ABSTRACT
E2, along with E(rns) and E1, is an envelope glycoprotein of Classical Swine Fever Virus (CSFV). E2 is involved in several virus functions: cell attachment, host range susceptibility and virulence in natural hosts. Here we evaluate the role of a specific E2 region, (818)CPIGWTGVIEC(828), containing a putative fusion peptide (FP) sequence. Reverse genetics utilizing a full-length infectious clone of the highly virulent CSFV strain Brescia (BICv) was used to evaluate how individual amino acid substitutions within this region of E2 may affect replication of BICv. A synthetic peptide representing the complete E2 FP amino acid sequence adopted a ß-type extended conformation in membrane mimetics, penetrated into model membranes, and perturbed lipid bilayer integrity in vitro. Similar peptides harboring amino acid substitutions adopted comparable conformations but exhibited different membrane activities. Therefore, a preliminary characterization of the putative FP (818)CPIGWTGVIEC(828) indicates a membrane fusion activity and a critical role in virus replication.
Subject(s)
Classical Swine Fever Virus/physiology , Viral Envelope Proteins/metabolism , Virus Internalization , Amino Acid Substitution , Animals , Cell Line , Cell Membrane/metabolism , Classical Swine Fever Virus/genetics , Liposomes/metabolism , Models, Molecular , Mutant Proteins/chemistry , Mutant Proteins/genetics , Mutant Proteins/metabolism , Protein Conformation , Reverse Genetics , Swine , Viral Envelope Proteins/chemistry , Viral Envelope Proteins/geneticsABSTRACT
Health care reform is also occurring in Australia and effects hospital chaplaincy programs. "Economic rationalism" is the philosophic foundation of this effort and its contrast with the values inherit in hospital chaplaincy are highlighted. Selected research results from the Australian system are described and the authors offer a perspective on the cost efficiency of hospital chaplaincy.
