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INTRODUÇÃO: A interação entre leucócitos e plaquetas é fundamental para o início e a progressão da reestenose e da aterosclerose. Recentemente foi evidenciado em estudos in vitro que a integrina Mac-1 dos leucócitos se liga à glicoproteína Ibalfa (GP Ibalfa) das plaquetas e que esta interação possui uma função central na firme adesão e transmigração de leucócitos em locais de deposição de plaquetas. Entretanto, não há estudos in vivo que avaliam a importância da interação entre a integrina Mac-1 dos leucócitos e a GP Ibalfa das plaquetas (alfaMbeta2-GP Ibalfa) em modelo experimental de lesão vascular. MÉTODO: Um peptídeo denominado M2 ou anticorpo anti-M2 foi desenvolvido para bloquear a interação da integrina Mac-1 dos leucócitos com a GP Ibalfa das plaquetas, visando, deste modo, inibir a adesão de leucócitos na superfície do vaso coberta por plaquetas, a proliferação celular e a hiperplasia neointimal. Este peptídeo foi injetado e comparado com anticorpo-controle em camundongos C57B1/6J submetidos à lesão vascular da artéria femoral com corda-guia. Um dia (controle: n= 6; anti-M2: n= 6), 5 dias (controle: n= 9; anti-M2: n= 9) ou 28 dias (controle: n= 9; anti-M2: n= 9) após a lesão vascular, as artérias femorais foram retiradas para a realização de morfometria e imunohistoquímica. RESULTADOS: O bloqueio da interação alfaMbeta2-GP Ibalfa promoveu redução estatisticamente significativa de 75% do número de leucócitos na camada média no primeiro dia após a lesão vascular (controle: 7,9 ± 5,0% do total de células na camada média; versus anti-M2: 2,0 ± 1,6%; p=0,021), bem como determinou diminuição estatisticamente significativa de 42% em 5 dias (controle: 42,3 ± 12,9% do total de células na neoíntima; versus anti-M2: 24,6 ± 10,8%; p=0,047) e de 58% em 28 dias do acúmulo de leucócitos na neoíntima em desenvolvimento (controle: 7,9 ± 3,0% versus anti-M2: 3,3 ± 1,3%; p=0,012). A proliferação celular na camada média do vaso em 5 dias pós-lesão vascular apresentou redução estatisticamente significativa de 64% com o bloqueio da interação alfaMbeta2-GP Ibalfa (controle: 5,0 ± 2,9% do total de células na camada média; versus anti-M2: 1,8 ± 0,5%; p=0,043), assim como houve diminuição significativa de 47% da proliferação celular na camada íntima do vaso em 28 dias (controle: 3,8 ± 1,7% do total de células na camada íntima; versus anti-M2: 2,0 ± 1,2%; p=0,047). O bloqueio da interação alfaMbeta2-GP Ibalfa também determinou redução estatisticamente significativa de 56% do espessamento intimal em 28 dias (controle: 10.395 ± 3.549um2; versus anti-M2: 4.561 ± 4.915um2; p=0,012). CONCLUSÕES: O recrutamento de leucócitos após a lesão vascular é dependente da interação alfaMbeta2-GP Ibalfa e a neutralização desta interação inibe a proliferação celular e a formação neointimal. / INTRODUCTION: The interaction between leukocytes and platelets is fundamental for the beginning and the progression of restenosis and atherosclerosis. Recent in vitro studies have shown that the leukocyte integrin Mac-1 binds to the platelet glycoprotein (GP) Ibalfa, and this interaction plays a central role in the leukocyte firm adhesion and transmigration at sites of platelet deposition. However, there is no in vivo study evaluating the importance of the integrin Mac-1 and GP Ibalfa (alfaMbeta2-GP Ibalfa) interaction in experimental models of vascular injury. METHODS: A peptide termed M2 or anti-M2 antibody was developed to block the leukocyte Mac-1 and platelet GP Ibalfa interaction, aiming to inhibit the adhesion of leukocytes to the platelet-coated surface of vessels as well as the cellular proliferation and the neointimal hyperplasia. The peptide was injected and compared with a control-antibody in C57B1/6J mice subjected to wire-induced femoral artery injury. One day (control: n= 6; anti-M2: n= 6), 5 days (control: n= 9; anti-M2: n= 9) or 28 days (control: n= 9; anti-M2: n= 9) after vascular injury, the femoral arteries were harvested for morphometry and immunohistochemistry. RESULTS: The alfaMbeta2-GP Ibalfa interaction blockade promoted a statistically significant 75% reduction in leukocytes in the medial layer on the first day after vascular injury (control: 7.9 ± 5.0% out of the total cells in the