Antibiotic Resistance Protocols 2nd Edition (Methods in Molecular Biology Vol. 642)

Several years have passed since the first edition of our book, and much has changed in the world of microbiology. The trend of increasing antibiotic resistance has continued with, for example, the emergence of extensively resistant M. tuberculosis. Our understanding of antibiotic resistance biology has increased. Genomic methods have become more accessible allowing antibiotic researchers to probe not only the sequence of antibiotic resistance determinants but also the mechanism whereby they are expressed and regulated. Array methodology has allowed a large number of genes to be studied simultaneously, while real time PCR has allowed detailed dissection of gene expression. We have also begun to understand the importance of biofilms and the physiological state of the organism in permitting organisms to survive in the face of antibiotics and the act of developing resistance.

The molecular revolution has provided the tools that allow us to detect and characterise resistance determinants, and these may be sequence based, nucleic acid amplification methods, or detection of the activity of efflux pumps. The ability of a resistant organism to survive and thrive in the clinical environment is fundamental to the evolution of resistance, and considerable progress has been made in the understanding of the role of fitness in the selection of resistant organisms. This understanding has been aided by an increasing range of physiological characteristics that are tested, and automated methods of testing have improved the reproducibility and robustness of these assays. The need to develop new antibiotic agents means that we have to develop new high throughput methods of evaluating susceptibility. The interaction between bacterial and human populations is an important area of investigation, and we need better techniques to characterise the population genetics of the organisms and better sampling tools to ensure that this data is representative.

The editors have sought to bring together examples of a diverse range of modern techniques applied in antibiotic research, and they hope that you find these useful in planning your own research.

- The Editors -

Contents

Part I Epidemiology and Population Genetics

• 1 The RM Test for Determining Methicillin-Resistant Staphylococcus aureus Lineages
• 2 Unravelling the Ecology of Antibiotic Resistant Bacteria in the Nasopharynx

Part II Genomics and Gene Expression

• 3 Methods to Determine Antibiotic Resistance Gene Silencing
• 4 Studying Gene Induction of Glycopeptide Resistance Using Gene Swapping
• 5 Real-Time PCR Methods to Study Expression of Genes Related to Hypermutability
• 6 Use of DNA Arrays to Study Transcriptional Responses to Antimycobacterial Compounds

Part III Fitness Mutation and Physiology

• 7 Assays of Sensitivity of Antibiotic-Resistant Bacteria to Hydrogen Peroxide and Measurement of Catalase Activity
• 8 Measurements of Heme Levels and Respiration Rate in Salmonella enterica Serovar Typhimurium
• 9 Methods to Determine Fitness in Bacteria
• 10 Application of Continuous Culture for Measuring the Effect of Environmental Stress on Mutation Frequency in Mycobacterium tuberculosis

Part IV Detection of Resistance

• 11 Identification of Efflux-Mediated Multi-drug Resistance in Bacterial Clinical Isolates by Two Simple Methods
• 12 Evaluation of Efflux Activity of Bacteria by a Semi-automated Fluorometric System
• 13 Measuring the Activity of Active Efflux in Gram-Negative Bacteria
• 14 Rapid Characterization of b-Lactamases by Multiplex PCR
• 15 Rapid Methods for Testing Inhibitors of Mycobacterial Growth
• 16 Use of Biofilm Model Systems to Study Antimicrobial Susceptibility
• 17 Rapid Analysis of Resistant Mutant Genotypes Using Pyrosequencing®

Index

About the Editors

• Stephen H. Gillespie, MD, UCL Medical School, London, UK. Email: sgillesp@medsch.ucl.ac.uk
• Timothy D. McHugh, Ph.D, UCL Medical School, London, UK. Email: t.mchugh@medsch.ucl.ac.uk

Product Details

• Hardcover: 248 pages
• Publisher: Humana Press; 2nd ed. edition (April 16, 2010)
• Language: English
• ISBN-10: 160327278X
• ISBN-13: 978-1603272780
• Product Dimensions: 9.9 x 7.1 x 0.8 inches

List Price: $119.00 

 

Giordano: Cell Cycle Regulation and Differentiation in Cardiovascular and Neural Systems

The brain and heart have been used as metaphors for thinking and feeling, for cognition and emotion. These ideas have been demonstrated to have a high scientific value. In fact, complex physiopathological relationships exist between these two organs. This book will deal with several aspects of the cardiovascular and nervous systems from a new viewpoint. It will describe the differences and similarities in their differentiation pathways with a peculiar emphasis on the role of cell cycle regulation and cell differentiation.

