Response of high-risk of recurrence/progression bladder tumours expressing sialyl-Tn and sialyl-6-T to BCG immunotherapy

High risk of recurrence/progression bladder tumours is treated with Bacillus Calmette-Guérin (BCG) immunotherapy after complete resection of the tumour. Approximately 75% of these tumours express the uncommon carbohydrate antigen sialyl-Tn (Tn), a surrogate biomarker of tumour aggressiveness. Such changes in the glycosylation of cell-surface proteins influence tumour microenvironment and immune responses that may modulate treatment outcome and the course of disease. The aim of this work is to determine the efficiency of BCG immunotherapy against tumours expressing sTn and sTn-related antigen sialyl-6-T (s6T). METHODS: In a retrospective design, 94 tumours from patients treated with BCG were screened for sTn and s6T expression. In vitro studies were conducted to determine the interaction of BCG with high-grade bladder cancer cell line overexpressing sTn. RESULTS: From the 94 cases evaluated, 36 had recurrence after BCG treatment (38.3%). Treatment outcome was influenced by age over 65 years (HR=2.668; (1.344-5.254); P=0.005), maintenance schedule (HR=0.480; (0.246-0.936); P=0.031) and multifocality (HR=2.065; (1.033-4.126); P=0.040). sTn or s6T expression was associated with BCG response (P=0.024; P<0.0001) and with increased recurrence-free survival (P=0.001). Multivariate analyses showed that sTn and/or s6T were independent predictive markers of recurrence after BCG immunotherapy (HR=0.296; (0.148-0.594); P=0.001). In vitro studies demonstrated higher adhesion and internalisation of the bacillus to cells expressing sTn, promoting cell death. CONCLUSION: s6T is described for the first time in bladder tumours. Our data strongly suggest that BCG immunotherapy is efficient against sTn- and s6T-positive tumours. Furthermore, sTn and s6T expression are independent predictive markers of BCG treatment response and may be useful in the identification of patients who could benefit more from this immunotherapy.

Culturas, políticas e novas identidades: uma leitura europeia sobre questões de género na Ásia

Este ensaio discute algumas leituras críticas de textos teóricos da área das ciências sociais e humanas sobre o estatuto de género em países asiáticos, tentando estabelecer quais as suas principais problemáticas e metodologias. Presta especial atenção à questão das vozes femininas silenciadas e das práticas ignoradas do quotidiano das mulheres, problematizando o que sucede – ou pode suceder – quando às mulheres é permitido não só possuir um espaço social próprio (“a room of their own”, para citar Virginia Woolf), mas também uma voz própria. Para Edward Said o conceito ocidental de orientalismo implicava uma concepção masculina particular do mundo, mais evidente em romances e diários de viagem, onde as mulheres eram geralmente criaturas da fantasia masculina de poder. Esta concepção masculina do mundo oriental tende a ser estática, construindo-se assim o estereótipo do “eterno oriental”. As mulheres, tal como o “oriental”, nunca falam de si mesmos, das suas verdadeiras emoções, desejos e histórias: têm de ser representados, alguém tem de falar por si. No âmbito deste estudo, analisam-se alguns processos ideológicos e retóricos através dos quais a identidade das mulheres é construída e representada, tanto pelas próprias mulheres, como por vozes substitutas. A etnografia, a antropologia, a historiografia, a ficção, a cultura popular, os media e todos os tipos de fontes textuais e visuais desempenham um papel de relevo na invenção e na reinvenção de antigas e de novas identidades femininas, e na circulação destas no tempo e no espaço.

“To laugh in tragic joy” – a linguagem poética de W. B. Yeats em Lapis Lazuli

Artigo em texto integral no link da versão do editor

Efeito da ligadura funcional na prevenção da recorrência da entorse do tornozelo numa equipa de basquetebol feminino

Introdução: O basquetebol é considerado um desporto de alto risco para a ocorrência de lesões, nomeadamente das entorses do tornozelo. Como a história de lesão anterior é um fator de risco para a ocorrência de entroses têm-se desenvolvido estratégias de prevenção para evitar as recidivas, um dos métodos possíveis é o recurso a ligaduras funcionais em “tape”. Objetivo: Neste trabalho vai-se verificar se as ligaduras funcionais em “tape” previnem a recorrência de entorse do tornozelo, numa equipa feminina de basquetebol sub-16. Métodos: A amostra é composta por 9 atletas de basquetebol sub-16 com história de entorse no tornozelo na época precedente, vão-se utilizar ligaduras funcionais no tornozelo. Resultados: A entorse teve 9 registos na época anterior. Não houve registo de qualquer lesão da tibiotársica nos treinos ou jogos, durante a aplicação das ligaduras. Conclusão: A utilização das ligaduras funcionais no grupo estudado, como método de prevenção foi eficaz, não se registando qualquer entorse no decorrer das atividades desportivas (treinos/jogos).