medial layer versus anti-M2: 2.0 ± 1.6%; p=0.021), as well as determined a statistically significant 42% decrease 5 days later (control: 42.3 ± 12.9% out of the total cells in the neointima versus anti-M2: 24.6 ± 10.8%; p=0.047), and a 58% decrease in leukocyte accumulation in the developing neointima 28 days later (control: 7.9 ± 3.0% versus anti-M2: 3.3 ± 1.3%; p=0.012). The cellular proliferation in the vessel medial layer 5 days after vascular injury presented a statistically significant 64% reduction by the alfaMbeta2-GP Ibalfa interaction blockade (control: 5.0 ± 2.9% out of the total cells in the medial layer versus anti-M2: 1.8 ± 0.5%; p=0.043), and there was also a significant 47% decrease in the vessel intimal layer cellular proliferation 28 days later (control: 3.8 ± 1.7% out of the total cells in the intimal layer versus anti-M2: 2.0 ± 1.2%; p=0.047). Furthermore, the alfaMbeta2-GP Ibalfa interaction blockade determined a statistically significant 56% reduction in the intimal thickening 28 days after vascular injury (control: 10,395 ± 3,549um2 versus anti-M2: 4,561 ± 4,915um2; p=0.012). CONCLUSIONS: The leukocyte recruitment after vascular injury depends on the alfaMbeta2-GP Ibalfa interaction, and its neutralization inhibits cellular proliferation and neointimal formation.

In einer prospektiven Studie wurden bei 28 kritisch kranken Patienten einer interdisziplinären Intensivstation die Plasmaspiegel von TNF[alpha], sCD14, Interleukin-6 (IL-6), sICAM-1 und sE-Selektin gemessen. Ziel der Studie war es, die genannten Parameter in ihrer Wertigkeit als Frühparameter der Sepsis zu untersuchen. Die Plasmaspiegel der Parameter TNF[alpha], IL-6, sCD14, sICAM-1 und sE-Selektin wurden mittels ELISA-Testkits bestimmt. Insgesamt wurde in einem Zeitraum von 11 Beobachtungstagen täglich 10 ml Blut entnommen, zentrifugiert und bis zur Verarbeitung tiefgefroren. Gruppe 1:Patienten mit einer mikrobiellen Infektion, die im Beobachtungszeitraum keine Sepsis entwickelten. Alle Patienten dieser Gruppe überlebten,Gruppe 2:Patienten mit einer mikrobiellen Infektion, die im Beobachtungszeitraum eine Sepsis mit Organdysfunktion entwickelt haben und überlebten, / In a prospective study, we determined the plasma levels of TNF[alpha], sCD14, Interleukin-6 (IL 6), sICAM-1 and sE-Selectin of 28 critically ill patients on our interdisciplinary intensive care unit. The aim of our study was to find out if these parameters are valuable for the early diagnosis of septicaemia. Plasma levels of TNF[alpha], IL-6, sCD14, sICAM-1 and sE-Selectin were measured with ELISA-test-kits. In a period of 11 days, we took 10 ml of blood daily which was refrigerated until examination. GROUP 1:patients with bacterial infections who did not develop septicaemia. All of these patients survived.GROUP 2:patients with bacterial infections who presented with symptoms of disturbed organic function within the examination period and survived.GROUP 3: patients with bacterial infections who developped symptoms of severe septicaemia and died because of multiple organic failure. Results: In patients with septicaemia, TNF[alpha]-levels were significantly higher than in patients without septicaemia. TNF[alpha]-levels can not be used as prognostic parameters in septicaemia because of the short half-life-time.sCD14-levels were significantly higher in patients with septicaemia in the first two days of observation. sCD14-levels can not be used as a prognostic criteria in septicaemia.In patients with septicaemia, we found significant higher Interleucin-6-levels compareed to patients without septicaemia. IL6 prooved to be a good marker for septicaemia. In combination with plasma levels of Se_Selectin, it is criteria for severity of the septicaemia and propable outcome of patients.Pathologically high plasma levels of sICAM-1 were measured in patients with septicaemia. S-ICAM-1 is an early indicator for activation of withe blood cells and danger of septicaemia. The exact blood level of s-ICAM-1 did not correlate with the outcome of patients.sE-Selectin-levels were significantly higher in patients with septicaemia than in patients without septicaemia. The persistence of high sE-Selectin levels indicates possible septicaemia early and is correlated with the outcome of patients.

