Introduction: Basal-like (BL) breast cancer is an aggressive sub-type of breast cancer for which there is no targeted systemic therapy. C-Met is a receptor tyrosine kinase implicated in breast cancer. Clinical trials assessing the efficacy of anti-c-Met therapy are underway, yet few studies have analysed the clinical significance of c-Met expression and/or activation in breast cancer, in particular whether there is a correlation with molecular sub-type. The aims of this study are: 1) to establish the clinical significance of c-Met expression in invasive breast cancer, 2) evaluate the novel proximity ligation assay (PLA) as a method of measuring c-Met activation and 3) address the effect of hepatocyte growth factor (HGF)-mediated c-Met phosphorylation on migration and protein expression in cell lines representative of the BL sub-type. Methods: Immunohistochemistry for c-Met was performed on 1455 cases of breast cancer using tissue microarray (TMA) technology. The PLA was performed on TMAs constructed from 181 breast cancers. C-Met expression and the PLA product were correlated with clinico-pathological parameters and survival. The effects of HGF on cell migration and protein expression were assessed using migration assays, western blots and immunofluorescent studies. Results: C-Met expression was independently associated with BL breast cancer (odds ratio = 6.44, 95% confidence interval (CI) = 1.74-23.78, p = 0.005) and reduced overall survival (hazard ratio = 1.81, 95% CI = 1.07-3.06), p = 0.026). The PLA signal was not associated with molecular sub-type or survival. HGF stimulation was associated with a significant increase in BL cell migration (p < 0.01) but no evidence of epithelial-mesenchymal transition was observed. Conclusion: My findings suggest BL breast cancer patients should be included in future trials of anti-c-Met therapy. Further work is necessary to establish the prognostic utility of the PLA as a measure of c-Met activation and the mechanisms driving HGF-mediated cell migration.

Methylation of cytosines within CpG dinucleotides is a feature of vertebrate DNA. The precise role of DNA methylation is unknown to date, although it has been implicated in several processes relating to transcriptional regulation. One approach to study DNA methylation is the characterization of proteins that bind specifically to methylated DNA. One such family of proteins is the methyl-CpG binding domain (MBD) containing family and MBD1 is a member of this family. MBD1 is implicated in transcriptional repression and various mechanisms by which it might bring about gene silencing have been proposed. These are mainly based on studies reporting interactions between MBD1 and various proteins that regulate chromatin structure. Also MBD1 function can be modified by PIAS proteins, which stimulate its conjugation to SUMO (small ubiquitinlike modifier).The original aim of this work was to address two questions about MBD1: (1) Does MBD1 form part of a stable complex with other factors, and if so, what are the identities of the other components? Purification of MBD1 revealed the presence of no stably bound interacting proteins. However, some evidence indicates MBD1 may interact with itself and form dimers, a finding which impacts on many aspects of the function of MBD1. Also a proteomics screen for transient interaction partners identified candidate binding partners for MBD1 and the related protein MeCP2, which may throw light on the function of these proteins. (2) Are there any activities which regulate MBD1 function by the removal of SUMO from this protein? No activities capable of removing SUMO from native MBD1 were found but it was demonstrated that this modification leads to the destabilization of MBD1 in vitro. The relevance of this finding in vivo is yet to be determined.

Educational games intend to make learning more enjoyable, but at the potential cost of compromising learning efficiency. Therefore, instead of creating educational games, we create learning environment with game-like elements: the elements of games that are engaging. Our approach is to assess each game-like element in terms of benefits such as enhancing engagement as well as its costs such as sensory or working memory overload, with a goal of maximizing both engagement and learning. We developed different four versions of a math tutor with different degree of being game-like such as adding narrative and visual feedback. Based on a study with 297 students, we found that students reported more satisfaction with more 'game-like' tutor but we were not able to detect any conclusive difference in learning among the different tutors. We collected student data of various types such as their attitude and enjoyment via surveys, performance within tutor via logging, and learning as measured by a pre/post-test. We created a causal model using software TETRAD and contrast the causal modeling approach to the results we achieve with traditional approaches such as correlation matrix and multiple regression. Relative to traditional approaches, we found that causal modeling did a better job at detecting and representing spurious association, and direct and indirect effects within variables. Causal model, augmented with domain knowledge about likely causal relationships, resulted in much more plausible and interpretable model. We propose a framework for blending exploratory results from causal modeling with randomized controlled studies to validate hypotheses.

