Keywords:
Intestines--Microbiology.
;
Electronic books.
Type of Medium:
Online Resource
Pages:
1 online resource (290 pages)
Edition:
1st ed.
ISBN:
9781536123388
Series Statement:
Bacteriology Research Developments
URL:
https://ebookcentral.proquest.com/lib/geomar/detail.action?docID=5014354
DDC:
612.33000000000004
Language:
English
Note:
Intro -- Contents -- Preface -- Chapter 1 -- Introduction -- 1.1. History and Development of Probiotic Bacteria -- 1.1.1. Definition of Probiotics and Probiotic-Active Substances -- 1.1.2. Probiotic Properties and Classification -- 1.2. Microencapsulation - Mechanisms to Protect Sensitive Probiotic Strains -- 1.2.1. Microencapsulation of Probiotic Bacteria -- 1.3. Gastrointestinal (GI) Tract -- 1.3.1. Functions of Microbiota in the GI Tract -- 1.4. Gut-Associated Immunological Tissue (GALT) -- 1.4.1. Structure and Function of the GALT -- 1.4.2. Probiotics and the GALT -- 1.5. Immune System -- 1.5.1. Innate Immunological Responses -- 1.5.2. Adaptive Immunological Responses -- 1.6. Probiotic Immunomodulatory Activities -- 1.6.1. Probiotic Effects on the Innate Immune System -- 1.6.2. Probiotic Effects on the Adaptive Immune System -- 1.7. Confocal Microscopy -- 1.7.1. Fluorescent Dyes -- 1.7.2. Fluorescent Proteins -- 1.7.2.1. Humanized Monster Green® Fluorescent Protein (hMGFP) -- 1.7.2.2. DsRed2 Fluorescent Protein -- 1.7.3. Intracellular Tracking of DNA, RNA and Proteins Using Image Analysis Software -- 1.7.3.1. Fluorescence Correlation Spectroscopy (FCS) -- 1.7.3.2. Raster Image Correlation Spectroscopy (RICS) -- 1.8. Overall Aim of Study -- Chapter 2 -- Growth of Probiotic Bacteria in Mammalian Cell Culture Conditions and Confocal Microscopy Analysis of Microencapsulated Probiotic Bacteria -- 2.1. Introduction -- 2.1.1. Microencapsulation of Probiotic Bacteria -- 2.1.2. Probiotic Bacteria and Pathogenic Bacteria Strains -- 2.2. Aims and Objectives -- 2.3. Materials and Methods -- 2.3.1. Bacterial Growth Curves -- 2.3.1.1. Bacteria Cells and Growing Media -- 2.3.1.2. Growth Curves for L. acidophilus, B. lactis and S. pyogenes -- 2.3.2. Bacteria Growth Curves in Mammalian Cell Culture Conditions.
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2.3.2.1. Bacteria and Mammalian Cell Culture Reagents -- 2.3.2.2. Bacteria Growth Curves in Mammalian Cell Culture Conditions -- 2.3.3. Viability Standard Curve -- 2.3.3.1. Reagents -- 2.3.3.2. Staining of Probiotic Bacteria with LIVE/DEAD® BacLight™ Bacterial Viability and Counting Kit (Invitrogen) and Development of a Viability Standard Curve -- 2.3.4. Microencapsulation of Bacteria and the Development of a Viability Standard Curve -- 2.3.4.1. Microencapsulation Materials -- 2.3.4.2. Microencapsulation of Probiotic Bacteria and Incubation on MRS Agar and in Mammalian Cell Culture Media -- 2.3.4.2.1. MRS Agar and Standard Anaerobic Bacteria Incubation Conditions -- 2.3.4.2.2. Mammalian Cell Culture Media and Conditions -- 2.3.4.3. Staining of Bacteria with LIVE/DEAD® BacLight™ Bacterial Viability and Counting Kit (Invitrogen) and Microencapsulation of Bacteria -- 2.3.4.4. Laser Scanning Confocal Microscopy (LSCM) Analysis of Microencapsulated Bacteria -- 2.3.4.5. Analysis of Microencapsulated Bacteria Viability Using the Bitplane Imaris Software -- 2.3.5. Statistics -- 2.4. Results -- 2.4.1. Bacteria Growth Curves in Broth and Mammalian Cell Culture Conditions -- 2.4.2. Viability Standard Curve -- 2.4.3. Microencapsulation of Bacteria -- 2.4.3.1. Laser Scanning Confocal Microscopy Analysis of Microencapsulated Probiotic Bacteria -- 2.4.3.2. Growth of Microencapsulated Probiotic Bacteria on MRS Agar and in Mammalian Cell Culture Media -- 2.4.3.3. Analysis and Development of a Viability Standard Curve for Microencapsulated Bacteria Using the Bitplane Imaris Software -- 2.5. Discussion -- 2.5.1. Bacterial Growth Curves in Broth and Mammalian Cell Culture Media and Conditions -- 2.5.2. Composition of Mammalian Cell Culture Media and Broth - Effects on Bacterial Growth -- 2.5.3. Microencapsulation of Probiotic Bacteria.
