Vesicular stomatitis virus (VSV) is considered a prototype for studying non-segmented negative-stranded ribonucleic acid (RNA) viruses. Livestock are naturally infected by VSV, causing severe economic impact due to lack of any effective treatment. RNA interference (RNAi)-based therapeutics are promising alternatives to control viral infections. Lentiviral vector systems deliver artificial short hairpin RNA (shRNA) into the genome of cells to activate the RNAi pathway. In this study, an RNAi-based approach to generate cell lines with reduced susceptibility to VSV (Indiana) infection was tested. First, eight shRNAs targeting either the nucleocapsid (N), phosphoprotein (P), or the polymerase (L) viral genes were designed and introduced into cell systems. To test the potency of the shRNAs for silencing the target viral transcripts, semi-quantitative polymerase chain reaction (PCR) analysis of viral N, P, and L transcripts was performed. Then, supernatants from infected groups were evaluated by microtitration and immunoblot. Finally, the effect of VSV genomic variability in the target region of shRNAs was predicted by partial sequencing field and laboratory-adapted strains. Viral transcripts were significantly reduced in cells stably expressing shRNAs targeting the N viral gene (nucleotides 67-97 or 1312-1332; p<0.05) or P gene (nucleotides 1772-1792; p&lt;0.05). Reduction in viral transcripts was not observed by other VSV-shRNAs tested. Reduction of viral transcripts by the N-shRNA (sh-1312) was accompanied by a decrease in viral protein. Also, a reduction in the viral particles shed from cells expressing N-shRNAs (nucleotides 67-97, p&lt;0.05) was noted. The results also showed complementarity of target gene sequences for shRNAs in the sequence from the laboratory-adapted strain and single base substitutions in the corresponding regions from VSV field isolates. However, these mismatches did not occur within the seed region of the shRNAs. In conclusion, partial silencing of viral transcripts by a single shRNA does not block VSIV replication; however, partial impairment of VSIV replication was observed in N-shRNAs expressing cells. During infection, the naturally high level of N gene transcription may have modulated the sh-RNA effect. The combination of the most potent shRNAs identified here into a multiple shRNA vector may result in further reduction of viral replication. These data contribute to ongoing development of effective RNAi-based technologies to combat viral diseases.

Thesis (Ph. D.)--University of Sydney, 1997. / Includes tables. Title from title screen (viewed Apr. 27, 2009) Submitted in fulfilment of the requirements for the degree of Doctor of Philosophy to the Discipline of Removable Prosthodontics, Faculty of Dentistry. Degree awarded 1997; thesis submitted 1996. Includes bibliography. Also available in print form.

Thesis (Ph.D.)--University of Tennessee Health Science Center, 2008. / Title from title page screen (viewed on September 9, 2008). Research advisor: Michael A. Whitt, Ph.D. Document formatted into pages (xi, 119 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 104-119).

Thesis (M.S.)--Northern Michigan University, 2008. / Includes bibliographical references (leaves 75-81).

Thesis (M.S.)--University of Wisconsin--Madison, 1970. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1953. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 127-142).

Made available in DSpace on 2014-06-11T19:35:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-05-26Bitstream added on 2014-06-13T19:44:49Z : No. of bitstreams: 1 sanita_pv_dr_arafo.pdf: 1953204 bytes, checksum: a2f3ce08a1a45c4435513d02ab367cba (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Este estudo in vivo teve como objetivos avaliar: 1. a prevalência das diferentes espécies de Candida em pacientes diabéticos portadores de estomatite protética e compará-la àquela de pacientes não diabéticos com/sem estomatite protética; 2. a eficiência de dois tratamentos para estomatite protética em pacientes diabéticos: terapia antifúngica com nistatina e desinfecção de próteses totais por micro-ondas. Para isso, 210 pacientes usuários de prótese totais foram selecionados e distribuídos em três grupos de estudo. O grupo controle - GC foi formado por 90 pacientes não diabéticos e com mucosa oral saudável; os grupos experimentais foram formados por portadores de estomatite protética, sendo 80 pacientes não diabéticos - EPND e 40 diabéticos do tipo 2 controlados - EPD. Uma avaliação intra-oral dos pacientes dos grupos experimentais foi realizada e a aparência clínica da mucosa inflamada foi classificada de acordo com os critérios de Newton em graus I, II ou III. Coletas microbiológicas das próteses totais foram realizadas com swab estéril e plaqueadas em meio CHROMagar Candida. Em seguida, os procedimentos de identificação foram realizados: análise de microcultivo, teste de triagem fenotípica em caldo hipertônico e testes bioquímicos (ID-32C). Os resultados obtidos foram analisados por Intervalo de Confiança de Bonferroni e testes Exato de Fisher e Qui Quadrado (α=0,05). Os 40 pacientes diabéticos foram, então, submetidos ao tratamento da estomatite protética. Para isso, foram aleatoriamente distribuídos em 2 grupos (n=20): grupo NIS – tratamento com antifúngico tópico nistatina (suspensão oral - 100.000UI/mL), 4 vezes ao dia, por 14 dias; grupo MIC – tratamento por meio da irradiação das totais por micro-ondas (650W / 3 minutos), 3 vezes por semana, por 14 dias. Dois parâmetros... / The aim of this in vivo study was to evaluate: 1. the prevalence of Candida spp. in well-controlled type 2 diabetic patients and compare it to that found in non-diabetics with/without denture stomatitis; 2. the efficacy of two treatments of denture stomatitis in diabetic patients: topical nystatin and denture microwave disinfection. Two hundred and ten denture wearer patients were divided into three groups of study. The control group - CG was formed by 90 individuals without diabetes or denture stomatitis and the experimental groups were formed by patients with denture stomatitis, being 80 non-diabetics - DSND and 40 well-controlled type 2 diabetics - DSD. Mucosal characteristics of denture stomatitis patients were classified in types I, II, and III, according to Newton’s criteria. Mycological samples were taken from the dentures and cultured in CHROMagar′s plates. Candida spp. were identified by micro-cultivation, hypertonic Sabouraud broth, and bioMérieux ID32C assays. Results were analyzed by means of Bonferroni corrected confidence interval, Fisher’s exact test, and Chi-square analysis of several proportions (α=0.05). The 40 diabetic patients were divided into two groups (n=20) and submitted to two treatments for denture stomatitis: NYS group – patients were treated with topical nystatin, 4 times a day, for 14 days; MW group – patients had their dentures microwaved (650W / 3 minutes), 3 times per week, for 14 days. Mycological samples were taken from the palates and the tissue surface of the dentures for quantification and identification of Candida spp. and standardized photographs of the palates were taken for the clinical analysis. Microbiological and clinical evaluations were repeated at baseline... (Complete abstract click electronic access below)

<p>The long-range aim of this thesis was to further the understanding of the mechanisms involved in virus interference. To this end, biochemical and immunological procedures were used to characterize and define the virus-specific proteins of VSV. Biological studies of homotypic and heterotypic ·interference were also carried out. </p> / Thesis / Doctor of Philosophy (PhD)

Thesis (Ph. D.)--Medical University of South Carolina, 1998. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M. Sc.)--Université Laval, 1999. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.