Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 3 uppsatser.

Finally, NF-kappaB and another transcriptional regulator p53 are usually tightly related in their control of cell survival. Opposite to NF-kappaB, p53 mediates cell death signals, usually activated under DNA damage and subsequently involved in cell growth control, DNA damage repair or apoptosis. It was found in this study, DNA damage and cell cycle arrest responses tended to participate with the anti-HSV-1 activity. Although in HEp-2 cells, TCS induced more DNA damage ratio and S and G2/M phase arrest proportion than HSV-1 infected cells, but more p53 was expressed and activated by phosphorylating at Ser 15 by TCS in HSV-1 infected HEp-2 cells than uninfected ones. In the same time the activation of BAX, which promotes the apoptotic function of p53, increased during TCS treatment when infected with HSV-1, the p53 therefore regulates apoptosis in HSV-1 infected cell during TCS treatment. / Firstly, we demonstrated that TCS reduced HSV-1 antigen and DNA content, The IC50 (half maximal inhibitory concentration) of TCS on HSV-1 replication was 2.5 +/- 0.23 mug/ml. The anti-HSV-1 effect of TCS was related to interfering with viral replication during 3 to 15 hours after infection which coincide with early to late viral replication period. TCS had no effect on HSV-1 attachment, penetration or immediate early gene expression. However, the expression of early gene, late gene and virion release were diminished. / Fourthly, the role of the nuclear factor-kappaB (NF-kappaB) in the anti-HSV-1 effect of TCS was explored. NF-kappaB initiates cell survival pathways. It is widely involved in viral infection and replication to make sure virus overcomes the host cells immune response. We found HSV-1 enhanced the activity of NF-kappaB in HEp-2 cells by triggering its translocation from cytoplasm to nuclear. However, during the anti-HSV-1 effect of TCS, TCS suppressed HSV-1-aroused NF-kappaB translocation in HEp-2 cells, the inhibition of NF-kappaB activity in HSV-1-infected cells by TCS treatment tend to abolish the anti-apoptosis effect developed by HSV-1, so that the host cells suffered more extracellular stress and showed more apoptosis ratio than uninfected ones. / Taken together, this study demonstrated TCS interfered with HSV-1 early to late infection period. TCS selectively induced more HSV-1 infected HEp-2 cells to apoptosis than uninfected ones, the selectivity of TCS was due to apoptotic signaling pathway switching from CD95 (Fas/Apo-1)-mediated type I to type II apoptotic pathway. Furthermore, during TCS induced-apoptosis in HSV-1 infected cells, TCS suppressed NF-kappaB activation that triggered by HSV-1 infection. At the meanwhile, p53 participated in the TCS-induced apoptosis regulation in infected cell. / TCS is toxic to cell because its RIP activity, killing of the viral host cells certainly inhibits virus expansion, only when it kills more infected cells, the material could be considered as an anti-viral agent. It was found TCS induced losing of cell viability and enhancing in apoptosis in HEp-2 cells and HSV-1 infected HEp-2 cells. The decrease of cell viability and increase of apoptosis ratio were enhanced when HEp-2 cells were infected with HSV-1 compared with uninfected ones. The 50% of effect concentration (EC50 ) in cytotoxicity and apoptosis were decreased from 24.64 +/- 1.17 mug/ml and 37.57 +/- 1.47 mug/ml in uninfected HEp-2 cells to 3.01 +/- 1.30 mug/ml and 3.89 +/- 1.31 mug/ml