Ischemia followed by reperfusion, as happens in myocardial infarction, or the development of acute respiratory distress syndrome, are associated with a exaggerated extravasation of leukocytes into the surrounding tissue , which may cause severe bystander damage. In animal models of these diseases, pharmacological blockage of the leukocyte adhesion molecule, L-selectin (CD62L), has been shown to be partially protective by reduction or inhibition of leukocyte-mediated secondary tissue damage. The aim of this project was the development of an in vitro assay to investigate the relative effectiveness of potential L-selectin antagonists. Ideally, the assay should require low sample volumes and allow for measurements of larger series of reagents. The assay system investigated was based on the homotypic granulocyte aggregation under shear stress, which mimicks the L-selectin-dependent adhesion of leukocytes to previously arrested neutrophils on vascular endothelium. After optimizing numerous variables of the aggregation assay, the requirement of L-selectin for the homotypic granulocyte aggregation induced was demonstrated by inhibition experiments using soluble L-selectin or monoclonal antibodies directed against the lectin domain of L-selectin. Several carbohydrate polymers with L-selectin binding properties, such as the seaweed-derived fucose polymer fucoidin, high-molecular-weight dextran sulfate or heparin, also inhibited neutrophil aggregation in a dose-dependent fashion. However, despite employing a flow cytometry-based read-out technique, the assay remained extremely labor intensive, precluding investigations of extended series. Therefore, the homotypic aggregation experiments with freshly isolated human granulocytes remains a useful tool to further evaluate specific questions of L-selectin dependent adhesion processes, but it is not apt for transfer into routine laboratories.

INTRODUÇÃO: Eventos vasculares e imunológicos são centrais na patogênese da granulomatose com poliangeíte (GPA). Moléculas de adesão celular tem papel fundamental no recrutamento de células inflamatórias do sangue para os tecidos. Diferentes leitos vasculares apresentam particularidades na expressão de moléculas de adesão celular explicando talvez parte da especificidade da GPA por determinados órgãos. A elevação no nível sérico de moléculas de adesão celular e aumento da expressão destas em amostras de biópsia renal já foram demonstrados em pacientes com vasculite ANCA associada. No entanto, o fenômeno ainda não foi estudado in situ no pulmão. OBJETIVO: O objetivo deste estudo foi analisar o padrão de expressão endotelial pulmonar de três moléculas de adesão celular na GPA, in situ: molécula de adesão intercelular 1 (ICAM-1), molécula e adesão vascular 1 (VCAM-1) e E-selectina. MÉTODOS: Examinou-se a expressão endotelial de ICAM-1, VCAM-1, E-selectina usando marcação imuno-histoquímica em secções de pulmão fixadas e parafinadas de lesões de GPA (n = 8 casos, 90 secções transversais de vasos analisados). Foram também analisados controles positivos: amostras de pulmão de doença intersticial associada à esclerodermia (SScl) (n = 8 casos, 96 secções transversais de vasos analisados) e controles negativos (n = 9 casos, 90 secções transversais de vasos analisados). A quantificação imuno-histoquímica foi realizada no aumento de 400x usando a técnica de point-counting. RESULTADOS: ICAM-1: A expressão endotelial mediana de ICAM-1 esteve aumentada de forma semelhante na GPA e na SScl (81% and 73%, respectivamente; p = 0.97). A comparação com o grupo controle (26.3%) revelou diferença estatisticamente significativa entre controle e GPA (p <0.001) quanto entre controle e SScl (p = 0.017). VCAM-1: A expressão mediana de VCAM-1 esteve significativamente aumentada na GPA se comparada a SScl (79.5% vs 41.4%; p = 0.012), no entanto, a expressão endotelial de VCAM-1 nos controles também esteve moderadamente aumentada (49.8%) e não houve diferença estatística entre SScl e controles ( p = 0.549) ou entre GPA e controles (p = 0.242). E-selectina: A expressão endotelial mediana de E-selectina esteve aumentada de forma semelhante na GPA e SScl (100% e 88.2%, respectivamente; p = 0.272). A comparação com o grupo controle (13.8%) revelou diferença estatisticamente significativa entre controle e GPA (p < 0.001) e controle e SScl (p = 0.045). CONCLUSÃO: Esses achados evidenciam o fenômeno de ativação endotelial pulmonar in situ em lesões de GPA. O perfil de expressão de moléculas de adesão parece ter particularidades em diferentes