Background. VAV interacting Krüppel-like factor (VIK) is a novel transcription factor. Previously our lab reported a series of breast cancer tumour samples where VIK methylation was associated with an increased risk of recurrence in tamoxifen-treated patients, indicating a role for VIK in ER positive breast cancer and endocrine resistance. Additionally VIK has been shown to be involved in cell cycle regulation, interacting with CDK4 and VAV1. The cyclin D-CDK4/6-Rb pathway is frequently dysregulated in ER positive breast cancer. Combined treatment of palbociclib, a highly selective CDK4/6 inhibitor, with endocrine therapy substantially improved outcome of patients with ER positive metastatic breast cancer. Increasing clinical use means acquired resistance to palbociclib is emerging as a major clinical challenge. Results. VIK was confirmed to be subject to regulation by DNA methylation in breast cancer. VIK methylation correlated to transcriptional silencing of mRNA in both cancer cell lines and primary tumours. To determine the functional significance of loss of VIK expression, VIK was knocked down in unmethylated breast cancer cell lines and a normal breast epithelial cell line. Knockdown resulted in cell death via induction of apoptosis. VIK knockdown altered cell cycle progression from G1 to S phase. Expression of multiple regulatory cell cycle proteins was altered, differentially in normal and tumour cells. Treatment with the CDK4/6 inhibitor, palbociclib, in cells with reduced VIK expression resulted in decreased sensitivity to the drug, inducing a shift in IC50 value towards resistance. In a model of acquired resistance, T47D cells were cultured long-term with palbociclib generating resistant clones. VIK was significantly downregulated in all resistant clones to barely detectable mRNA levels, suggesting a role for VIK in resistance to CDK4/6 inhibition. Conclusion. This PhD has confirmed VIK is a novel epigenetically regulated gene in breast cancer. VIK expression is essential to both normal and tumour breast cell survival and is involved in the regulation of the G1/S phase transition in the cell cycle. Loss of VIK expression potentially contributes to the development of acquired resistance to CDK4/6 inhibitors.

Breast cancer is the most common cancer in the UK and Basal-like breast cancer (a highly aggressive subtype) accounts for approximately 8-22% of all cases depending on ethnicity. Unlike most human malignancies and indeed other PAM50 breast cancer subtypes, the vast majority of Basal-like tumours are positive for wild type p16. This p16 signature is associated with a particularly poor prognosis and p16-positive Basal-like breast cancer remains the most clinically challenging subtype and is the focus of this project. Pro-senescence therapies are gaining momentum as attractive strategies for the treatment of those breast cancers with current unmet clinical need. To identify targets for pro-senescence therapy in p16-positive Basal-like breast cancer, a genome‐wide siRNA screen and two subsequent validation screens using two p16-positive cancer cell lines were performed. Screening revealed 20 siRNAs that induced senescence within both cancer cell lines. Strikingly, 11 of these 20 siRNAs targeted ribosomal proteins, implicating disrupted ribosomal biosynthesis in senescence activation in p16-positive Basal-like breast cancer. Importantly, subsequent experiments in normal human mammary epithelial cells established that specific ribosomal protein knockdown is well tolerated by normal cells. Analysis of the METABRIC data set showed a high degree of ribosomal dysregulation in Basal-like tumours and revealed that all 11 ribosomal hits identified were frequently overexpressed in p16-positive Basal-like breast cancers. Kaplan Meier analysis confirmed that elevated expression of six of the 11 ribosomal proteins correlates with a reduced overall survival in these women, further supporting a role for these proteins as drivers of disease. These six ribosomal hits, associated with the poorest patient survival, were prioritised for further validation. Senescence induction was found to be highly stable, and associated with dramatic changes to nucleolar morphology, reminiscent of the nucleolar signature observed upon premature senescence induction in normal human mammary epithelial cells. In addition, siRNA rescue experiments indicated that senescence initiation is dependent on p16 and p21 expression and is accompanied by p16 nuclear translocation and p21 degradation. Further, ribosomal protein silencing in MDA-MB-231 cells (p16-null Basal-like breast cancer cell line) resulted in a 'death-like' phenotype, partially dependent on p21 expression suggesting that, within a cancer context, ribosomal protein silencing may induce a differential response depending on the status of p16. In conclusion, it is proposed that these six ribosomal candidates may form the basis of a novel pro-senescence therapy for p16-positive Basal-like breast cancer. They may also represent novel prognostic biomarkers for this disease subset and may help to improve disease stratification and future directed personalised therapies.

