Thesis (Ph. D.)--University of Chicago, Pritzker School of Medicine, Committee on Genetics, March 2003. / Includes bibliographical references. Also available on the Internet.

Thesis (M.D.)--University of Hong Kong, 1992. / Includes bibliographical references (leaves 251-295) Also available in print.

Thesis (M. Phil.)--University of Hong Kong, 2005. / Title proper from title frame. Also available in printed format.

Thesis (Ph. D.)--University of Texas at Austin, 2007. / Vita. Includes bibliographical references.

Thesis (Ph. D.)--Ohio State University, 2007. / Title from first page of PDF file. Includes bibliographical references (p. 148-166).

Mammalian cell culture is typically operated at the physiological temperature of 37°C. Low temperature cell culture at 30-33°C, in particular for CHO cells, increased the specific productivity of many recombinant proteins amongst many other benefits. However, the cell density is lower, thus reducing the total protein yield. Of the 17 mammalian cold-stress genes reported to be up- or down-regulated at low temperature, CIRP shows potential as a gene target for improving recombinant protein production, as its expression levels were reported to affect both growth and specific productivity. In this study, it was shown that over-expression of the cold-stress gene CIRP did not cause growth arrest in CHO cells, in contrast to previous reports. However, over-expression of CIRP successfully improved the specific productivity and total yield of a recombinant interferon-γ CHO cell-line at 37°C by 25%. / Singapore-MIT Alliance (SMA)

abstract: Host organisms have evolved multiple mechanisms to defend against a viral infection and likewise viruses have evolved multiple methods to subvert the host's anti-viral immune response. Vaccinia virus (VACV) is known to contain numerous proteins involved in blocking the cellular anti-viral immune response. The VACV E3L protein is important for inhibiting the anti-viral immune response and deletions within this gene lead to a severe attenuation. In particular, VACV containing N-terminal truncations in E3L are attenuated in animal models and fail to replicate in murine JC cells. Monkeypox virus (MPXV) F3L protein is a homologue of the VACV E3L protein, however it is predicted to contain a 37 amino acid N-terminal truncation. Despite containing an N-terminal truncation in the E3L homologue, MPXV is able to inhibit the anti-viral immune response similar to wild-type VACV and able to replicate in JC cells. This suggests that MPXV has evolved another mechanism(s) to counteract host defenses and promote replication in JC cells. MPXV produces less dsRNA than VACV during the course of an infection, which may explain why MPXV posses a phenotype similar to VACV, despite containing a truncated E3L homologue. The development of oncolytic viruses as a therapy for cancer has gained interest in recent years. Oncolytic viruses selectively replicate in and destroy cancerous cells and leave normal cells unharmed. Many tumors possess dysregulated anti-viral signaling pathways, since these pathways can also regulate cell growth. Creating a mutation in the N-terminus of the VACV-E3L protein generates an oncolytic VACV that depends on dysregulated anti-viral signaling pathways for replication allowing for direct targeting of the cancerous cells. VACV-E3Ldel54N selectively replicates in numerous cancer cells lines and not in the normal cell lines. Additionally, VACV-E3Ldel54N is safe and effective in causing tumor regression in a xenograph mouse model. Lastly, VACV-E3Ldel54N was capable of spreading from the treated tumors to the untreated tumors in both a xenograph and syngeneic mouse model. These data suggest that VACV-E3Ldel54N could be an effective oncolytic virus for the treatment of cancer. / Dissertation/Thesis / Ph.D. Microbiology 2010

