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  • 1
    Description / Table of Contents: The first book of its kind, devoted exclusively to the assessment of DNA damage by the Comet assay in modern toxicology, The vast number of chemicals existing or being added into the environment, have globally aroused great concern regarding their adverse effects in human population. Development and validation of sensitive and better test systems which can assess the adverse effects of chemicals at an early stage for intervention strategies to be implemented in time is currently in progress. This book documents the latest research and showcases the versatile, state-of-the-art technique - the Comet assay - in the field of modern toxicology. The assay is a simple, sensitive rapid and visual technique for the quantitative and qualitative assessment of DNA damage in single cells. The Comet Assay in Toxicology is the first book of its kind to be devoted exclusively to the Comet assay and its applications as an important tool in modern toxicology. This multi-author book will serve as both a reference and a guide to investigations in the biomedical, biochemical and pharmaceutical sciences. Written by investigators from the fields of genetic toxicology and human epidemiology, the authors have first-hand knowledge from their chosen sub-specialities and are active contributors to the peer-reviewed scientific literature. The book is divided into five major sections, reflecting the range of interest in the exploitation of this assay. The book's scope begins with an introduction section reviewing its genesis for those new to the technique and the current knowledge of the various fields in which it finds wide acceptance. This section sets the scene by explaining why the assay has become the most sensitive and sought after assay in modern toxicology. Next is a whole section that considers various procedures being followed to assess different types of DNA damage in various cell types and is contributed by experts in the respective fields. The third section puts together the specific applications of the assay in the diverse fields ranging from genetic toxicity testing to human monitoring, and environmental toxicology. The fourth section consists of the guidelines and recommendations for the conduct of the assay in in vitro and in vivo systems, based on the recommendations of the International Workgroups on genotoxicity test procedures. Finally, the book draws to a close with an assessment of the statistics used for the understanding of the data generated by the assay. This is a unique reference book as it provides the scientific community with the advances in Comet assay as well as its applications. It also incorporates a detailed section with instant and comprehensive information on the procedure of the assay and the latest protocols being used worldwide as well as statistical analyses to be followed. The book is aimed at students as well as scientists in the area of molecular epidemiology and genetic toxicology
    Type of Medium: Online Resource
    Pages: 374 p , Online-Ressource , 43 b&w, ill
    Edition: RSC eBook Collection 1968-2009
    Series Statement: Issues in toxicology v. 5
    Language: English
    Note: Ebook , Preface-- Part I: Genesis of Comet assay-- Part II: Various procedures for the Comet assay-- Oxidative damage-- Water disinfection by-products-- Comet assay in Plants-- Double Strand-- Breaks in bacteria-- Comet - FISH-- Double strand breaks (DSBs) - radiation/challenge assay - Radiation-- Apoptosis-- Multiple mouse organs-- Part III: Applications of Comet assay-- Dietary intervention studies-- Comet assay in Mussels-- Exercise-- Arsenic-- DSBs - radiation/challenge assay - Radiation-- In vivo Comet assay-- In vitro parallelogram approaches-- Photobiology-- Comet assay in sperm-- Comet assay in Human monitoring-- Part IV: Guidelines for comet assay-- In Vitro And In Vivo Guidelines for the Comet Assay-- In vivo Guidelines for Comet-- Part V: Image analysis and Statistics-- Models for image analysis-- Statistics in comet assay.
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  • 2
    Online Resource
    Online Resource
    La Vergne :Royal Society of Chemistry, The,
    Keywords: Environmental Exposure - adverse effects. ; Electronic books.
    Description / Table of Contents: Male germ line mutagenesis and the effects on developmental defects in the next generation.
    Type of Medium: Online Resource
    Pages: 1 online resource (317 pages)
    Edition: 1st ed.
    ISBN: 9781847557643
    Series Statement: Issn Series
    DDC: 572.838
    Language: English
    Note: Male-mediated Developmental Toxicology -- Contents -- Foreword -- Introductory Scientific Presentations -- CHAPTER 1 Epigenetic Transgenerational Actions of Endocrine Disruptors through the Male Germ-Line -- 1.1 Review -- Acknowledgments -- References -- Heritable Effects in Humans -- CHAPTER 2 Reproductive Outcomes among Men Treated for Cancer -- 2.1 Why Study Cancer Survivors? -- 2.2 Endpoints -- 2.3 Decreased Fertility -- 2.4 The Five-Center Study -- 2.4.1 Design -- 2.4.2 Results -- 2.4.3 The Male Experience -- 2.5 The Childhood Cancer Survivor Study -- 2.5.1 Design -- 2.5.2 Results -- 2.6 Future Directions -- 2.7 Counseling -- References -- CHAPTER 3 Cancer in Siblings of Children with Cancer in the Nordic Countries: A Population-Based Cohort Study Paediatric Cancer: An Indicator of Familial Cancer Risk? -- 3.1 Epidemiological Research in the Nordic Countries -- 3.1.1 Unique Personal Identification Numbers -- 3.1.2 Danish Health Registries: Valuable Tools in MedicalResearch -- 3.1.3 The Danish Cancer Registry -- 3.2 Role of Dominant and Recessive Conditions in Cancer Causation -- 3.3 Cancer in Siblings of Children with Cancer in the Nordic Countries: A Population-Based Cohort Study -- 3.3.1 Patients, Procedures and Analyses -- 3.3.2 Results -- 3.4 Cancer in Parents of Children with Cancer in Denmark -- 3.5 Cancer in Off spring of Cancer Survivors: A Population-Based Cohort Study in the Nordic Countries -- 3.6 Paediatric Cancer: An Indicator of Familial Cancer Risk? -- References -- CHAPTER 4 What Harms the Developing Male Reproductive System? -- 4.1 Introduction -- 4.2 Descriptive Epidemiology of Conditions Affectingthe Male Reproductive System -- 4.2.1 Semen Quality and Fertility -- 4.2.2 Testicular Cancer -- 4.2.3 Anomalies of the Male Genitalia -- 4.2.4 The Question of Linkage -- 4.3 Can Endocrine Disruption Explain these Observations?. , 4.3.1 Oestrogens -- 4.3.2 Anti-androgens -- 4.3.3 The Expected Spectrum of Effects -- 4.3.4 Interpretation -- 4.4 Genetic Epidemiology -- 4.4.1 Semen Quality and Fertility -- 4.4.2 Testicular Cancer -- 4.4.3 Anomalies of the Male Genitalia -- 4.5 Can Genetic Damage Explain these Observations? -- 4.5.1 Can Genetic Disorders Increase Rapidly Over Time? -- 4.5.2 Is Heritable Infertility Possible? -- 4.5.3 So What is Going on? -- 4.6 Conclusions -- References -- CHAPTER 5 Links between Paternal Smoking and Childhood Cancer -- 5.1 Introduction -- 5.2 The Oxford Survey of Childhood Cancers -- 5.2.1 OSCC Data: 1977-1981 -- 5.2.2 OSCC Data: 1953-1955 -- 5.2.3 OSCC Data: 1971-1976 -- 5.3 Inter Regional Epidemiological Survey of Childhood Cancers -- 5.4 Other Studies -- 5.5 Discussion -- Acknowledgments -- References -- CHAPTER 6 Feasibility Study of Metal Effects on the X:Y Ratio in Human Sperm -- 6.1 Introduction -- 6.2 Research Methods -- 6.3 Findings -- 6.4 Discussion -- 6.5 Conclusion -- Acknowledgments -- References -- CHAPTER 7 Use of the Sperm Chromatin Structure Assay (SCSAs) as a Diagnostic Tool in the Human Infertility Clinic -- 7.1 Introduction -- 7.2 Materials and Methods -- 7.3 SCSA Data and Clinical Results -- 7.4 Conclusions -- References -- CHAPTER 8 Safety of Sperm for Use in Intra-Cytoplasmic Sperm Injection -- 8.1 Introduction -- 8.2 Incidence of Sperm Chromosome Aneuploidy and Implications on Reproduction -- 8.3 Epigenetic Effects and Their Relation to Intra-Cytoplasmic Sperm Injection -- 8.4 The Impact of Sperm Nuclear DNA Strand Breakson Reproductive Outcome -- 8.5 Improving the Safety of Intra-Cytoplasmic Sperm Injection -- 8.6 Conclusion -- References -- Animal Models -- CHAPTER 9 Male-Mediated F1 Effects in Mice Exposed to Di(2-ethylhexyl)phthalate (DEHP) -- 9.1 Introduction -- 9.2 Methods -- 9.2.1 Effects in Exposed Generation. , 9.2.2 Effects in the Offspring of Exposed Males -- 9.2.3 Statistical Analysis -- 9.3 Results -- 9.4 Discussion -- Acknowledgments -- References -- CHAPTER 10 Prevention of Adverse Effects of Cancer Treatment on the Germline -- 10.1 Infertility Resulting from Cancer Treatment -- 10.2 Block to Spermatogonial Differentiation -- 10.3 Approaches to Prevention of Adverse Effects on the Germline -- 10.4 Protection or Stimulation of Spermatogenic Recovery by Hormonal Suppression -- 10.5 Mechanisms of Hormonal Stimulation of Spermatogenic Recovery in Rat -- 10.6 Potential for Clinical Application -- 10.7 Application in Conjunction with Spermatogonial Transplantation -- Acknowledgments -- References -- CHAPTER 11 Molecular Changes in Spermand Early Embryos after Paternal Exposure to a Chemotherapeutic Agent -- 11.1 Introduction -- 11.2 Cyclophosphamide Induces a Stage-Specific Response in Male Germ Cells -- 11.2.1 Altered Gene Expression during Spermatogenesis -- 11.2.2 Increased Chromosomal Aberrations -- 11.2.3 Effects on Cell-Cycle Progression -- 11.2.4 Disturbances in Chromatin Structure -- 11.3 Paternal Cyclophosphamide Exposure Disrupts Programming in the Early Embryo -- Acknowledgments -- References -- CHAPTER 12 Transmissible Genetic Risk Causing Tumours in Mice and Humans -- 12.1 Introduction -- 12.2 Experimental Evidences -- 12.2.1 Embryonic Deaths and Malformations -- 12.2.2 Induction of Tumours in the Offspring -- 12.2.3 Possible Mechanism and Molecular Studies -- 12.3 Relevant Human Studies -- 12.4 Summary -- Acknowledgments -- References -- CHAPTER 13 Heritable Effects on DNA Damage Following Paternal F0 Germline Irradiation -- 13.1 Introduction -- 13.2 Materials and Methods -- 13.2.1 Animals -- 13.2.2 Irradiation and Dosimetry -- 13.2.3 Sample Collection -- 13.2.4 Assessment of Sperm Motility and Morphology. , 13.2.5 Testicular and Epididymal Sperm Counts -- 13.2.6 Sperm Comet Assay -- 13.2.7 Statistical Analyses -- 13.3 Results -- 13.3.1 F0 Males -- 13.3.2 Breeding Results for Acutely Irradiated F0 Males -- 13.3.3 F2 Generation Male Offspring -- 13.3.4 F3 Generation Male Offspring -- 13.4 Discussion -- 13.4.1 Overview -- 13.4.2 Delayed Effects of Acute Spermatogonial Irradiation onSperm from F0 Male Mice -- 13.4.3 Selection vs. Genome Modification -- 13.4.4 Heritable Effects of Paternal F0 Irradiation on F2 and F3Generation Sperm -- 13.5 Conclusion -- Acknowledgments -- References -- CHAPTER 14 Influence of DNA Methylationand Genomic Imprinting in the Male Germ Line on Pregnancy Outcome -- 14.1 Introduction -- 14.2 DNA Methylation and DNA Methyltransferases:Roles and Importance -- 14.3 Acquisition of DNA Methylation Patterns During Spermatogenesis -- 14.4 DNA Methyltransferase Enzymes and their Regulation in the Male Germ Line -- 14.5 Perturbing DNA Methylation in the MaleGermline: Consequences for the Progeny -- 14.5.1 DNA Methyltransferase Deficiency -- 14.5.2 Cytidine Analogues -- 14.5.3 Folate Pathway Enzymes -- 14.6 Conclusions and Implications -- Acknowledgments -- References -- CHAPTER 15 Information Content of Ejaculate Spermatozoa and its Potential Utility in Toxicologyand Infertility Based Research Programmes -- 15.1 Introduction -- 15.2 Composition of Spermatozoal RNA -- 15.3 Utility of Spermatozoal RNA -- 15.4 Spermatozoal Chromatin -- 15.5 DNA Damage Susceptibility -- 15.6 Transmission of the Paternal Genome -- 15.7 Conclusions -- References -- Germline Mutagenesis -- CHAPTER 16 Origin of Paternal Mutations -- 16.1 Historical -- 16.2 Male Excess and Paternal Age Effect for BaseSubsitutions -- 16.3 Exceptions to the Sex and Paternal Age Effect -- 16.4 Hot Spots -- 16.5 Sperm Analysis -- 16.6 Premeiotic Selection or Sperm Competition. , 16.7 Conclusion -- References -- CHAPTER 17 Redox Regulation of DNA Damage in the Male Germ Line -- 17.1 Introduction -- 17.2 The Central Hypothesis -- 17.3 Causes of DNA Damage in Spermatozoa -- 17.4 Oxidative Stress -- 17.5 Origins of Oxidative Stress -- 17.6 Conclusions -- Acknowledgments -- References -- CHAPTER 18 Advances in the Direct Measurements of Partial Chromosomal Duplication, Deletions and Breaks in Humanand Murine Sperm by Sperm FISH -- 18.1 Introduction -- 18.2 FISH Methods for Detecting Human Sperm Carrying Structural Chromosomal Aberrations -- 18.3 FISH Methods for Detecting Mouse Sperm Carrying Chromosome Structural Aberrations -- 18.4 Mechanisms of Induction of Chromosomal Aberrations Detectable by Sperm FISH -- 18.5 Interspecies Comparison of Baseline Frequencies of Chromosomal Aberrations -- 18.6 Applications of the Novel Sperm FISH Methods for Detecting Sperm Carrying Chromosomal Aberrations -- 18.7 Implications for Future Studies -- Acknowledgments -- References -- CHAPTER 19 Radiation-induced Transgenerational Instability in Mice -- 19.1 Introduction -- 19.2 Somatic Effects -- 19.3 Germline Effects -- 19.4 Mechanisms -- 19.5 Conclusions -- Acknowledgments -- References -- CHAPTER 20 New Genetic Information Generated by Endogenous Reverse Transcription in Sperm Cells -- 20.1 Introduction -- 20.2 Mouse Sperm Chromatin Contains Domains withan ''Active-Like'' Open Conformation and Enriched in RT-Coding Elements -- 20.3 Exogenous RNA Incubated with MurineSpermatozoa is Reverse Transcribed into cDNA Copies that are Transferred to Embryos at Fertilisation -- 20.4 cDNA Copies are Mosaic Propagated in Adult F0 and Mosaic Transmitted to F1 Progeny -- 20.5 cDNA Copies are Transcriptionally Competent and are Expressed in Various Tissues -- 20.6 Risks for Human-Assisted Reproduction. , 20.7 Genesis and Non-Mendelian Inheritance of Extrachromosomal ''Retrogenes''.
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  • 3
    Keywords: Electronic books.
    Description / Table of Contents: By addressing considerations of efficacy and safety of drugs and chemicals used to combat COVID-19, virtually in real-time, this book documents and highlights the advances in science and place the toxicology, pharmaceutical science, public health and medical community in a better position to advise in future epidemics.
    Type of Medium: Online Resource
    Pages: 1 online resource (594 pages)
    Edition: 1st ed.
    ISBN: 9781839163647
    Language: English
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  • 4
    Online Resource
    Online Resource
    La Vergne :Royal Society of Chemistry, The,
    Keywords: DNA damage -- Testing. ; Gel electrophoresis. ; Electronic books.
    Description / Table of Contents: The first book of its kind, devoted exclusively to the assessment of DNA damage by the Comet assay in modern toxicology.
    Type of Medium: Online Resource
    Pages: 1 online resource (479 pages)
    Edition: 1st ed.
