Expression and analysis of a non-glycosylated varicella-zoster virus subunit vaccine

Includes bibliographical references (pages [42]-46)Varicella-zoster virus (VZV) encodes at least five glycoproteins designated gpl, gpn, gpm, gpIV, and gpV. gpl is encoded by gene 68 and is the most abundant and immunogenic of the VZV glycoproteins. A VZV subunit vaccine, expressing a truncated VZV gpl gene encoding 511 amino acids (TgpI-511) has been developed. Further investigation was carried out to express and analyze a nonglycosylated form of the VZV subunit vaccine. A prokaryotic expression system was utilized to subclone the truncated gene 68, encoding the non-glycosylated subunit vaccine (nTgpI), into the pFUS vector creating the recombinant vector, pFUSl. Protein analysis showed the expression of a fusion protein in Escherichia coli with a size of 70 kDa containing nTgpI (58 kDa) and thioredoxin (th) of 12 kDa. Western blot analysis demonstrated: (1) the reactivity of nTgpI-th with monoclonal antibodies specific for native VZV gpl; and (2) the biological activity of nTgpI-th through its recognition by VZV polyclonal antibodies from a zoster patient. In addition, rabbit immunization of nTgpI-th elicited the production of antibodies which recognized native VZV glycoproteins. These results indicated that the denatured nTgpI-th produced a humoral response in a rabbit which was reactive to native VZV gpl. However, these studies revealed that this prokaryotic expression system was not suitable to produce large quantities of the nTgpI-th vaccine possibly due to the lack of glycosylation of nTgpI-th in E. coli.M.S. (Master of Science

Continuous epidural ropivacaine 0.2% for analgesia after lower abdominal surgery

The purpose of this study was to determine whether a lumbar epidural infusion of ropivacaine 0.2% would provide effective analgesia with an acceptably low incidence of motor blockade and side effects after lower abdominal surgery. After combined general and epidural anesthesia and surgery, 125 patients were randomly assigned to receive either saline or ropivacaine 0.2% at a rate 6, 8, 10, 12, or 14 mL/h (Groups R6, R8, R10, R12, and R14, respectively) for 21 h. Supplemental analgesia, if required, was provided with intravenous patient-controlled analgesia with morphine. Data were collected at 4, 8, and 21 h, and included morphine consumption, pain scores at rest and with coughing, motor and sensory block, and adverse events. Cumulative morphine consumption was less in Groups R10, R12, and R14 compared with the saline group. At 4 h analgesia was better among patients receiving ropivacaine, but at 21 h pain scores were identical. Sensory blockade at 8 and 21 h was greater in the ropivacaine groups compared with the saline group. Approximately 30% of R8, R10, and R12 patients, and 63% of R14 patients had demonstrable motor block of the lower limbs at 21 hours. We conclude that lumbar epidural ropivacaine 0.2% reduces parenteral morphine requirements but has little effect on pain scores and may be associated with motor blockade