doenças a exemplo da maior expressão de VCAM-1 na GPA em relação à SScl. Essas observações contribuem para o conhecimento fisiopatogênico na GPA. / INTRODUCTION: Vascular and immunologic processes are central to the pathogenesis of granulomatosis with polyangiitis (GPA). Endothelial cellular adhesion molecules have a central role in recruiting leukocytes to sites of inflammation. Moreover, different vascular beds are phenotypically and functionally distinct with regard to expression of cellular adhesion molecules. They have been shown to be elevated in sera and in renal biopsies of patients with active ANCA-associated vasculitis. Despite of that, the expression of cellular adhesion molecules has not been studied in situ in the lungs. OBJECTIVE: Within this context, the aim of this study was to analyze the in situ pulmonary endothelial immunohistochemical pattern of expression of three cellular adhesion molecules in GPA: intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and E-selectin . METHODS: For such, we examined formalin-fixed, paraffin-embedded sections of lung lesions of GPA (n = 8 cases, 90 transverse sections of pulmonary vessels), negative controls which were obtained from autopsies ( n = 9 cases, 90 transverse sections of pulmonary vessels analyzed) and positive controls which were cases of interstitial lung disease associated with systemic sclerosis (SScl) (n = 8 cases, 96 transverse sections of pulmonary vessels). The quantification of the immunohistochemical staining was performed at x400 magnification using the technique of point-counting, previously described. RESULTS: ICAM-1: Median endothelial expression of ICAM-1 was similarly enhanced in GPA and SScl (81% and 73%, respectively; p = 0.97). When compared to controls (26.3%), both GPA (p <0.001) and SScl (p = 0.017) had significantly higher endothelial expression of ICAM-1. VCAM-1: Median endothelial expression of VCAM-1 was significantly enhanced in GPA when compared to SScl (79.5% vs 41.4%; p = 0.012), however the endothelial expression of VCAM-1 in the controls was also moderately enhanced (49.8%) and no statistically significant difference was found between SScl and controls ( p = 0.549) or between GPA and controls (p = 0.242). E-selectin: Median endothelial expression of E-selectin was similarly enhanced in GPA and SScl (100% and 88.2%), respectively; p = 0.272. When compared to controls (13.8%), both GPA (p < 0.001) and SScl (p = 0.045) had significantly higher endothelial expression of E-selectin. CONCLUSION: These observations are evidence of in situ pulmonary endothelial activation in lesions of GPA. The profile of expression of cellular adhesion molecules seems to be particular for each disease state and timing as evidenced by the enhanced expression of VCAM-1 in GPA when compared to SScl. These observations add information to the pathogenetic knowledge of GPA

INTRODUÇÃO: Eventos vasculares e imunológicos são centrais na patogênese da granulomatose com poliangeíte (GPA). Moléculas de adesão celular tem papel fundamental no recrutamento de células inflamatórias do sangue para os tecidos. Diferentes leitos vasculares apresentam particularidades na expressão de moléculas de adesão celular explicando talvez parte da especificidade da GPA por determinados órgãos. A elevação no nível sérico de moléculas de adesão celular e aumento da expressão destas em amostras de biópsia renal já foram demonstrados em pacientes com vasculite ANCA associada. No entanto, o fenômeno ainda não foi estudado in situ no pulmão. OBJETIVO: O objetivo deste estudo foi analisar o padrão de expressão endotelial pulmonar de três moléculas de adesão celular na GPA, in situ: molécula de adesão intercelular 1 (ICAM-1), molécula e adesão vascular 1 (VCAM-1) e E-selectina. MÉTODOS: Examinou-se a expressão endotelial de ICAM-1, VCAM-1, E-selectina usando marcação imuno-histoquímica em secções de pulmão fixadas e parafinadas de lesões de GPA (n = 8 casos, 90 secções transversais de vasos analisados). Foram também analisados controles positivos: amostras de pulmão de doença intersticial associada à esclerodermia (SScl) (n = 8 casos, 96 secções transversais de vasos analisados) e controles negativos (n = 9 casos, 90 secções transversais de vasos analisados). A quantificação imuno-histoquímica foi realizada no aumento de 400x usando a técnica de point-counting. RESULTADOS: ICAM-1: A expressão endotelial mediana de ICAM-1 esteve aumentada de forma semelhante na GPA e na SScl (81% and 73%, respectivamente; p = 0.97). A comparação com