Counterfeit medicines: relevance, consequences and strategies to combat the global crisis

Counterfeiting of medicines, also known as “falsification” or “adulteration”, is the process in which the identity, origin, or history of genuine medicines are intentionally modified. Currently, counterfeit medicines are a global crisis that affects and is mostly caused by developing countries in Asia, Africa and Latin America. These countries lack strict law enforcement against this practice and have low-income populations with medicinal needs. Lately, the crisis has escalated, impacting developed countries as well, e.g., the US and the EU, mainly via the Internet. Despite this extension, some current laws aim to control and minimize the crisis’ magnitude. Falsification of medicines maintains an illegitimate supply chain that is connected to the legitimate one, both of which are extremely complex, making such falsification difficult to control. Furthermore, political and economic causes are related to the crisis’ hasty growth, causing serious consequences for individuals and public health, as well as for the economy of different countries. Recently, organizations, technologies and initiatives have been created to overcome the situation. Nevertheless, the development of more effective measures that could aggregate all the existing strategies into a large functioning network could help prevent the acquisition of counterfeit medicines and create awareness among the general population

Effect of Fas/FasL interaction on CD8+ T cell differentiation in vitro

Uma abordagem científica essencial que tem sido central para o estudo da biologia e do papel das células T CD8 + durante as respostas imunes é a melhor compreensão dos mecanismos de morte/sobrevivência dessas células. Durante a fase de contração, a morte de células T antígenoespecíficas pode ser alcançada por morte celular induzida por ativação (AICD) que ocorre após a ligação de Fas e FasL, ambos expressos por células T ativadas. Essa interação trimeriza o Fas, resultando no recrutamento da proteína adaptadora Fas-associated death domain (FADD) e das caspases-8 e/ou -10, criando o complexo de sinalização induzido pela morte (DISC). Então, o DISC ativa a caspase-8 ou 10 que inicia uma cascata levando à apoptose. O conhecimento e a manipulação dos mecanismos de morte/sobrevivência podem melhorar as habilidades de matar células T, otimizando as imunoterapias contra o câncer e os métodos de prevenção de infecções por vírus. Seguindo essa ideia, o projeto estudou o efeito de moléculas de morte/sobrevivência, como Fas e FasL, na ativação e diferenciação de células T CD8 + in vitro. Neste estudo, primeiro padronizamos o isolamento de células T CD8 + frescas de baços de camundongos de tipo selvagem (WT) e analisamos a ativação e diferenciação dessas células para os subconjuntos Tc0 (controle somente de ativação), Tc1 e Tc2 por Multicolor Flow Citometria (MFC) e Reação em Cadeia da Polimerase em Tempo Real (qPCR), posteriormente aplicando a mesma avaliação em camundongos deficientes em FasL (gld). Por fim, avaliamos a ativação de Tc0, Tc1 e Tc2 e a diferenciação entre camundongos WT e gld por MFC. Nossos resultados mostraram que isolamento, ativação e diferenciação eficientes em ambas as linhagens de camundongos foram alcançados. No geral, a deficiência de FasL não interfere na ativação, diferenciação e atividade efetora das células T CD8 +.One scientific essential approach that has been central to the study of CD8 + T cell biology and role during immune responses, is the better understanding of death/survival mechanisms of these cells. During the contraction phase, death of antigen-specific T cells can be achieved by activation-induced cell death (AICD) that occurs upon ligation of Fas and FasL, both expressed by activated T cells. This interaction trimerizes Fas, resulting in the recruitment of Fas-associated death domain (FADD) adaptor protein and the caspases-8 and/or -10, creating the Death-induced signaling complex (DISC). Then, DISC activates caspase-8 or 10 that initiate a cascade leading to apoptosis. The knowledge and manipulation of death/survival mechanisms could improve T cells killing skills, optimizing cancer immunotherapies and prevention methods of virus infections. Following this idea, the project studied the effect of death/survival molecules, such as Fas and FasL, in CD8 + T cell activation and differentiation in vitro. In this study, we first standardized fresh CD8 + T cell isolation from spleens of wild-type (WT) mice and analyzed the activation, and differentiation of these cells towards the Tc0 (activation-only control), Tc1, and Tc2 subsets by Multicolor Flow Cytometry (MFC) and Real-Time Polymerase Chain Reaction (qPCR), later applying the same evaluation on FasL-deficient mice (gld). Last, we assessed Tc0, Tc1, and Tc2 activation, and differentiation betweenWT and gld mice by MFC. Our results showed that efficient isolation, activation and differentiation in both mice lineages were achieved. Overall, FasLdeficiency does not interfere with the activation, differentiation, and effector activity of CD8 + T cells