Cell cycle exit represents the fundamental step to trigger differentiation of cells and induction of a novel program of gene expression leading to the elaboration of a specialized phenotype. Moreover, there is evidence demonstrating that several components of the cell cycle machinery play a major role also in cell specification and differentiation both in neural and cardiovascular systems. The differentiation process will be evaluated starting from the most early cell precursor, i.e., stem cells. The attention of readers will be focused also on transcription factors with differentiating properties and on their relationship with cell cycle regulators.

In summary, this book will offer an in-depth analysis of the differentiation process in two systems that have profound relationships with one another and, therefore, will help us to better understand their biology by providing the tools to dissect the molecular basis of pathological conditions. The book will prompt the scientific community to perceive cell cycle regulation and differentiation under a novel and more comprehensive light.

Contents

Part I Introduction to Cell Cycle and Differentiation

• 1 Short Introduction to the Cell Cycle
• 2 The Basic Helix-Loop-Helix Transcription Factors in Neural Differentiation
• 3 Transcription Factors and Muscle Differentiation

Part II Stem Cells

• 4 The Neural Stem Cells
• 5 Cardiac Stem and Progenitor Cells
• 6 Muscle Stem Cells
• 7 Marrow Stromal Mesenchymal Stem Cells

Part III Cell Cycle and Differentiation in the Nervous System

• 8 Neurogenesis in the Central Nervous System: Cell Cycle Progression/Exit and Differentiation of Neuronal Progenitors

Part IV Cell Cycle and Differentiation in the Cardiovascular System

• 9 Cell Cycle and Differentiation in the Cardiovascular System
• 10 Cell Cycle and Differentiation in Vessels

Part V Cell Cycle and Differentiation in the Muscle

• 11 Cell Cycle Regulation in Myogenesis

Subject Index

About the Editors

• Antonio Giordano, College of Science & Technology, Sbarro Institute for Cancer Research, Temple University, N. 12th Street 1900, 19122 Philadelphia, Pennsylvania, USA. Email: giordano@temple.edu
• Umberto Galderisi, Dipto. Medicina Sperimentale, Università Napoli Federico II, Sezione di Biotecnologia e Biologia Molecolare, Via Costantinopoli 16, 80138 Napoli, Italy. Email: umberto.galderisi@unina2.it

Product Details

• Hardcover: 267 pages
• Publisher: Springer; 1st edition (August 30, 2010)
• Language: English
• ISBN-10: 160327152X
• ISBN-13: 978-1603271523
• Product Dimensions: 9.1 x 6.2 x 0.8 inches

List Price: $189.00 

 

Schwartzbach: Immunoelectron Microscopy: Methods and Protocols (Methods in Molecular Biology Vol. 657)

Cell biology is the science of correlating cell structure and function. The electron microscopist obtains high-resolution pictures of the intricate structures found in cells. Electron tomography and serial sections can be used to reconstruct the three-dimensional structure of the cell and its organelles. Pictures are worth a thousand words, but they are unable to provide information regarding the function of the intricate structures found in cells and their macromolecular composition.

The biochemist and molecular biologist determine the functions of the molecules, macromolecular complexes, and organelles found within cells. They isolate individual cellular constituents and reconstruct vital cellular processes. These in vitro experiments provide a detailed understanding of cellular function. Organelle isolation provides a method to place macromolecular functions within a structural context. Understanding structure–function relationships through organelle isolation has restricted utility because organelles cannot be isolated from every organism, not every organelle can be isolated free of contamination by other organelles, suborganellular compartments often cannot be purified for biochemical characterization, and, when a protein is recovered in multiple organelles, it is often difficult to distinguish true localization from contamination artifacts. 

