genes may precede the development of overt toxicity. The microarray approach is used in conjunction with proteomic techniques to identify specific proteins that may serve as signature biomarkers. The longer-range goal of these efforts is to develop a reference relational database of chemical effects in biological systems (CEBS) that can be used to define common mechanisms of toxicity, chemical and drug actions, to define cellular pathways of response, injury and, ultimately, disease. In order to implement this strategy, the NCT has created a consortium of research organizations and private sector companies to actively collaborative in populating the database with high quality primary data. The evolution of discrete databases to a knowledge base of toxicogenomics will be accomplished through establishing relational interfaces with other sources of information on the structure and activity of chemicals such as that of the National Toxicology Program (NTP) and with databases annotating gene identity, sequence, and function.     

Environmental proteomics and metallomics

Monitoring environmental pollution using biomarkers requires detailed knowledge about the markers, and many only allow a partial assessment of pollution. New proteomic methods (environmental proteomics) can identify proteins that, after validation, might be useful as alternative biomarkers, although this approach also has its limitations, derived mainly from their application to non-model organisms. Initial studies using environmental proteomics were carried out in animals exposed to model pollutants, and led to the concept of protein expression signatures. Experiments have been carried out in model organisms (yeast, Arabidopsis, rat cells, or mice) exposed to model contaminants. Over the last few years, proteomics has been applied to organisms from ecosystems with different pollution levels, forming the basis of an environmental branch in proteomics. Another focus is connected with the presence of metals bound to biomolecules, which adds an additional dimension to metal-biomolecule and metalloprotein characterization - the field of metallomics. The metallomic approach considers the metallome: a whole individual metal or metalloid species within a cell or tissue. A metallomic analytical approach (MAA) is proposed as a new tool to study and identify metalloproteins.     

Role of biomarkers in monitoring exposures to chemicals: present position,future prospects

Biomarkers are becoming increasingly important in toxicology and human health. Many research groups are carrying out studies to develop biomarkers of exposure to chemicals and apply these for human monitoring. There is considerable interest in the use and application of biomarkers to identify the nature and amounts of chemical exposures in occupational and environmental situations. Major research goals are to develop and validate biomarkers that reflect specific exposures and permit the prediction of the risk of disease in individuals and groups. One important objective is to prevent human cancer. This review presents a commentary and consensus views about the major developments on biomarkers for monitoring human exposure to chemicals. A particular emphasis is on monitoring exposures to carcinogens. Significant developments in the areas of new and existing biomarkers, analytical methodologies, validation studies and field trials together with auditing and quality assessment of data are discussed. New developments in the relatively young field of toxicogenomics possibly leading to the identification of individual susceptibility to both cancer and non-cancer endpoints are also considered. The construction and development of reliable databases that integrate information from genomic and proteomic research programmes should offer a promising future for the application of these technologies in the prediction of risks and prevention of diseases related to chemical exposures. Currently adducts of chemicals with macromolecules are important and useful biomarkers especially for certain individual chemicals where there are incidences of occupational exposure. For monitoring exposure to genotoxic compounds protein adducts, such as those formed with haemoglobin, are considered effective biomarkers for determining individual exposure doses of reactive chemicals. For other organic chemicals, the excreted urinary metabolites can also give a useful and complementary indication of exposure for acute exposures. These methods have revealed ‘backgrounds’ in people not knowingly exposed to chemicals and the sources and significance of these need to be determined, particularly in the context of their contribution to background health risks.     

Role of biomarkers in monitoring exposures to chemicals: present position, future prospects