o grupo controle (26.3%) revelou diferença estatisticamente significativa entre controle e GPA (p <0.001) quanto entre controle e SScl (p = 0.017). VCAM-1: A expressão mediana de VCAM-1 esteve significativamente aumentada na GPA se comparada a SScl (79.5% vs 41.4%; p = 0.012), no entanto, a expressão endotelial de VCAM-1 nos controles também esteve moderadamente aumentada (49.8%) e não houve diferença estatística entre SScl e controles ( p = 0.549) ou entre GPA e controles (p = 0.242). E-selectina: A expressão endotelial mediana de E-selectina esteve aumentada de forma semelhante na GPA e SScl (100% e 88.2%, respectivamente; p = 0.272). A comparação com o grupo controle (13.8%) revelou diferença estatisticamente significativa entre controle e GPA (p < 0.001) e controle e SScl (p = 0.045). CONCLUSÃO: Esses achados evidenciam o fenômeno de ativação endotelial pulmonar in situ em lesões de GPA. O perfil de expressão de moléculas de adesão parece ter particularidades em diferentes doenças a exemplo da maior expressão de VCAM-1 na GPA em relação à SScl. Essas observações contribuem para o conhecimento fisiopatogênico na GPA. / INTRODUCTION: Vascular and immunologic processes are central to the pathogenesis of granulomatosis with polyangiitis (GPA). Endothelial cellular adhesion molecules have a central role in recruiting leukocytes to sites of inflammation. Moreover, different vascular beds are phenotypically and functionally distinct with regard to expression of cellular adhesion molecules. They have been shown to be elevated in sera and in renal biopsies of patients with active ANCA-associated vasculitis. Despite of that, the expression of cellular adhesion molecules has not been studied in situ in the lungs. OBJECTIVE: Within this context, the aim of this study was to analyze the in situ pulmonary endothelial immunohistochemical pattern of expression of three cellular adhesion molecules in GPA: intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and E-selectin . METHODS: For such, we examined formalin-fixed, paraffin-embedded sections of lung lesions of GPA (n = 8 cases, 90 transverse sections of pulmonary vessels), negative controls which were obtained from autopsies ( n = 9 cases, 90 transverse sections of pulmonary vessels analyzed) and positive controls which were cases of interstitial lung disease associated with systemic sclerosis (SScl) (n = 8 cases, 96 transverse sections of pulmonary vessels). The quantification of the immunohistochemical staining was performed at x400 magnification using the technique of point-counting, previously described. RESULTS: ICAM-1: Median endothelial expression of ICAM-1 was similarly enhanced in GPA and SScl (81% and 73%, respectively; p = 0.97). When compared to controls (26.3%), both GPA (p <0.001) and SScl (p = 0.017) had significantly higher endothelial expression of ICAM-1. VCAM-1: Median endothelial expression of VCAM-1 was significantly enhanced in GPA when compared to SScl (79.5% vs 41.4%; p = 0.012), however the endothelial expression of VCAM-1 in the controls was also moderately enhanced (49.8%) and no statistically significant difference was found between SScl and controls ( p = 0.549) or between GPA and controls (p = 0.242). E-selectin: Median endothelial expression of E-selectin was similarly enhanced in GPA and SScl (100% and 88.2%), respectively; p = 0.272. When compared to controls (13.8%), both GPA (p < 0.001) and SScl (p = 0.045) had significantly higher endothelial expression of E-selectin. CONCLUSION: These observations are evidence of in situ pulmonary endothelial activation in lesions of GPA. The profile of expression of cellular adhesion molecules seems to be particular for each disease state and timing as evidenced by the enhanced expression of VCAM-1 in GPA when compared to SScl. These observations add information to the pathogenetic knowledge of GPA