    ISBN: 9781847559746
    Series Statement: Issn Series
    DDC: 572.8/77
    Language: English
    Note: The Comet Assay in Toxicology -- Contents -- Section I: Genesis of Comet Assay -- Chapter 1 The Comet Assay: A Versatile Tool for Assessing DNA Damage -- 1.1 Introduction -- 1.2 Bacteria -- 1.3 Plant Models -- 1.3.1 The Comet Assay in Lower Plants -- 1.3.2 The Comet Assay in Higher Plants -- 1.4 Animal Models -- 1.4.1 Lower Animals -- 1.5 Higher Animals -- 1.5.1 Vertebrates -- 1.6 The Specificity, Sensitivity and Limitations of the Comet Assay -- 1.7 Conclusions -- Acknowledgements -- References -- Section II: Various Procedures for the Comet Assay -- Chapter 2 Detection of Oxidised DNA Using DNA Repair Enzymes -- 2.1 Introduction -- 2.2 Methods for Measuring DNA Oxidation Damage -- 2.3 Enzyme Specificity -- 2.4 Applications -- 2.5 Protocol -- 2.5.1 Equipment -- 2.5.2 Supplies -- 2.5.3 Reagents, Buffers and Enzymes -- 2.5.4 Procedure -- Acknowledgment -- References -- Chapter 3 Microplate-Based Comet Assay -- 3.1 Introduction -- 3.2 Microplate Comet Assay -- 3.3 Drinking-Water Disinfection Byproducts -- 3.4 Chinese Hamster Ovary Cells -- 3.5 CHO Cell Microplate Comet Assay Protocol -- 3.5.1 CHO Cell Treatment -- 3.5.2 Preparation of Comet Microgels -- 3.5.3 Comet Microscopic Examination -- 3.5.4 Normalisation of CHO Cell Comet Data and Statistical Analysis -- 3.6 Utility of the Microplate Comet Assay in Comparing Classes of DBPs -- 3.6.1 Microplate Comet Analysis of the Haloacetonitriles -- 3.6.2 Microplate Comet Analysis of the Haloacetamides -- 3.6.3 Comparison of SCGE Genotoxic Potency Values of the Haloacetonitriles and Haloacetamides -- 3.7 Advantages of the Mammalian CellMicroplate Comet Assay -- Acknowledgements -- References -- Chapter 4 The Use of Higher Plants in the Comet Assay -- 4.1 Introduction -- 4.2 Differences between the Animal and Plant Comet Assay -- 4.3 Cultivation and Treatment of Plants for the Comet Assay. , 4.3.1 Onion (Allium cepa) -- 4.3.2 Tobacco (Nicotiana tabacum) -- 4.3.3 Broad Bean (Vicia faba) -- 4.3.4 Plants used for In-Situ Studies -- 4.4 Isolation of Nuclei from Plant Tissues -- 4.4.1 Isolation of Nuclei via Protoplast Formation -- 4.4.2 Isolation of Nuclei by Mechanical Destruction of the Cell Wall -- 4.5 Preparation of Comet Assay Slides -- 4.6 DNA Unwinding and Electrophoresis -- 4.7 DNA Staining -- 4.8 Reading the Slides, Expressing DNA Damage, Statistics -- 4.9 Comet Assay Procedure -- 4.10 Reagents, Media, Buffers -- 4.11 Equipment and Software -- 4.12 Determination of Toxicity -- 4.13 Correlation between the DNA Damage Evaluated by the Comet Assay and Other Genetic Endpoints in Plants -- 4.14 The Utility of the Comet Assay for Genotoxic Studies in the Laboratory -- 4.15 The Utility of the Comet Assay as an In Situ Marker -- 4.16 Comet Assay with Irradiated Food of Plant Origin -- 4.17 Recommendations for Plant Comet Assay Users -- Abbreviations -- References -- Chapter 5 Methods for Freezing Blood Samples at -80°C for DNA Damage Analysis in Human Leukocytes -- 5.1 Introduction -- 5.2 Materials and Methods -- 5.2.1 Protocol I -- 5.2.2 Protocol II -- 5.2.3 Fresh Blood -- 5.2.4 Fresh Blood Stored on Ice Prior to Freezing -- 5.2.5 Image and Data Analysis -- 5.3 Results and Discussion -- References -- Chapter 6 Development and Applications of the Comet-FISH Assay for the Study of DNA Damage and Repair -- 6.1 Introduction -- 6.2 The Comet-FISH Assay Procedure -- 6.3 Applications of the Comet-FISH Assay -- 6.3.1 Discovery of the Comet-FISH Assay -- 6.3.2 Using Comet-FISH to Measure DNA Damage -- 6.3.3 Using Comet-FISH to Quantify DNA Repair -- 6.3.4 Summary of Studies -- 6.4 Limitations of Comet-FISH Assay -- 6.4.1 Practical Difficulties -- 6.4.2 Imaging Difficulties -- 6.4.3 Interpretation of Results -- 6.5 Conclusion -- References. , Chapter 7 Detection of DNA Damage in Drosophila and Mouse -- 7.1 General Protocol for the Assessment of DNA Damage Using the Alkaline Comet Assay -- 7.1.1 Chemicals and Materials -- 7.1.2 Preparation of Reagents -- 7.1.3 Preparation of Agarose-Coated (Base) Slides for the Comet Assay -- 7.1.4 Preparation of Microgel Slides for the Comet Assay -- 7.1.5 Electrophoresis of Microgel Slides -- 7.1.6 Evaluation of DNA Damage -- 7.2 The Alkaline Comet Assay in Multiple Organs of Mouse -- 7.2.1 Chemicals and Materials -- 7.2.2 Methodology -- 7.3 The Alkaline Comet Assay in Drosophila melanogaster -- 7.3.1 Chemicals and Materials -- 7.3.2 Methodology -- 7.4 Conclusions -- Acknowledgements -- References -- Section III: Applications of Comet Assay -- Chapter 8 Clinical Applications of the Comet Assay -- 8.1 Introduction -- 8.2 The Comet Assay Methodology -- 8.3 Clinical Studies -- 8.4 Discussion and Conclusions -- References -- Chapter 9 Applications of the Comet Assay in Human Biomonitoring -- 9.1 Biomonitoring and Biomarkers - An Introduction -- 9.2 The (Modified) Comet Assay -- 9.3 Guidelines for Biomonitoring Studies -- 9.4 Biomonitoring with the Comet Assay: Special Considerations -- 9.4.1 Surrogate and Target Cells -- The Use of White Blood Cells -- 9.4.2 Sampling Time and Transport -- 9.4.3 Reference Standards -- 9.4.4 What Affects the Background Level of DNA Damage? -- 9.5 DNA Damage as a Marker of Environmental Exposure and Risk -- 9.6 DNA Repair as a Biomarker of Individual Susceptibility -- 9.7 Protocols -- 9.7.1 Protocol for Blood Sample Collection and Long-Term Storage of Lymphocytes for the Measurement of DNA Damage and Repair -- 9.7.2 Comet Assay - Determination of DNA Damage (Strand Breaks and Oxidised Bases) -- 9.7.3 In Vitro Assays for DNA Repair -- 9.8 Solutions, etc. -- 9.8.1. Lysis Solution. , 9.8.2. Buffer F (Enzyme Reaction Buffer for FPG, Endonuclease III, and In Vitro BER Assay) -- 9.8.3 Buffer F+Mg (Used for In Vitro NER Assay) -- 9.8.4 Buffer A (Used in In Vitro Repair Assays) -- 9.8.5 Triton Solution -- 9.8.6 Ro 19-8022 (Photosensitiser) -- 9.8.7 Electrophoresis Solution -- 9.8.8 Neutralising Buffer -- 9.8.9 Agarose -- 9.8.10 Enzymes -- 9.9 Analysis and Interpretation of Results -- 9.9.1 Quantitation -- 9.9.2 Calculation of Net Enzyme-Sensitive Sites -- 9.9.3 Calibration -- 9.9.4 How to Deal with Comet Assay Data Statistically -- 9.10 Conclusions -- Acknowledgements -- References -- Chapter 10 The Comet Assay in Human Biomonitoring -- 10.1 Introduction -- 10.2 Human Monitoring -- 10.3 Environmental Exposure -- 10.4 Lifestyle Exposure -- 10.5 Occupational Exposure -- 10.6 Reviews -- 10.7 Usefulness of the Comet Assay in Human Monitoring -- 10.8 Conclusions -- References -- Chapter 11 Comet Assays in Dietary Intervention Trials -- 11.1 Introduction -- 11.2 Experimental Design of Human Studies -- 11.3 Indicator Cells and Media -- 11.4 Conventional SCGE Trials with Complex Foods and Individual Components - The Current State of Knowledge -- 11.5 Use of SCGE Trials to Detect Protection Against DNA-Reactive Carcinogens -- 11.6 Use of SCGE Experiments to Monitor Alterations of the DNA-Repair Capacity -- 11.7 What Have We Learned from Intervention Studies so Far? -- 11.8 Future Perspectives -- References -- Chapter 12 The Comet Assay for the Evaluation of Genotoxic Exposure in Aquatic Species -- 12.1 Introduction -- 12.2 Protocols, Cell Types and Target Organs -- 12.3 Application of the Comet Assay to Invertebrate Species -- 12.3.1 Freshwater Invertebrates -- 12.3.2 Marine Invertebrates -- 12.4 Application of the Comet Assay to Vertebrate Species -- 12.4.1 Freshwater Vertebrates -- 12.4.2 Marine Vertebrates -- 12.5 Conclusions -- References. , Chapter 13 The Alkaline Comet Assay in Prognostic Tests for Male Infertility and Assisted Reproductive Technology Outcomes -- 13.1 Introduction -- 13.2 Sites of DNA Damage in Sperm -- 13.2.1 Oxidative Stress, a Major Cause of DNA Damage -- 13.2.2 Oxidative Stress, Antioxidant Therapies -- 13.2.3 Sperm DNA Damage Tests -- 13.2.4 Modifications to the Alkaline Comet Assay for Use with Sperm -- 13.2.5 Sperm DNA Adducts and their Relationship with DNA Fragmentation -- 13.3 Can Sperm DNA Integrity Predict Success? Relationships with Assisted Conception Outcomes -- 13.4 Clinically Induced DNA Damage -- 13.4.1 Cryopreservation -- 13.4.2 Vasectomy -- 13.5 A Major Barrier to Progress -- 13.6 Opportunities and Challenges - The Establishment of Clinical Thresholds and the Integration of DNA Testing into Clinical Practice -- Acknowledgements -- References -- Chapter 14 The Comet Assay in Sperm - Assessing Genotoxins in Male Germ Cells -- 14.1 Introduction -- 14.2 Single-Cell Gel Electrophoresis -- 14.3 The Use of Sperm with the Comet Assay -- 14.3.1 Human Sperm -- 14.3.2 Modifying Existing Comet Protocols for the Use of Sperm -- 14.3.3 Sperm DNA and the Comet Assay -- 14.3.4 The Sperm Comet Assay and the Use of Repair Enzymes -- 14.3.5 Assessing the Sperm Comet -- 14.3.6 Comet-FISH on Sperm -- 14.3.7 Cryopreserved versus Fresh Sperm -- 14.3.8 Viability Considerations -- 14.3.9 Statistical Analysis -- 14.4 Utilising Male Germ Cells with the Comet Assay -- 14.4.1 In Vivo Comet Assay -- 14.4.2 In Vitro Comet Assay -- 14.5 The Sperm Comet Assay versus Other Assays Used in Reproductive Toxicology -- 14.6 Conclusions -- Acknowledgements -- References -- Section IV: Regulatory, Imaging and Statistical Considerations -- Chapter 15 Comet Assay - Protocols and Testing Strategies -- 15.1 Introduction -- 15.2 Applications of the In Vivo Comet Assay for Regulatory Purposes. , 15.3 Recommendations for Test Performance.
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  • 5
    Online Resource
    Online Resource
    La Vergne :Royal Society of Chemistry, The,
    Keywords: Gel electrophoresis. ; DNA damage--Testing. ; Electronic books.
    Description / Table of Contents: This updated edition of The Comet Assay in Toxicology provides the latest information on an important and versatile tool for assessing DNA damage.
    Type of Medium: Online Resource
    Pages: 1 online resource (611 pages)
    Edition: 2nd ed.
    ISBN: 9781782622895
    Series Statement: Issn Series
    DDC: 572.8/77
    Language: English
    Note: Cover -- Preface -- Contents -- Section I: Genesis of Comet Assay -- Chapter 1 The Comet Assay: A Versatile Tool for Assessing DNA Damage -- 1.1 Introduction -- 1.1.1 Bacteria -- 1.2 Plant Models -- 1.2.1 The Comet Assay in Lower Plants and Fungi -- 1.2.2 The Comet Assay in Higher Plants -- 1.3 Animal Models -- 1.3.1 Lower Animals -- 1.4 Higher Animals -- 1.4.1 Vertebrates -- 1.5 The Specificity, Sensitivity and Limitations of the Comet Assay -- 1.6 Conclusions -- References -- Section II: Various Procedures for the Comet Assay -- Chapter 2 High-throughput Measurement of DNA Breaks and Oxidised Bases with the Comet Assay -- 2.1 Introduction -- 2.2 Methods for Measuring DNA Oxidation Damage -- 2.3 Enzyme Specificity -- 2.4 Applications -- 2.5 Protocol -- 2.5.1 Equipment -- 2.5.2 Supplies -- 2.5.3 Reagents, Buffers and Enzymes -- 2.5.4 Procedure -- Acknowledgments -- References -- Chapter 3 Microplate-based Comet Assay -- 3.1 Introduction -- 3.2 Microplate Comet Assay -- 3.3 Drinking-water Disinfection Byproducts -- 3.4 Chinese Hamster Ovary Cells -- 3.5 CHO Cell Microplate Comet Assay Protocol -- 3.5.1 CHO Cell Treatment -- 3.5.2 Preparation of Comet Microgels -- 3.5.3 Comet Microscopic Examination -- 3.5.4 Normalisation of CHO Cell Comet Data and Statistical Analysis -- 3.6 Utility of the Microplate Comet Assay in Comparing Classes of DBPs -- 3.6.1 Microplate Comet Analysis of the Haloacetonitriles -- 3.6.2 Microplate Comet Analysis of the Haloacetamides -- 3.6.3 Comparison of SCGE Genotoxic Potency Values of the Haloacetonitriles and Haloacetamides -- 3.7 Advantages of the Mammalian Cell Microplate Comet Assay -- Acknowledgments -- References -- Chapter 4 The Use of Higher Plants in the Comet Assay -- 4.1 Introduction -- 4.2 Differences between the Animal and Plant Comet Assay -- 4.3 Cultivation and Treatment of Plants for the Comet Assay. , 4.3.1 Onion (Allium cepa) -- 4.3.2 Tobacco (Nicotiana tabacum) -- 4.3.3 Broad Bean (Vicia faba) -- 4.3.4 Plants Used for In situ Studies -- 4.4 Isolation of Nuclei from Plant Tissues -- 4.4.1 Isolation of Nuclei via Protoplast Formation -- 4.4.2 Isolation of Nuclei by Mechanical Destruction of the Cell Wall -- 4.5 Preparation of Comet Assay Slides -- 4.6 DNA Unwinding and Electrophoresis -- 4.7 DNA Staining -- 4.8 Reading the Slides, Expressing DNA Damage, Statistics -- 4.9 Comet Assay Procedure -- 4.10 Reagents, Media, Buffers -- 4.11 Equipment and Software -- 4.12 Determination of Toxicity -- 4.13 Correlation between the DNA Damage Evaluated by the Comet Assay and Other Genetic Endpoints in Plants -- 4.14 The Utility of the Comet Assay for Genotoxic Studies in the Laboratory -- 4.15 The Utility of the Comet Assay as an In situ Marker -- 4.16 Comet Assay with Irradiated Food of Plant Origin -- 4.17 Recommendations for Plant Comet Assay Users -- Abbreviations -- References -- Chapter 5 Methods for Freezing Blood Samples at -80°C for DNA Damage Analysis in Human Leukocytes -- 5.1 Introduction -- 5.2 Materials and Methods -- 5.2.1 Protocol I -- 5.2.2 Protocol II -- 5.2.3 Fresh Blood -- 5.2.4 Fresh Blood Stored on Ice Prior to Freezing -- 5.2.5 Image and Data Analysis -- 5.3 Results and Discussion -- References -- Chapter 6 Development and Applications of the Comet-FISH Assay for the Study of DNA Damage and Repair -- 6.1 Introduction -- 6.2 The Comet-FISH Assay Procedure -- 6.3 Applications of the Comet-FISH Assay -- 6.3.1 Discovery of the Comet-FISH Assay -- 6.3.2 Using Comet-FISH to Measure DNA Damage -- 6.3.3 Using Comet-FISH to Quantify DNA Repair -- 6.3.4 Summary of Studies -- 6.4 Limitations of Comet-FISH Assay -- 6.4.1 Practical Difficulties -- 6.4.2 Imaging Difficulties -- 6.4.3 Interpretation of Results -- 6.5 Conclusion -- References. , Chapter 7 Detection of DNA Damage in Different Organs of the Mouse -- 7.1 Introduction -- 7.2 The Alkaline Comet Assay in Multiple Organs of Mice -- 7.2.1 Chemicals and Materials -- 7.2.2 Methodology -- 7.3 Conclusions -- Acknowledgments -- References -- Chapter 8 Detection of DNA Damage in Drosophila -- 8.1 Introduction -- 8.2 General Protocol for the Assessment of DNA Damage Using the Alkaline Comet Assay -- 8.2.1 Chemicals and Materials -- 8.2.2 Preparation of Reagents -- 8.2.3 Preparation of Agarose Coated (Base) Slides for the Comet Assay -- 8.2.4 Preparation of Microgel Slides for the Comet Assay -- 8.2.5 Electrophoresis of Microgel Slides -- 8.2.6 Evaluation of DNA Damage -- 8.3 The Alkaline Comet Assay in Drosophila melanogaster -- 8.3.1 Chemicals and Materials -- 8.3.2 Methodology -- 8.4 Conclusion -- Acknowledgments -- References -- Section III: Applications of Comet Assay -- Chapter 9 The Comet Assay: Clinical Applications -- 9.1 Introduction -- 9.2 The Comet Assay Methodology -- 9.3 Clinical Studies -- 9.4 Discussion and Conclusions -- References -- Chapter 10 Applications of the Comet Assay in Human Biomonitoring -- 10.1 Biomonitoring and Biomarkers - An Introduction -- 10.2 The (Modified) Comet Assay -- 10.3 Guidelines for Biomonitoring Studies -- 10.4 Biomonitoring with the Comet Assay: Special Considerations -- 10.4.1 Surrogate and Target Cells -- The Use of White Blood Cells -- 10.4.2 Sampling Time and Transport -- 10.4.3 Reference Standards -- 10.4.4 What Affects the Background Level of DNA Damage? -- 10.5 DNA Damage as a Marker of Environmental Exposure and Risk -- 10.6 DNA Repair as a Biomarker of Individual Susceptibility -- 10.7 Protocols -- 10.7.1 Protocol for Blood Sample Collection and Long-term Storage of Lymphocytes for the Measurement of DNA Damage and Repair. , 10.7.2 Comet Assay - Determination of DNA Damage (Strand Breaks and Oxidised Bases) -- 10.7.3 In vitro Assays for DNA Repair -- 10.8 Solutions, etc. -- 10.8.1 Lysis Solution -- 10.8.2 Buffer F (Enzyme Reaction Buffer for FPG, End on uclease III, and In vitro BER Assay) -- 10.8.3 Buffer F+Mg (Used for In vitro NER Assay) -- 10.8.4 Buffer A (Used in In vitro Repair Assays) -- 10.8.5 Triton Solution -- 10.8.6 Ro 19-8022 (Photosensitiser) -- 10.8.7 Electrophoresis Solution -- 10.8.8 Neutralising Buffer -- 10.8.9 Agarose -- 10.8.10 Enzymes -- 10.9 Analysis and Interpretation of Results -- 10.9.1 Quantitation -- 10.9.2 Calculation of Net Enzyme-sensitive Sites -- 10.9.3 Calibration -- 10.9.4 How to Deal with Comet Assay Data Statistically -- 10.10 Conclusions -- Acknowledgments -- References -- Chapter 11 Comet Assay in Human Biomonitoring -- 11.1 Introduction -- 11.2 Human Monitoring -- 11.3 Environmental Exposure -- 11.4 Lifestyle Exposure -- 11.5 Occupational Exposure -- 11.6 Reviews -- 11.7 Usefulness of the Comet Assay in Human Monitoring -- 11.8 Conclusions -- References -- Chapter 12 Use of Single-cell Gel Electrophoresis Assays in Dietary Intervention Trials -- 12.1 Introduction -- 12.2 Different Endpoints -- 12.3 Experimental Design of Human Studies -- 12.4 Indicator Cells and Media -- 12.5 Conventional SCGE Trials With Complex Foods and Individual Components-The Current State of Knowledge -- 12.5.1 Definition of the Quality Score (QS) -- 12.6 Use of SCGE Trials to Detect Protection Against DNA-reactive Carcinogens -- 12.7 Use of SCGE-experiments to Monitor Alterations of the DNA-repair Capacity (Base- and Nucleotide-excision Repair) -- 12.8 What Have We Learned From Intervention Studies So Far? -- 12.9 Future Perspectives -- 12.9.1 Hot Topics -- 12.9.2 Detection of Antioxidants -- 12.9.3 Standardization. , 12.9.4 Search for Mechanistic Explanations -- 12.9.5 Interpretation Problems -- References -- Chapter 13 The Application of the Comet Assay in Aquatic Environments -- 13.1 Introduction -- 13.2 Protocols, Cell Types and Target Organs -- 13.3 Application of the Comet Assay to Invertebrate Species -- 13.3.1 Freshwater Invertebrates -- 13.3.2 Marine Invertebrates -- 13.4 Application of the Comet Assay to Vertebrate Species -- 13.4.1 Freshwater Vertebrates -- 13.4.2 Marine Vertebrates -- 13.5 Conclusions -- References -- Chapter 14 The Alkaline Comet Assay in Prognostic Tests for Male Infertility and Assisted Reproductive Technology Outcomes -- 14.1 Introduction -- 14.2 Sites of DNA Damage in Sperm -- 14.2.1 Oxidative Stress, a Major Cause of DNA Damage -- 14.2.2 Oxidative Stress, Antioxidant Therapies -- 14.2.3 Sperm DNA Damage Tests -- 14.2.4 Modifications to the Alkaline Comet Assay for Use with Sperm -- 14.2.5 Sperm DNA Adducts and their Relationship with DNA Fragmentation -- 14.3 Can Sperm DNA Integrity Predict Success? Relationships with Assisted Conception Outcomes -- 14.4 Clinically Induced DNA Damage -- 14.4.1 Cryopreservation -- 14.4.2 Vasectomy -- 14.5 A Major Barrier to Progress -- 14.6 Opportunities and Challenges - The Establishment of Clinical Thresholds and the Integration of DNA Testing into Clinical Practice -- Acknowledgments -- References -- Chapter 15 The Comet Assay in Sperm-Assessing Genotoxins in Male Germ Cells -- 15.1 Introduction -- 15.2 The Comet Assay (Single-cell Gel Electrophoresis) -- 15.3 The Use of Sperm with the Comet Assay -- 15.3.1 Human Sperm -- 15.3.2 Sperm DNA and the Comet Assay -- 15.3.3 Modifying Existing Comet Protocols for Somatic Cells for Use with Sperm -- 15.3.4 The Two-tailed Sperm Comet Assay -- 15.3.5 The Sperm Comet Assay and the Use of Repair Enzymes -- 15.3.6 Assessing the Sperm Comet. , 15.3.7 Comet-FISH on Sperm.
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  • 6
    Keywords: Vaccines. ; Electronic books.
    Description / Table of Contents: By addressing considerations of efficacy and safety of drugs and chemicals used to combat COVID-19, virtually in real-time, this book documents and highlights the advances in science and place the toxicology, pharmaceutical science, public health and medical community in a better position to advise in future epidemics.
    Type of Medium: Online Resource
    Pages: 1 online resource (594 pages)
    Edition: 1st ed.
