This investigation was concerned with characterizing a tumor line induced and maintained in this laboratory. Various chemical assays, cell counts, and electron microscopy were the methods employed to characterize the blood of mice bearing the tumor at days 3, 6, 9, and 12 after injection of the 1.2 x 10^8 tumor cells.

The purpose of this investigation was (1) to isolate and purify glyoxalase I from the livers of normal DBA/lJ mice and the livers from mice bearing a lymphosarcoma tumor; and (2) to determine, at least with respect to glyoxalase I, if the tumor has an effect on the chemical properties or structure of macromolecules in an organ removed from tumor locale and not histologically affected by its presence.

The problem with which this investigation is concerned is that of determining the nature of events leading to the change. of normal cells into malignant cells. The design of the study is multi-phasic: (A) to establish the presence or absence of an oncogenic virion, (B) to demonstrate by use of the electron microscopy any ultracellular alteration in malignant or transformed tissues, (C) to investigate the nature of the transforming agent in the murine lymphosarcoma, and (D) to employ various methods to demonstrate cellular transformations in vivo and in vitro. It is concluded that the transforming and tumorinducing agent in this' investigation was not a virion, but an infectious ribonucleic acid genome or a segment of a viral genome which had become integrated into the genome of the mouse cells. The vision has lost its ability to form a protein coat; therefore it is not demonstrable as a virion. But the ribonucleic acid is able to infect other cells and transform them from normal to neoplastic tissues.

Orientador: Deilson Elgui de Oliveria / Banca: Lígia Niéro-Melo / Banca: Daniel Onofre Vidal / Banca: Alexandrona Sartori / Banca: Otávio Cesar Carvalho Guimarães Baiocchi / Resumo: Linfoma de Hodgkin clássico (LHc) é uma neoplasia maligna que apresenta um padrão aberrante de expressão de citocinas, incluindo IL-10 e TNF-a. Polimorfismos de nucleotídeo único (SNPs) nos genes codificadores de IL-10 e TNF-a ou de suas proteínas regulatórias, como NFkB, podem interferir na patobiologia do LHc. No presente estudo, SNPs nas regiões promotoras dos genes de IL-10 (SNP/pIL-10 -592, rs1800872; e SNP/pIL-10 -1082, rs1800896) e TNF-a (SNP/pTNF-a -238, rs361525; e SNP/pTNF-a -862, rs1800630), assim como na região intrônica do gene NFkB1 (SNP/iNFkB1, rs1585215) foram genotipados em 73 pacientes com LHc e avaliados em relação à sua associação com parâmetros prognósticos clínicos e laboratoriais da doença. SNPs/pIL-10 AA foram significativamente associados a maiores contagens de leucócitos e menores valores de linfócitos ao diagnóstico. No caso de TNF-a, SNP/pTNF-a -238 AG foi associado à presença de infecção pelo EBV enquanto SNP/pTNF-a -862 CC apresentou-se mais frequentemente com leucocitose. SNP/iNFkB1 AA associaramse à doença em estádio IV e padrão extranodal de apresentação. Entretanto, nenhum dos SNPs avaliados teve efeito no desfecho do tratamento. Esse estudo mostrou que alguns genótipos de SNPs nos genes de IL-10 e TNF-a estão associados a parâmetros prognósticos no LHc. Adicionalmente, pela primeira vez foram descritas associações entre SNP/iNFkB1 (rs1585215) e aspectos clínicos da doença / Abstract: Classic Hodgkin lymphoma (cHL) is a malignant lymphoid neoplasia that shows aberrant expression of cytokines, including IL-10 and TNF-a. Single nucleotide polymorphisms (SNPs) in genes encoding IL-10 and TNF-a or their regulatory proteins, such as NFkB, may impact cHL pathobiology. In this study, SNPs in the promoter regions of genes encoding for IL-10 (SNP/pIL-10 -592, rs1800872; and SNP/pIL-10 -1082, rs1800896), and TNF-a (SNP/pTNF-a -238, rs361525; and SNP/pTNF-a -862, rs1800630), as well as in the intronic region of the NFkB1 gene (SNP/iNFkB1, rs1585215) were genotyped in 73 patients with cHL and evaluated against clinical and laboratory prognostic parameters for the disease. SNPs/pIL-10 AA were significantly associated with higher leukocyte and lower lymphocyte counts at diagnosis. In case of TNF-a, SNP/pTNF-a -238 AG was associated with EBV infection while SNP/pTNF-a -862 CC presented more frequently with leukocytosis. SNP/iNFkB1AA generally had stage IV and extranodal disease at diagnosis. Nonetheless, none of the studied SNPs had effect on on treatment outcome. This study shows that some SNPs genotypes for IL-10 and TNF-a genes are associated with prognostic parameters in cHL; furthermore, this is the first time that the SNP/iNFkB1 (rs1585215) was implicated in clinical features of the disease / Doutor

