No description available.

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Proefschrift Universiteit van Amsterdam. / Auteursnaam op omslag: Anneloor ten Asbroek. Met lit. opg. - Met samenvatting in het Nederlands.

Zugl.: Frankfurt (Main), Universiẗat, Diss., 2008.

Univ., Diss., 2009--Marburg.

Streptococcus pyogenes is a re-emerging pathogen that produces superficial and life threatening invasive diseases. One important virulence factor in S. pyogenes is hyaluronic acid capsule which has been shown to increase expression at sub-body temperatures in certain strains. This study showed that thermoregulation is common in invasive clinical isolates. Regulation was shown to occur independent of the CovRS two-component regulator in a post transcription manner and before protein level regulation. The endoribonuclease, CvfA, was also confirmed to be required for capsule thermoregulation. The search for a regulator lead to the discovery of the 1st antisense RNAs in S. pyogenes found opposite the capsule synthesis genes. Its role if any in capsule has not been discovered. Finally, a group of sRNAs were characterized adding to the knowledge of this layer of regulation in S. pyogenes.

The complex heterogeneity of schizophrenia has proved difficult to replicate in preclinical animal models. Of the many molecular targets implicated with schizophrenia, this thesis focuses on synapsin II - a pre-synaptic protein critical for neurotransmission and synaptogenesis; and parvalbumin - a calcium-binding protein found in interneurons of the dorsolateral prefrontal cortex (DLPFC) and the striatum (STR). Patients with schizophrenia display reduced levels of synapsin II mRNA in the DLPFC, while decreased activation of parvalbumin neurons in the same region has resulted in schizophrenia-like cognitive deficits. Knockdown of synapsin II in the medial prefrontal cortex (mPFC) of neonate and adult rats has previously induced schizophrenia-like alterations. However, there are concerns that must be addressed before novel animal models of schizophrenia can be developed using reductions in synapsin II. This thesis was designed to 1) eliminate maternal separation (MS) between post-natal days (PD) 14-23, which correlates with a neurodevelopmental synapsin II model, as a means of inducing schizophrenia-like behaviours; 2) reassess the use of fully and partially phosphorothioated first-generation antisense oligonucleotides to reduce synapsin II levels, and 3) evaluate parvalbumin expression in the STR following synapsin II knockdown. Results from this study indicate 1) a 36 hour MS regimen during PD 14-23 did not cause behavioural changes bearing resemblance to schizophrenia; 2) oligonucleotide sequences stabilized completely with phosphorothioate bonds were insufficient in reducing synapsin II levels and caused localized necrosis, while partially modified sequences induced a slight knockdown effect without cell death; and 3) levels of striatal parvalbumin expression were decreased in rats receiving the partially, but not fully, modified antisense sequences. The findings strengthen the face validity and safety profile of the synapsin II knockdown model. Novel evidence has also been provided for the role of parvalbumin in the striatum and suggests its influence on cognitive dysfunction in schizophrenia. / Thesis / Master of Science (MSc)

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No description available.