INTRODUÇÃO: A crescente incidência de câncer de mama, que cada vez mais acomete mulheres jovens, tem despertado muito interesse no diagnóstico precoce e na busca do tratamento mais eficaz e minimamente agressivo. Da mesma forma, vem aumentando a parcela da população com alto risco para câncer de mama, para a qual as atenções têm se voltado no sentido de se encontrar um caminho que permita prevenir o surgimento da neoplasia. Inspirados por Peyrat, que primeiro associou o câncer de mama com a presença de fator de crescimento insulina like tipo 1 (IGF-1), vários autores se lançaram nesta procura e, apesar de controversa, a literatura internacional tende a mostrar uma relação direta entre IGF-1 e câncer de mama na pré-menopausa. A proteína carreadora de IGF do tipo 3, ou IGFBP-3, também foi adicionada ao rol de sustâncias com possibilidade de promover câncer, porém, não tem demonstrado clara regularidade em seu mecanismo de ação. Tendo como alvo a população com alto risco para câncer de mama, quer histopatológico, quer familiar, e direcionada pelos cálculos matemáticos de Gail e Tyrer-Cuzick, essa tese objetivou avaliar a relação entre IGF-1 e IGFBP-3 em mulheres brasileiras atendidas pelo Setor de Mastologia da Disciplina de Ginecologia do Departamento de Obstetrícia e Ginecologia da Faculdade de Medicina da Universidade de São Paulo. MÉTODOS: estudo transversal em que foram comparados os níveis séricos de IGF-1 e IGFBP-3, dosados pelo método imunométrico quimioluminescente, em 3 diferentes grupos: mulheres com câncer de mama não tratado (n= 51), mulheres com risco populacional (n= 66) e mulheres com alto risco para câncer de mama (n= 108). Foram considerados fatores de comparação: idade, estado menopausal e índice de massa corpórea. RESULTADOS: 1) não houve diferenças entre os grupos com respeito ao IMC; 2) foi estatisticamente significativa a diferença das médias de idades dos grupos, sendo as mulheres com alto risco para câncer de mama mais jovens (p<0,001); 3) quanto ao estado menopausal, houve significativa diferença entre os grupos, com predomínio de mulheres pós-menopausa no grupo de pacientes com câncer de mama (58,8%), com p <0,001; 4) não foi encontrada relação estatisticamente significante entre percentual de IGF-1 em relação à mediana, percentual de IGFBP-3 em relação à mediana e razão dos percentuais de IGF-1 e IGFPB-3 em relação à mediana para os 3 grupos estudados;4) não houve variações nas dosagens de acordo com o tipo de situação determinante do alto risco para câncer de mama (familiar ou histopatológico); 5) não houve alteração estatisticamente significativa das variáveis estudadas, mesmo após subdivisão do grupo de alto risco (risco vitalício entre 20 e 29% X risco vitalício superior a 29%). CONCLUSÃO: Não foi observada variação dos níveis séricos de IGF-I e IGFBP-3 em mulheres com câncer de mama, com alto risco para câncer de mama ou com risco populacional / INTRODUCTION: The rising incidence of breast cancer, a disease that is increasingly affecting young women, has aroused a great deal of interest in early diagnosis and the search for a more efficient and minimally aggressive treatment. Likewise, the population at high risk for breast cancer has been growing, and it is now the focus of research efforts in the struggle to prevent neoplasia. Inspired by Peyrat, who was the first to associate breast cancer with insulin-like growth factor type 1 (IGF-1), several authors have taken on the challenge, and the international literature now leans towards a direct relationship, albeit still controversial, between IGF-1 and breast cancer in