Biomarkers are becoming increasingly important in toxicology and human health. Many research groups are carrying out studies to develop biomarkers of exposure to chemicals and apply these for human monitoring. There is considerable interest in the use and application of biomarkers to identify the nature and amounts of chemical exposures in occupational and environmental situations. Major research goals are to develop and validate biomarkers that reflect specific exposures and permit the prediction of the risk of disease in individuals and groups. One important objective is to prevent human cancer. This review presents a commentary and consensus views about the major developments on biomarkers for monitoring human exposure to chemicals. A particular emphasis is on monitoring exposures to carcinogens. Significant developments in the areas of new and existing biomarkers, analytical methodologies, validation studies and field trials together with auditing and quality assessment of data are discussed. New developments in the relatively young field of toxicogenomics possibly leading to the identification of individual susceptibility to both cancer and non-cancer endpoints are also considered. The construction and development of reliable databases that integrate information from genomic and proteomic research programmes should offer a promising future for the application of these technologies in the prediction of risks and prevention of diseases related to chemical exposures. Currently adducts of chemicals with macromolecules are important and useful biomarkers especially for certain individual chemicals where there are incidences of occupational exposure. For monitoring exposure to genotoxic compounds protein adducts, such as those formed with haemoglobin, are considered effective biomarkers for determining individual exposure doses of reactive chemicals. For other organic chemicals, the excreted urinary metabolites can also give a useful and complementary indication of exposure for acute exposures. These methods have revealed 'backgrounds' in people not knowingly exposed to chemicals and the sources and significance of these need to be determined, particularly in the context of their contribution to background health risks.     

Proteomic analysis of methyl parathion-responsive proteins in Sparus latus liver

The contamination of marine ecosystems by organophosphate pesticides is of great concern. The use of protein expression profiles may provide a good method to help us understand the methyl parathion (MP) toxicity to aquatic organisms. In this study, Sparus latus, was selected as the target organism. The toxicological effects of MP were investigated after 24 h exposure using proteomics to analyze their liver tissues. Certain enzyme activity parameters of the liver extracts were also examined, including CAT. After analyzing the proteomic profile of the liver using 2D gel electrophoresis, we found that the protein expression levels of 25 spots increased or decreased significantly in the exposed groups. Sixteen of the 25 protein spots were successfully identified using MALDI-TOF MS/MS. These proteins were roughly categorized into diverse functional classes such as cell redox homeostasis, metabolic processes and cytoskeleton system. These data demonstrated that proteomics was a powerful tool to provide valuable insights into the possible mechanisms of toxicity of MP contaminants in aquatic species. Additionally, these data may provide novel biomarkers for evaluation of MP contamination in the environment.     

RNA‑seq技术在水生生物生态毒理学中的应用进展

水域是地球环境的重要组成部分,也是最易受污染的生态系统之一。水生态系统中不同营养级别的水生生物可通过摄食、接触等多种途径摄入水体中的污染物。因此,监测水域污染物对水生生物和生态系统的影响,解析污染物对不同水生生物的毒性机制,筛选敏感、有效的生物标志物对生态毒理学研究和环境风险评价具有重要意义。RNA测序（RNA sequencing,RNA?seq）技术因所需样品量少,且不需参考序列,可在整体水平上鉴定基因差异表达,成为水生生物生态毒理学研究的最佳方法之一。基于此,介绍了RNA?seq技术的基本流程与数据分析过程,对该技术在不同生态位的水生生物（如鱼类、两栖类、贝类、甲壳类等）生态毒理学中的应用展开综述,并对RNA?seq技术面临的不足、挑战及发展趋势进行探讨,以期为该技术在水生生物生态毒理学研究中的应用,尤其是水生态环境中污染物胁迫水生生物机制的阐明及污染水域生态环境恢复提供参考。     

To label or not to label: applications of quantitative proteomics in neuroscience research

Proteomics has provided researchers with a sophisticated toolbox of labeling-based and label-free quantitative methods. These are now being applied in neuroscience research where they have already contributed to the elucidation of fundamental mechanisms and the discovery of candidate biomarkers. In this review, we evaluate and compare labeling-based and label-free quantitative proteomic techniques for applications in neuroscience research. We discuss the considerations required for the analysis of brain and central nervous system specimens, the experimental design of quantitative proteomic workflows as well as the feasibility, advantages, and disadvantages of the available techniques for neuroscience-oriented questions. Furthermore, we assess the use of labeled standards as internal controls for comparative studies in humans and review applications of labeling-based and label-free mass spectrometry approaches in relevant model organisms and human subjects. Providing a comprehensive guide of feasible and meaningful quantitative proteomic methodologies for neuroscience research is crucial not only for overcoming current limitations but also for gaining useful insights into brain function and translating proteomics from bench to bedside.     

