Explore the vast range of titles available.

Japanese encephalitis (JE) virus (JEV) is a mosquito-borne flavivirus found across Asia that is closely related to West Nile virus. There is no known antiviral treatment for any flavivirus. Results from in vitro studies and animal models suggest intravenous immunoglobulin (IVIG) containing virus-specific neutralizing antibody may be effective in improving outcome in viral encephalitis. IVIG's anti-inflammatory properties may also be beneficial. We performed a pilot feasibility randomized double-blind placebo-controlled trial of IVIG containing anti-JEV neutralizing antibody (ImmunoRel, 400mg/kg/day for 5 days) in children with suspected JE at two sites in Nepal; we also examined the effect on serum neutralizing antibody titre and cytokine profiles. 22 children were recruited, 13 of whom had confirmed JE; 11 received IVIG and 11 placebo, with no protocol violations. One child (IVIG group) died during treatment and two (placebo) subsequently following hospital discharge. Overall, there was no difference in outcome between treatment groups at discharge or follow up. Passive transfer of anti-JEV antibody was seen in JEV negative children. JEV positive children treated with IVIG had JEV-specific neutralizing antibody titres approximately 16 times higher than those treated with placebo (p=0.2), which was more than could be explained by passive transfer alone. IL-4 and IL-6 were higher in the IVIG group. A trial of IVIG for JE in Nepal is feasible. IVIG may augment the development of neutralizing antibodies in JEV positive patients. IVIG appears an appealing option for JE treatment that warrants further study. ClinicalTrials.gov NCT01856205.

The double subgenomic Sindbis (dsSIN) virus expression system was used to deliver RNA transcripts complementary to endogenous mosquito mRNA to inhibit gene expression in vivo. dsSIN has been used to stably express genes in Aedes aegypti salivary glands and to inhibit flavivirus replication by expression of RNA transcripts complementary to the flavivirus genome. The dsSIN viruses TE/3′2J and MRE/3′ 2J were assessed for their ability to inhibit the expression of endogenous mosquito genes expressed exclusively in the salivary glands. Experiments were conducted to inhibit expression of a reporter gene in a germline transformed Ae. aegypti line (APY-LUC43). These mosquitoes constituitively express luciferase from the mosquito apyrase promoter. APY-LUC43 mosquitoes were intrathoracically inoculated with TE/3′ 2J/α-luc virus, which transcribed RNA complementary to the 5 ′ end of the luciferase mRNA. Luciferase activity was monitored over time in mosquitoes infected with either TE/3′2J/α-luc or control dsSIN viruses expressing unrelated antisense RNAs. Mosquitoes infected with TE/3′2J/α-luc virus exhibited 90% reduction in luciferase activity compared to uninfected and control dsSIN-infected mosquitoes at 5 and 9 days postinoculation. Apyrase is a salivary gland protein with anti-platelet aggregating properties. TE/3′2J/α-apyrase virus was engineered to contain 375 bases from the 5′ coding region of the Ae. aegypti apyrase gene, inserted in antisense orientation. TE/3 ′2J/α-apyrase was intrathoracically inoculated into female adult mosquitoes, and apyrase activity and protein production was assessed. TE/3′2J/α-apyrase virus effectively infected salivary gland tissue; however, no diminishment of apyrase production was observed. An orally infective dsSIN chimeric virus was engineered that contained the structural genes of URE16 and the nonstructural genes and second subgenomic promoter of TE/3′2J/α-apyrase. MRE/3′ 2J/α-apyrase was tested for its ability to infect larvae per os and inhibit expression of apyrase in adults. Infection rates of up to 42% were obtained, but inhibition of apyrase expression was not observed. These studies support the overall hypothesis of this dissertation, that expression of an endogenous mosquito gene can be specifically inhibited by RNA delivered by the SIN virus expression system. The dsSIN antisense RNA expression system provides an important tool for studying gene expression in vivo, and may lead to defining genetic determinants of mosquito vector competence.

Neurodegenerative disorders with high brain iron include Parkinson disease, Alzheimer disease and several childhood genetic disorders categorized as neuroaxonal dystrophies. We mapped a locus for infantile neuroaxonal dystrophy (INAD) and neurodegeneration with brain iron accumulation (NBIA) to chromosome 22q12-q13 and identified mutations in PLA2G6, encoding a calcium-independent group VI phospholipase A sub(2), in NBIA, INAD and the related Karak syndrome. This discovery implicates phospholipases in the pathogenesis of neurodegenerative disorders with iron dyshomeostasis.

Neurodegenerative disorders with high brain iron include Parkinson disease, Alzheimer disease and several childhood genetic disorders categorized as neuroaxonal dystrophies. We mapped a locus for infantile neuroaxonal dystrophy (INAD) and neurodegeneration with brain iron accumulation (NBIA) to chromosome 22q12-q13 and identified mutations in PLA2G6 , encoding a calcium-independent group VI phospholipase A 2 , in NBIA, INAD and the related Karak syndrome. This discovery implicates phospholipases in the pathogenesis of neurodegenerative disorders with iron dyshomeostasis.