    ISBN: 9781839163654
    DDC: 016.35960973
    Language: English
    Note: Cover -- The Coronavirus Pandemic and the Future Volume 1: Virology, Epidemiology, Translational Toxicology and Therapeutics -- Dedication -- Foreword -- Preface -- Contents -- Contents -- Chapter 1 - What We Know About the Life- threatening Novel Human Coronavirus -- 1.1 Introduction -- 1.2 The Chronology of the Outbreak -- 1.3 Initial Transmission -- 1.3.1 Worldwide Spread of the Virus -- 1.4 SARS-CoV- 2 and Its Etiology -- 1.5 Classification and Grouping -- 1.5.1 Alpha- CoVs -- 1.5.2 Beta- CoVs -- 1.5.3 Gamma- CoVs -- 1.5.4 Delta- CoVs -- 1.6 Epidemiological History of Human Coronaviruses -- 1.6.1 Severe Acute Respiratory Syndrome -- 1.6.2 Middle East Respiratory Syndrome -- 1.6.3 Detailed Documentation of SARS- CoV- 2 and COVID- 19 -- 1.7 Introductory Virology and Pathophysiology of SARS- CoV- 2 -- 1.7.1 Viral Genome -- 1.7.2 Interaction of the Spike and Receptor for Entry -- 1.7.3 Genomic Differences -- 1.7.4 Pathophysiology -- 1.8 Executive Summary and Conclusions -- List of Abbreviations -- Acknowledgements -- References -- Chapter 2 - Severe Acute Respiratory Syndrome Coronavirus- 2 (SARS-CoV- 2): Structure, Lifecycle, and Replication Machinery -- 2.1 Introduction -- 2.2 Classification of Coronaviruses -- 2.3 Genomic Structure of SARS- CoV-Ł2 -- 2.4 Life Cycle of SARS- CoV- 2 -- 2.4.1 Attachment and Entry -- 2.4.2 Replicase Protein Expression -- 2.4.3 Replication and Transcription -- 2.4.4 Assembly and Release -- 2.5 SARS- CoV- 2 RNA Replication Machinery -- References -- Chapter 3 - Accumulated Epidemiological Lessons and China's COVID- 19 Response -- 3.1 Introduction -- 3.2 Accumulated Lessons from China's Response to SARS in 2003 -- 3.3 Epidemiologicial Perspective on China's COVID- 19 Response -- 3.4 China's Control and Prevention Measures in COVID- 19 -- 3.4.1 Mask- wearing and its Comforting Effect. , 3.4.2 Epidemiological Source- tracking and Slow- down of Community Transmission -- 3.5 Comparing China's Response Against SARS and COVID- 19 -- 3.6 Concluding Remarks -- 3.7 Executive Summary -- References -- Chapter 4 - Clinical Presentation, Pathophysiology and Histopathology -- 4.1 Introduction -- 4.1 Introduction -- 4.2 The Distinct Clinical Trajectory and Disease Spectrum of COVID- 19 -- 4.3 A Novel Application of Smartphone Technology -- 4.4 COVID- 19 and Comorbidities in Severe Cases -- 4.5 ARDS and its Sequelae in COVID- 19 -- 4.5.1 ARDS in COVID- 19 -- 4.5.2 Sepsis -- 4.5.3 Septic Shock -- 4.6 Pathophysiology -- 4.6.1 Mechanism of SARS-Cov- 2 Invasion into Host Cells -- 4.7 Histopathology and Disease Manifestations: Introduction -- 4.7.1 Histopathology -- 4.7.2 Pulmonary Manifestations -- 4.7.3 Cardiovascular Manifestations -- 4.7.4 Gastrointestinal Manifestations -- 4.7.5 Renal Manifestations -- 4.7.6 Neurological Manifestations -- 4.7.7 Musculocutaneous Manifestations -- 4.7.8 Hematological Manifestations -- 4.8 Executive Summary -- Acknowledgements -- References -- Chapter 5 - Evaluation of the Disease, Sample Collection and Diagnostics -- 5.1 Introduction -- 5.1.1 Symptoms of COVID- 19 -- 5.1.2 Detection of Disease Based on Symptoms -- 5.1.3 Evaluation Criteria for COVID- 19 Based on ICMR and WHO Guidelines -- 5.2 Sample Collection -- 5.2.1 Overview of Sample Collection Guidelines from the WHO -- 5.2.1.1 Nasopharyngeal or Oropharyngeal Swabs -- 5.2.1.2 Nasal Mid- turbinate Swab -- 5.2.1.3 Anterior Nares Specimen -- 5.2.1.4 Nasopharyngeal Wash/Aspirate or Nasal Wash/Aspirate -- 5.2.1.5 Bronchoalveolar Lavage, Tracheal Aspirate, Pleural Fluid, Lung Biopsy -- 5.2.1.6 Sputum (Phlegm) -- 5.2.1.7 Saliva -- 5.2.1.8 Other -- 5.3 Recent Diagnostics -- 5.3.1 Molecular Diagnostics -- 5.3.2 Laboratory Examinations (Modified from Cascella et al.17). , 5.3.3 Imaging -- 5.3.3.1 Chest X- ray -- 5.3.3.2 Chest CT -- 5.3.3.3 Lung Ultrasound -- 5.3.3.4 Scanning Techniques -- 5.3.4 Blood Parameter Levels (Laboratory Diagnostics) -- 5.3.5 Genetic Susceptibility to COVID- 19 -- 5.3.6 Importance of Real-time RT-PCR in the Detection of COVID- 19 -- 5.3.6.1 Viral Nucleic Acid Detection -- 5.3.7 Serological Assays -- 5.3.8 Enzyme- linked Immunosorbent Assay -- 5.3.9 Magnetic Chemiluminescence Enzyme Immunoassay -- 5.3.10 Colloidal Gold Immunochromatographic Assay -- 5.3.11 Lateral Flow Immunoassay -- 5.3.12 Rapid Antigen Test -- 5.3.13 Neutralization Antibody Assay -- 5.3.14 Other Useful Diagnostics for COVID- 19 -- 5.3.15 False Negatives and Positives -- 5.4 Advantages and Disadvantages of Different Diagnostic Approaches -- 5.4.1 Detection Based on Patient History -- 5.4.2 Detection of Disease Based on Symptoms -- 5.4.3 Scanning Techniques -- 5.4.3.1 CT Scan -- 5.4.3.2 Chest X- ray -- 5.4.3.3 18F- FDG PET/CT -- 5.4.4 Laboratory Serum Diagnostics (e.g. WBC, Inflammatory Markers) -- 5.4.5 Molecular Diagnosis -- 5.4.5.1 RT- PCR -- 5.4.5.2 RT- LAMP -- 5.4.5.3 Transcription- mediated Amplification -- 5.4.6 CRISPR- based Diagnosis -- 5.4.7 Serological Assays -- 5.4.7.1 ELISA -- 5.4.7.2 GICA -- 5.4.7.3 MCLIA -- 5.4.7.4 Lateral Flow Immunoassay -- 5.4.7.5 Neutralization Bioassay -- 5.4.7.6 Rapid Antigen Test -- 5.5 Executive Summary -- Acknowledgements -- References -- Chapter 6 - Therapeutic Options Initially Available for COVID- 19 Patients and Initial Clinical Trials -- 6.1 Introduction to Therapeutic Options Initially Available for COVID- 19 Patients -- 6.1.1 General Guidelines for the Treatments Provided to COVID- 19 Patients -- 6.1.2 Common Treatment Regimens Provided to COVID- 19 Patients -- 6.1.3 Medicinal Plants as Therapy for COVID- 19 -- 6.1.4 Convalescent Plasma Therapy. , 6.1.5 Mesenchymal Stem Cell-Derived Exosomes as Therapy for COVID- 19 -- 6.1.6 Other Treatment Options -- 6.1.6.1 Corticosteroids -- 6.2 Introduction to Ongoing Clinical Trials -- 6.2.1 Ongoing Trials on Various Antiviral Agents -- 6.2.2 Immunosuppressants/Immunomodulators -- 6.2.3 Current Trials on MSCs for COVID- 19 -- 6.3 Executive Summary -- Acknowledgements -- References -- Chapter 7 - Clinical Epidemiology of Coronavirus Disease 2019: Infectivity, Clinical Spectrum and Presentation, and Population Distribution -- 7.1 Introduction -- 7.2 Infection and Tests -- 7.2.1 Viral Tests -- 7.2.2 Population- based Infection Rate -- 7.2.3 Antibody Tests -- 7.3 Clinical Spectrum of COVID- 19 -- 7.3.1 Classification by Clinical Type -- 7.3.2 Asymptomatic Cases -- 7.3.3 Severe Cases and Comorbidity -- 7.3.4 Clinical and Immunological Response -- 7.4 Regional Variation in Clinical Presentation -- 7.4.1 Clinical Manifestation -- 7.4.2 Chemosensory Dysfunction -- 7.4.3 Common Symptoms -- 7.4.3.1 Asia -- 7.4.3.2 USA -- 7.4.3.3 Europe -- 7.5 Population Distribution -- 7.5.1 Pregnancy and Pregnant Women -- 7.5.2 Infants and Pediatric Patients -- 7.5.3 Asymptomatic Cases and Clinical Severity in Children -- 7.5.4 Older Adults -- 7.5.5 Sex Differences -- 7.5.6 Occupation -- 7.6 Case- fatality Rate -- 7.7 Executive Summary