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30 years of the Comet Assay: an overview with some new insights

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Book Series: Frontiers Research Topics ISSN: 16648714 ISBN: 9782889196494 Year: Pages: 174 DOI: 10.3389/978-2-88919-649-4 Language: English
Publisher: Frontiers Media SA
Subject: Science (General) --- Genetics
Added to DOAB on : 2016-08-16 10:34:25
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By means of this ‘Frontiers in Genetics’ research topic, we are celebrating 30 years of the Comet Assay. The first paper on this single-cell gel electrophoresis assay was published in 1984 by O. Ostling and K.J. Johanson (Biochem. Biophys. Res. Commun. Vol.123: 291-298). The comet assay is a versatile and sensitive method for measuring single - and double-strand breaks in DNA. By including lesion-specific enzymes in the assay, its range and sensitivity are greatly increased, but it is important to bear in mind that their specificity is not absolute. The comet assay (with and without inclusion of lesion-specific enzymes) is widely used as a biomarker assay in human population studies - primarily to measure DNA damage, but increasingly also to assess the capacity of cells for DNA repair. Ostling and Johanson (Biochem. Biophys. Res. Commun., 1984) were also the first to report experiments to measure DNA repair, by simply following the decrease of DNA damage over time after challenging cells with ionising radiation. However, this approach is time-consuming and laborious as it requires an extended period of cell culture and is therefore not ideal for biomonitoring studies, which typically require high-throughput processing of many samples. As an alternative approach, the in vitro comet-based repair assay was developed: a cell extract is incubated with a DNA substrate containing specific lesions, and DNA incisions accumulate. The in vitro comet-based repair assay has been modified and improved over the past decade: it was first devised to measure base excision repair of oxidised purines in lymphocytes (Collins et al., Mutagenesis, 2001), but has since been adapted for other lesions and thus other repair pathways, as well as being applied to tissue samples in addition to cell suspensions. Even after 30 years, the comet assay is still in a growth phase, with many new users each year. Many questions are repeatedly raised, which may seem to have self-evident answers, but clearly, it is necessary to reiterate them for the benefit of the new audience, and sometimes being forced to think again about old topics can shed new light. Different applications of the comet assay are discussed in this special issue, including: genotoxicity testing in different organisms, human biomonitoring, DNA repair studies, environmental biomonitoring and clinical studies. Furthermore, we consider and where possible answer questions, including the ones raised by Raymond Tice at the 8th International Comet Assay Workshop in Perugia (Italy 2009): What is the spectrum of DNA damage detected by the various versions of the comet assay?; What are the limitations associated with each application?; What should be done to standardize the assay for biomonitoring studies?; Can the comet assay be used to monitor changes in global methylation status?; What cell types are suitable for detecting genotoxic substances and their effects in vivo and in vitro?; Can the assay be fully automated?; and more. So this ‘Frontiers in Genetics’ research topic is written for the beginner as well as for the experienced users of the comet assay.

ADN forense: problemas éticos y jurídicos

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Los análisis genéticos afectan a derechos fundamentales, por lo que el uso de esta información debe estar supeditado a vías de control democrático. En la actualidad, la lucha contra grandes delitos, como por ejemplo el terrorismo, aparentemente legitima a invadir derechos antes considerados intangibles. La presente obra, fruto del trabajo multidisciplinar llevado a cabo por juristas, filósofos, biólogos, técnicos y médicos, tiene por objetivo poner de manifiesto cuáles son los problemas ético-jurídicos derivados de la obtención, el análisis y el almacenamiento del ADN, así como sus usos judiciales y extrajudiciales. Ante el difícil equilibrio entre libertad individual y seguridad colectiva, este libro ayuda a comprender los conflictos que subyacen en el manejo de una herramienta informativa tan poderosa como son las muestras y los perfiles del ADN.

Advances in DNA Repair

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ISBN: 9789535122098 9789535142133 Year: Pages: 496 DOI: 10.5772/58716 Language: English
Publisher: IntechOpen
Subject: Genetics
Added to DOAB on : 2019-10-03 07:51:49

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This book edition is intended to provide a concise summary for select topics in DNA repair, a field that is ever-expanding in complexity and biologic significance. The topics reviewed ranged from fundamental mechanisms of DNA repair to the interface between DNA repair and a spectrum on cellular process to the clinical relevance of DNA repair in oncologic paradigms. The information in this text should provide a foundation from which one can explore the various topics in depth. The book serve as a supplementary text in seminar courses with focus on DNA repair as well as a general reference for scholars with an interest in DNA repair.