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2.5.3.1. Distinguishing Viable and Non-Viable Cells Using Fluorescent Stains -- 2.5.3.2. Viability Standard Curve for Microencapsulated Bacteria -- 2.5.3.3. Microencapsulation Ingredients - Effects on the Diffusion of Soluble Factors and Mammalian Cell Culture Media -- 2.5.3.4. Viability and Distribution of Microencapsulated Bacteria -- 2.5.3.5. Release of Microencapsulated Bacteria -- 2.6. Conclusion -- Chapter 3 -- Development of a Fluorescent Immune Cell Model -- 3.1. Introduction -- 3.1.1. Fluorescent Proteins as Reporter Molecules of Gene and Protein Expression -- 3.1.2. Fluorescent Dyes to Confirm Localization of Fluorescent Protein Expression -- 3.1.3. The L45 and L23 Progenitor Immune Cell Lines -- 3.2. Aims and Objectives -- 3.3. Materials and Methods -- 3.3.1. Transfection of the Mammalian Immune Cells, L23 and L45, with the Fluorescent Protein Plasmids, phMGFP and pCIneo-DsRed2 -- 3.3.1.1. Transfection Reagents -- 3.3.1.2. Fluorescent Protein Plasmids -- 3.3.1.3. Porcine Progenitor Immune Cell Lines -- 3.3.1.4. Transfection Procedure -- 3.3.1.5. Laser Scanning Confocal Microscopy (LSCM) Analysis of Transfected Progenitor Immune Cells -- 3.3.1.6. Comparison of the Growth-Rates of Immune Cells Expressing Fluorescent Proteins and not Expressing Fluorescent Proteins Cell -- 3.3.2. Identification of Transfected Cellular Fluorescence Expression by Comparative Analysis with Nucleic Acid and Lysosomal-Specific Fluorescent Probes -- 3.3.2.1. Fluorescent Probes -- 3.3.2.2. Cell Suspension Preparation and Staining with SYTO 59® and LysoSensor™ Blue DND-167 -- 3.3.2.3. Laser-Scanning Confocal Microscopy (LSCM) Settings and Wavelength Scans -- 3.3.2.4. Laser-Scanning Confocal Microscopy (LSCM) Analysis of DNA, RNA and Lysosomal Regions in Transfected Cells Using SYTO 59® and LysoSensor™ Blue DND-167.