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Submitted by Hiolanda Rêgo (hiolandar@gmail.com) on 2013-08-13T16:46:26Z No. of bitstreams: 1 Dissertação_ICS_Heidiane Alves dos Santos.pdf: 1654611 bytes, checksum: d1c1496a80a371c6d05868103f946b63 (MD5) / Made available in DSpace on 2013-08-13T16:46:26Z (GMT). No. of bitstreams: 1 Dissertação_ICS_Heidiane Alves dos Santos.pdf: 1654611 bytes, checksum: d1c1496a80a371c6d05868103f946b63 (MD5) / A linfadenite caseosa é uma doença infecto-contagiosa crônica, de ocorrência mundial que acomete pequenos ruminantes, causando grandes perdas econômicas. Esta doença tem como agente etiológico a bactéria Corynebacterium pseudotuberculosis e caracteriza-se pela formação de granulomas em gânglios linfáticos superficiais, podendo também acometer órgãos e linfonodos internos, como forma de resposta do sistema imune do hospedeiro à penetração deste agente que resiste a ação bactericida das células fagóciticas. Este patógeno se relaciona filogeneticamente ao Mycobacterium tuberculosis. A avaliação da resposta imune é uma importante ferramenta para localização de animais possivelmente infectados, dificultando desta forma a disseminação do agente. Estudos têm demonstrado a importância de citocinas na defesa de patógenos intracelulares facultativos, inclusive para C. pseudotuberculosis, destacando-se o interferon-gama (IFN-) citocina característica de células Th1, que pode está envolvida na proteção à patógenos intracelulares. O presente estudo avaliou aspectos da resposta imunológica de camundongos da linhagem C57Black/6 selvagem e knockout para IFN-gama, durante a infecção por Corynebacterium pseudotuberculosis. Os grupos foram acompanhados ao longo de 7 e 14 dias após infecção intraperitoneal com 107 UFC, durante os quais foram avaliados a disseminação bacteriana, a frequência de granulomas, a variação de peso do baço, o padrão de migração celular para a cavidade peritoneal, além da resposta imune humoral através da dosagem de IgG e subclasses e resposta imune celular através da dosagem de citocinas e imunofenotipagem. Os camundongos knockout para IFN-gama se mostraram mais susceptíveis à infecção por Corynebacterium pseudotuberculosis apresentando maior disseminação bacteriana nos linfonodos mesentéricos, maior frequência de granulomas, aumento considerável no peso do baço, intensa migração celular para a cavidade peritoneal, principalmente neutrófilos e macrófagos e expressão de linfócitos T CD8+ nas células esplênicas, e consequentemente menor expressão de citocinas pró-inflamatórias e regulatórias. Estes achados apontam para a importância do IFN-γ na resposta imune a patógenos intracelulares a exemplo de C. pseudotuberculosis, ao tempo em que demonstra a resposta deficiente do sistema imune dos animais nocauteados à infecção. / Salvador

