O objetivo do estudo foi avaliar a importância da expressão das proteínas topoisomerase II alfa (topo II) e HER-2 como fatores preditvos da resposta à quimioterapia neoadjuvante e prognóstico em pacientes com câncer de mama nos estádio clínico II e III. Pacientes e métodos: 99 pacientes receberam quimioterapia neoadjuvante com docetaxel (75mg /m²) e epirrubicina (50 mg/m²) em infusão endovenosa no dia 1 a cada 3 semanas após terem sido submetidas a biópsia incisional. Foi complementado tratamento sistêmico com quimioterapia adjuvante com CMF ou FEC de acordo com o estado axilar avaliada após a cirurgia definitiva e/ou hormonioterapia de acordo com a avaliacãodos receptores hormonais. Avaliamos a taxa de resposta ao tratamento neoadjuvante e a influência da topo II alfa e do HER-2 na taxa de resposta à quimioterapia neoadjuvante bem comona sobrevida livre de doença e sobrevida global. Também foram avaliadas a expressão dos receptores hormonais. Resultados: a taxa de resposta clínica objetiva foi de 80,8 % com 9,1 % de resposta patológica completa. A expressão da topo II alfa nao apresentou significância nas taxas de resposta ou na sobrevida das pacietnes e nao houve correlação entre a expressão desta proteína e de HER-2. A superexpressão da proteína HER-2 foi associada com uma redução significante nas taxas de sobrevida livre de doença e sobrevida global (p= 0,04 e p= 0,004, respectivamente). Conclusão: a expressão da topo II alfa não demonstrou, em nosso estudo, ser fator preditivo ou prognóstico nas pácientes submetidas a quimioterapia neoadjuvante com docetaxel e epirrubicina. / The objective of this study is to evaluate the importance of the expression of the proteins topoisomerase II alpha (topo II) and HER-2 as predictive factors to response to neoadjuvant chemotherapy and the prognosis of patients diagnosed with clinical stage II and stage III breast cancer. Patients and methods: 99 patients have received neoadjuvant chemotherapy with docetaxel (75mg /m²) and epirrubicine (50 mg/m²) through intravenous infusion on D1 q3 weeks, after submitted to pathologic specimen harvest. Systemic treatment was then complemented with CMF or FEC according to the status of axilla involvement after surgical staging and/or hormone therapy according tohormone receptor status. We evaluated the response rate to neoadjuvant treatment and the influence of topo II alpha and HER-2 expression on the response rate and disease free survival and overall survival. The expression of hormone receptors was also evaluated. Results: Objective clinical response was 78,8%, with 8,2% of complete pathological response.Topo II alpha expression did not correlate to response to chemotherapy or survival and there was no correlation between topo II alpha expression and HER-2 expression. Superexpression of HER-2 protein was associated to a significant reduction in disease free survival and overall survival (p=0,04 and p=0,004, respectively). Conclusion: topo II alpha expression did not demonstrate, in our study, to be a predictive nor prognostic factor to the patientssubmitted to neoadjuvant with docetaxel and epirrubicin.