by Liang Yiu-hon. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1995. / Includes bibliographical references (leaves 104-117). / ACKNOWLEDGMENTS --- p.i / ABSTRACT --- p.ii / ABBREVIATIONS --- p.iii / AMINO ACIDS SHORTHAND --- p.v / TABLE OF CONTENTS --- p.vi-x / Chapter CHAPTER 1 --- INTRODUCTION / Chapter 1.1 --- General Introduction --- p.1 / Chapter 1.2 --- The Discovery of IGFs --- p.1 / Chapter 1.3 --- The Growth Promoting Actions of IGFs --- p.3 / Chapter 1.4 --- Molecular Biology of IGFs in Mammals --- p.6 / Chapter 1.4.1 --- IGF Genes and Transcripts --- p.6 / Chapter 1.4.2 --- Regulation of IGF Gene Expression --- p.8 / Chapter 1.5 --- IGF Binding Proteins --- p.11 / Chapter 1.5.1 --- Regulation of IGF Action by IGF Binding Proteins --- p.11 / Chapter 1.6 --- The Insulin and IGF Receptors --- p.13 / Chapter 1.6.1 --- IGF-I Receptor --- p.13 / Chapter 1.6.2 --- IGF-II Receptor --- p.13 / Chapter 1.6.3 --- Insulin/IGF-I Hybrid Receptor --- p.15 / Chapter 1.7 --- IGF in Mammalian Fetal Growth --- p.17 / Chapter 1.8 --- The Role of IGFs in Fish --- p.19 / Chapter 1.9 --- Aims of the Present Study --- p.26 / Chapter CHAPTER 2 --- GENERAL METHODOLOGY / Chapter 2.1 --- Materials --- p.27 / Chapter 2.2 --- Methods --- p.32 / Chapter 2.2.1 --- Gene Clean --- p.32 / Chapter 2.2.1A --- Gene Clean by Glassmilk Method --- p.32 / Chapter 2.2.1B --- Gene Clean by Sephaglas´ёØ BandPrep Kit --- p.32 / Chapter 2.2.2 --- Preparation of Radioactive Nucleic Acid Probes --- p.33 / Chapter 2.2.3 --- Sephadex G-50 Spun-column Chromatography --- p.33 / Chapter 2.2.4 --- Small Scale Alkali Preparation of Plasmid DNA --- p.34 / Chapter 2.2.5 --- Large Scale Preparation of Plasmid DNA 36 - using Wizard Maxiprep Kit (Promega) / Chapter 2.2.6 --- DNA Sequencing using T7 DNA Polymerase Sequencing Kit (Pharmacia) --- p.37 / Chapter 2.2.7 --- Restriction Enzyme Digestion --- p.38 / Chapter 2.2.8 --- Agarose Gel Electrophoresis --- p.39 / Chapter 2.2.9 --- Dephosphorylation of Linearized Plasmid DNA --- p.39 / Chapter 2.2.10 --- Ligation of Foreign DNA with Linearized Plasmid --- p.40 / Chapter 2.2.11 --- Transformation of Plasmid Vector into Competent Cell (Heat Shock Method) --- p.40 / Chapter 2.2.12 --- Blotting : Transfer of DNA to Nylon Membrane --- p.41 / Chapter 2.2.12A --- Capillary Transfer of DNA to Nylon Membrane in 10X SSC --- p.41 / Chapter 2.2.12B --- Capillary Transfer of DNA to Nylon Membrane under Alkaline Condition --- p.42 / Chapter CHAPTER 3 --- SCREENING OF COMMON CARP LIVER cDNA LIBRARY / Chapter 3.1 --- Introduction --- p.44 / Chapter 3.2 --- Materials and Methods --- p.45 / Chapter 3.2.1 --- Materials --- p.45 / Chapter 3.2.2 --- Methods --- p.48 / Chapter 3.2.2.1 --- Preparation of the Plating Host --- p.48 / Chapter 3.2.2.2 --- Phage Titering --- p.48 / Chapter 3.2.2.3 --- Primary Screening of Common Carp Liver cDNA Library --- p.48 / Chapter 3.2.2.4 --- Purification of the Positive Clone --- p.49 / Chapter 3.2.2.5 --- Checking the Insert Size of the Positive Clone --- p.50 / Chapter 2.2.2.6 --- In vivo Excision to Release Phagemid from the Phage vector --- p.51 / Chapter 3.2.2.7 --- Plasmid Minipreparation of the Positive Clone --- p.52 / Chapter 3.2.2.8 --- Restriction Enzyme Digestion to Release the Insert --- p.52 / Chapter 3.2.2.9 --- Large Scale Plasmid Preparation of the Positive Clone --- p.53 / Chapter 3.2.2.10 --- DNA Sequencing of the Positive Clone --- p.53 / Chapter 3.2.2.11 --- Restriction Mapping of the Positive Clone --- p.53 / Chapter 3.2.2.12 --- Subcloning of the Positive Clone into Plasmid Vectors --- p.53 / Chapter 3.2.2.13 --- DNA Sequencing of the Subclones --- p.54 / Chapter 3.3 --- Results and Discussion --- p.55 / Chapter CHAPTER 4 --- RNA ASSAY USING REVERSE TRANSCRIPTION- POLYMERASE CHAIN REACTION / Chapter 4.1 --- Introduction --- p.70 / Chapter 4.2 --- Materials and Methods --- p.71 / Chapter 4.2.1 --- Materials --- p.71 / Chapter 4.2.2 --- Methods --- p.72 / Chapter 4.2.2.1 --- Tissue Preparation --- p.72 / Chapter 4.2.2.2 --- Total RNA Extraction --- p.72 / Chapter 4.2.2.3 --- Electrophoresis of RNA in Agarose Gel Containing Formaldehyde --- p.73 / Chapter 4.2.2.4 --- First Strand cDNA Synthesis --- p.74 / Chapter 4.2.2.5 --- IGF-I Specific PCR --- p.75 / Chapter 4.2.2.6 --- Preparation of Carp IGF Conserve Region --- p.75 / Chapter 4.2.2.7 --- Southern Hybridization of PCR Products --- p.76 / Chapter 4.3 --- Results and Discussion --- p.76 / Chapter CHAPTER 5 --- GENOMIC SOUTHERN ANALYSIS / Chapter 5.1 --- Introduction --- p.80 / Chapter 5.2 --- Materials and Methods --- p.81 / Chapter 5.2.1 --- Materials --- p.81 / Chapter 5.2.2 --- Methods --- p.82 / Chapter 5.2.2.1 --- Preparation of Genomic DNA from Carp Testis --- p.82 / Chapter 5.2.2.2 --- Restriction Enzyme Digestion of Genomic DNA --- p.82 / Chapter 5.2.2.3 --- Southern Blotting of the Digested Genomic DNA --- p.83 / Chapter 5.2.2.4 --- Preparation of the Carp IGF-I Specific Probe --- p.83 / Chapter 5.2.2.5 --- Genomic Southern Hybridization --- p.83 / Chapter 5.3 --- Results and Discussion --- p.84 / Chapter CHAPTER 6 --- THE SEARCH OF OTHER IGF cDNA SUBTYPES IN COMMON CARP / Chapter 6.1 --- Introduction --- p.88 / Chapter 6.2 --- Materials and Methods --- p.89 / Chapter 6.2.1 --- Materials --- p.89 / Chapter 6.2.2 --- Methods --- p.91 / Chapter 6.2.2.1 --- Screening using a Conserve Region cDNA Probe of Carp IGF-I --- p.91 / Chapter 6.2.2.2 --- PCR using IGF-I Specific Primers --- p.92 / Chapter 6.2.2.3 --- PCR Using T3 and T7 Primers --- p.92 / Chapter 6.2.2.4 --- Southern Blot Analysis of T3 and T7 PCR Products of cDNA Insert --- p.93 / Chapter 6.2.2.5 --- DNA Sequencing of Positive Clones --- p.94 / Chapter 6.3 --- Results and Discussion --- p.94 / Chapter CHAPTER 7 --- GENERAL DISCUSSION AND CONCLUSION --- p.99 / REFERENCES --- p.104-117