Immunoelectron microscopy is the technique that bridges the information gap between biochemistry, molecular biology, and ultrastructural studies placing macromolecular functions within a cellular context. Immunoelectron microscopy can be used on virtually every unicellular and multicellular organism. The only requirements are suitable fixation protocols and the availability of an antibody to the molecule whose structural location is to be determined. Structure–function relationships can be determined even when it is impossible to purify the organelle or suborganellular compartment containing the macromolecules being studied. Most importantly, immunoelectron microscopy is a totally objective procedure that is not dependent on conjectures as to where the protein is localized and thus which organelles to isolate for biochemical studies.

The successful application of immunoelectron microscopy requires combining the tools of the molecular biologist with those of the microscopist. From the molecular biology toolbox, this volume will present methods for antigen production by protein expression in bacterial cells and by expression of epitope tagged proteins in plant and animal cells. Methods for production of anti-peptide, monoclonal, and polyclonal antibodies will be presented. From the microscopy toolbox, this volume will present methods for cryoultramicrotomy and rapid freeze-replacement fixation which have the advantage of retaining protein antigenicity at the expense of ultrastructural integrity as well as chemical fixation methods that maintain structural integrity while sacrificing protein antigenicity. Plants and algae contain cell walls, vacuoles, and other structures which present barriers to antibody penetration and complicate fixation. Due to these problems, separate chapters will discuss fixation and immunolabeling protocols for animals, plants, and algae. Preand post-embedding immunogold labeling protocols will be presented. Pre-embedding methods perform immunogold labeling before ultrathin sections are prepared from resinembedded samples resulting in greater sensitivity and better microstructure preservation. Post-embedding methods perform immunolabeling after ultrathin sections are prepared from resin-embedded samples resulting in decreased antigenicity. The detailed methods and notes will facilitate choosing the best method for the antibody and biological material to be studied. Finally, methods will be presented for immunogold labeling of two antigens for protein colocalization studies, for three-dimensional reconstruction of intracellular antigen distribution, for immunogold labeling of DNA, and for immunogold scanning electron microscopy. The toolbox created by this volume will facilitate an increased understanding of structure–function relationships.

Contents

PART I MOLECULAR TOOLBOX

• 1. Protein Antigen Expression in Escherichia coli for Antibody Production
• 2. Expression of Epitope-Tagged Proteins in Plants
• 3. Expression of Epitope-Tagged Proteins in Arabidopsis Leaf Mesophyll Protoplasts
• 4. Transient Expression of Epitope-Tagged Proteins in Mammalian Cells
• 5. Production and Purification of Polyclonal Antibodies
• 6. Production and Purification of Monoclonal Antibodies
• 7. Production of Antipeptide Antibodies
• 8. Preparation of Colloidal Gold Particles and Conjugation to Protein A, IgG, F(ab’)2, and Streptavidin