premenopause. The IGF binding protein 3 (IGFBP-3) has also been added to the list of cancer-causing agents, but its mechanism of action has not shown to be clearly regular. Targeting a population at high risk for breast cancer for histopathologic or familial reasons, and guided by the mathematical calculations of Gail and Tyrer-Cuzick, this thesis aimed at evaluating the relationship between IGF-1 and IGFB-3 in Brazilian female outpatients at the Mastology Sector of the Gynecology Discipline of the Department of Obstetrics and Gynecology of the School of Medicine of the University of São Paulo. METHODS: Transverse study to compare the serum levels of IGF-1 and IGFBP- 3 measured by the chemiluminescent immunometric method. The patients were divided into 3 different groups: women with untreated breast cancer (n=51), women with a population-based risk of breast cancer (n=66), and women at high risk for breast cancer (n=108). The comparison factors were age, menopausal state, and body mass index. RESULTS: 1) there were no differences among the groups with respect to BMI; 2) but there were statistically significant differences among the groups regarding mean age, and the younger women were those at a higher risk for breast cancer (p<0.001); 3) as for menopausal state, the groups were significantly different, and the postmenopausal women were prevalent among the patients with breast cancer (58.8%; p<0.001); 4) no statistically significant relationship was found between the IGF-1 percentage and the median, the IGFBP-3 percentage and the median, or the IGF-1 and IGFBP-3 ratio and the median in any of the groups; 5) measurements did not vary according to the determinant reason for high breast cancer risk (familial or histopathologic); 6) statistically significant changes in the variables under consideration did not take place, even after subdivision of the high-risk group (lifelong risk between 20% and 29% X lifelong risk over 29%). CONCLUSION: The serum levels of IGF-I and IGFBP-3 showed no variation among women with breast cancer, at high risk for cancer, or with populationbased risk

The focus of this thesis is to examine posts on library’s Facebook pages and to investigate possible connections between posts and user response. The study examines posts published from thirty different libraries Facebook pages over a two week period to examine if any correlation between different subjects or types of posts affects the degree and type of response from the users.The findings show that photos stand for the majority of the published post concerning its type and that activities at the library was the most common subject of posts. After using eta as measure of association a tenuous connection between degree of response and the posts subject and between the degree of response and the type of post was found. The standard deviation, however, was high in the majority of the variables used. The data also showed that posts containing videos had a very high mean of user response as did the subject culture news, although either of them constituted a sufficiently large part of the examined material in order to prove a connection. Previous research also supports the connection between posting of multimedia like videos and the engagement from the users. / Program: Bibliotekarie