SOUSA, J. C. Fatores Reguladores de Interferon (IRFs) em pacientes com Síndrome Mielodisplásica. 2015. 200 f. Tese (Doutorado em Ciências Médicas) - Faculdade de Medicina, Universidade Federal do Ceara, Fortaleza, 2015. / Submitted by Erika Fernandes (erikaleitefernandes@gmail.com) on 2016-09-26T12:46:52Z No. of bitstreams: 1 2015_tese_jcsousa.pdf: 1726859 bytes, checksum: b83c00c082a73dcf53401b31ef53acda (MD5) / Approved for entry into archive by Erika Fernandes (erikaleitefernandes@gmail.com) on 2016-09-26T12:47:01Z (GMT) No. of bitstreams: 1 2015_tese_jcsousa.pdf: 1726859 bytes, checksum: b83c00c082a73dcf53401b31ef53acda (MD5) / Made available in DSpace on 2016-09-26T12:47:01Z (GMT). No. of bitstreams: 1 2015_tese_jcsousa.pdf: 1726859 bytes, checksum: b83c00c082a73dcf53401b31ef53acda (MD5) Previous issue date: 2015-07-20 / Myelodysplastic Syndrome (MDS) is characterized by peripheral cytopenias, defects in hematopoiesis, increased apoptosis and intramedullary risk of transformation to AML. Various advances have been made in understanding the pathogenesis of MDS and there is evidence that bone marrow failure that occurs in MDS is mediated by abnormal activation of signaling in the innate immune system. Interferon regulatory factors (IRF-Interferon Regulatory Factor) form a family of transcription factors that play a central role in regulating immune responses, differentiation and proliferation of hematopoietic cells, cell cycle regulation, apoptosis, and oncogenesis. It is believed that IRFs may have an important role in the pathogenesis of MDS. Therefore, the objective of this study was to evaluate the profile of methylation and gene expression of IRFs in bone marrow (BM) of patients with MDS. From samples of 119 patients diagnosed with MDS was conducted the study of gene expression by real-time PCR and methylation by QMSP (quantitative methilation specific PCR) of the nine family members of IRFs. The expression of genes IRF2, IRF3, and IRF7 IRF8 were different between BM cells of MDS patients and healthy individuals (P = 0.002, 0.002, 0.028 and 0.016, respectively). IRF1 a higher level of expression was associated in patients with hypocellular marrow (p = 0.018). The IRF3, and IRF7 IRF8 genes have been associated with patients in peripheral blood cytopenias (p = 0.028; 0.001; 0.008, respectively). Furthermore, methylation of IRF1, IRF2, IRF3, IRF6 and IRF8 was associated with higher risk characteristics in SMD, such as advanced forms, unfavorable karyotype, cytopenias, blast above 5% and high-risk categories established by IPSS, IPSS-R and WPSS. Multivariate analysis revealed that patients with higher expression of IRF3 had higher overall survival (p = 0.001), whereas patients with higher expression of IRF5 had 5.4 more likely to progress to AML (p <0.001 95% CI = 1098-26829 ) and 9 times more likely to come to death (p = 0.001, 95% CI 2.39 to 32.69). Ambiguously, patients with methylated IRF5 had 4 times more likely to come to death (p = 0.041, 95% CI 1.066 to 20,192). We can conclude that the expression and methylation of IRFs can have great impact prognosis in this disease. The expression of IRF3 is a favorable prognostic marker, while expression and methylation IRF5 are unfavorable prognostic markers in MDS. / A síndrome mielodisplásica (SMD) é caracterizada por citopenias periféricas, defeitos na hematopoese, aumento da apoptose intramedular e risco de transformação para LMA. Vários avanços têm sido realizados para o entendimento da patogênese da SMD e há evidências de que a falha na medula óssea que ocorre na SMD seja mediada pela ativação anormal da sinalização do sistema imune inato. Os fatores reguladores de interferon (IRF-Interferon Regulatory Factor) formam uma família de fatores de transcrição que possuem um papel central na regulação de respostas imunes, diferenciação e proliferação de células hematopoéticas, regulação do ciclo celular, apoptose e oncogênese. Acredita-se que os IRFs possam ter um papel importante da patogênese da SMD. Portanto, o objetivo do presente trabalho foi avaliar o perfil de metilação e expressão dos genes dos IRFs em células da medula óssea (MO) de pacientes com SMD. A partir de amostras de 119 pacientes com diagnóstico de SMD foi realizado o estudo da expressão gênica, por PCR em tempo real e da metilação por QMSP (quantitative methilation specific PCR) dos nove membros da família dos IRFs. A expressão dos genes IRF2, IRF3, IRF7 e IRF8 foram diferentes entre células de MO de pacientes com SMD e indivíduos saudáveis (p=0,002; 0,002; 0,028 e 0,016, respectivamente). Um maior nível de expressão do IRF1 foi associado a pacientes com medula hipocelular (p=0,018). Os genes IRF3, IRF7 e IRF8 foram associados a pacientes com citopenias no sangue periférico (p= 0,028; 0,001; 0,008, respectivamente). Por outro lado, a metilação dos IRF1, IRF2, IRF3, IRF6 e IRF8 foi associada a características de maior risco em SMD, tais como, formas avançadas, cariótipo desfavorável, citopenias, blastos acima de 5% e com categorias de alto risco estabelecidas pelo IPSS, R-IPSS e WPSS. A análise multivariada revelou que pacientes com maior expressão do IRF3 apresentaram maior sobrevida global (p=0,001), enquanto os pacientes com uma maior expressão do IRF5 apresentaram 5,4 mais chances de evoluir para LMA (p<0,001 IC95%=1.098-26.829) e 9 vezes mais chance de vir a óbito (p=0,001; IC95%=2,39-32,69). Ambiguamente, pacientes com o IRF5 metilado possuíam 4 vezes mais chance de vir a óbito (p=0,041; IC95%=1,066-20.192). Podemos concluir que a expressão e a metilação dos IRFs podem ter grande impacto prognóstico nessa doença. A expressão do IRF3 é um marcador de prognóstico favorável, enquanto a expressão e a metilação do IRF5 são marcadores de prognóstico desfavorável em SMD.