PART II MICROSCOPY TOOLBOX

• 9. Immunoelectron Microscopy of Chemically Fixed Developing Plant Embryos
• 10. Pre-embedding Immunogold Localization of Antigens in Mammalian Brain Slices
• 11. Pre-embedding Immunoelectron Microscopy of Chemically Fixed Mammalian Tissue Culture Cells
• 12. Immunoelectron Microscopy of Cryofixed and Freeze-Substituted Plant Tissues
• 13. In Vivo Cryotechniques for Preparation of Animal Tissues for Immunoelectron Microscopy
• 14. Immunoelectron Microscopy of Cryofixed Freeze-Substituted Mammalian Tissue Culture Cells
• 15. Immunoelectron Microscopy of Cryofixed Freeze-Substituted Saccharomyces cerevisiae
• 16. High-Resolution Molecular Localization by Freeze-Fracture Replica Labeling
• 17. Pre-embedding Electron Microscopy Methods for Glycan Localization in Chemically Fixed Mammalian Tissue Using HorseradishPeroxidase-Conjugated Lectin
• 18. Pre-embedding Nanogold Silver and Gold Intensification
• 19. The Post-embedding Method for Immunoelectron Microscopy of Mammalian Tissues: A Standardized Procedure Based on Heat-Induced Antigen Retrieval
• 20. Double-Label Immunoelectron Microscopy for Studying the Colocalization of Proteins in Cultured Cells
• 21. Serial Section Immunoelectron Microscopy of Algal Cells
• 22. Freeze-Etch Electron Tomography for the Plasma Membrane Interface
• 23. Localization of rDNA at Nucleolar Structural Components by Immunoelectron Microscopy
• 24. Immunogold Labelling for Scanning Electron Microscopy
• 25. Horseradish Peroxidase as a Reporter Gene and as a Cell-Organelle-Specific Marker in Correlative Light-Electron Microscopy
• 26. Monitoring Rapid Endocytosis in the Electron Microscope via Photoconversion of Vesicles Fluorescently Labeled with FM1-43

Subject Index

From the Back Cover

Immunoelectron microscopy is a key technique that bridges the information gap between biochemistry, molecular biology, and ultrastructural studies placing macromolecular functions within a cellular context. In Immunoelectron Microscopy: Methods and Protocols, expert researchers combine the tools of the molecular biologist with those of the microscopist. From the molecular biology toolbox, this volume presents methods for antigen production by protein expression in bacterial cells, methods for epitope tagged protein expression in plant and animal cells allowing protein localization in the absence of protein specific antibodies as well as methods for the production of anti-peptide, monoclonal, and polyclonal antibodies. From the microscopy toolbox, sample preparation methods for cells, plant, and animal tissue are presented. Both cryo-methods, which have the advantage of retaining protein antigenicity at the expense of ultrastructural integrity, as well as chemical fixation methods that maintain structural integrity while sacrificing protein antigenicity have been included, with chapters examining various aspects of immunogold labeling. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and essential, Immunoelectron Microscopy: Methods and Protocols seeks to facilitate an increased understanding of structure function relationships.

Product Details

• Hardcover: 352 pages
• Publisher: Humana Press; 1st edition (July 20, 2010)
• Language: English
• ISBN-10: 160761782X
• ISBN-13: 978-1607617822
• Product Dimensions: 10.1 x 6.9 x 1 inches

List Price: $139.00 

 

Pecorini: Molecular Biology of Cancer 2nd edition: Mechanisms, Targets & Therapeutics

Many of the cells in our body undergo tightly regulated replication to enable growth to occur and to ensure that damaged or worn out tissues are replaced. Regulation is mediated via many different mechanisms. It is when these mechanisms fail, and regulation is lost, that cancer can result. Recent advances in molecular cell biology have greatly expanded our understanding of the processes and mechanisms which underpin the regulation of cell replication, and what is happening at the molecular level when these fail.

Now in its second edition, The Molecular Biology of Cancer: Mechanisms, Targets, and Therapeutics gives a fresh approach to the study of the molecular basis of cancer. Written with the undergraduate student in mind, Lauren Pecorino focuses on how our understanding of the defective mechanisms which drive cancer is leading to the development of new targeted therapeutic agents.

Opening with an overview of the key hallmarks of cancer as a disease state, the book then leads us through the principal components of the regulatory machinery through which cell division is controlled and corrupted with the onset of cancer. Within each chapter, students are shown how this molecular knowledge is being applied to develop new targeted therapeutic strategies.

Ideal for courses in the molecular biology of cancer and cancer biology, The Molecular Biology of Cancer, Second Edition, is an invaluable resource for any student wishing to gain insight into the molecular basis of the disease and formulate possible solutions for its effective control.