and Conclusions -- References -- Chapter 8 - SARS-CoV-2 Genomics and Host Cellular Susceptibility Factors of COVID- 19 -- 8.1 Introduction -- 8.2 Coronavirus and Human Diseases -- 8.2.1 Classification of Coronaviruses -- 8.2.2 Human Coronaviruses and Diseases -- 8.2.3 The Natural Origin of HCoVs -- 8.3 Genomic Structure and Genetics -- 8.3.1 Viral Genomic Structure -- 8.3.2 The Replicase Locus and Proteins -- 8.3.2.1 Nonstructural Proteins -- 8.3.2.2 Structural Proteins -- 8.3.2.3 Accessory Proteins -- 8.3.3 Genomic Mutations. , 8.4 Viral Replication -- 8.4.1 Attachment and Cell Entry -- 8.4.2 Translation and RNA Replication -- 8.4.3 Assembly and Release of Virion -- 8.5 Host Cellular Factors -- 8.5.1 ACE2 -- 8.5.2 TMPRSS2 -- 8.5.3 Furin -- 8.5.4 DPP4 -- 8.5.5 CD147 -- 8.5.6 GRP78 -- 8.5.7 Cathepsins -- 8.5.8 Ly6E -- 8.5.9 L- SIGN and DC- SIGN -- 8.5.10 Sialic Acid -- 8.5.11 Plasmin and Other Proteases -- 8.6Executive Summary -- References -- Chapter 9 - Infection and Pathogenesis of SARS-CoV- 2: an Immunological Perspective -- 9.1 Introduction -- 9.2 SARS-CoV- 2 Virology -- 9.2.1 Structure -- 9.2.2 Genome -- 9.2.3 Proteome -- 9.2.4 Mutation -- 9.2.5 Transmission -- 9.3 Pathogenesis -- 9.3.1 Attachment to the Host Cell -- 9.3.2 Penetration of SARS-CoV- 2 Inside the Host Cell -- 9.3.3 Assembly of the RTC -- 9.3.4 Genome Replication, Transcription, and Translation -- 9.3.5 Virion Assembly and Release -- 9.3.6 Incubation Period -- 9.4 Disease Progression -- 9.5 Immunology -- 9.5.1 Innate Immune Response -- 9.5.1.1 Evasion of Innate Sensing by SARS-CoV- 2 -- 9.5.1.1.1 Evasion of PRR Sensing.Coronavirus- mediated antagonism starts with the evasion of PRR recognition. The ssRNA of coronaviruses c... -- Inhibition of Signalling Cascades. As described, TLR and RLR activation induces signalling cascades, which leads to the release ... -- Inhibition of IFN-1396983920I. Based on previous studies, various mechanisms have been adopted by coronaviruses to inhibit IFN-1... -- 9.5.1.2 Imbalanced Antiviral Defense and Pro- inflammatory Cytokine Production -- 9.5.1.3 Macrophages -- Macrophage Responses to SARS-CoV- 2 Infection.Macrophages and monocytes accumulate and trigger cytokine secretion, leading to c... -- 9.5.1.4 Alveolar type II ATII Cells in the Innate Immune Response -- 9.5.1.5 Neutrophils -- 9.5.1.6 Dendritic Cells -- 9.5.1.7 Natural Killer Cells. , 9.5.1.7.1 Modification of NK Cells Upon SARS-CoV- 2 Infection.Studies on peripheral blood samples of COVID- 19 patients have shown that i.
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  • 7
    Keywords: Electronic books.
    Description / Table of Contents: By addressing considerations of efficacy and safety of drugs and chemicals used to combat COVID-19, virtually in real-time, this book documents and highlights the advances in science and place the toxicology, pharmaceutical science, public health and medical community in a better position to advise in future epidemics.
    Type of Medium: Online Resource
    Pages: 1 online resource (694 pages)
    Edition: 1st ed.
    ISBN: 9781839166839
    Language: English
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  • 8
    Online Resource
    Online Resource
    La Vergne :RSC,
    Keywords: Electronic books.
    Description / Table of Contents: This book is a must for newcomers to the field who wish to gain an understanding of what toxicology is all about.
    Type of Medium: Online Resource
    Pages: 1 online resource (582 pages)
    Edition: 2nd ed.
    ISBN: 9781847550798
    Language: English
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  • 9
    Online Resource
    Online Resource
    La Vergne :RSC,
    Keywords: Electronic books.
    Description / Table of Contents: The first book of its kind, devoted exclusively to the assessment of DNA damage by the Comet assay in modern toxicology.
    Type of Medium: Online Resource
    Pages: 1 online resource (479 pages)
    Edition: 1st ed.
    ISBN: 9781847559746
    Series Statement: ISSN Series
    Language: English
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  • 10
    Keywords: Vaccines. ; Electronic books.
    Description / Table of Contents: By addressing considerations of efficacy and safety of drugs and chemicals used to combat COVID-19, virtually in real-time, this book documents and highlights the advances in science and place the toxicology, pharmaceutical science, public health and medical community in a better position to advise in future epidemics.
    Type of Medium: Online Resource
    Pages: 1 online resource (694 pages)
    Edition: 1st ed.
    ISBN: 9781839166846
    DDC: 016.35960973
    Language: English
    Note: Cover -- The Coronavirus Pandemic and the Future Volume 2: Virology, Epidemiology, Translational Toxicology and Therapeutics -- Dedication -- Foreword -- Preface -- Contents -- Contents -- Chapter 1 - In Silico Approaches for Drug Repurposing for SARS-CoV- 2 Infection -- 1.1 Introduction -- 1.1.1 What is Drug Repurposing -- 1.1.2 Why Drug Repurposing -- 1.1.2.1 Regulatory Considerations -- 1.1.3 Why Drug Repurposing for COVID-19? -- 1.1.4 Why In Silico Approaches for Drug Repurposing -- 1.1.4.1 Gene Profiling -- 1.1.4.2 In Vitro Screening -- 1.1.4.3 Phenotypic Screening -- 1.1.4.4 Binding Assays for Target Interactions -- 1.1.4.5 Cellular Thermal Shift Assay -- 1.1.4.6 The In Silico Effort -- 1.2 Understanding SARS-CoV- 2 from an In Silico Perspective -- 1.2.1 Structural Proteins of SARS-CoV- 2 -- 1.2.1.1 Surface Spike Protein (S-protein) -- 1.2.1.2 Nucleocapsid Protein (N-protein) -- 1.2.1.3 Envelope Protein (E-protein) -- 1.2.1.4 Membrane Protein (M-protein) -- 1.2.2 Non-structural and Accessory Proteins of SARS-CoV- 2 -- 1.2.3 Structure of SARS-CoV- 2 Proteins -- 1.3 Structure-based Approaches for Drug Repurposing -- 1.3.1 Docking Studies in the Main Protease (Mpro/3CLpro) -- 1.3.1.1 Crystal Structures Used in Docking -- 1.3.1.2 Methods or Programs Used in Docking -- 1.3.1.3 Ligands and Databases Used for Screening -- 1.3.1.4 Molecular Dynamics Programs Used -- 1.3.1.5 Duration of MD Simulation -- 1.3.1.6 Free-energy Estimation Method Used -- 1.3.1.7 Analysis of Hit Drugs from Docking -- 1.3.1.8 Some Noteworthy Studies on Mpro -- 1.3.1.8.1 Study Reporting Good Correlation of Predicted Binding Affinity with Experiments.Huynh et al. performed MD simulation of apo and ... -- 1.3.1.8.2 Studies Using Pharmacophore or Shape-based Methods for Screening of Drugs. Arun. , 1.3.1.8.3 Novel Methods of Screening. Sencanski et al. reported a novel two-step approach to identify -- 1.3.1.8.4 Studies in which Important Observations are Reported.Jin et al. reported identification of lead compounds by combining structure... -- 1.3.2 Docking Studies in RNA-dependent RNA Polymerase -- 1.3.3 Docking Studies in Papain-like Protease -- 1.3.4 Docking Studies in the Nucleocapsid Protein (N-protein) -- 1.3.5 Docking Studies in the Spike Glycoprotein (S-protein) -- 1.3.6 Docking