肥大細胞是過敏和炎症的主要效應細胞，其激活機制包括了IgE依賴性和非IgE依賴性的激活。IgE依賴性激活是指抗原與IgE的高親和力受體FcεRI上的IgE結合，促使FcεRI受體交聯而引起變態反應。其它的肥大細胞促分泌素如神經肽P物質，能夠激活百日咳毒素（PTX）敏感性的G蛋白而介導非IgE依賴性的細胞激活。最近的研究指出，肥大細胞表達Toll樣受體家族，提示肥大細胞也積極參與固有免疫反應。本研究主要探討Toll樣受體2激動劑肽聚糖（PGN）和合成激動劑Pam3CSK4對人類肥大細胞的影響，及其對抗原和P物質引起的肥大細胞激活的調控。 / Toll樣受體2激動劑本身不引起人類肥大細胞脫顆粒，但抑制抗原和P物質引起的肥大細胞脫顆粒。鈣動員是引起肥大細胞脫顆粒的關鍵因素。Pam3CSK4通過抑制抗原和P物質鈣動員來抑制肥大細胞脫顆粒。PGN只抑制抗原鈣動員，卻對P物質沒有影響。 / PGN和Pam3CSK4皆刺激人類肥大細胞釋放白細胞介素8(IL-8)和腫瘤壞死因子α（TNF-α）。Pam3CSK4通過激活G₀蛋白，Erk，Ca²⁺/calcineurin/NFAT和TAK信號通路引起肥大細胞釋放IL-8。其間，Go蛋白的激活介導Erk和Ca²⁺/calcineurin/NFAT信號通路的活化。與Pam3CSK4不同，PGN通過激活JNK, Erk, PI3K和TAK信號通路引起肥大細胞釋放IL-8。此外，雖然PTX敏感性G蛋白不影響PGN刺激引起的IL-8釋放，它卻抑制PGN刺激引起的Erk激活。 / Pam3CSK4與抗原協同作用刺激肥大細胞釋放IL-8和TNF-α，PGN與抗原卻並無協同作用。PGN與P物質協同作用刺激肥大細胞釋放IL-8和TNF-α，Pam3CSK4卻幹擾P物質的作用。在Pam3CSK4與抗原的協同作用中，Erk，Ca²⁺/calcineurin/NFAT和TAK信號通路起重要作用。PGN與P物質的協同作用則通過Erk, Ca²⁺/calcineurin/NFAT，NF-κB，PI3K和TAK這五條信號通路。 / 本研究表明，不同的Toll樣受體2激動劑能通過不同的作用機制介導和調控人類肥大細胞的反應。同時，我們首次發現G₀蛋白參與人類肥大細胞Toll樣受體2信號的激活。由於Toll樣受體2與感染和炎症息息相關，繼續研究Toll樣受體2激活對人類肥大細胞的調控機制，有助於促進開發抗感染和炎症藥物，意義深遠。 / Mast cells are activated by IgE-dependent and -independent mechanisms and play a pivotal role in both allergic and inflammatory responses. The classical IgE-dependent mechanism involves the binding of antigens to the receptor-bound IgE and crosslinking of the high-affinity receptor for IgE (FcεRI). For the poly-basic secretagogues, such as the neuropeptide substance P, they can directly stimulate pertussis toxin (PTX)-sensitive G proteins in mast cells in an IgE-independent manner. Recent studies also discover the expression of the Toll-like receptors on mast cells, indicating that mast cells are active players in innate immunity against a wide variety of pathogens. In this study, we investigated the effects of Toll-like receptor 2 (TLR2) ligands peptidoglycan (PGN) and Pam3CSK4 on human mast cell line LAD2 cells activation and the modulatory effects of these TLR2 ligands on LAD2 cells activities in response to anti-IgE and substance P. / TLR2 ligands did not cause significant degranulation on their own, but inhibited anti-IgE and substance P induced degranulation. Pam3CSK4 acted through TLR2, while the inhibitory effect of PGN involved other non-TLR2 related mechanisms. Pretreatment of Pam3CSK4 inhibited calcium mobilization induced by anti-IgE and substance P. However, pretreatment of PGN only inhibited calcium mobilization induced by anti-IgE, but failed to demonstrate similar effect on substance P. / Both TLR2 ligands triggered the release of IL-8 and TNF-α from LAD2 cells in TLR2-dependent manner. G protein, MAPKs, Ca²⁺/calcineurin/NFAT, PI3K/Akt and TAK pathways were differentially activated by PGN and Pam3CSK4. Release of IL-8 induced by Pam3CSK4 required the involvement of G₀ protein, Erk, Ca²⁺/calcineurin/ NFAT and TAK signaling pathways, but not PI3K/Akt and NF-κB. Meanwhile, G₀ protein was required for the upstream regulation of Erk and Ca²⁺/calcineurin/NFAT signaling cascades activated by Pam3CSK4. In contrast to Pam3CSK4, IL-8 release induced by PGN required the activation of JNK, Erk, PI3K and TAK signaling pathways, but not Ca²⁺ /calcineurin/NFAT and NF-κB. PTX-sensitive Gi/o protein was also involved in PGN induced Erk phosphorylation without influencing IL-8 release. / Pam3CSK4 acted in synergy with anti-IgE to augment the release of IL-8 and TNF-α, but PGN failed to demonstrate similar effect. In contrast, PGN acted in synergy with substance P, while co-stimulation of Pam3CSK4 with substance P failed to demonstrate similar synergism. Erk, Ca²⁺/calcineurin/NFAT and TAK signaling pathways were required for the synergistic action of Pam3CSK4 combined with anti-IgE, while synergistic release of IL-8 induced by PGN and substance P required the activation of Ca²⁺/calcineurin/NFAT, Erk, NF-κB, PI3K, and TAK signaling networks and was enhanced by Ca²⁺/calcineurin/NFAT and NF-κB signaling cascades in LAD2 cells, although NF-κB was not required for IL-8 release induced by PGN or substance P. / These ndings suggest that activation of human mast cells LAD2 can be differentially modified by different TLR2 ligands via distinct signaling pathways. We identify for the first time the involvement of G₀ protein in TLR2 signaling transduction in human mast cells. Further studies of the regulation of mast cells by Toll-like receptors will provide important opportunities for the therapeutic manipulation of infection and allergic diseases. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Yu, Yangyang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 205-233). / Abstract also in Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iv / Acknowledgements --- p.vi / Publication --- p.vii / Abbreviations --- p.viii / Contents --- p.x / Chapter 1 --- Introduction --- p.1 / Origin of mast cells --- p.1 / Cytokines and growth factors required for mast cells development --- p.3 / Mediators release from mast cell --- p.7 / Mast cells activation by classical IgE-dependent pathway --- p.13 / Substance P and mast cells --- p.20 / Mast cells in host defense --- p.23 / Toll-like receptors and mast cells --- p.25 / Aims --- p.31 / Chapter 2 --- Materials and Methods --- p.33 / Materials --- p.33 / Methods --- p.42 / LAD2 mast cells culture --- p.42 / Degranulation assay --- p.43 / IL-8 and TNF-α measurement --- p.44 / Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) --- p.44 / Western Blotting --- p.46 / Calcium mobilization assay --- p.47 / Flow cytometry assay --- p.48 / siRNA Transfection --- p.48 / Statistical analysis --- p.49 / Chapter 3 --- Functional Studies of Toll-Like Receptor 2 on Human Mast Cells Activation --- p.51 / Experimental conditions --- p.56 / Results --- p.57 / Discussions --- p.62 / Chapter 4 --- Modulatory Effects of Toll-Like Receptor 2 on Human Mast Cells in Response to Anti-IgE and the Signaling Pathways Involved in the Events --- p.80 / Experimental conditions --- p.92 / Results --- p.93 / Discussions --- p.102 / Chapter 5 --- Modulatory Effects of Toll-Like Receptor 2 on Human Mast Cells Activation in Response to Substance P and Signaling Pathways Involved in the Event --- p.136 / Experimental conditions --- p.140 / Results --- p.141 / Discussions --- p.152 / Chapter 6 --- General Discussion --- p.188 / Chapter 7 --- References --- p.205

INTRODUÇÃO: A crescente incidência de câncer de mama, que cada vez mais acomete mulheres jovens, tem despertado muito interesse no diagnóstico precoce e na busca do tratamento mais eficaz e minimamente agressivo. Da mesma forma, vem aumentando a parcela da população com alto risco para câncer de mama, para a qual as atenções têm se voltado no sentido de se encontrar um caminho que permita prevenir o surgimento da neoplasia. Inspirados por Peyrat, que primeiro associou o câncer de mama com a presença de fator de crescimento insulina like tipo 1 (IGF-1), vários autores se lançaram nesta procura e, apesar de controversa, a literatura internacional tende a mostrar uma relação direta entre IGF-1 e câncer de mama na pré-menopausa. A proteína carreadora de IGF do tipo 3, ou IGFBP-3, também foi adicionada ao rol de sustâncias com possibilidade de promover câncer, porém, não tem demonstrado clara regularidade em seu mecanismo de ação. Tendo como alvo a população com alto risco para câncer de mama, quer histopatológico, quer familiar, e direcionada pelos cálculos matemáticos de Gail e Tyrer-Cuzick, essa tese objetivou avaliar a relação entre IGF-1 e IGFBP-3 em mulheres brasileiras atendidas pelo Setor de Mastologia da Disciplina