Syngeneic graft-versus-host disease (SGVHD) is induced by reconstituting lethally irradiated mice with syngeneic BM cells followed by a 21 day treatment with the immunosuppressive agent cyclosporine A (CsA). Clinical symptoms of the disease appear 2-3 weeks following cessation of CsA therapy and disease-associated inflammation occurs primarily in the colon and liver. The development of SGVHD is a complex process resulting from the cooperative interaction of multiple effector cell populations including NK cells, T cells and macrophages. TH1 cytokines (IL-12, TNF-α, IFN- γ), produced by these effector cells, serve as inflammatory mediators contributing to the pathogenesis of SGVHD. The SGVHD conditioning agents, irradiation and CsA, are both required for the development of disease and contribute to the production of oxidative stress. Time course studies revealed increased reactive oxygen and nitrogen species (ROS/RNS), as well as, increased colon mRNA levels for TNF-α and iNOS in CsA-treated versus control BMT animals. Since ROS/RNS are known to mediate CsA toxicity, studies were undertaken to determine the effect of oxidative stress on the induction of SGVHD. In vivo treatment with the antioxidant MnTBAP caused a reduction in colon mRNA levels for iNOS and TNF-α, as well as delayed disease development, suggesting a role for oxidative stress in the development of SGVHD. In addition, CD4+ T cells have been shown to play an important role in the inflammatory response observed in the gut of SGVHD mice. Time course studies revealed significant increases in the migration of CD4+ T cells as early as day 14 post- BMT into the colon of CsA mice as well as significant elevated mRNA levels of cell adhesion molecules. Homing studies revealed that a labeled CD4+ T cell line, generated from SGVHD mice, migrated in larger numbers into the gut of CsA-treated mice compared to control animals. This study demonstrated that CD4+ T cells responsible for the pathogenesis observed in murine SGVHD are present early after BMT in colons of CsA-treated mice, suggesting that during the 21 days of immunosuppression therapy functional mechanisms are in place that result in increased homing of effector cells to colons of CsA-treated mice.

Introdução: O Câncer de próstata (CaP) é o tumor mais comum e a segunda causa de óbito por câncer no homem. O diagnóstico é feito pelo toque retal, nível sérico do antígeno prostático específico (PSA) e biópsia. A avaliação prognóstica é definida pelo PSA, estádio e escore de Gleason, porém, nenhum destes fatores clássicos, mesmo quando avaliados em conjunto, tem excelente performance na determinação prognóstica desta neoplasia. O CD44 é uma glicoproteína transmembrana envolvida em múltiplos processos celulares entre eles a adesão, migração, transmissão de sinal e apoptose. Tem como seu principal ligante o ácido hialurônico sendo assim um importante elemento na interação entre a célula e a matriz e por isso tem sido estudada como potencial marcador em várias neoplasias. Além da forma padrão (CD44s), apresenta ainda dez isoformas resultantes de splicing. São poucos os estudos de CD44 no CaP, principalmente relacionados a expressão de suas isoformas. Objetivo: Analisar e correlacionar a expressão de CD44s e de suas isoformas CD44v2, CD44v3, CD44v4, CD44v5, CD44v6, CD44v7, CD44v8, CD44v9 e CD44v10 no câncer localizado de próstata com os fatores prognósticos clássicos. Material e métodos: Analisamos retrospectivamente os espécimes cirúrgicos de 94 pacientes submetidos a prostatectomia radical (PR) entre outubro de 1997 e abril 2002. A expressão da forma standard, assim como das suas oito variantes resultado de splicing alternativo foi analisada através da reação em cadeia da polimerase quantitativa em tempo real (qRT-PCR). O grupo controle foi constituído pelos espécimes cirúrgicos de 14 pacientes com hiperplasia prostática benigna (HPB) submetidos a cirurgia aberta. Correlacionamos o perfil de expressão da forma padrão e suas variantes com escore de Gleason, estadiamento patológico, nível de PSA pré-operatório e recidiva bioquímica definida por níveis de PSA acima de 0,2ng/ml em seguimento médio de 53,3 meses. Resultados: Encontramos subexpressão de CD44s e superexpressão de todas as isoformas resultado de splicing alternativo no CaP localizado quando comparado a HPB. Observamos que a média de expressão da maioria das variantes foi maior nos pacientes que não recidivaram, porém apenas a maior expressão de CD44v2 esteve significativamente associada a menor chance de recidiva bioquímica (p=0,044). Conclusão: No CaP, existe uma alteração na expressão de CD44, com perda de CD44s e aumento de expressão de todas as suas variantes. No entanto, a recorrência tumoral mostrou-se significativamente associada a perda da expressão de CD44v2 / Introduction: Prostate cancer (PC) is the most common tumor and the second leading cause of cancer death in men. The diagnosis is made by rectal examination, serum prostate-specific antigen (PSA) and biopsy. The prognosis is defined by the PSA, tumor stage and Gleason escore, however, none of these classical factors, even when combined, have excellent accuracy in determining tumor behavior. The CD44 is a