Studies in NSP1 -- 1.3.7 Docking Studies in NSP13/Helicase -- 1.3.8 Docking Studies in NSP15/Endonucleases -- 1.3.9 Docking Studies in NSP16 -- 1.3.10 Docking in Main Protease and Spike Glycoprotein -- 1.3.11 Docking in Multiple Structural Proteins -- 1.3.12 Docking in Proteases -- 1.3.13 Docking in Multiple Targets -- 1.3.13.1 Studies Involving Large Numbers of Targets -- 1.3.13.2 Studies Involving Main Protease and Other Targets -- 1.3.13.3 Studies on Mixed Targets -- 1.3.14 Discussion and Consensus Screening Protocol from the Reviewed Literature -- 1.4 Ligand-based Approaches for Drug Repurposing -- 1.4.1 QSAR-based Approaches -- 1.4.2 Pharmacophore-based Approaches -- 1.5 Other Approaches for Drug Repurposing -- 1.5.1 Machine Learning-based Methods -- 1.5.1.1 Machine Learning Using Molecular Descriptors -- 1.5.1.2 Machine Learning Using Docking Interactions -- 1.5.2 Pharmacology-based Network Analysis Methods -- 1.5.2.1 Protein-Protein Interactions -- 1.5.2.2 Expression Profiling -- 1.6 Understanding Human Targets in COVID-19 From an In Silico Perspective -- 1.6.1 Host Proteins Involved in the SARS-CoV- 2 Life Cycle -- 1.6.2 Host Response to SARS-CoV- 2 Infection -- 1.6.2.1 SARS-CoV- 2 Induced Immune Response -- 1.6.3 Structural Information of Human Proteins in COVID-19 -- 1.7 Structure-based Approaches for Drug Repurposing Using Human Proteins. , 1.7.1 Docking Studies in Angiotensin Converting Enzyme-2 -- 1.7.1.1 Targeting the ACE-2 Receptor -- 1.7.1.2 Targeting ACE-2 and Spike Receptor -- 1.7.1.3 Targeting ACE-2 and Spike Receptor in Addition to Network-based Associations -- 1.7.2 Docking Studies in Transmembrane Protease, Serine 2 (TMPRSS2) -- 1.7.3 Docking Studies in Glucose-Regulated Protein 78 (GRP78) -- 1.7.4 Docking Studies in Furin -- 1.7.5 Docking Studies in ARDS Targets -- 1.7.5.1 TNF-­α -- 1.8 Summary of Hits from Reviewed Literature -- 1.9 Concluding Remarks -- 1.10 Executive Summary -- Author Contributions -- Acknowledgements -- References -- Chapter 2 - Vaccination and Vaccines for COVID-19 -- 2.1 Introduction -- 2.2 Vaccination in the Context of a Pandemic Outbreak -- 2.3 COVID-19 -- 2.3.1 Specific Strategies for COVID-19 -- 2.3.1.1 DNA Vaccines -- 2.3.1.2 mRNA Vaccines -- 2.3.1.3 Attenuated Vaccines -- 2.4 Mental Health Aspects of Immunization and Vaccination -- 2.5 Vaccines for COVID-19 -- 2.5.1 SARS-CoV-2 -- 2.5.2 Immune Response to SARS-CoV- 2 and Previous Coronavirus Infections -- 2.5.3 Platforms for COVID-19 Vaccine Development -- 2.5.3.1 DNA-based Vaccines -- 2.5.3.1.1 Inovio.Inovio Pharmaceuticals is an American company based in Plymouth Meeting, Pennsylvania, USA, that specializes in manufactu... -- 2.5.3.2 RNA-based Vaccines -- 2.5.3.2.1 Moderna/NIAID. Moderna is an American company based in Cambridge, Massachusetts, that has developed an mRNA -based vaccine, mRNA -1273. The mRNA vaccine -- 2.5.3.2.2 BioNTech/Fosun/Pfizer.BioNTech, a German company, together with Pfizer, an American company, have developed another mRNA- based ... -- 2.5.3.3 Non-replicating Viral Vector Vaccines -- 2.5.3.3.1 AstraZeneca/University of Oxford. The University of Oxford has formed a partnership with the -- 2.5.3.4 Inactivated Vaccines. , 2.5.3.4.1 Wuhan Institute of Biological Products/Beijing Institute of Biological Products.The above institutions and the pharmaceutical co... -- 2.5.3.4.2 Indian Vaccine Development.There are several institutions including academic/research and vaccine manufacturing companies in Ind... -- 2.5.3.5 Challenges in COVID-19 Vaccine Development -- 2.6 Executive Summary -- References -- Chapter 3 - Understanding the Emergence of SARS-CoV- 2 Viral Variants From a Genomic Perspective -- 3.1 Introduction -- 3.2 A History of Pandemics -- 3.3 Origin and Classification of SARS-CoV-2 -- 3.4 Genomic Architecture of SARS-CoV- 2 -- 3.4.1 Overview -- 3.4.2 Structural Proteins -- 3.4.3 Non-structural Proteins -- 3.5 The Emergence of SARS-CoV- 2 Variants -- 3.6 Antigenic Variations and Immune Escape Mutations -- 3.7 Leading Vaccines Across the Globe -- 3.8 Conclusion -- References -- Chapter 4 - Susceptibility and Spread of SARS-CoV- 2 in Animals -- 4.1 Introduction -- 4.2 Wild Animals -- 4.3 Laboratory Animals -- 4.4 Companion Animals -- 4.5 Farmed Animals -- 4.6 Conclusions -- References -- Chapter 5 - Profiling Some Plant-based Immunomodulatory Bioactive Compounds for COVID-19 Prophylaxis and Treatment Based on Indian Traditional Medicine -- 5.1 Introduction -- 5.2 Prevention of SARS-CoV- 19 Infection: Stage 1 -- 5.3 Incubation to Non-severe Symptomatic Phase - Stage 2 -- 5.4 Severe Respiratory Symptomatic Phase with Hyperinflammation - Stage 3 -- 5.5 Multi-organ Response to COVID-19 Infection - Stage 4 -- 5.6 Summary and Outlook -- 5.7 Executive Summary -- List of Abbreviations -- Authors' Contributions -- Declaration of Competing Interest -- Acknowledgements -- References -- Chapter 6 - The Potential Therapeutic Effects of Natural Products, Herbs, and Mushrooms Against COVID-19 -- 6.1 Origins and Pathogenesis of COVID-19. , 6.2 The Immune Profile in COVID-19Infection -- 6.3 Natural Marine Compounds as Potent Inhibitors Against SARS-CoV- 2 -- 6.4 Potential Use of Mushrooms and Herbs Against COVID-19 Infection -- 6.5 Conclusion -- References -- Chapter 7 - Application of Chinese Herbal Medicine in COVID-19 -- 7.1 Introduction -- 7.2 Representative Prescriptions -- 7.2.1 Qingfei Paidu Decoction (QFPDD, 清 排毒汤) -- 7.2.2 Huashi Baidu Formula (HSBDF, 化湿 毒方) -- 7.2.3 XuanFei Baidu Formula (XFBDF, 宣 毒方) -- 7.3 Finished Patent Medicines -- 7.3.1 Lianhua Qingwen Capsules (LH-­C, 清瘟 囊) -- 7.3.2 Jinhua Qingan Granules (JH-­G, 清感 粒) -- 7.3.3 Xuebijing Injection (XBJ, 必净注射液) -- 7.4 Other Chinese Herbs -- 7.4.1 Ephedra Herba (Ma Huang, ) -- 7.4.2 Honeysuckle (Jin Yin Hua, ) -- 7.4.3 Scutellariae (Huang Qin, ) -- 7.4.4 Glycyrrhizae Radix (Gan Cao, 甘 ) -- 7.4.5 Armeniacae Semen (Ku Xing Ren, 杏仁) -- 7.4.6 Sophorae flavescentis Radix (Ku Shen, 参) -- 7.4.7 Curcuma longa (Jiang Huang, 姜 ) -- 7.5 Conclusion -- List of Abbreviations -- References -- Chapter 8 - COVID-19 Interventional and Therapeutic Clinical Trials: Small Molecules, Interactions, Outcomes and Opportunities -- 8.1 Introduction -- 8.2 Removal-of- treatment Preventative Clinical Trials -- 8.2.1 Trial Aims -- 8.2.2 Trial Populations -- 8.2.3 Potential Therapeutics -- 8.2.4 Trial Designs -- 8.2.5 Primary Outcomes -- 8.2.6 Secondary Outcomes -- 8.2.7 Toxicities -- 8.2.8 Opportunities -- 8.3 Addition-of- treatment Preventative and/or Curative Trials -- 8.3.1 Trial Aims -- 8.3.2 Trial Populations -- 8.3.3 Potential Therapeutics -- 8.3.4 Trial Designs -- 8.3.5 Primary Outcomes -- 8.3.6 Secondary Outcomes -- 8.3.7 Toxicity -- 8.3.7.1 Population-specific Toxicity -- 8.3.7.2 Interactions -- 8.3.8 Opportunities -- 8.4 Supportive Clinical Trials -- 8.4.1 Trial Aims -- 8.4.2 Trial Populations -- 8.4.3 Potential Therapeutics. , 8.4.4 Trial Designs.
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