FASL polymorphism is associated with response to bacillus Calmette-Guérin immunotherapy in bladder cancer

Objective Deregulation of FAS/FASL system may lead to immune escape and influence bacillus Calmette-Guérin (BCG) immunotherapy outcome, which is currently the gold standard adjuvant treatment for high-risk non–muscle invasive bladder tumors. Among other events, functional promoter polymorphisms of FAS and FASL genes may alter their transcriptional activity. Therefore, we aim to evaluate the role of FAS and FASL polymorphisms in the context of BCG therapy, envisaging the validation of these biomarkers to predict response. Patients and methods DNA extracted from peripheral blood from 125 patients with bladder cancer treated with BCG therapy was analyzed by Polymerase Chain Reaction—Restriction Fragment Length Polymorphism for FAS-670 A/G and FASL-844 T/C polymorphisms. FASL mRNA expression was analyzed by real-time Polymerase Chain Reaction. Results Carriers of FASL-844 CC genotype present a decreased recurrence-free survival after BCG treatment when compared with FASL-844 T allele carriers (mean 71.5 vs. 97.8 months, P = 0.030) and have an increased risk of BCG treatment failure (Hazard Ratio = 1.922; 95% Confidence Interval: [1.064–3.471]; P = 0.030). Multivariate analysis shows that FASL-844 T/C and therapeutics scheme are independent predictive markers of recurrence after treatment. The evaluation of FASL gene mRNA levels demonstrated that patients carrying FASL-844 CC genotype had higher FASL expression in bladder tumors (P = 0.0027). Higher FASL levels were also associated with an increased risk of recurrence after BCG treatment (Hazard Ratio = 2.833; 95% Confidence Interval: [1.012–7.929]; P = 0.047). FAS-670 A/G polymorphism analysis did not reveal any association with BCG therapy outcome. Conclusions Our results suggest that analysis of FASL-844 T/C, but not FAS-670 A/G polymorphisms, may be used as a predictive marker of response to BCG immunotherapy.

Visualização e manipulação de informação em ambientes 3D imersivos

Ainda antes da invenção da escrita, o desenho foi utilizado para descrever a realidade, tendo evoluído ao longo dos tempos, ganhando mais qualidade e pormenor e recorrendo a suportes cada vez mais evoluídos que permitissem a perpetuação dessa imagem: dessa informação. Desde as pinturas rupestres, nas paredes de grutas paleolíticas, passando pelos hieróglifos, nos templos egípcios, nas gravuras das escrituras antigas e nos quadros sobre tela, a intenção sempre foi a de transmitir a informação da forma mais directa e perceptível por qualquer indivíduo. Nos dias de hoje as novas tecnologias permitem aceder à informação com uma facilidade nunca antes vista ou imaginada, estando certamente ainda por descobrir outras formas de registar e perpetuar a informação para as gerações vindouras. A fotografia está na origem das grandes evoluções da imagem, permitindo capturar o momento, tornando-o “eterno”. Hoje em dia, na era da imagem digital, além de se mostrar a realidade, é possível incorporar na imagem informação adicional, de modo a enriquecer a experiência de visualização e a maximizar a aquisição do conhecimento. As possibilidades da visualização em três dimensões (3D) vieram dar o realismo que faltava ao formato de fotografia original. O 3D permite a imersão do espectador no ambiente que, a própria imagem retrata, à qual se pode ainda adicionar informação escrita ou até sensorial como, por exemplo, o som. Esta imersão num ambiente tridimensional permite ao utilizador interagir com a própria imagem através da navegação e exploração de detalhes, usando ferramentas como o zoom ou ligações incorporados na imagem. A internet é o local onde, hoje em dia, já se disponibilizam estes ambientes imersivos, tornando esta experiência muita mais acessível a qualquer pessoa. Há poucos anos ainda, esta prática só era possível mediante o recurso a dispositivos especificamente construídos para o efeito e que, por isso, apenas estavam disponíveis a grupos restritos de utilizadores. Esta dissertação visa identificar as características de um ambiente 3D imersivo e as técnicas existentes e possíveis de serem usadas para maximizar a experiência de visualização. Apresentar-se-ão algumas aplicações destes ambientes e sua utilidade no nosso dia-a-dia, antevendo as tendências futuras de evolução nesta área. Serão apresentados exemplos de ferramentas para a composição e produção destes ambientes e serão construídos alguns modelos ilustrativos destas técnicas, como forma de avaliar o esforço de desenvolvimento e o resultado obtido, comparativamente com formas mais convencionais de transmitir e armazenar a informação. Para uma avaliação mais objectiva, submeteram-se os modelos produzidos à apreciação de diversos utilizadores, a partir da qual foram elaboradas as conclusões finais deste trabalho relativamente às potencialidades de utilização de ambientes 3D imersivos e suas mais diversas aplicações.