in HSV-1 infected HEp-2 cells respectively. / The reason of type I to type II apoptosis pathway transition might due to the activity change of death receptor on HEp-2 cells. The type I apoptotic pathway induced by TCS was related to CD95 (Fas/Apo-1) system activation and signaling pathway. When HEp-2 cells were infected with HSV-1, the CD95 (Fas/Apo-1) was suppressed by HSV-1 infection. As a result, TCS triggered a less CD95 (Fas/Apo-1) dependent type II apoptotic pathway in the infected cells. / Thirdly, TCS activated different apoptotic pathways, namely type I and type II apoptotic pathways, between uninfected and infected cells. The type I apoptotic pathway bypasses the dependence on the mitochondrial but quickly activates a large amount of caspase-8 at the CD95 (Fas/Apo-1) formed death inducing signaling complex (DISC), which amplifies the signal. By contrast, the formation of the DISC in the type II apoptotic pathway is strongly reduced. It depends on loss of the mitochondrial transmembrane potential (DeltaPsi m) and release of cytochrome c and capase-9 activation to mediate apoptosis signal transduction. We found in HSV-1 infected an uninfected HEp-2 cells, TCS induced the loss of DeltaPsim, this DeltaPsim losing was increased when HEp-2 cells were infected with HSV-1. Furthermore, when there were no HSV-1 infection, TCS induced caspase-dependent type I apoptosis pathway that quickly activated large amount of caspase-8 after TCS treatment. However, when infected with HSV-1, this pathway turned into mitochondrial dependent type II pathway involving caspase-9 response, whose apoptosis ratio was diminished by over expressed Bcl-2, which is a hallmark defining type I or type II apoptosis. / Trichosanthin (TCS) is a type I ribosome inactivating protein (RIP), it was found to inhibit human simplex virus type 1 (HSV-I) but the anti-HSV-I mechanism is unclear. HSV-1 is a widely distributed DNA virus, it causes large range of human diseases. During the lytic life cycle of HSV-1, highly regulated cascade of genes are expressed to interfere with host cell metabolism and immune response. In this context the anti-HSV-1 mechanism of TCS in human epithelial carcinoma HEp-2 cells was studied. / He, Dongxu. / Advisers: Wing Ho Yung; Siu Cheung Tam. / Source: Dissertation Abstracts International, Volume: 73-06, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 124-138). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Yau, Kwok Hei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 64-71). / Abstracts in English and Chinese. / Chapter Chapter 1: --- Introduction --- p.1 / Trichosanthin --- p.2 / Apoptosis --- p.10 / Herpes Simplex Virus --- p.16 / Conclusion --- p.28 / Chapter Chapter 2: --- Materials and Methods --- p.29 / Cell lines and virus --- p.30 / Infectivity assay --- p.30 / Treatment of cells and virus infection --- p.32 / MTT assay for cytotoxicity --- p.34 / Preparation of cell lysate --- p.35 / Bradford assay for protein concentration --- p.36 / Western blot analysis --- p.37 / ELISA for quantification of HSV-1 antigen --- p.38 / Statistical analyses --- p.39 / Chapter Chapter 3: --- Results --- p.40 / Cytotoxicity and anti-herpetic activity of TCS and CHX --- p.41 / Selective cytotoxicity of TCS toward HSV-1 infected cells --- p.44 / Selective cytotoxicity is implicated in the antiviral activity of TCS --- p.50 / The effect of selective cytotoxicity on TI value --- p.53 / Chapter Chapter 4: --- Discussion --- p.55 / References --- p.64