de Ginecologia do Departamento de Obstetrícia e Ginecologia da Faculdade de Medicina da Universidade de São Paulo. MÉTODOS: estudo transversal em que foram comparados os níveis séricos de IGF-1 e IGFBP-3, dosados pelo método imunométrico quimioluminescente, em 3 diferentes grupos: mulheres com câncer de mama não tratado (n= 51), mulheres com risco populacional (n= 66) e mulheres com alto risco para câncer de mama (n= 108). Foram considerados fatores de comparação: idade, estado menopausal e índice de massa corpórea. RESULTADOS: 1) não houve diferenças entre os grupos com respeito ao IMC; 2) foi estatisticamente significativa a diferença das médias de idades dos grupos, sendo as mulheres com alto risco para câncer de mama mais jovens (p<0,001); 3) quanto ao estado menopausal, houve significativa diferença entre os grupos, com predomínio de mulheres pós-menopausa no grupo de pacientes com câncer de mama (58,8%), com p <0,001; 4) não foi encontrada relação estatisticamente significante entre percentual de IGF-1 em relação à mediana, percentual de IGFBP-3 em relação à mediana e razão dos percentuais de IGF-1 e IGFPB-3 em relação à mediana para os 3 grupos estudados;4) não houve variações nas dosagens de acordo com o tipo de situação determinante do alto risco para câncer de mama (familiar ou histopatológico); 5) não houve alteração estatisticamente significativa das variáveis estudadas, mesmo após subdivisão do grupo de alto risco (risco vitalício entre 20 e 29% X risco vitalício superior a 29%). CONCLUSÃO: Não foi observada variação dos níveis séricos de IGF-I e IGFBP-3 em mulheres com câncer de mama, com alto risco para câncer de mama ou com risco populacional / INTRODUCTION: The rising incidence of breast cancer, a disease that is increasingly affecting young women, has aroused a great deal of interest in early diagnosis and the search for a more efficient and minimally aggressive treatment. Likewise, the population at high risk for breast cancer has been growing, and it is now the focus of research efforts in the struggle to prevent neoplasia. Inspired by Peyrat, who was the first to associate breast cancer with insulin-like growth factor type 1 (IGF-1), several authors have taken on the challenge, and the international literature now leans towards a direct relationship, albeit still controversial, between IGF-1 and breast cancer in premenopause. The IGF binding protein 3 (IGFBP-3) has also been added to the list of cancer-causing agents, but its mechanism of action has not shown to be clearly regular. Targeting a population at high risk for breast cancer for histopathologic or familial reasons, and guided by the mathematical calculations of Gail and Tyrer-Cuzick, this thesis aimed at evaluating the relationship between IGF-1 and IGFB-3 in Brazilian female outpatients at the Mastology Sector of the Gynecology Discipline of the Department of Obstetrics and Gynecology of the School of Medicine of the University of São Paulo. METHODS: Transverse study to compare the serum levels of IGF-1 and IGFBP- 3 measured by the chemiluminescent immunometric method. The patients were divided into 3 different groups: women with untreated breast cancer (n=51), women with a population-based risk of breast cancer (n=66), and women at high risk for breast cancer (n=108). The comparison factors were age, menopausal state, and body mass index. RESULTS: 1) there were no differences among the groups with respect to BMI; 2) but there were statistically significant differences among the groups regarding mean age, and the younger women were those at a higher risk for breast