Contents

• 1: Introduction
• 2: DNA structure and stability: mutations vs. repair
• 3: Regulation of gene expression
• 4: Growth factor signaling and oncogenes
• 5: The cell cycle
• 6: Growth inhibition and tumor suppressor genes
• 7: Apoptosis
• 8: Stem cells and differentiation
• 9: Metastasis
• 10: Infections and inflammation
• 11: Nutrients, hormones, and gene interactions
• 12: The Cancer Industry: drug development and clinical trial design
• 13: Cancer in the future: focus on diagnostics and immunotherapy

Key Features

• Show how an understanding of the molecular processes underpinning cancer can be directly applied to the development of new therapeutics, giving students a clear appreciation of how the theory relates to real life scenarios.
• Provides a carefully structured overview of the molecular mechanisms that regulate cell replication, and relates defects in these mechanisms to the onset of cancer, so that students can clearly see the link between the two.
• Draws on the latest research in this dynamic field but presents it in a manner readily accessible to the student; clarity is not sacrificed to achieve a contemporary account of the field.
• Online Resource Centre features additional resources for both lecturers and students, enhancing the value of this text as a teaching and learning tool.

New to this Edition

• Three new chapters. 'The Cell Cycle' gives extended treatment of this central aspect of cancer biology. 'Infections and Inflammation' focuses on the relationship between infectious agents and inflammation, and the onset of cancer. 'The Cancer Industry' gives an overview of the process through which drugs are developed and tested, setting the therapeutics theme that runs through the book in a professional context.
• Enhanced coverage of apoptosis.
• Over 25 new figures plus a new colour plate section, to further support and illustrate concepts presented in the written text.
• A new 'How do we know that?' feature, in which experimental methods and results are discussed, giving students an insight into how our understanding of cancer biology is developed.
• An extended glossary, to help those new to the subject grasp the terminology used.

Product Details 

• Paperback: 320 pages
• Publisher: Oxford University Press, USA; 2 edition (May 6, 2008)
• Language: English
• ISBN-10: 0199211485
• ISBN-13: 978-0199211487
• Product Dimensions: 9.5 x 7.4 x 0.6 inches 

List Price: $59.95 

Mendelsohn: The Molecular Basis of Cancer 3rd Edition (Expert Consult Series)

The Molecular Basis of Cancer: with Expert Consult 3rd Edition by John Mendelsohn, Peter M. Howley, Mark A. Israel, Joe W. Gray and Craig B. Thompson explores the scientific basis for our current understanding of malignant transformation and the pathogenesis and treatment of cancer. A team of leading experts thoroughly explain the molecular biologic principles that underlie the diagnostic tests and therapeutic interventions now being used in clinical trials and practice. Incorporating cutting-edge advances and the newest research, the book provides thorough descriptions of everything from molecular abnormalities in common cancers to new approaches for cancer therapy. A new full-color format expedites reference, and now as an Expert Consult title, it enables you to conveniently search the entire text online, download all of the books images for your personal use, and review periodic updates.
 


Contents

I - CARCINOGENESIS AND CANCER GENETICS

• 1 - Cancer: A Genetic Disorder
• 2 - Oncogenes and Signal Transduction
• 3 - Tumor Suppressor Genes
• 4 - DNA Repair Pathways and Human Cancer
• 5 - Epigenetics and Cancer
• 6 - Infectious Agents and Cancer
• 7 - Environmental Carcinogenesis
• 8 - Animal Models: Flies, Fish, and Yeast
• 9 - Genetic Mouse Models of Cancer

II - CANCER BIOLOGY

• 10 - Cancer Stem Cells
• 11 - Signal Transduction by Growth Factor Receptors
• 12 - Cell Growth
• 13 - Regulation of the Cell Cycle
• 14 - Metabolism of Cell Growth and Proliferation
• 15 - Apoptosis, Autophagy, and Necrosis
• 16 - Cellular Senescence
• 17 - The Tumor Microenvironment in Cancer Progression
• 18 - Tumor Angiogenesis
• 19 - Invasion and Metastasis

III - MOLECULAR PATHOLOGY AND DIAGNOSTICS

• 20 - Cancer Genomics
• 21 - Transcriptome Analysis
• 22 - Mass Spectrometry in Cancer Biology
• 23 - Imaging and Cancer
• 24 - Bioinformatics
• 25 - Early Detection of Cancer: Molecular Screening

