Springer protocols feed by Infectious Diseases 
This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.
This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Assessment of Neutrophil Apoptosis
Timely neutrophil apoptosis and cell clearance by surrounding phagocytes are essential components of the resolution phase of acute inflammation. Programmed cell death by apoptosis occurs with maintenance of an intact cell membrane in order to prevent the release of histotoxic intracellular products such as proteases and reactive oxidant species into the extracellular surroundings as occurs during necrosis. Macrophage phagocytosis results in attenuation of toll-like receptor-driven proinflammatory mediator production further promoting inflammation resolution. Failures in this cascade of events can result in tissue damage, c...
Source: Springer protocols feed by Infectious Diseases - February 28, 2014 Category: Infectious Diseases Source Type: news
Subcellular Fractionation of Human Neutrophils and Analysis of Subcellular Markers
The neutrophil has long been recognized for its impressive number of cytoplasmic granules that harbor proteins indispensable for innate immunity. Analysis of isolated granules has provided important information on how the neutrophil grades its response to match the challenges it meets on its passage from blood to tissues. Nitrogen cavitation was developed as a method for disruption of cells on the assumption that sudden reduction of the partial pressure of nitrogen would lead to aeration of nitrogen dissolved in the lipid bilayer of plasma membranes. We find that cells are broken by the shear stress that is associated with...
Source: Springer protocols feed by Infectious Diseases - February 28, 2014 Category: Infectious Diseases Source Type: news
Collection of In Vivo Transmigrated Neutrophils from Human Skin
A wealth of knowledge on the life and death of human neutrophils has been obtained by the in vitro study of isolated cells derived from peripheral blood. However, neutrophils are of main importance, physiologically as well as pathologically, after they have left circulation and transmigrated to extravascular tissues. The journey from blood to tissue is complex and eventful, and tissue neutrophils are in many aspects distinct from the cells left in circulation. Here we describe how to obtain human tissue neutrophils in a controlled experimental setting from aseptic skin lesions created by the application of negative pressur...
Source: Springer protocols feed by Infectious Diseases - February 28, 2014 Category: Infectious Diseases Source Type: news
Neutrophil Isolation from Nonhuman Species
We present methods here for reproducibly isolating highly purified neutrophils from large animals (bovine, equine, ovine), small animals (murine and rabbit), and nonhuman primates (cynomolgus macaques), and describe optimized details for obtaining the highest cell purity, yield, and viability. We also describe methods to verify phagocytic capacity in the purified cell populations using a flow cytometry-based phagocytosis assay. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - February 28, 2014 Category: Infectious Diseases Source Type: news
Isolation of Human Neutrophils from Venous Blood
Venous blood provides a ready source of large numbers of unstimulated granulocytes and mononuclear cells. Exploiting the differences in the relative densities of the leukocytes circulating in venous blood, one can separate leukocytes from erythrocytes as well as isolate the individual leukocyte populations in high purity for use in ex vivo studies. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - February 28, 2014 Category: Infectious Diseases Source Type: news
The Role of Neutrophils in the Immune System: An Overview
Neutrophils, also known as polymorphonuclear leukocytes (PMNs), have long been considered as the short-lived, nonspecific white cells that form pus—and also happen to kill invading microbes. Indeed, neutrophils were often neglected (and largely not considered) as immune cells. This historic view of neutrophils has changed considerably over the past several decades, and we know now that, in addition to playing the predominant role in the clearance of bacteria and fungi, they play a major role in shaping the host response to infection and immune system homeostasis. The change in our view of the role of neutrophils in t...
Source: Springer protocols feed by Infectious Diseases - February 28, 2014 Category: Infectious Diseases Source Type: news
Isolation of Chromosomal and Plasmid DNA from Staphylococcus epidermidis
The following describes noncommercial methods for the purification of genomic and plasmid DNA from S. epidermidis. These include both large-scale, high molecular weight and quick, small-scale chromosomal DNA extractions, and also a standard alkaline lysis method of plasmid preparation. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
Genetic Manipulation of Staphylococci
The ability to genetically manipulate bacteria is essential to understanding gene/protein function in these organisms. While basic cloning has become routine in molecular biology, many still view the ability to make directed mutations as a daunting or intimidating task. To aid the staphylococcal research community, the goal of this treatise is to describe the method of allelic exchange using temperature-sensitive plasmids that we have used to successfully produce a variety of mutations including single nucleotide changes in the Staphylococcus aureus chromosome. In addition, this chapter provides extensive summaries to aid ...