Despite improved treatments for Non-Hodgkin’s Lymphoma (NHL) and Multiple Myeloma (MM), most patients eventually relapse and these diseases remain largely incurable. This has precipitated recent research into more clinically relevant in vitro models to enable development of more effective therapies. We have validated and standardised two in vitro primary culture models using tumour samples derived from patients with NHL, Chronic Lymphocytic Leukaemia (CLL) and MM. Several novel findings have been demonstrated. In vitro sensitivity of primary NHL cells cocultured in a CD40L model predicted clinical response to bortezomib in patients receiving the drug in a phase II trial. In vitro sensitivity correlated with CD40 expression, identifying a potential surrogate biomarker for response to bortezomib. The novel HDAC inhibitor, UCL67022 was 10-fold more potent than vorinostat in NHL and produced synergy when combined with bortezomib. UCL67022 maintained its potency in primary MM samples grown in an HS-5 stromal model. It modulated cytokine secretion resulting in downregulation of cytokine-induced signalling pathways (JAK/STAT3). A novel Hsp90 inhibitor, KW-2478 maintained activity in the HS-5 model and enhanced the activity of bortezomib and melphalan. Hsp70 was identified as a potential surrogate biomarker to monitor the combinatorial effect in future clinical trials. A highly synergistic and schedule-dependent cytotoxic effect occurred when primary MM cells were pre-treated with melphalan followed by bortezomib, with important implications for future clinical trial design. IL-6, IL-8 and VEGF levels correlated with resistance to bortezomib and melphalan and were associated with activation of JAK/STAT, MAPK and PI3K/Akt signalling pathways. Antibody neutralization of IL-6, IL-8 and VEGF resulted in restoration of drug sensitivity. We have therefore demonstrated the ability of primary culture models to predict response to chemotherapy, to identify therapeutically beneficial novel agents and to enable study of tumour microenvironmental interactions responsible for drug resistance in patients with haematological malignancies.

Mantle Cell Lymphoma (MCL) is a rare but aggressive Non-Hodgkin Lymphoma (NHL). Although t(11;14) is a hallmark of MCL, it is insufficient for lymphomagenesis. The phosphoinositide-3 kinase (PI3K) pathway is thought to play an important role in MCL pathogenesis and the PI3K p110δ isoform is enriched in leucocytes making it an attractive target. Early phase trials evaluating the p110δ selective inhibitor GS-1101 however demonstrate inferior responses in MCL compared to chronic lymphocytic leukaemia and indolent NHL. The relative contribution of the class Ia PI3K isoforms p110α (PIK3CA), p110β (PIK3CB) and p110δ (PIK3CD) was therefore evaluated in MCL. Immunohistochemistry on MCL tissue microarrays revealed that while p110δ was highly expressed, p110α showed wide variation and p110β expression was the weakest. A significant increase in p110α expression was found with disease progression. Although GS-1101 was sufficient to abolish B-cell receptor mediated PI3K activation, additional p110α inhibition was necessary to abolish constitutive PI3K activation in MCL exhibiting high p110α expression. Compared to GS-1101, GDC-0941 (p110α and p110δ inhibitor) had greater in vitro toxicity and a high PIK3CA/PIK3CD mRNA ratio (> twice ratio in healthy B-cells) was able to identify primary MCL samples that were resistant to GS-1101 but significantly more sensitive to GDC—0941. This ratio also increased with disease progression. No PIK3CA or PIK3R1 activating mutations were found. In summary, blockade of both p110α and p110δ appears to be necessary for effective PI3K inhibition in MCL, particularly with relapse. The PIK3CA/PIK3CD mRNA ratio may help identify those patients that are most likely to respond. Finally, a disseminated xenograft model of human primary MCL was established in NSG mice. Engraftment of primary MCL was demonstrated by peripheral blood flow cytometry, tissue immunohistochemistry and FISH for t(11;14). This model is potentially valuable for pre-clinical in vivo testing of novel drugs for this incurable disease.