Macrophages as biomarkers in BCG treatment response in bladder cancer

Bladder cancer is a common urologic cancer and the majority has origin in the urothelium. Patients with intermediate and high risk of recurrence/progression bladder cancer are treated with intravesical instillation with Bacillus Calmette-Guérin, however, approximately 30% of patients do not respond to treatment. At the moment, there are no accepted biomarkers do predict treatment outcome and an early identification of patients better served by alternative therapeutics. The treatment initiates a cascade of cytokines responsible by recruiting macrophages to the tumor site that have been shown to influence treatment outcome. Effective BCG therapy needs precise activation of the Th1 immune pathway associated with M1 polarized macrophages. However, tumor-associated macrophages (TAMs) often assume an immunoregulatory M2 phenotype, either immunosuppressive or angiogenic, that interfere in different ways with the BCG induced antitumor immune response. The M2 macrophage is influenced by different microenvironments in the stroma and the tumor. In particular, the degree of hypoxia in the tumors is responsible by the recruitment and differentiation of macrophages into the M2 angiogenic phenotype, suggested to be associated with the response to treatment. Nevertheless, neither the macrophage phenotypes present nor the influence of localization and hypoxia have been addressed in previous studies. Therefore, this work devoted to study the influence of TAMs, in particular of the M2 phenotype taking into account their localization (stroma or tumor) and the degree of hypoxia in the tumor (low or high) in BCG treatment outcome. The study included 99 bladder cancer patients treated with BCG. Tumors resected prior to treatment were evaluated using immunohistochemistry for CD68 and CD163 antigens, which identify a lineage macrophage marker and a M2-polarized specific cell surface receptor, respectively. Tumor hypoxia was evaluated based on HIF-1α expression. As a main finding it was observed that a high predominance of CD163+ macrophage counts in the stroma of tumors under low hypoxia was associated with BCG immunotherapy failure, possibly due to its immunosuppressive phenotype. This study further reinforces the importance the tumor microenvironment in the modulation of BCG responses.

The predominance of M2-polarized macrophages in the stroma of low-hypoxic bladder tumors is associated with BCG immunotherapy failure

OBJECTIVE: Bacillus Calmette-Guérin (BCG) immunotherapy is the gold standard treatment for superficial bladder tumors with intermediate/high risk of recurrence or progression. However, approximately 30% of patients fail to respond to the treatment. Effective BCG therapy needs precise activation of the type 1 helper cells immune pathway. Tumor-associated macrophages (TAMs) often assume an immunoregulatory M2 phenotype and may directly interfere with the BCG-induced antitumor immune response. Thus, we aim to clarify the influence of TAMs, in particular of the M2 phenotype in stroma and tumor areas, in BCG treatment outcome. PATIENTS AND METHODS: The study included 99 patients with bladder cancer treated with BCG. Tumors resected before treatment were evaluated using immunohistochemistry for CD68 and CD163 antigens, which identify a lineage macrophage marker and a M2-polarized specific cell surface receptor, respectively. CD68+ and CD163+ macrophages were evaluated within the stroma and tumor areas, and high density of infiltrating cells spots were selected for counting. Hypoxia, an event known to modulate macrophage phenotype, was also assessed through hypoxia induced factor (HIF)-1α expression. RESULTS: Patients in whom BCG failed had high stroma-predominant CD163+ macrophage counts (high stroma but low tumor CD163+ macrophages counts) when compared with the ones with a successful treatment (71% vs. 47%, P = 0.017). Furthermore, patients presenting this phenotype showed decreased recurrence-free survival (log rank, P = 0.008) and a clear 2-fold increased risk of BCG treatment failure was observed in univariate analysis (hazard ratio = 2.343; 95% CI: 1.197-4.587; P = 0.013). Even when adjusted for potential confounders, such as age and therapeutic scheme, multivariate analysis revealed 2.6-fold increased risk of recurrence (hazard ratio = 2.627; 95% CI: 1.340-5.150; P = 0.005). High stroma-predominant CD163+ macrophage counts were also associated with low expression of HIF-1α in tumor areas, whereas high counts of CD163+ in the tumor presented high expression of HIF-1α in tumor nests. CONCLUSIONS: TAMs evaluation using CD163 is a good indicator of BCG treatment failure. Moreover, elevated infiltration of CD163+ macrophages, predominantly in stroma areas but not in the tumor, may be a useful indicator of BCG treatment outcome, possibly owing to its immunosuppressive phenotype.