IV - MOLECULAR PATHOGENESIS OF SPECIFIC MALIGNANCIES

• 26 - Lymphoma/Myeloma
• 27 - Molecular Genetics of ALL
• 28 - Biology of Chronic and Acute Myeloid Leukemia
• 29 - Molecular Biology of Childhood Neoplasms
• 30 - Molecular Basis of Lung Cancer
• 31 - Molecular Abnormalities in Colon and Rectal Cancer
• 32 - Breast Cancer
• 33 - Molecular Basis of Prostate Cancer
• 34 - Molecular Pathogenesis of Epithelial Ovarian Cancer
• 35 - Molecular Pathogenesis of Pancreatic Adenocarcinoma
• 36 - Molecular Biology of Melanoma
• 37 - Soft-Tissue Sarcomas
• 38 - Head and Neck Squamous Cell Carcinoma
• 39 - Primary Brain Tumors
• 40 - Epithelial Skin Cancer1
• 41 - Endocrine Cancer
• 42 - Gastric and Gastroesophageal Adenocarcinoma

V - MOLECULAR BASIS OF CANCER THERAPY

• 43 - From Bench to Bedside with Targeted Therapies
• 44 - Screening Strategies for Targeted Therapeutics
• 45 - Animal Models to Predict Anticancer Efficacy
• 46 - Pharmacokinetics, Pharmacodynamics, and Pharmacogenetics1
• 47 - Phase 1 Trials Today
• 48 - Cancer Therapeutics
• 49 - Natural and Acquired Resistance to Cancer Therapies
• 50 - Molecular Basis of Radiation Therapy
• 51 - Hematopoietic Growth Factors and Cytokines
• 52 - Interferons
• 53 - Cancer and the Cellular Immune Response
• 54 - Cancer-Specific Vaccines
• 55 - Monoclonal Antibody Therapy of Cancer
• 56 - Gene Therapy and Oncolytic Viruses
• 57 - RNA as a Therapeutic Molecule
• 58 - Heat Shock Protein 90 and the Proteasome
• 59 - Cancer Chemoprevention 

Book Features 

• Offers online access the book's complete contents fully searchable along with periodic updates, and artwork from the text for your personal use.
  
• Features sweeping updates throughout, including molecular targets for the development of anti-cancer drugs, gene therapy, and vaccines... keeping you on the cutting edge of your specialty.
  
• Offers a new, more user-friendly full-color format so the information that you need is easier to find.
• Presents abundant figures all redrawn in full color illustrating major concepts for easier comprehension.
• Features numerous descriptions of the latest clinical strategies helping you to understand and take advantage of todays state-of-the-art biotechnology advances.

Website Features

• Consult the book from any computer at home, in your office, or at any practice location.
• Instantly locate the answers to your clinical questions via a simple search query.
• Quickly find out more about any bibliographical citation by linking to its MEDLINE abstract.
• Images: Browse a Library of all book images. Easily select, organize, and download your images into a presentation. 

About the Author

• John Mendelsohn, MD, President, The University of Texas, MD Anderson Cancer Center, Houston, TX; 
• Peter Howley, MD, Professor and Chairman, Department of Pathology, Harvard Medical School, Boston, MA; 
• Mark A. Israel, MD, Director, Preuss Laboratory of Molecular Neuro-Oncology Brain Tumor Research Center, University of California, San Francisco, San Francisco, CA; 
• Lance A. Liotta, MD., PhD, Chief, Laboratory of Pathology, National Cancer Institute, National Institutes of Health, San Francisco, CA.

Product Details 
 

• Hardcover: 704 pages
  
• Publisher: Saunders; 3 edition (March 26, 2008)
  
• Language: English
  
• ISBN-10: 1416037039
  
• ISBN-13: 9781416037033
  
• Dimensions: 11.1 x 8.8 x 1.4 inches
  

List Price: $179.00