Herpes simplex virus type 1 (HSV-1) is a common human pathogen that establishes life-long latent infection in sensory neurones. This makes it potentially useful as a gene therapy vector to target neuronal cells. HSV-1 enters cells by membrane fusion, the viral envelope and most tegument proteins dissociate, and the capsid is transported to the cell nucleus to establish infection. There is increasing evidence that the retrograde transport of HSV-1 along sensory axons is mediated by cytoplasmic dynein, but the viral and cellular proteins involved are not known. Cytoplasmic dynein is the major molecular motor involved in minus-end-directed cellular transport along microtubules. It is a large complex molecule, with heavy chains providing motility, while intermediate and light chains are involved in specific cargo binding. A library of HSV-1 capsid and tegument structural genes was constructed and tested for interaction with dynein subunits in a yeast two-hybrid system. A strong interaction was demonstrated between the HSV-1 outer capsid protein VP26 (UL35), as well as the tegument protein VP11/12 (UL46), with the homologous 14 kDa dynein light chains rp3 and Tctex1. In vitro pull-down assays confirmed binding of VP26 to rp3, Tctex1 and cytoplasmic dynein complexes. Recombinant HSV-1 capsids +/- VP26 were used in similar pull-down assays. Only VP26+ capsids bound to rp3. Recombinant HSV-1 capsids were microinjected into living cells and incubated at 37�C. After 1 h capsids were observed to co-localise with rp3, Tctex1 and microtubules. After 2 or 4 h VP26+ capsids had moved closer to the cell nucleus, while VP26- capsids remained in a random distribution. Our results suggest that the HSV-1 outer capsid protein VP26 mediates binding of incoming capsids to the retrograde motor cytoplasmic dynein during cellular infection, through interactions with dynein light chains. It is hoped that these findings will help in the development of a synthetic viral vector, which may allow targeted gene therapy in patients with neurological diseases.

Thesis (Ph. D.)--University of Sydney, 2005. / Title from title screen (viewed 20 May 2008). Submitted in fulfilment of the requirements for the degree of Doctor of Philosophy to the Faculty of Medicine. Degree awarded 2005; thesis originally submitted 2004, corrected version submitted April 2005. Includes bibliographical references. Also available in print form.

Thesis (Ph.D.)--Kent State University, 2006. / Title from PDF t.p. (viewed Mar. 11, 2009). Advisor: John J. Docherty. Keywords: herpes simplex, virus, resveratrol, NF-kappaB, NSAID Includes bibliographical references (p. 100-105).

Thesis (Ph. D.)--Duke University, 2008. / Includes bibliographical references.

Thesis (D.P.H.)--University of Michigan.

Thesis (M.S.W.)--Smith College School for Social Work, Northampton, Mass., 2009. / Includes bibliographical references (p. 65-68).

Thesis (M.S.)--Brigham Young University. Dept. of Microbiology and Molecular Biology, 2005. / Includes bibliographical references.

Dissertation (D.P.H.)--University of Michigan.

Thesis (Ph.D.)--McMaster University, 2004. / Advisor: John P. Capone. Includes bibliographical references (p. 179-[208]). Available also online.

Ocorrências comuns de infecções virais graves, aparecimento de cepas resistentes e um limitado número de quimioterápicos antivirais disponíveis mostram a necessidade da busca por novas substâncias ativas como antivirais. Muitas substâncias derivadas de plantas são candidatas ao estudo do seu potencial na terapia sistêmica e/ou profilaxia de herpes simplex virus 1 (HSV-1). Dentre as plantas com atividade antiviral, destacam-se as do gênero Baccharis. O objetivo geral do trabalho foi o estudo da composição química e avaliação da atividade antiviral das folhas de Baccharis anomala D.C. através de fracionamento bioguiado. Utilizando precipitação com etanol e fracionamento por permeação molecular foi possível separar os constituintes químicos ativos contra o vírus HSV-1 presentes no extrato aquoso de Baccharis anomala. Os testes de prospecção de constituintes químicos indicaram a presença de taninos, catequinas e saponinas no extrato aquoso da espécie. Através da análise cromatográfica foi possível detectar a presença de compostos fenólicos, utilizando-se coloração com cloreto férrico e reagente natural. Em relação à atividade antiviral, a fração ativa denominada AQ PPT FR 4-5 apresentou pronunciada atividade antiviral, inibindo a replicação viral em 100 % nas concentrações de 1,25, 0,625, 0,312 e 0,156 mg/mL, contra as cepas ATCC-VR733 e Aciclovir-resistente 29-R do HSV-1. Em relação ao mecanismo de ação, observou-se atividade virucida da fração AQ PPT FR 4-5. Estes resultados são muito importantes, pois, de acordo com a literatura, ainda não foram relatados compostos com atividade virucida úteis clinicamente para o tratamento de infecções por HSV-1. Conclui-se, portanto, que Baccharis anomala possui potente atividade antiviral contra o vírus HSV-1 e é promissora para estudos posteriores que visem ao isolamento, identificação e estudo do mecanismo de ação antiviral de compostos ativos da espécie, considerando a emergência de cepas resistentes e a necessidade de compostos com novos mecanismos de ação. / Common occurrences of severe viral infections, emergence of resistant strains and a limited number of available antiviral chemotherapeutics show the need to search for new active substances as antiviral. Many compounds derived from plants are candidates for the study of their potential in systemic therapy and/or prophylaxis of herpes simplex virus type 1 (HSV-1). Among the plants with antiviral activity, those from the genus Baccharis are remarkable. The main objective of the work was the study of the chemical composition and evaluation of the antiviral activity of extracts from Baccharis anomala D.C., using bioactivity guided fractionation. Through precipitation with ethanol and fractionation by molecular permeation it was achieved the separation of the active chemical constituents against HSV-1 virus in the aqueous extract of Baccharis anomala. Phytochemical tests indicated the presence of tannins, catechins and saponins in the aqueous extract. By thin layer chromatography it was detected the presence of phenolic compounds using ferric chloride and natural reagent. Concerning the antiviral activity, the active fraction named AQ PPT FR 4-5 showed pronounced antiviral activity, represented by 100 % inhibition of viral replication at concentrations of 1.25, 0.625, 0.312 and 0.156 mg/mL, against the strains ATCC-VR733 and Acyclovir-resistant 29-R of HSV-1 virus. Regarding the mechanism of action, virucidal activity on the fraction AQ PPT FR 4-5 was detected, which is also very important because, as far as we know, compounds with virucidal activity for clinical use in the treatment of HSV-1 infections were not reported yet. In conclusion, Baccharis anomala displayed pronounced antiviral activity against HSV-1 virus and it is promising for further studies aimed to the isolation, identification and mechanism of action of antiviral active compounds of the species, considering the emergence of resistant strains and the need for compounds with new mechanisms of action.