cancer (p<0.001); 3) as for menopausal state, the groups were significantly different, and the postmenopausal women were prevalent among the patients with breast cancer (58.8%; p<0.001); 4) no statistically significant relationship was found between the IGF-1 percentage and the median, the IGFBP-3 percentage and the median, or the IGF-1 and IGFBP-3 ratio and the median in any of the groups; 5) measurements did not vary according to the determinant reason for high breast cancer risk (familial or histopathologic); 6) statistically significant changes in the variables under consideration did not take place, even after subdivision of the high-risk group (lifelong risk between 20% and 29% X lifelong risk over 29%). CONCLUSION: The serum levels of IGF-I and IGFBP-3 showed no variation among women with breast cancer, at high risk for cancer, or with populationbased risk

A doença de Behçet tem sua fisiopatologia caracterizada por hiperatividade neutrofílica, particularmente em relação à quimiotaxia, e períodos de atividade da doença podem ser desencadeados por exposição a estreptococos. Uma vez que células do sistema imune inato são ativadas pelo ácido lipoteicoico (LTA) de bactérias gram-positivas via receptor toll-like (TLR) 2 e CD14, cujas expressões são reguladas pelos fatores estimulantes de colônias de granulócitos (G-CSF) e granulócitos-macrófagos (GM-CSF), o objetivo principal deste estudo foi determinar se há hiperexpressão de TLR2 em neutrófilos de DB ativa e se a quimiotaxia de polimorfonucleares (PMN) neutrófilos na DB poderia ser hiperestimulada pelo LTA. Além do TLR2, foram medidas as expressões de TLR4, CD14, CD114 (receptor de G-CSF) e CD116 (receptor de GM-CSF) nos neutrófilos e nos monócitos de pacientes com doença de Behçet (DB), as concentrações séricas de CD14 solúvel (CD14s) e as respostas quimiotáxicas dos PMNs de DB sob diferentes estímulos. A expressão dos receptores foi medida pela citometria de fluxo, as concentrações séricas por ELISA e as respostas quimiotáxicas foram avaliadas em câmara de Boyden. Nos PMNs, os receptores foram igualmente expressos nos dois grupos e, estimulados com LTA, suas respostas quimiotáxicas também foram similares. Somente à incubação com plasma os PMNs de DB desenvolveram hiperquimiotaxia em relação aos PMNs controles. A expressão do TLR2 foi maior em monócitos de DB em relação aos controles, e a concentração de CD14s sérica, de origem monocitária, foi maior nos pacientes com DB ativa. Em conjunto, os resultados demonstram que PMNs de DB, isoladamente, não reagem exacerbadamente ao LTA, e suas respostas migratórias são estritamente dependentes de fatores estimulantes solúveis. / Expressions of toll-like receptor (TLR) 2, TLR4, CD14, CD114 and CD116 were assessed on polymorphonuclear (PMN) neutrophils and monocytes of patients with Behçets disease (BD). PMN chemotactic responses under different stimulations were also measured. The objective was to determine if BD PMN chemotaxis may be overstimulated by lipoteichoic acid (LTA) from gram-positive bacteria. Receptor expressions were measured by flow cytometry and PMN chemotaxis was assessed in a Boyden chamber. Only TLR2 expression was higher on monocytes of the BD group than in control group. On PMNs, however, TLR2 expression was similar in both groups and, when stimulated with LTA, BD PMN cells showed chemotactic responses similar to the controls. These cells only exhibited increased chemotaxis when incubated with plasma. In conclusion, isolated BD PMN did not overreact to LTA, and its hyperchemotaxis is strictly dependent on soluble stimulating factors