Made available in DSpace on 2017-07-10T19:24:05Z (GMT). No. of bitstreams: 1 Marcelo Alan Primo.pdf: 3445063 bytes, checksum: 29c3b04fea727923f8cfb2b19dba08d7 (MD5) Previous issue date: 2007-07-06 / The uniformity of the application in the soil of fertilizers is an important factor in the efficiency of use of the fertilizers and his effect stop plants. The distributors of fertilizers to I throw demand her some basic tunings in what concerns to the flow and work width. The rehearsals of work width and distribution uniformity are of difficult execution for the farmer for they demand equipments, resources and complex methods. Therefore, this work had as objective evaluates alternative collectors to turn more accessible the tuning and evaluation of distributors of fertilizers solid her throw. The work was accomplished in the Agrarian Foundation of Agricultural Research - FAPA, located in the district of Among Rivers, municipal district of Guarapuava - Paraná. Four different groups of collectors were used: i) Collectors built according to the norm ISO 5690/1; ii) Collectors of polyethylene (plastic trays) with soubrette to avoid the material rebound; iii) Collectors formed by boxes of milk long life; iv) Collectors formed by vases of polyethylene (Number 3,5). To the evaluation of the strip of traverse distribution, the collectors were distributed side by side parallel amongst themselves, forming a traverse line to the displacement of the tractor and distributor. A tractor was used Ford, model 7630, in which distributors were coupled: Jan, model Lancer Master 1200; Incomagri, model Sembra 1300-P AH; and Nogueira, model Rota Flow. The alternative collectors presented a larger retention than the established collectors. This relative difference among the collections was larger in the extremities of the distribution profile. The width executes of work obtained for the coefficients of variation of 12,5, 15, 17,5 and 20% varied among the collectors in the different courses. The collector that more it resembled each other the norm were the vases of polyethylene. It was observed that the height of the collectors went more important for his/her efficiency of what his/her collection area. / A uniformidade da aplicação no solo de fertilizantes é um importante fator na eficiência de uso dos fertilizantes e seu efeito para as plantas. Os distribuidores de fertilizantes a lanço exigem algumas regulagens básicas no que concerne à vazão e largura de trabalho. Os ensaios de largura de trabalho e uniformidade de distribuição são de difícil execução para o agricultor por exigirem equipamentos, recursos e métodos complexos. Portanto, este trabalho teve como objetivo avaliar coletores alternativos para tornar mais acessível a regulagem e avaliação de distribuidores de fertilizantes sólidos a lanço. O trabalho foi realizado na Fundação Agrária de Pesquisa Agropecuária FAPA, localizada no distrito de Entre Rios, município de Guarapuava - Paraná. Foram utilizados quatro diferentes conjuntos de coletores: i) Coletores construídos segundo a norma ISO 5690/1; ii) Coletores de polietileno (bandejas plásticas) com sombrite para evitar o ricochete de material; iii) Coletores formados por caixas de leite longa vida; iv) Coletores formados por vasos de polietileno (Número 3,5). Para a avaliação da faixa de distribuição transversal, os coletores foram distribuídos lado a lado paralelos entre si, formando uma linha transversal ao deslocamento do trator e distribuidor. Foi utilizado um trator Ford, modelo 7630, no qual foram acoplados distribuidores: Jan, modelo Lancer Master 1200; Incomagri, modelo Sembra 1300-P AH; e Nogueira, modelo Rota Flow. Os coletores alternativos apresentaram uma retenção maior que os coletores normatizados. Esta diferença relativa entre as coletas foi maior nas extremidades do perfil de distribuição. A largura efetiva de trabalho obtida para os coeficientes de variação de 12,5, 15, 17,5 e 20% variou entre os coletores nos diferentes percursos. O coletor que mais se assemelhou a norma foram os vasos de polietileno. Observou-se que a altura dos coletores foi mais importante para sua eficiência do que a sua área de coleta.

Made available in DSpace on 2017-05-12T14:47:21Z (GMT). No. of bitstreams: 1 Marcelo Alan Primo.pdf: 3445063 bytes, checksum: 29c3b04fea727923f8cfb2b19dba08d7 (MD5) Previous issue date: 2007-07-06 / The uniformity of the application in the soil of fertilizers is an important factor in the efficiency of use of the fertilizers and his effect stop plants. The distributors of fertilizers to I throw demand her some basic tunings in what concerns to the flow and work width. The rehearsals of work width and distribution uniformity are of difficult execution for the farmer for they demand equipments, resources and complex methods. Therefore, this work had as objective evaluates alternative collectors to turn more accessible the tuning and evaluation of distributors of fertilizers solid her throw. The work was accomplished in the Agrarian Foundation of Agricultural Research - FAPA, located in the district of Among Rivers, municipal district of Guarapuava - Paraná. Four different groups of collectors were used: i) Collectors built according to the norm ISO 5690/1; ii) Collectors of polyethylene (plastic trays) with soubrette to avoid the material rebound; iii) Collectors formed by boxes of milk long life; iv) Collectors formed by vases of polyethylene (Number 3,5). To the evaluation of the strip of traverse distribution, the collectors were distributed side by side parallel amongst themselves, forming a traverse line to the displacement of the tractor and distributor. A tractor was used Ford, model 7630, in which distributors were coupled: Jan, model Lancer Master 1200; Incomagri, model Sembra 1300-P AH; and Nogueira, model Rota Flow. The alternative collectors presented a larger retention than the established collectors. This relative difference among the collections was larger in the extremities of the distribution profile. The width executes of work obtained for the coefficients of variation of 12,5, 15, 17,5 and 20% varied among the collectors in the different courses. The collector that more it resembled each other the norm were the vases of polyethylene. It was observed that the height of the collectors went more important for his/her efficiency of what his/her collection area. / A uniformidade da aplicação no solo de fertilizantes é um importante fator na eficiência de uso dos fertilizantes e seu efeito para as plantas. Os distribuidores de fertilizantes a lanço exigem algumas regulagens básicas no que concerne à vazão e largura de trabalho. Os ensaios de largura de trabalho e uniformidade de distribuição são de difícil execução para o agricultor por exigirem equipamentos, recursos e métodos complexos. Portanto, este trabalho teve como objetivo avaliar coletores alternativos para tornar mais acessível a regulagem e avaliação de distribuidores de fertilizantes sólidos a lanço. O trabalho foi realizado na Fundação Agrária de Pesquisa Agropecuária FAPA, localizada no distrito de Entre Rios, município de Guarapuava - Paraná. Foram utilizados quatro diferentes conjuntos de coletores: i) Coletores construídos segundo a norma ISO 5690/1; ii) Coletores de polietileno (bandejas plásticas) com sombrite para evitar o ricochete de material; iii) Coletores formados por caixas de leite longa vida; iv) Coletores formados por vasos de polietileno (Número 3,5). Para a avaliação da faixa de distribuição transversal, os coletores foram distribuídos lado a lado paralelos entre si, formando uma linha transversal ao deslocamento do trator e distribuidor. Foi utilizado um trator Ford, modelo 7630, no qual foram acoplados distribuidores: Jan, modelo Lancer Master 1200; Incomagri, modelo Sembra 1300-P AH; e Nogueira, modelo Rota Flow. Os coletores alternativos apresentaram uma retenção maior que os coletores normatizados. Esta diferença relativa entre as coletas foi maior nas extremidades do perfil de distribuição. A largura efetiva de trabalho obtida para os coeficientes de variação de 12,5, 15, 17,5 e 20% variou entre os coletores nos diferentes percursos. O coletor que mais se assemelhou a norma foram os vasos de polietileno. Observou-se que a altura dos coletores foi mais importante para sua eficiência do que a sua área de coleta.