Thesis (Ph. D.)--University of Missouri--Columbia, 2003. / "May 2003." Typescript. Vita. Includes bibliographical references (leaves 139-154).

Thesis (Ph. D.)--University of Missouri--Columbia, 2002. / Typescript. Vita. Includes bibliographical references (leaves 89-103). Also available on the Internet.

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

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

Orientador: Silvia Regina Brandalise / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-11T14:31:49Z (GMT). No. of bitstreams: 1 Mendonca_ReginaMariaHolandade_D.pdf: 3329753 bytes, checksum: 332276846d50abe7bcfd9a72c1a6bb9c (MD5) Previous issue date: 2008 / Resumo: Introdução: Dentre os efeitos secundários à quimioterapia, a mucosite oral é um dos mais freqüentes. Entre os fatores que contribuem para esta susceptibilidade estão a alta taxa de renovação celular e a presença dos microorganismos, que encontram na cavidade oral condição propícia para sua instalação. Objetivos: Analisar a associação entre grau de mucosite e a presença de HSV-1 (herpes simplex vírus tipo 1), Candida spp e bactérias da cavidade oral, em pacientes com Leucemia Linfóide Aguda, em quimioterapia. O segundo objetivo foi calcular o risco para os diferentes graus de mucosite, relacionados à presença do HSV-1, Candida spp e bactérias da cavidade oral nesta população. Método: Estudo prospectivo, unicêntrico, realizado no período de agosto de 2005 a outubro de 2006. Foram estudados 71 casos consecutivos de crianças com diagnóstico de Leucemia Linfóide Aguda, tratados de acordo com o Protocolo GBTLI LLA-99. A avaliação do grau de mucosite foi feita segundo critérios do National Cancer Institute. As análises da microbiota oral foram realizadas no 14° dia da terapia de Indução (D14) e 56° dia, na fase de Intensificação. A identificação do HSV-1 foi realizada pela técnica de reação em cadeia de polimerase (PCR). A identificação de bactérias e fungos, por testes bacterioscópicos e de cultivo. A análise da associação entre os elementos estudados e a intensidade de mucosite foi feita pelo teste exato de Fisher. O odds ratio foi calculado por regressão logística. O nível de significância foi de 5%. Resultados: No D14 foi encontrada associação estatisticamente significativa entre o grau da mucosite e a presença de HSV-1 (p<0,0001), bem como Candida spp (p=0,0078) e total de colônias de Candida spp (p=0,0012). No D56 foi encontrada associação significativa entre o grau da mucosite e a positividade do HSV-1 no D14 (p<0,0001), a presença de HSV-1 no D56 (p<0,0001) e o número de colônias de Candida spp no D56 (p=0,0247). No D14 foram considerados fatores de risco para aumento do grau da mucosite, a presença do HSV-1 (OR=65; IC=95%) e Candida spp (OR=15; IC=95%). No D56 os fatores de risco foram a presença do HSV-1 no D14 (OR=54; IC=95%) e a presença de HSV-1 no D56 (OR=57; IC=95%). Dentre os diferentes grupos das bactérias encontradas nos D14 e D56, o Streptococcus viridans foi identificado em 100% dos pacientes, em ambos momentos. Conclusões: A presença do HSV-1 e da Candida spp esteve associada ao grau da mucosite, nos momentos avaliados. Não se observou associação entre a presença dos distintos grupos bacterianos e o agravamento da mucosite oral / Abstract: Introduction: Oral mucositis is the most common side-effect related to chemotherapy. Oral microbiota and high cellular turn over of oral epithelium are contributing factors for mucositis. Objective: This study was designed to verify the associations of HSV-1(herpes simplex virus), Candida spp and oral bacterial presence in children and adolescents with Acute Lymphoblastic Leukemia (ALL) while on chemotherapy, and oral mucositis. The second objective was to calculate the odds ratio (OR) for oral mucositis grade in this population. Methods: A prospective study was conducted from August 2005 to October 2006 in one single center. Seventy one consecutive cases of children diagnosed with ALL were studied. The mucositis severity degree was done according to NCI Criteria. Oral microbiota analyses were performed on D14 and D56 of cancer therapy. HSV-1 identification was performed by PCR. Bacteria and fungy identification were obtained by standard bacteriology and cultivation tests. Fisher¿s exact test was used for analysis of the association analysis among the microorganisms and the mucositis grade. The OR was calculated by logistic regression. The significance level was 5%. Results: On D14 there was a significant association between mucositis grade and the HSV-1 presence (p<0.0001), as well as, Candid spp (p= 0.0078) and Candida spp colony numbers (p=0.0012). On D56 there was a significant association between mucositis grade and HSV-1 presence at D14 (p<0.0001), HSV-1 presence at D56 (p<0.0001) and Candida spp colony numbers at D56 (p=0,0247). On D14 the risks factors associated with an increase of the mucositis grade were HSV-1 presence (OR=65; IC=95%) and Candida spp (OR=15; IC=95%. On D56 the risk factors for mucositis were HSV-1 presence at D14 (OR=54; IC=95%) and HSV-1 presence at D56 (OR=57; IC=95%). Among the different groups of bacteria evaluated on D14 and D56, Streptococcus viridans occurred in 100% of all cases in both time points. Conclusion: HSV-1 and Candida spp presence was associated with mucositis grade on the evaluated time treatment intervals. There was no association between bacteria groups and oral mucositis severity / Doutorado / Saude da Criança e do Adolescente / Doutor em Saude da Criança e do Adolescente

