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Visualisation of Signalling in Immune Cells

Currently, a great number of approaches are employed in investigation of the immune system. These range from experiments in live animals and biochemical techniques to investigate whole organs or cell populations down to single cell and molecular techniques to look at dynamics in specific ...

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Imaging Interactions Between the Immune and Cardiovascular Systems In Vivo by Multiphoton Microscopy

Several recent studies in immunology have used multiphoton laser-scanning microscopy to visualise the induction of an immune response in real time in vivo. These experiments are illuminating the cellular and molecular interactions involved in the induction, maintenance and reg ...

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Applying an Adaptive Watershed to the Tissue Cell Quantification During T-Cell Migration and Embryonic Development

Cell and particle quantification is one of the frequently used techniques in biology and clinical study. Variations of cell/particle population and/or protein expression level can provide information on many biological processes. In this chapter, we propose an image-based automa ...

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Identifying Homing Interactions in T-Cell Traffic in Human Disease

Description of the molecular mechanisms which regulate the traffic of lymphocyte populations over recent years has significantly enhanced our understanding of the processes underlying acquired immunity and also permitted the development of therapies targeted at specific l ...

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Tracking Antigen-Experienced Effector T Cells In Vitro and In Vivo

The TCR complex is a multisubunit complex, comprising at least eight transmembrane units. The clonotypic TCR α and β chains are responsible for antigen recognition, whilst the invariant chains of the CD3 complex (δ, ɛ and γ) and two zeta (ζ) polypeptides couple antigen recognition to downstream s ...

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Preclinical Testing of Strategies for Therapeutic Targeting of Human T-Cell Trafficking In Vivo

Naive T cells are migratory cells that continuously recirculate between blood and lymphoid tissues. Antigen-specific stimulation of T cells within the lymph nodes reprograms the trafficking properties of T cells by inducing a specific set of adhesion molecules and chemokine recept ...

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Fluorescence and Electron Microscopy Methods for Exploring Antimicrobial Peptides Mode(s) of Action

Due to the increasing resistance of microbial pathogens to the available drugs, the identification of new antimicrobial agents with a new mechanism of action is urgently needed. In this context, cationic antimicrobial peptides (AMPs) are considered promising candidates. Although ...

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Use of Atomic Force Microscopy as a Tool to Understand the Action of Antimicrobial Peptides on Bacteria

Atomic force microscopy (AFM) has been extensively used to image the three-dimensional surface morphology of a broad range of biological samples, including Gram-negative bacteria, imaged in the presence of antimicrobial peptides (AMPs). Although this technique provides high mo ...

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Solid-State NMR Investigations of Membrane-Associated Antimicrobial Peptides

Solid-state NMR and other biophysical investigations have revealed many mechanistic details about the interactions of antimicrobial peptides with membranes. These studies have shaped our view on how these peptides cause the killing of bacteria, fungi, or tumour cells and how they per ...

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Dynamic Transitions of Membrane-Active Peptides

Membrane-active peptides or protein segments play an important role in many biological processes at the cellular interface to the environment. They are involved, e.g., in cellular fusion or host defense, where they can cause not only merging but also the destabilization of cell membranes. Ma ...

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Comparing Bacterial Membrane Interactions of Antimicrobial Peptides and Their Mimics

Interactions with bacterial membranes are integral to the mechanisms of action of all antimicrobial peptides (AMPs), regardless of their final cellular targets. Here, we describe in detail two biophysical techniques that can be used to measure the membrane activities of AMPs and antimi ...

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Binding and Permeabilization of Model Membranes by Amphipathic Peptides

Measurement of binding and activity of antimicrobial and cytolytic amphipathic peptides on membranes is essential to understanding their function and cell specificity. The use of model systems has provided a wealth of information on the interactions of amphipathic peptides with m ...

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Antimicrobial Peptides: The LPS Connection

An expanding body of evidence is rendering manifest that many cationic antimicrobial peptides are endowed with different properties and activities, well beyond their direct action on microbes. One of the most interesting and potentially important research avenue on the alternati ...

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High-Throughput Screening for Antimicrobial Peptides Using the SPOT Technique

The SPOT technique provides a fast, cost-efficient, and highly parallel method to synthesize peptide arrays on cellulose. Peptides synthesized on cellulose can be easily cleaved from the support and used directly in a screening assay for antimicrobial activity. Depending on the equip ...

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Synthesis of Antimicrobial Peptides Using the SPOT Technique

Developing new lead structures for drugs against multiresistant bacteria is an urgent need for modern medicine. Antimicrobial peptides are a class of drugs that can be used to discover such structures. In order to support development of this research, a fast, easy, and inexpensive method to sy ...

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Synthesis and Thermodynamic Characterization of Small Cyclic Antimicrobial Arginine and Tryptophan-Rich Peptides with Selectivity for Gram-Negative Ba

One promising strategy to combat the proliferation of bacteria resistance toward current antibiotics is the development of peptide-based drug. Among these compounds is a group of small cyclic peptides rich in arginine (Arg) and tryptophan (Trp) residues with selective toxicity towa ...

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Characterization of the Leishmanicidal Activity of Antimicrobial Peptides

This chapter describes the basic methodology to assay the activity of antimicrobial peptides (AMPs) on Leishmania, a human protozoan parasite. The protocols included can be methodologically divided into two major blocks. The first one addresses the basic technology for growth of the di ...

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Antimicrobial and Antibiofilm Activity of Quorum Sensing Peptides and Peptide Analogues Against Oral Biofilm Bacteria

Widespread antibiotic resistance is a major incentive for the investigation of novel ways to treat or prevent infections. Much effort has been put into the discovery of peptides in nature accompanied by manipulation of natural peptides to improve activity and decrease toxicity. The ever i ...

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Measuring Antimicrobial Peptide Activity on Epithelial Surfaces in Cell Culture

To more accurately assess the activity and role of epithelial cell-derived antimicrobial peptides in their native settings, it is essential to perform assays at the surfaces under relevant conditions. In order to carry this out, we utilize three-dimensional cultures of airway and gingi ...

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Ex Vivo Skin Infection Model

Over the past decade, the emergence of bacterial strains resistant to conventional antibiotics has necessitated the discovery and development of new antimicrobial therapies. This chapter describes a skin infection model that is based on the use of excised skin derived from the domestic ...

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