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
The Isolation and Analysis of Phenol-Soluble Modulins of Staphylococcus epidermidis
Phenol-soluble modulins (PSMs) are multifunctional peptide toxins produced by many staphylococcal strains. PSMs have received much recent attention, owing to multiple reports underscoring their importance for staphylococcal pathogenesis. Members of the PSM family may be strongly cytolytic to neutrophils and other cell types; promote inflammatory, receptor-mediated responses in several human cell types; and contribute to biofilm structuring and detachment. Here we describe biochemical methods to isolate, purify, and quantitatively analyze Staphylococcus epidermidis PSMs. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
Growth and Preparation of Staphylococcus epidermidis for NMR Metabolomic Analysis
The “omics” era began with transcriptomics and this progressed into proteomics. While useful, these approaches provide only circumstantial information about carbon flow, metabolic status, redox poise, etc. To more directly address these metabolic concerns, researchers have turned to the emerging field of metabolomics. In our laboratories, we frequently use NMR metabolomics to acquire a snapshot of bacterial metabolomes during stressful or transition events. Irrespective of the “omics” method of choice, the experimental outcome depends on the proper cultivation and preparation of bacterial samples. I...
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
Multilocus Sequence Typing of Staphylococcus epidermidis
Multilocus sequence typing (MLST) is a genotyping method that is well suited for studying the population genetics and evolution of Staphylococcus epidermidis. The central MLST database for S. epidermidis continues to grow, and new analysis methods for extracting historical information from MLST data continue to be developed. Even in this era of whole-genome sequencing, MLST provides a reference genotyping method, and the central MLST database provides a unique catalog of genetic variants. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
Pulsed Field Gel Electrophoresis of Staphylococcus epidermidis
Pulsed field gel electrophoresis (PFGE) is one of the older methods for the molecular characterization and comparison of microorganisms including bacteria. Nevertheless, PFGE continues to be recognized as the gold standard for molecular typing due to output spanning >90 % of the bacterial genome and standardized protocols and reagents applicable to a wide range of organisms including S. epidermidis. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
Identification of Staphylococcus epidermidis in the Clinical Microbiology Laboratory by Molecular Methods
Biochemical assays for the phenotypic identification of coagulase-negative staphylococci in the clinical microbiology laboratory have been well described in previous publications (Becker and Von Eiff Manual of Clinical Microbiology, ASM Press, Washington, pp. 308–330, 2011; Kloos and Wolfshohl J Clin Microbiol 16:509–516, 1982). This discussion focuses on identification of Staphylococcus epidermidis through molecular and proteomic methods. Molecular assays have been shown to be more discriminatory between the coagulase-negative staphylococcal species than are phenotypic assays (Zadoks and Watts Vet Microbiol 13...
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
Staphylococcus epidermidis Pathogenesis
Staphylococcus epidermidis is the most frequently encountered member of the coagulase-negative staphylococci on human epithelial surfaces. It has emerged as an important nosocomial pathogen, especially in infections of indwelling medical devices. The mechanisms that S. epidermidis uses to survive during infection are in general of a passive nature, reflecting their possible origin in the commensal life of this bacterium. Most importantly, S. epidermidis excels in forming biofilms, sticky agglomerations that inhibit major host defense mechanisms. Furthermore, S. epidermidis produces a series of protective surface polymers a...
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
Clinical Characteristics of Infections in Humans Due to Staphylococcus epidermidis
Staphylococcus epidermidis is the most common cause of primary bacteremia and infections of indwelling medical devices. The ability to cause disease is linked to its natural niche on human skin and ability to attach and form biofilm on foreign bodies. This review focuses on the S. epidermidis clinical syndromes most commonly encountered by clinicians and future potential treatment modalities. (Source: Springer protocols feed by Infectious Diseases)
Source: Springer protocols feed by Infectious Diseases - November 18, 2013 Category: Infectious Diseases Source Type: news