Na radioterapia, o planejamento é parte fundamental do tratamento de um paciente. Para que o planejamento seja bem sucedido, o físico deve utilizar parâmetros físicos baseados na interação da radiação com a matéria para determinar a dose no volume alvo. Esses parâmetros são obtidos a partir de campos com dimensões bem definidas (geralmente quadrados), porém, na radioterapia é comum aparecer campos irregulares. Para resolver este problema, utilizam-se métodos que convertem esses campos irregulares em equivalentes de campos quadrados. Quando se tem campos pequenos, a aproximação de Sterling (algoritmo que permite a obtenção do campo quadrado equivalente a partir da razão entre quatro vezes a área efetiva de irradiação pelo perímetro que a envolve) é bem sucedida, porém quando o campo é muito grande e irregular, esta relação mais tão eficiente. Para tal caso, tem-se um método conhecido por Método de Clarkson. O método de Clarkson é um algoritmo que utiliza valores obtidos a partir de funções que relacionam a interação da radiação com a matéria, tais como, Razão Espalhamento Ar (SAR) e Razão Tecido Ar (TAR), para determinar especialmente campos quadrados equivalentes de campos irregulares e de grandes dimensões ou para calcular a dose de radiação em um ponto do paciente. A utilização deste método é muito trabalhosa, pois exige muitas horas de trabalho manual de um profissional especializado. Este trabalho desenvolveu um programa que analisa uma magem digital de um campo irregular, como os campos encontrados frequentemente no tratamento de linfomas do tipo Hodgking (manto), e determina um campo regular equivalente a este. As incertezas entre o método de Clarkson manual e o programado foram de ( -0,52 ± 0,60 )% e validam o programa desenvolvido. / Radiotherapy has a fundamental part that is the patient treatment planning. For its success, the medical physicist has to use physical parameters based on the radiation interaction with the matter, in this case the patient; in order to determine the radiation absorbed dose in the selected target volume. The physical parameters normally are related to regular radiation field sizes, although irregular fields can be also used in some treatments. To overcome this problem there are methods that convert irregular fields in regular fields, such as a square one. For small fields the Sterling\'s Method (technique to calculate equivalent square fields making use of ratio four times the effective area of irradiation by is perimeter) can be used. However, this method is not adequate for large irregular field. In this case, the Clarkson\'s Method is used. The Clarkson\'s Method uses values functions, such as Scattering-Air Ratio (SAR) and Tissue-Air Ratio (TAR), related with radiation interaction of with matter, to determine equivalent square fields from those large irregular ones or to calculate the radiation absorbed dose in the point of interest in a patient The application this method takes time, once it needs a considerable time from the medical physicist attention and also from the equipment. In this work, a software was developed to analyse digital images of large irregular fields, such as fields found in treatment of Hodgkin\'s disease (mantle) correlating them to regular equivalent fields. The uncertainty between the manual Clarkson and the programmed was inferred as ( -0,52 ± 0,60 )%, what can validate the program developed.

Made available in DSpace on 2016-06-07T17:12:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-08-25. Added 1 bitstream(s) on 2016-06-07T17:17:00Z : No. of bitstreams: 1 000864285.pdf: 2759439 bytes, checksum: 7135a8f61ee29b84b93a0f8f1e4fb0b9 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Linfoma de Hodgkin clássico (LHc) é uma neoplasia maligna que apresenta um padrão aberrante de expressão de citocinas, incluindo IL-10 e TNF-a. Polimorfismos de nucleotídeo único (SNPs) nos genes codificadores de IL-10 e TNF-a ou de suas proteínas regulatórias, como NFkB, podem interferir na patobiologia do LHc. No presente estudo, SNPs nas regiões promotoras dos genes de IL-10 (SNP/pIL-10 -592, rs1800872; e SNP/pIL-10 -1082, rs1800896) e TNF-a (SNP/pTNF-a -238, rs361525; e SNP/pTNF-a -862, rs1800630), assim como na região intrônica do gene NFkB1 (SNP/iNFkB1, rs1585215) foram genotipados em 73 pacientes com LHc e avaliados em relação à sua associação com parâmetros prognósticos clínicos e laboratoriais da doença. SNPs/pIL-10 AA foram significativamente associados a maiores contagens de leucócitos e menores valores de linfócitos ao diagnóstico. No caso de TNF-a, SNP/pTNF-a -238 AG foi associado à presença de infecção pelo EBV enquanto SNP/pTNF-a -862 CC apresentou-se mais frequentemente com leucocitose. SNP/iNFkB1 AA associaramse à doença em estádio IV e padrão extranodal de apresentação. Entretanto, nenhum dos SNPs avaliados teve efeito no desfecho do tratamento. Esse estudo mostrou que alguns genótipos de SNPs nos genes de IL-10 e TNF-a estão associados a parâmetros prognósticos no LHc. Adicionalmente, pela primeira vez foram descritas associações entre SNP/iNFkB1 (rs1585215) e aspectos clínicos da doença / Classic Hodgkin lymphoma (cHL) is a malignant lymphoid neoplasia that shows aberrant expression of cytokines, including IL-10 and TNF-a. Single nucleotide polymorphisms (SNPs) in genes encoding IL-10 and TNF-a or their regulatory proteins, such as NFkB, may impact cHL pathobiology. In this study, SNPs in the promoter regions of genes encoding for IL-10 (SNP/pIL-10 -592, rs1800872; and SNP/pIL-10 -1082, rs1800896), and TNF-a (SNP/pTNF-a -238, rs361525; and SNP/pTNF-a -862, rs1800630), as well as in the intronic region of the NFkB1 gene (SNP/iNFkB1, rs1585215) were genotyped in 73 patients with cHL and evaluated against clinical and laboratory prognostic parameters for the disease. SNPs/pIL-10 AA were significantly associated with higher leukocyte and lower lymphocyte counts at diagnosis. In case of TNF-a, SNP/pTNF-a -238 AG was associated with EBV infection while SNP/pTNF-a -862 CC presented more frequently with leukocytosis. SNP/iNFkB1AA generally had stage IV and extranodal disease at diagnosis. Nonetheless, none of the studied SNPs had effect on on treatment outcome. This study shows that some SNPs genotypes for IL-10 and TNF-a genes are associated with prognostic parameters in cHL; furthermore, this is the first time that the SNP/iNFkB1 (rs1585215) was implicated in clinical features of the disease

