Table of Contents
PART I ... Error! Bookmark not defined. GENERAL INTRODUCTION ... Error! Bookmark not defined. 1.1. The Onchocerciasis Problem ... Error! Bookmark not defined. 1.2. The Life Cycle of O. volvulus and C. elegans ... Error! Bookmark not defined. 1.3. Classification/Phylogeny of Nematodes ... Error! Bookmark not defined. 1.4. Pathogenesis of selected nematode infections ... Error! Bookmark not defined. 1.5. Trans-splicing in Nematodes ... Error! Bookmark not defined. 1.6. Nematode Body Plan ... Error! Bookmark not defined. 1.7. Clinical Manifestations of Onchocerciasis ... Error! Bookmark not defined. 1.8. Diagnosis of Onchocerciasis ... Error! Bookmark not defined. 1.9. Characterization of Onchocerca Antigens ... Error! Bookmark not defined. 1.10. Omics Research in Onchocerca ... Error! Bookmark not defined. 1.11. Helminths Excretory Secretory Products and Interaction with the Immune System ... Error! Bookmark not defined.
1.12. Onchocerca models of infections ... Error! Bookmark not defined. 1.13. Gangliosides ... Error! Bookmark not defined. 1.14. GM2 Activator Proteins ... Error! Bookmark not defined. 1.14. Rationale of The Study ... Error! Bookmark not defined. 2.0. Manuscript 1... Error! Bookmark not defined. 2.1. Objectives of the Study: ... Error! Bookmark not defined. 2.2. Specific objectives of the study were: ... Error! Bookmark not defined. 2.3. Main Results ... Error! Bookmark not defined. 2.4. Conclusions ... Error! Bookmark not defined. 3.0. Manuscript 2... Error! Bookmark not defined. 3.1. Overall Objective: ... Error! Bookmark not defined. 3.2. Specific Objectives: ... Error! Bookmark not defined. 3.3. Main Results ... Error! Bookmark not defined. 3.4. Conclusions ... Error! Bookmark not defined. PART II ... Error! Bookmark not defined. IDENTIFICATION AND CHARACTERIZATION OF THE ONCHOCERCA VOLVULUS
EXCRETORY SECRETORY PRODUCT OV28CRP, A PUTATIVE GM2 ACTIVATOR
Abstract ... Error! Bookmark not defined. Author Summary ... Error! Bookmark not defined. Introduction ... Error! Bookmark not defined. Materials and Methods ... Error! Bookmark not defined. Ethics Statement... Error! Bookmark not defined.
Onchocerca Parasite Material ... Error! Bookmark not defined. Serum Samples ... Error! Bookmark not defined. Sequence Analysis and phylogenetic Analysis ... Error! Bookmark not defined. Cloning, Expression and Purification of OvGM2AP ... Error! Bookmark not defined. Enzymatic deglycosylation of OvGM2AP and Mass spectrometry ... Error! Bookmark not defined. Peptide synthesis and generation of Polyclonal antibodies (GenScript, USA) .. Error! Bookmark not defined.
Stage specific expression of OvGM2AP ... Error! Bookmark not defined. Serological Characterization of OvGM2AP ... Error! Bookmark not defined. GM2AP activity assay ... Error! Bookmark not defined. MUG and MUGS competitive inhibition assay ... Error! Bookmark not defined. Statistical Analyses ... Error! Bookmark not defined. Accession Numbers ... Error! Bookmark not defined. Results ... Error! Bookmark not defined.
Structural features of OvGM2AP, a putative Onchocerca volvulus GM2AP Activator protein ... Error! Bookmark not defined. Transcriptional and translational expression patterns of OvGM2AP gene ... Error! Bookmark not defined.
Mass expression and purification of recombinant OvGM2AP from insect cells .... Error! Bookmark not defined.
The asparagine at position 173 of the recombinant OvGM2AP is glycosylated Error! Bookmark not defined.
The humoral immune response to OvGM2AP divulges its immunogenicity .... Error! Bookmark not defined.
Acknowledgements ... Error! Bookmark not defined. Supporting Information ... Error! Bookmark not defined. PART III ... Error! Bookmark not defined. MOLECULAR AND FUNCTIONAL CHARACTERIZATION OF A PHOSPHOLIPID BINDING NEMATODE GM2 ACTIVATOR PROTEIN ... Error! Bookmark not defined. INTRODUCTION ... Error! Bookmark not defined. RESULTS ... Error! Bookmark not defined. Spatiotemporal expression of CPP-1 in C. elegans reporter transgenics ... Error! Bookmark not defined.
Hoechst 33342 Cuticle permeability assessment ... Error! Bookmark not defined. Rescue experiments ... Error! Bookmark not defined. Bioinformatics analysis of OvGM2AP and CPP-1 sequence ... Error! Bookmark not defined. Electron Microscopy Analysis... Error! Bookmark not defined. Production of OvGM2AP and CPP-1 in Sf21 insect cells ... Error! Bookmark not defined. Characterization of post translational modification of OvGM2AP and CPP-1 Error! Bookmark not defined.
Gel filtration analysis of OvGM2AP and CPP-1 ... Error! Bookmark not defined. Analysis of Lipid binding properties of recombinant OvGM2AP and CPP-1 .. Error! Bookmark not defined.
Localization of disulphide bridges in OvGM2AP and CPP-1 ... Error! Bookmark not defined. DISCUSSION ... Error! Bookmark not defined. Lipid binding ability: A role in worm survival? ... Error! Bookmark not defined. Localization of the GM2AP homologues in the worm ... Error! Bookmark not defined. Pathophysiological function of OvGM2AP ... Error! Bookmark not defined. MATERIALS AND METHODS ... Error! Bookmark not defined.
C. elegans Strains ... Error! Bookmark not defined.
Molecular Biology ... Error! Bookmark not defined.
Transgenics ... Error! Bookmark not defined.
Cuticle sensitivity and permeability assay ... Error! Bookmark not defined.
Hoechst staining ... Error! Bookmark not defined.
Recombinant protein production ... Error! Bookmark not defined.