transmembrane glycoprotein related to many cell processes including adhesion, migration, signaling transduction and apoptosis, being considered a potential marker in many tumors. Besides the standard form (CD44s) there are ten isoforms resulted from alternative splicing. There is scarce data in the literature regarding the role of CD44 and its isoforms in PC prognosis. Objective: To analyze the expression of the CD44s and its isoforms CD44v2, CD44v3, CD44v4, CD44v5, CD44v6, CD44v7, CD44v8, CD44v9 and CD44v10 in in surgical specimens of localized PC correlating the expression profile with the classical prognostic factors and biochemical recurrence. Methods: We retrospectively analyzed the surgical specimens of 94 patients that underwent radical prostatectomy between October 1997 and April 2002. The standard form as well as its nine variants were analyzed by quantitative, real time polymerase chain reaction (qRT-PCR). The control group consisted of 14 specimens of patients with benign prostatic hyperplasia (BPH) submitted to open surgery. We correlated the expression of the variants with the Gleason escore, pathological stage, pre-operatory PSA and the occurrence of biochemical recurrence defined as PSA higher than 0.2 ng/ml in a mean follow up of 53.3 months. Results: CD44s was underexpressed, and all isoforms were overexpressed in PC when compared with BPH. However, the mean expression of most variants was higher in patients without recurrence, while a higher expression of CD44v2 correlated with higher recurrence free survival rate (p=0.044). Conclusion: In PC, there is a change in CD44 expression with a loss of CD44s and an overexpression of all of the CD44 variants. We also found that biochemical recurrence was significantly associated with loss of CD44v2 expression.

RESUMO STOCCO, B. Avaliação do efeito de contraceptivos hormonais sobre a hemostasia. 2011. 92f. Dissertação (Mestrado). Faculdade de Ciências Farmacêuticas de Ribeirão Preto Universidade de São Paulo, Ribeirão Preto, 2011. Introdução: Mais de 100 milhões de mulheres no mundo fazem uso de métodos contraceptivos hormonais. Apesar de seu desejado efeito na contracepção, sua metabolização ocorre no fígado estimulando a síntese de proteínas plasmáticas, dentre elas, as que controlam os sistemas de coagulação e fibrinólise, conferindo um estado de hipercoagulabilidade em suas usuárias. A literatura aponta também alterações no metabolismo dos lipídios e carboidratos, além de modular a expressão de moléculas de adesão presentes no endotélio. Para combater os efeitos indesejáveis destes medicamentos, baixas concentrações de estrógenos e diferentes progestágenos foram introduzidos, pois, a estes últimos é conferida a atividade anti estrogênica destas formulações. Estudos independentes sugerem que progesteronas de terceira e quarta geração possuem atividade anti-estrogênica menor em relação as de segunda geração. Objetivos: avaliar a ocorrência de alterações hemostáticas, analisar o perfil lipídico e quantificar moléculas de adesão no soro ou plasma da população feminina brasileira usuária de contraceptivos orais de segunda e quarta gerações. Materiais e Métodos: 70 participantes distribuídas em quatro grupos a saber : grupo I (controle- 20 pacientes); grupo II (DRSP/30EE- 20 pacientes); grupo III (DRSP/20EE- 16 pacientes) e grupo IV (LNG/30EE-14 pacientes). Foram avaliados os seguintes parâmetros: TP, Fator VII, TTPA, Fator XII, Fibrinogênio, Fator 1+2, Proteína C, Proteína S, Antitrombina, D dímeros, PAI 1, VCAM, ICAM, E- selectina, HDL, LDL,VLDL, Colesterol total e Triglicérides. Resultados e discussões: grupo II promoveu diminuição em TP e Prot. S; e aumento em HDL,VLDL,Col. total e TG; grupo III diminuiu TP, TTPA, Prot.S, ICAM e VCAM ; e aumentou Fibrinogênio, D-dímeros, HDL,VLDL,Col. total e TG e grupo IV diminuiu TP, Prot.C e aumentou ICAM e VCAM. Conclusões: Dos medicamentos estudados apontamos que: o grupo II promoveu alterações significativas no perfil lipídico caracterizando um estado pró-trombótico, apesar de apresentar o maior aumento nos valores de HDL e poucas alterações associadas à hipercoagulabilidade; o grupo III promoveu o maior número de alterações hipercoagulantes, alterou também perfil lipídico contribuindo para um estado pró- trombótico, embora tenha apresentado proteção endotelial e o grupo IV foi o medicamento que promoveu melhor proteção endotelial, não alterou perfil lipídico e ocasionou poucas alterações associadas à hipercoagulabilidade / Introduction: More than 100 million women in the world