Overexpression of tumour-associated carbohydrate antigen sialyl-Tn in advanced bladder tumours

Little is known on the expression of the tumour-associated carbohydrate antigen sialyl-Tn (STn), in bladder cancer. We report here that 75% of the high-grade bladder tumours, presenting elevated proliferation rates and high risk of recurrence/progression expressed STn. However, it was mainly found in non-proliferative areas of the tumour, namely in cells invading the basal and muscle layers. STn was also found in tumour-adjacent mucosa, which suggests its dependence on a field effect of the tumour. Furthermore, it was not expressed by the normal urothelium, demonstrating the cancer-specific nature of this antigen. STn expression correlated with that of sialyltransferase ST6GalNAc.I, its major biosynthetic enzyme. The stable expression of ST6GalNAc.I in the bladder cancer cell line MCR induced STn expression and a concomitant increase of cell motility and invasive capability. Altogether, these results indicate for the first time a link between STn expression and malignancy in bladder cancer. Hence, therapies targeting STn may constitute new treatment approaches for these tumours.

A theoretical study on modelling the respiratory tract with ladder networks by means of intrinsic fractal geometry

Fractional order modeling of biological systems has received significant interest in the research community. Since the fractal geometry is characterized by a recurrent structure, the self-similar branching arrangement of the airways makes the respiratory system an ideal candidate for the application of fractional calculus theory. To demonstrate the link between the recurrence of the respiratory tree and the appearance of a fractional-order model, we develop an anatomically consistent representation of the respiratory system. This model is capable of simulating the mechanical properties of the lungs and we compare the model output with in vivo measurements of the respiratory input impedance collected in 20 healthy subjects. This paper provides further proof of the underlying fractal geometry of the human lungs, and the consequent appearance of constant-phase behavior in the total respiratory impedance.

The role of functional polymorphisms in immune response genes as biomarkers of BCG Immunotherapy outcome in bladder cancer: Establishment of a predictive profile in a Southern Europe population

OBJECTIVE: To evaluate the predictive value of genetic polymorphisms in the context of BCG immunotherapy outcome and create a predictive profile that may allow discriminating the risk of recurrence. MATERIAL AND METHODS: In a dataset of 204 patients treated with BCG, we evaluate 42 genetic polymorphisms in 38 genes involved in the BCG mechanism of action, using Sequenom MassARRAY technology. Stepwise multivariate Cox Regression was used for data mining. RESULTS: In agreement with previous studies we observed that gender, age, tumor multiplicity and treatment scheme were associated with BCG failure. Using stepwise multivariate Cox Regression analysis we propose the first predictive profile of BCG immunotherapy outcome and a risk score based on polymorphisms in immune system molecules (SNPs in TNFA-1031T/C (rs1799964), IL2RA rs2104286 T/C, IL17A-197G/A (rs2275913), IL17RA-809A/G (rs4819554), IL18R1 rs3771171 T/C, ICAM1 K469E (rs5498), FASL-844T/C (rs763110) and TRAILR1-397T/G (rs79037040) in association with clinicopathological variables. This risk score allows the categorization of patients into risk groups: patients within the Low Risk group have a 90% chance of successful treatment, whereas patients in the High Risk group present 75% chance of recurrence after BCG treatment. CONCLUSION: We have established the first predictive score of BCG immunotherapy outcome combining clinicopathological characteristics and a panel of genetic polymorphisms. Further studies using an independent cohort are warranted. Moreover, the inclusion of other biomarkers may help to improve the proposed model.