Advances in Farm Animal Genomic Resources

Authors: --- --- --- --- et al.
Book Series: Frontiers Research Topics ISSN: 16648714 ISBN: 9782889197354 Year: Pages: 293 DOI: 10.3389/978-2-88919-735-4 Language: English
Publisher: Frontiers Media SA
Subject: Genetics --- Science (General)
Added to DOAB on : 2016-04-07 11:22:02
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The history of livestock started with the domestication of their wild ancestors: a restricted number of species allowed to be tamed and entered a symbiotic relationship with humans. In exchange for food, shelter and protection, they provided us with meat, eggs, hides, wool and draught power, thus contributing considerably to our economic and cultural development. Depending on the species, domestication took place in different areas and periods. After domestication, livestock spread over all inhabited regions of the earth, accompanying human migrations and becoming also trade objects. This required an adaptation to different climates and varying styles of husbandry and resulted in an enormous phenotypic diversity. Approximately 200 years ago, the situation started to change with the rise of the concept of breed. Animals were selected for the same visible characteristics, and crossing with different phenotypes was reduced. This resulted in the formation of different breeds, mostly genetically isolated from other populations. A few decades ago, selection pressure was increased again with intensive production focusing on a limited range of types and a subsequent loss of genetic diversity. For short-term economic reasons, farmers have abandoned traditional breeds. As a consequence, during the 20th century, at least 28% of farm animal breeds became extinct, rare or endangered. The situation is alarming in developing countries, where native breeds adapted to local environments and diseases are being replaced by industrial breeds. In the most marginal areas, farm animals are considered to be essential for viable land use and, in the developing world, a major pathway out of poverty. Historic documentation from the period before the breed formation is scarce. Thus, reconstruction of the history of livestock populations depends on archaeological, archeo-zoological and DNA analysis of extant populations. Scientific research into genetic diversity takes advantage of the rapid advances in molecular genetics. Studies of mitochondrial DNA, microsatellite DNA profiling and Y-chromosomes have revealed details on the process of domestication, on the diversity retained by breeds and on relationships between breeds. However, we only see a small part of the genetic information and the advent of new technologies is most timely in order to answer many essential questions. High-throughput single-nucleotide polymorphism genotyping is about to be available for all major farm animal species. The recent development of sequencing techniques calls for new methods of data management and analysis and for new ideas for the extraction of information. To make sense of this information in practical conditions, integration of geo-environmental and socio-economic data are key elements. The study and management of farm animal genomic resources (FAnGR) is indeed a major multidisciplinary issue. The goal of the present Research Topic was to collect contributions of high scientific quality relevant to biodiversity management, and applying new methods to either new genomic and bioinformatics approaches for characterization of FAnGR, to the development of FAnGR conservation methods applied ex-situ and in-situ, to socio-economic aspects of FAnGR conservation, to transfer of lessons between wildlife and livestock biodiversity conservation, and to the contribution of FAnGR to a transition in agriculture (FAnGR and agro-ecology).

Advances in Genomics and Epigenomics of Social Insects

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Book Series: Frontiers Research Topics ISSN: 16648714 ISBN: 9782889450800 Year: Pages: 155 DOI: 10.3389/978-2-88945-080-0 Language: English
Publisher: Frontiers Media SA
Subject: Genetics --- Science (General)
Added to DOAB on : 2017-07-06 13:27:36
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Social insects are among the most successful and ecologically important animals on earth. The lifestyle of these insects has fascinated humans since prehistoric times. These species evolved a caste of workers that in most cases have no progeny. Some social insects have worker sub-castes that are morphologically specialized for discrete tasks. The organization of the social insect colony has been compared to the metazoan body. Males in the order Hymenoptera (bees, ants and wasps) are haploid, a situation which results in higher relatedness between female siblings. Sociality evolved many times within the Hymenoptera, perhaps spurred in part by increased relatedness that increases inclusive fitness benefits to workers cooperating to raise their sisters and brothers rather than reproducing themselves. But epigenetic processes may also have contributed to the evolution of sociality. The Hymenoptera provide opportunities for comparative study of species ranging from solitary to highly social. A more ancient clade of social insects, the termites (infraorder Isoptera) provide an opportunity to study alternative mechanisms of caste determination and lifestyles that are aided by an array of endosymbionts. This research topic explores the use of genome sequence data and genomic techniques to help us explore how sociality evolved in insects, how epigenetic processes enable phenotypic plasticity, and the mechanisms behind whether a female will become a queen or a worker.

Advances in Research on Down Syndrome

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ISBN: 9789535137627 9789535137634 9789535140412 Year: Pages: 148 DOI: 10.5772/intechopen.68260 Language: English
Publisher: IntechOpen
Subject: Genetics
Added to DOAB on : 2019-10-03 07:51:50

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This book provides a concise yet comprehensive source of current information on Down syndrome. It focuses on exciting areas of research on chromosome editing, neurogenomics and diseases associated with Down syndrome. Research workers, scientists, medical graduates and physicians will find this book as an excellent source for consultation and references. Key features of this book are chromosome engineering in Down syndrome, mental retardation and cognitive disability, prenatal diagnosis and diseases associated with Down syndrome. Although aimed primarily for research workers on Down syndrome, we hope that the appeal of this book will extend beyond the narrow confines of academic interest and be exciting to wider audience, especially parents, relatives and health care providers who work with infants and children with Down syndrome.