,
3.3.2.5. Co-Localization Analysis Using Bitplane Imaris Software -- 3.3.3. Anisomycin Treatment of Transfected Cells to Evaluate the Relationship between Apoptosis and Fluorescence Expression -- 3.3.3.1. Anisomycin Preparation -- 3.3.3.2. Anisomycin Treatment of Transfected Cells -- 3.3.3.3. Fluorescent Probes -- 3.3.3.4. Laser Scanning Confocal Microscopy Analysis of Apoptosis-Induced Immune Cells -- 3.3.3.5. Co-Localization Analysis Using Bitplane Imaris Software -- 3.3.4. Mitogen Treatment of Transfected Cells to Evaluate the Relationship between Increased Cellular Activity and Fluorescence Expression -- 3.3.4.1. Concanavalin A (Con A) and Lipopolysaccharide (LPS) Preparation -- 3.3.4.2. Mitogen Treatment of Transfected Cells -- 3.3.4.3. Fluorescent Probes -- 3.3.4.4. Laser-Scanning Confocal Microscopy Analysis of Mitogen-Stimulated Immune Cells -- 3.3.4.5. Co-Localization Analysis Using Bitplane Imaris Software -- 3.3.5. Statistical Analysis -- 3.4. Results -- 3.4.1. Fluorescence Expression of Immune Cells Following Transfection -- 3.4.1.1. Growth-Rate of Transfected and Non-Transfected Cells -- 3.4.1.1.1. Growth Rate of L45, L45hMGFP and L45pCIneo-DsRed2 -- 3.4.1.1.2. Growth Rate of L23, L23hMGFP and L23pCIneo-DsRed2 -- 3.4.2. Identification of Transfected Cellular Fluorescence Expression by Comparative Analysis with Nucleic Acid and Lysosomal-Specific Fluorescent Probes -- 3.4.3. Co-Localization Analysis of Apoptotic-induced Intracellular Fluorescence, Hoechst 33342 and PI Expression Using Bitplane Imaris Software -- 3.4.3.1. Co-Localization Analysis of Mitogen-Stimulated Intracellular Fluorescence, Hoechst 33342 and PI Expression -- 3.5. Discussion -- 3.5.1. Fluorescence Expression of Progenitor Immune Cells Following Transfection with Fluorescent Protein Plasmids -- 3.5.1.1. Fluorescent Protein Vectors.
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3.5.1.2. Fluorescence Protein Expression by the Transfected L45 and L23 Cells -- 3.5.1.3. Effects on Fluorescence Expression of the Transfected Immune Cells Following Mitogen Treatments -- 3.5.1.4. Growth Rates of Transfected Immune Cells -- 3.5.2. Identification of Transfected Cellular Fluorescence Expression by Comparative Analysis with Nucleic Acid and Lysosomal-Specific Fluorescent Probes -- 3.5.2.1. Lysosomal and Nucleic Acid Staining of Transfected Immune Cells -- 3.5.2.2. Co-Localization of DsRed2 Fluorescent Protein with Lysosomal and Nucleic Acid Stains -- 3.5.2.3. Co-Localization of hMGFP Fluorescent Protein with Lysosomal and Nucleic Acid Stains -- 3.5.3. Anisomycin Treatment of Transfected Mammalian Immunological Cells to Evaluate the Relationship between Fluorescence Expression and Apoptosis -- 3.5.3.1. Anisomycin Treatment of Transfected Cells -- 3.5.3.2. Mitogen-Stimulation of Transfected Cells -- 3.6. Conclusion -- Chapter 4 -- Free and Microencapsulated Probiotic Bacteria-Effects on Immune Cell Proliferation -- 4.1. Introduction -- 4.1.1. Immune Cell Interactions with Probiotic Bacteria -- 4.1.2. Immune Response to Microencapsulated Probiotics -- 4.2. Aims and Objectives -- 4.3. Materials and Methods -- 4.3.1. Treatment of Porcine Progenitor Immune Cells with Free and Microencapsulated Bacteria Conditioned Mammalian Cell Culture Media -- 4.3.1.1. Bacteria-Conditioning of Mammalian Cell Culture Media -- 4.3.1.2. Treatment of Porcine Progenitor Immune Cells with Bacteria-Conditioned Media -- 4.3.1.3. Conditioning of Mammalian Cell Culture Media by Microencapsulated Bacteria -- 4.3.1.4. Treatment of Porcine Progenitor Immune Cells with Microencapsulated Bacteria-Conditioned Media -- 4.3.2. Statistical Analysis -- 4.4. Results.
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4.4.1. Free- and Microencapsulated Bacteria-Conditioned Mammalian Cell Culture Media Treatment of Porcine Progenitor Immune Cells.
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