make use of hormonal contraceptives. In spite of its desired effect on contraception, its metabolization occurs at liver, estimulating the synthesis of the plasmatic proteins, among them, the ones that control the coagulation system and fibrinolysis, confering a hypercoagulability state in their users. The literature also points changes in the metabolism of lipids and carbohydrates, besides modulate the expression of adhesion molecules present in the endothelium. Trying to combat undesirable effects of these drugs, low concentrations of estrogen and different progestogens were introduced, because, the antiestrogenic activity of these drugs is assigned to the progesterone used. Independent papers suggest that third and fourth generation progesterones have a smaller antiestrogenic activity in relation to the second generation one. Objectives: evaluate the occurrence of hemostatic alterations, analyze the lipid profile and quantify adhesion molecules in serum or plasma on the Brazilian female population who is user of contraceptives from second and fourth generation. Material and Methods: 70 participants were distributed into four groups: group I (control- 20 patients); group II (DRSP/30EE- 20 patients); group III (DRSP/20EE- 16 patients) and group IV (LNG/30EE- 14 patients). From these were assessed the following parameters: PT, Factor VII, APTT, Factor XII, Fibrinogen, Factor 1+2, Protein C, Protein S, Antithrombin, Ddimmers, PAI 1,VCAM, ICAM, E- selectin, HDL, LDL, VLDL, Total Cholesterol and Triglycerides.Results and discussion: group II promoted a decrease in PT and Prot.S; and an increase in HDL,VLDL, Total cholesterol and TG; group III decreased PT, APTT, Prot.S, ICAM and VCAM ; and increased Fribrinogen, D- dimers, HDL,VLDL,Total Cholesterol and TG and group IV decreased PT, Prot.C and increased ICAM and VCAM.Conclusions: among the drugs studied we aim that: the group II promoted significant changes in lipid profile featuring a pro- thrombotic state, in spite of presented the highest increase in the levels of HDL and few alterations associated to hypercoagulability; group III promoted the biggest number of hypercoagulability alterations, this drug also changed the lipid profile contributing to a pro-thrombotic state, although it has presented endothelial protection and the group IV it was the drug that promoted the best endothelial protection, did not change lipid profile and caused few alterations associated with hypercoagulability

O Glioblastoma multiforme (GBM) é o mais maligno tumor cerebral e apresenta uma alta capacidade proliferativa, invasiva e angiogênica. Neste estudo foi avaliada a distribuição espacial de moléculas envolvidas nesses processos num modelo de GBM de rato in vivo (C6). Por imunohistoquímicas observou-se que o tumor foi capaz de expressar diferentes elementos de matriz extracelular (colágenos I, III e IV, fibronectina, tenascinas C e R, vitronectina e proteoglicanos), moléculas de adesão (RHAMM, CD44 e integrinas) e a enzima proteolítica MMP-2. Esses elementos parecem se organizar num padrão pericelular nas células invasivas das bordas do tumor e nas células C6 migrando pelos vasos. A expressão de moléculas de adesão mostrou que o RHAMM pode ter um papel mais importante que o CD44 no reconhecimento do ácido hialurônico. A expressão de flt-1 e flk-1 pelas células tumorais sugere um papel desses receptores do VEGF não apenas na proliferação endotelial, mas também na proliferação tumoral. Todas as moléculas aqui analisadas são potenciais alvos terapêuticos futuros. / Glioblastoma multiforme (GBM) is the most malignant brain tumour and presents high proliferative, invasive and angiogenic capacities. In the present study the spatial distribution of molecules involved with these processes was analyzed in a rat GBM (C6) in vivo. Immunohistochemical analysis showed that tumours expressed extracellular matrix elements (collagens I, III and IV, fibronectin, tenascin C and R, vitronectin and proteoglycans), adhesion molecules (RHAMM, CD44 and integrins) and the proteolytic enzyme MMP-2. At the tumour borders and around the cells migrating along blood vessels these molecules seemed to organize in a pericellular pattern. The adhesion molecules analysis showed that RHAMM could have a pivotal role in hyaluronic acid recognition. Flt-1 and flk-1 expression by tumour cells suggests a role for these VEGF receptors not only in endotheliocyte metabolism, but also in tumour proliferation. All of the studied molecules are potential targets for anti-cancer therapies.