Proteomic applications in ecotoxicology

Within the growing body of proteomics studies, issues addressing problems of ecotoxicology are on the rise. Generally speaking, ecotoxicology uses quantitative expression changes of distinct proteins known to be involved in toxicological responses as biomarkers. Unlike these directed approaches, proteomics examines how multiple expression changes are associated with a contamination that is suspected to be detrimental. Consequently, proteins involved in toxicological responses that have not been described previously may be revealed. Following identification of key proteins indicating exposure or effect, proteomics can potentially be employed in environmental risk assessment. To this end, bioinformatics may unveil protein patterns specific to an environmental stress that would constitute a classifier able to distinguish an exposure from a control state. The combined use of sets of marker proteins associated with a given pollution impact may prove to be more reliable, as they are based not only on a few unique markers which are measured independently, but reflect the complexity of a toxicological response. Such a proteomic pattern might also integrate some of the already established biomarkers of environmental toxicity. Proteomics applications in ecotoxicology may also comprise functional examination of known classes of proteins, such as glutathione transferases or metallothioneins, to elucidate their toxicological responses.     

Clinical proteomics: present and future prospects

Advances in proteomics technology offer great promise in the understanding and treatment of the molecular basis of disease. The past decade of proteomics research, the study of dynamic protein expression, post-translational modifications, cellular and sub-cellular protein distribution, and protein-protein interactions, has culminated in the identification of many disease-related biomarkers and potential new drug targets. While proteomics remains the tool of choice for discovery research, new innovations in proteomic technology now offer the potential for proteomic profiling to become standard practice in the clinical laboratory. Indeed, protein profiles can serve as powerful diagnostic markers, and can predict treatment outcome in many diseases, in particular cancer. A number of technical obstacles remain before routine proteomic analysis can be achieved in the clinic; however the standardisation of methodologies and dissemination of proteomic data into publicly available databases is starting to overcome these hurdles. At present the most promising application for proteomics is in the screening of specific subsets of protein biomarkers for certain diseases, rather than large scale full protein profiling. Armed with these technologies the impending era of individualised patient-tailored therapy is imminent. This review summarises the advances in proteomics that has propelled us to this exciting age of clinical proteomics, and highlights the future work that is required for this to become a reality.     

膜整合蛋白质的“鸟枪法”蛋白质组学分析策略

疾病状态下生物膜表面蛋白质分子标记的表达量和修饰状态会发生改变。但由于其低丰度和不易溶解等特性,制约了膜蛋白质组学的研究,同时也制约了相关药物靶标的设计。近年来,为克服这些困难,学者们提出了"鸟枪法"的膜蛋白质组学研究策略。基于此,本文论述了"鸟枪法"的蛋白质组学分析的基本过程及其后续的部分改进工作。随着新的策略不断被采用,更多膜蛋白质的拓扑学特征和功能的相关研究一定会走上新的台阶。     

Utility and relevance of aquatic oligochaetes in Ecological Risk Assessment

Ecological risk assessment (EcoRA) provides both a process and a framework to evaluate the potential for adverse ecological effects occurring as a result of exposure to contaminants or other stressors. EcoRA begins with problem formulation/hazard identification, progresses to effects and exposure assessment, and culminates with risk characterization (an estimate of the incidence and severity of any adverse effects likely to occur). Key components of EcoRA include determining: stressors/contaminants of concern; sensitive, exposed biota; and, appropriate tests and organisms for evaluating effects. Aquatic oligochaetes are not generally used directly in EcoRA because of three major perceptions. First, EcoRA personnel are generally not familiar with or comfortable using this group of organisms. Second, there is believed to be a paucity of widely accepted toxicity tests with these organisms. Third, their taxonomy is considered difficult and uncertain. In fact, aquatic oligochaetes potentially have great utility and relevance to EcoRAs because of factors including: their importance in the aquatic food chain (e.g. prey to fauna including fish and waterfowl; as a vector for contaminant movement through the food chain from bacteria); many species are widely distributed and well studied; representatives include fresh, estuarine and marine species; as a group, they range from sensitive to insensitive over a wide range of environmental insults; they have a long history of use in pollution monitoring and assessment; and, relevant toxicity and biaccumulation tests exist. Toxicity testing under defined conditions is appropriate for problem formulation while more realistic testing for effects assessment (e.g. microcosms) is logistically easier with this group of organisms than with others due to their relatively small size. The importance of aquatic oligochaetes for EcoRA, in particular of sediments, is particularly compelling.     