Na radioterapia, o planejamento é parte fundamental do tratamento de um paciente. Para que o planejamento seja bem sucedido, o físico deve utilizar parâmetros físicos baseados na interação da radiação com a matéria para determinar a dose no volume alvo. Esses parâmetros são obtidos a partir de campos com dimensões bem definidas (geralmente quadrados), porém, na radioterapia é comum aparecer campos irregulares. Para resolver este problema, utilizam-se métodos que convertem esses campos irregulares em equivalentes de campos quadrados. Quando se tem campos pequenos, a aproximação de Sterling (algoritmo que permite a obtenção do campo quadrado equivalente a partir da razão entre quatro vezes a área efetiva de irradiação pelo perímetro que a envolve) é bem sucedida, porém quando o campo é muito grande e irregular, esta relação mais tão eficiente. Para tal caso, tem-se um método conhecido por Método de Clarkson. O método de Clarkson é um algoritmo que utiliza valores obtidos a partir de funções que relacionam a interação da radiação com a matéria, tais como, Razão Espalhamento Ar (SAR) e Razão Tecido Ar (TAR), para determinar especialmente campos quadrados equivalentes de campos irregulares e de grandes dimensões ou para calcular a dose de radiação em um ponto do paciente. A utilização deste método é muito trabalhosa, pois exige muitas horas de trabalho manual de um profissional especializado. Este trabalho desenvolveu um programa que analisa uma magem digital de um campo irregular, como os campos encontrados frequentemente no tratamento de linfomas do tipo Hodgking (manto), e determina um campo regular equivalente a este. As incertezas entre o método de Clarkson manual e o programado foram de ( -0,52 ± 0,60 )% e validam o programa desenvolvido. / Radiotherapy has a fundamental part that is the patient treatment planning. For its success, the medical physicist has to use physical parameters based on the radiation interaction with the matter, in this case the patient; in order to determine the radiation absorbed dose in the selected target volume. The physical parameters normally are related to regular radiation field sizes, although irregular fields can be also used in some treatments. To overcome this problem there are methods that convert irregular fields in regular fields, such as a square one. For small fields the Sterling\'s Method (technique to calculate equivalent square fields making use of ratio four times the effective area of irradiation by is perimeter) can be used. However, this method is not adequate for large irregular field. In this case, the Clarkson\'s Method is used. The Clarkson\'s Method uses values functions, such as Scattering-Air Ratio (SAR) and Tissue-Air Ratio (TAR), related with radiation interaction of with matter, to determine equivalent square fields from those large irregular ones or to calculate the radiation absorbed dose in the point of interest in a patient The application this method takes time, once it needs a considerable time from the medical physicist attention and also from the equipment. In this work, a software was developed to analyse digital images of large irregular fields, such as fields found in treatment of Hodgkin\'s disease (mantle) correlating them to regular equivalent fields. The uncertainty between the manual Clarkson and the programmed was inferred as ( -0,52 ± 0,60 )%, what can validate the program developed.

No description available.