Advances in Systems Immunology and Cancer

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Book Series: Frontiers Research Topics ISSN: 16648714 ISBN: 9782889193134 Year: Pages: 108 DOI: 10.3389/978-2-88919-313-4 Language: English
Publisher: Frontiers Media SA
Subject: Genetics --- Biology --- Biotechnology --- General and Civil Engineering --- Psychiatry --- Medicine (General) --- Physiology --- Neurology --- Science (General)
Added to DOAB on : 2016-02-05 17:24:33
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Aims and Scope: The Research Topic is designed to feature the latest innovative and leading-edge research, reviews and opinions on the study of complex and dynamic processes related to the mammalian immune system and cancer. All papers were meticulously selected to present our readers the multidisciplinary approach to tackle the existing challenges faced in these important fields. From high throughput experimental methodologies to computational and theoretical approaches, the articles are intended to introduce physicists, chemists, computer scientists, biologists and immunologists the idea of systems biology approach to the understanding of mammalian immune system and cancer processes. Attention was given to works that developed more effective approaches to the treatment of proinflammatory disease and cancer. The strong interdisciplinary focus will discuss biological systems at the level from a few molecules to the entire organism. Specific focus domain includes: Innate and adaptive immunity, cancer and cancer stem cell, genomic, proteomic and metabolic analysis, imaging, biophysics of immune and cancer response, computational modeling, non-linear analysis, statistical analysis, translational and disease models Types of articles: Viewpoint, commentaries, research letters, research articles, review and methodologies

Alterations of Epigenetics and MicroRNAs in Cancer and Cancer Stem Cell

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Book Series: Frontiers Research Topics ISSN: 16648714 ISBN: 9782889193455 Year: Pages: 79 DOI: 10.3389/978-2-88919-345-5 Language: English
Publisher: Frontiers Media SA
Subject: Biology --- Genetics --- Science (General)
Added to DOAB on : 2016-03-10 08:14:32
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Studies have shown that alterations of epigenetics and microRNAs (miRNAs) play critical roles in the initiation and progression of human cancer. Epigenetic silencing of tumor suppressor genes in cancer cells is generally mediated by DNA hypermethylation of CpG island promoter and histone modification such as methylation of histone H3 lysine 9 (H3K9) and tri-methylation of H3K27. MiRNAs are small non-coding RNAs that regulate expression of various target genes. Specific miRNAs are aberrantly expressed and play roles as tumor suppressors or oncogenes during carcinogenesis. Important tumor suppressor miRNAs are silenced by epigenetic alterations, resulting in activation of target oncogenes in human malignancies. Stem cells have the ability to perpetuate themselves through self-renewal and to generate mature cells of various tissues through differentiation. Accumulating evidence suggests that a subpopulation of cancer cells with distinct stem-like properties is responsible for tumor initiation, invasive growth, and metastasis formation, which is defined as cancer stem cells. Cancer stem cells are considered to be resistant to conventional chemotherapy and radiation therapy, suggesting that these cells are important targets of cancer therapy. DNA methylation, histone modification and miRNAs may be deeply involved in stem-like properties in cancer cells. Restoring the expression of tumor suppressor genes and miRNAs by chromatin modifying drugs may be a promising therapeutic approach for cancer stem cells. In this Research Topic, we discuss about alterations of epigenetics and miRNAs in cancer and cancer stem cell and understand the molecular mechanism underlying the formation of cancer stem cell, which may provide a novel insight for treatment of refractory cancer.

Antibody Engineering

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ISBN: 9789535138259 9789535138266 9789535139751 Year: Pages: 312 DOI: 10.5772/65238 Language: English
Publisher: IntechOpen
Subject: Genetics
Added to DOAB on : 2019-10-03 07:51:50

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Antibody Engineering comprises in vitro selection and modification of human antibodies including humanization of mouse antibodies for therapy, diagnosis, and research. This book comprises an overview about the generation of antibody diversity and essential techniques in antibody engineering: construction of immune, naive and synthetic libraries, all available in vitro display methods, humanization by chain shuffling, affinity maturation techniques, de novo synthesis of antibody genes, colony assays for library screening, construction of scFvs from hybridomas, and purification of monoclonal antibodies by exclusion chromatography. In addition, other topics that are discussed in this book are application and mechanism of single domain antibodies, structural diversity of antibodies, immune-mediated skin reactions induced by TNF-alpha recombinant antibodies, and bioinformatic approaches to select pathogen-derived peptide sequences for antibody targets.

Applications of RNA-Seq and Omics Strategies - From Microorganisms to Human Health

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ISBN: 9789535135036 9789535135043 Year: Pages: 328 DOI: 10.5772/66063 Language: English
Publisher: IntechOpen
Subject: Genetics
Added to DOAB on : 2019-10-03 07:51:50

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The large potential of RNA sequencing and other ""omics"" techniques has contributed to the production of a huge amount of data pursuing to answer many different questions that surround the science's great unknowns. This book presents an overview about powerful and cost-efficient methods for a comprehensive analysis of RNA-Seq data, introducing and revising advanced concepts in data analysis using the most current algorithms. A holistic view about the entire context where transcriptome is inserted is also discussed here encompassing biological areas with remarkable technological advances in the study of systems biology, from microorganisms to precision medicine.

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