Human Epidermal Growth Factor Receptor 2 (HER-2/neu) is a breast tumor antigen (Ag) commonly overexpressed in 30% of breast cancer cases. Both HER-2/neu-targeted DNA-based and transgene modified dendritic cell (DC)-based vaccines are potent elements in eliciting HER-2/neu specific antitumor immune responses; however, there has been no side-by-side comparison of these two different immunization methods. We utilized an in vivo murine tumor model expressing the rat neu Ag to compare the immunization efficacy between DC transduced with replication-deficient adenovirus containing neu (AdVneu), to form DCneu, and plasmid DNA (pcDNA) vaccine. DCneu displayed an upregulation of immunologically important molecules and inflammatory cytokines expression such as IL-6 that partially mediated conversion of the regulatory T (Tr) cell suppression. Wildtype FVB/N mice immunized with DCneu induced stronger HER-2/neu-specific humoral and cellular immune responses compared to plasmid DNA immunized mice. Furthermore, mice immunized with DCneu remained completely protected from tumor challenge compared to partial or no protection observed in DNA immunized mice in two tumor animal models. In FVBneuN transgenic mice, which develop spontaneous breast tumors at 4-8 months of age, DCneu significantly delayed tumor onset when immunization conducted in mice at a younger age. Taken together, we demonstrated that a HER-2/neu-gene modified DC vaccine is more potent than a plasmid DNA vaccine in inducing neu specific immune responses resulting in greater protective and preventative effects in the tumor models examined. <p>In another study, we examined the use of a DC-based cancer vaccine involving the phagocytosis of apoptotic tumor cells expressing heat shock protein 70 (HSP70). The dual role of HSP70, as an antigenic peptide chaperone and danger signal, makes it especially important in DC-based vaccination. In this study, we investigated the impacts of apoptotic transgenic MCA/HSP tumor cells expressing HSP70 on DC maturation, T cell stimulation and overall vaccine efficacy. We found that DC with phagocytosis of MCA/HSP in the early phase of apoptosis expressed more peptide-major histocompatibility class (pMHC) I complexes, stimulated stronger cytotoxic T lymphocytes (CTL) responses and induced greater immune protection against MCA tumor cell challenge, compared to mice immunized with DC that phagocytosed MCA/HSP cells in the late phase of apoptosis. Taken together, our data demonstrated that HSP70 expression on apoptotic tumor cells stimulated DC maturation and DC with phagocytosis of apoptotic tumor cells expressing HSP70 in early phase of apoptosis more efficiently induced tumor-specific CTL responses and immunity than DC with phagocytosis of apoptotic tumor cells in late phase of apoptosis. <p>Overall, we have examined variations in designing DC-based cancer vaccines in two completely different model systems. Taken together, our results may have an important impact in designing DC-based antitumor vaccines.