Orientadores: Clarice Weis Arns, Luciana Konecny Kohn / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T23:34:00Z (GMT). No. of bitstreams: 1 Bianchi_BiancaReal_M.pdf: 1619930 bytes, checksum: 92df5dc0e4cba5e4314f506e39ff0be1 (MD5) Previous issue date: 2012 / Resumo: O vírus herpes simples do tipo 1 (HSV-1), agente etiológico do herpes labial em humanos, é facilmente transmitido e têm o grande problema de causar infecções latentes, sendo que uma vez infectado, o indivíduo passa a ser o portador do vírus por toda a vida. O medicamento mais apropriado contra este tipo de vírus deve ter ação inibitória em qualquer estágio de sua replicação, além de baixa toxicidade, para que as células do hospedeiro não sejam afetadas. Os organismos marinhos representam uma vasta biodiversidade que inclui cerca de 80% de todas as espécies do planeta, o que nos leva a uma grande quantidade de informações que ainda poderão ser descobertas, inclusive acerca de compostos com atividade antiherpética, já que atualmente temos poucos medicamentos disponíveis e nem sempre de total eficácia. Para o estudo com HSV-1 foi escolhida a cepa KOS, por ser resistente ao medicamento considerado mais eficaz para o herpes humano, o aciclovir. Foi utilizada a linhagem celular VERO para o estudo da atividade antiviral de extratos de organismos marinhos. Inicialmente foi realizada uma triagem com 129 extratos. Utilizou-se a concentração de 50?g/ml para todos os extratos, considerando ativos aqueles que apresentaram 97% de inibição do crescimento viral. Dentro dos grupos analisados foram identificados 6 extratos brutos de fungos e 7 extratos brutos de esponjas marinhas como possíveis antivirais. O cálculo do Índice de Seletividade foi realizado para as amostras de fungos Demateaceous (grupo) e Trichoderma sp., apresentando os valores 0,03 e 0,3, com atividade nas fases de adsorção e inativação viral, respectivamente e, para as amostras de esponjas, Monanchora arbuscula e Hemimycale sp., ambas apresentando o valor 0,1, com atividade também nas fases de adsorção e inativação viral, respectivamente / Abstract: Herpes simplex virus type 1 (HSV-1), the etiologic agent of herpes labialis in humans, is transmitted easily and have great problem to cause latent infections, and once infected, the individual becomes the carrier of the virus by life. The most suitable medicament against such virus should have inhibitory action at any stage of its replication as well as low toxicity to the host cells is not impaired. Marine organisms represent a wide biodiversity that includes about 80% of all species on the planet, which leads to a large amount of information that can still be found, including about compounds that may help a possible treatment of symptoms caused by this virus, since currently available medicines have few and not always fully effective. For the study of HSV-1, the KOS strain was chosen because it is resistant to the drug considered most effective for the human herpes, the acyclovir. Was used the cell lines VERO (African Green Monkey - ATCC CCL 81) for the study the antiviral activity of extracts of marine organisms. Initially was realized a screening with 129. We used the concentration of 50?g/ml for all the extracts, whereas those with active 97% inhibition of viral growth. Within the groups analyzed were identified 6 extracts of fungus and 7 crude extracts of marine sponges as possible antiviral. The calculation of the Selectivity Index was conducted for samples of fungi Demateaceous (group) and Trichoderma sp., presenting the values 0.03 and 0.3 with activity phases of adsorption and viral inactivation, respectively, and for samples of sponges, Monanchora arbuscula and Hemimycale sp. both presenting the value 0.1, with activity also in the phases of adsorption and viral inactivation, respectively / Mestrado / Clinica Medica / Mestra em Ciências

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2006. / Härtill 4 uppsatser.