Diagnostics and correction of batch effects in large‐scale proteomic studies: a tutorial

Advancements in mass spectrometry‐based proteomics have enabled experiments encompassing hundreds of samples. While these large sample sets deliver much‐needed statistical power, handling them introduces technical variability known as batch effects. Here, we present a step‐by‐step protocol for the assessment, normalization, and batch correction of proteomic data. We review established methodologies from related fields and describe solutions specific to proteomic challenges, such as ion intensity drift and missing values in quantitative feature matrices. Finally, we compile a set of techniques that enable control of batch effect adjustment quality. We provide an R package, "proBatch", containing functions required for each step of the protocol. We demonstrate the utility of this methodology on five proteomic datasets each encompassing hundreds of samples and consisting of multiple experimental designs. In conclusion, we provide guidelines and tools to make the extraction of true biological signal from large proteomic studies more robust and transparent, ultimately facilitating reliable and reproducible research in clinical proteomics and systems biology.     

微塑料污染对浮游生物影响研究进展

微塑料作为一种新型的环境污染物,大量存在于水环境中,给水生生物带来了极大的危害。浮游生物是水生食物链的基础,是水生生态系统物质循环和能量流动的重要环节;同时,浮游生物也是对各种环境污染物最敏感的类群。了解微塑料对浮游生物的影响是评价其生态风险的重要依据。本文介绍了环境中微塑料来源、特征及水生态系统微塑料污染现状,阐述了微塑料对水生生物的直接和间接危害,并重点聚焦于浮游植物和浮游动物,从个体、种群和群落的层次详细总结了微塑料的影响及其作用机制。最后,本文指出当前针对浮游生物微观基因和蛋白质组学,以及宏观种群和群落响应等方面的研究还非常缺乏,为今后开展微塑料危害研究提供参考。     

Label-free quantification in clinical proteomics

Nowadays, proteomic studies no longer focus only on identifying as many proteins as possible in a given sample, but aiming for an accurate quantification of them. Especially in clinical proteomics, the investigation of variable protein expression profiles can yield useful information on pathological pathways or biomarkers and drug targets related to a particular disease. Over the time, many quantitative proteomic approaches have been established allowing researchers in the field of proteomics to refer to a comprehensive toolbox of different methodologies. In this review we will give an overview of different methods of quantitative proteomics with focus on label-free proteomics and its use in clinical proteomics.     

Helminths and protozoans of aquatic organisms as bioindicators of chemical pollution

There is no doubt that the aquatic environments receive large quantities of chemicals as consequence of human activities and that those substances have a detrimental effect on human health. Despite the obvious need for effective disposal of these substances, we need to understand and prevent the outcome of harmful environmental exposures. Thus, we need biomarkers and bioindicators to advance our understanding to these harmful exposures and their biological effects. In the last three decades a large number of publications has suggested that aquatic organisms and their parasites (mainly helminths and ciliate protozoans) are useful bioindicators of chemical pollution. However, the main weakness of this approach is that after exposure the population size of these parasites can increase or decrease without a consistent pattern. I suggest that this is in part due to the lack of focus on the correct spatial or temporal scales at which the environment is acting over our study object. Thus, I propose to use spatially explicit (= georeferenced) data for determining whether there is spatial structure in our study area. Spatial structure is the tendency of nearby samples to have attribute values more similar than those farther apart. These attributes are shaped by environmental variables acting at specific spatial and temporal scales. Thus, I suggest to consider these tools for determining the correct spatial or temporal scales of study, but also to record pollutant concentrations, bioindicators, biomarkers and parasites at individual host level. Combining this information with long-term monitoring programs is likely to improve our understanding of the effects of chemical pollutants over the aquatic environments.     

Development and application of proteomics technologies in Saccharomyces cerevisiae

Proteomics research focuses on the identification and quantification of "all" proteins present in cells, organisms or tissue. Proteomics is technically complicated because it encompasses the characterization and functional analysis of all proteins that are expressed by a genome. Moreover, because the expression levels of proteins strongly depend on complex regulatory systems, the proteome is highly dynamic. This review focuses on the two major proteomics methodologies, one based on 2D gel electrophoresis and the other based on liquid chromatography coupled to mass spectrometry. The recent developments of these methodologies and their application to quantitative proteomics are described. The model system Saccharomyces cerevisiae is considered to be the optimal vehicle for proteomics and we review studies investigating yeast adaptation to changes in (nutritional) environment.