Human Epidermal Growth Factor Receptor 2 (HER-2/neu) is a breast tumor antigen (Ag) commonly overexpressed in 30% of breast cancer cases. Both HER-2/neu-targeted DNA-based and transgene modified dendritic cell (DC)-based vaccines are potent elements in eliciting HER-2/neu specific antitumor immune responses; however, there has been no side-by-side comparison of these two different immunization methods. We utilized an in vivo murine tumor model expressing the rat neu Ag to compare the immunization efficacy between DC transduced with replication-deficient adenovirus containing neu (AdVneu), to form DCneu, and plasmid DNA (pcDNA) vaccine. DCneu displayed an upregulation of immunologically important molecules and inflammatory cytokines expression such as IL-6 that partially mediated conversion of the regulatory T (Tr) cell suppression. Wildtype FVB/N mice immunized with DCneu induced stronger HER-2/neu-specific humoral and cellular immune responses compared to plasmid DNA immunized mice. Furthermore, mice immunized with DCneu remained completely protected from tumor challenge compared to partial or no protection observed in DNA immunized mice in two tumor animal models. In FVBneuN transgenic mice, which develop spontaneous breast tumors at 4-8 months of age, DCneu significantly delayed tumor onset when immunization conducted in mice at a younger age. Taken together, we demonstrated that a HER-2/neu-gene modified DC vaccine is more potent than a plasmid DNA vaccine in inducing neu specific immune responses resulting in greater protective and preventative effects in the tumor models examined. <p>In another study, we examined the use of a DC-based cancer vaccine involving the phagocytosis of apoptotic tumor cells expressing heat shock protein 70 (HSP70). The dual role of HSP70, as an antigenic peptide chaperone and danger signal, makes it especially important in DC-based vaccination. In this study, we investigated the impacts of apoptotic transgenic MCA/HSP tumor cells expressing HSP70 on DC maturation, T cell stimulation and overall vaccine efficacy. We found that DC with phagocytosis of MCA/HSP in the early phase of apoptosis expressed more peptide-major histocompatibility class (pMHC) I complexes, stimulated stronger cytotoxic T lymphocytes (CTL) responses and induced greater immune protection against MCA tumor cell challenge, compared to mice immunized with DC that phagocytosed MCA/HSP cells in the late phase of apoptosis. Taken together, our data demonstrated that HSP70 expression on apoptotic tumor cells stimulated DC maturation and DC with phagocytosis of apoptotic tumor cells expressing HSP70 in early phase of apoptosis more efficiently induced tumor-specific CTL responses and immunity than DC with phagocytosis of apoptotic tumor cells in late phase of apoptosis. <p>Overall, we have examined variations in designing DC-based cancer vaccines in two completely different model systems. Taken together, our results may have an important impact in designing DC-based antitumor vaccines.

This research uses the interpretive framework of the Psychological Grotesque to address a protagonist's response when she is unable to integrate the experience of interpersonal trauma into her psyche. The framework reveals a survival mechanism, identity incorporation, with roots in the transgressive and evolutionary nature of the grotesque as discussed in the work of Mikhail Bakhtin, Mary Douglas and Leonard Cassuto. The psychological grotesque is explicated using Stvgo by Laura Hendrie, The Life Before Her Eyes, by Laura Kasischke, and Linden Hills by Gloria Naylor.The psychological grotesque reveals how the protagonist within each of these novels, when positioned within a specific matrix of contributing factors engages in flawed survival strategies to reconcile psychic fragmentation. Drawing on theories of trauma from the work of Bessel Van Der Kolk, Dori Laub, Sigmund Freud, Mardi Horowitz, Ronnie Janoff-Bulman, Laurie Vickroy, Cathy Caruth, Peter Woods and Tim Middleton, the definition of the matrix includes: interpersonal trauma, prior history of emotional problems, no social support network, and self-perceived complicity in the violence. Where these criteria are present, the protagonist is incapable of achieving the repair of her damaged psyche in order to reintegrate into society and relief from the pain of trauma. In an effort to repair her brokeness through the incorporation of parts of the identity of others, the protagonist creates a grotesque mental hybrid living in fractured time. The protagonist experiences destabilization of time due to incorporating the temporal perspective of the other identities. A flawed survival strategy, identity incorporation leads to further psychic fragmentation.The psychological grotesque takes up the challenge of communicating the effects of trauma and addresses the lack of a literary interpretive mechanism for trauma literature in which a critical component of the narrative is the story of female victims of interpersonal violence. This framework confronts the fact that representations of women and trauma are problematic due to how trauma resists linguistic representation and because women have historically been denied a voice in the canon. Therefore, by drawing on elements of the grotesque, specifically hybridity and transgression, this interpretive framework recuperates the experiences of traumatized protagonists. / Department of English

Dendritic cells (DCs) are promising mediators of anti-tumour immune responses. Unfortunately, a major hindrance to the development of highly effective DC vaccines is their short lifespan. Tumour antigen presentation may also not be optimal. We hypothesize that the introduction of exogenous survival factors (SFs) would prolong DC longevity and that modulation of TAA glycosylation will improve antigen presentation. To this end, we have constructed bicistronic lentivectors (LVs) encoding the xeno Tumour-Associated-Antigen (TAA), rHER-2/neu, and one of five candidate SFs. We demonstrated that our LVs can effectively protect transduced DCs from apoptosis when subjected to apoptosis-inducing conditions. TAA glycosylation has been proposed to obstruct the processing and presentation of peptides on MHC molecules. To address this second issue, we have engineered a LV that encodes a partially deglycosylated rHER-2/neu. Overall, we have generated the tools to alter intrinsic DC properties, which we believe will be integral to improving DC vaccine efficacy.

Dendritic cells (DCs) are promising mediators of anti-tumour immune responses. Unfortunately, a major hindrance to the development of highly effective DC vaccines is their short lifespan. Tumour antigen presentation may also not be optimal. We hypothesize that the introduction of exogenous survival factors (SFs) would prolong DC longevity and that modulation of TAA glycosylation will improve antigen presentation. To this end, we have constructed bicistronic lentivectors (LVs) encoding the xeno Tumour-Associated-Antigen (TAA), rHER-2/neu, and one of five candidate SFs. We demonstrated that our LVs can effectively protect transduced DCs from apoptosis when subjected to apoptosis-inducing conditions. TAA glycosylation has been proposed to obstruct the processing and presentation of peptides on MHC molecules. To address this second issue, we have engineered a LV that encodes a partially deglycosylated rHER-2/neu. Overall, we have generated the tools to alter intrinsic DC properties, which we believe will be integral to improving DC vaccine efficacy.

Non-small cell lung carcinoma is a leading cause of cancer mortality worldwide. There are gender and smoking associated differences both in tumour types and clinical outcome. Squamous cell carcinomas (SCC) are more frequent among smoking men while females develop adenocarcinomas (ADCA). NSCLC among never smokers are mainly ADCA, and occurs mostly in females. The present thesis elucidates the role of estrogen receptor (ER) and epidermal growth factor receptor family (EGFR/HER2-4) in NSCLC in the perspective of gender and histology as well as the influence of smoking on those biomarkers. A recently developed technique, tissue micro array (TMA), was employed.The question of how much of a tumour tissue that needed to be included in a TMA for biomarker analysis was analyzed by a statistical approach. Data indicates a sample size of three cylinders of tumour tissue with a diameter of 0.6 mm each as being appropriate and cost-effective. In order to optimally use the up to thousands of different tumour samples within a TMA, it would be optimal to serially cut and store slides before performing in situ detection of proteins and nucleic acids. Applying up to date methodology, and by evaluation with image analysis, data are presented that shows that such handling of TMA slides would be possible without any loss of biomarker information. ERα is more frequently observed in ADCA and in females and a local estradiol synthesis is supported by the presence of aromatase. ERβ is identified as a positive prognostic marker in ADCA. Smoking is associated to increased levels of ERβ mRNA. EGFR over expression is associated with a ligand. Independent phosporylation of ERα. HER-4 intracellular domain may also act as a co-activator to ERα in ADCA, especially among neversmokers. The question of ER and EGFR family signalling crosstalk as a potential target for combined targeted therapy is raised.

Title: Combinatorial Games Theory Author: Tomáš Valla Department / Institute: IUUK MFF UK Supervisor: Prof. RNDr. Jaroslav Nešetřil, DrSc., IUUK MFF UK Abstract: In this thesis we study the complexity that appears when we consider the competitive version of a certain environment or process, using mainly the tools of al- gorithmic game theory, complexity theory, and others. For example, in the Internet environment, one cannot apply any classical graph algorithm on the graph of connected computers, because it usually requires existence of a central authority, that manipu- lates with the graph. We describe a local and distributed game, that in a competitive environment without a central authority simulates the computation of the weighted vertex cover, together with generalisation to hitting set and submodular weight func- tion. We prove that this game always has a Nash equilibrium and each equilibrium yields the same approximation of optimal cover, that is achieved by the best known ap- proximation algorithms. More precisely, the Price of Anarchy of our game is the same as the best known approximation ratio for this problem. All previous results in this field do not have the Price of Anarchy bounded by a constant. Moreover, we include the results in two more fields, related to the complexity of competitive...