ESSL - Posters
URI permanente para esta coleção:
Navegar
Entradas recentes
- NOX: innovating sickle cell disease genetic diagnosis with microfluidic disposable chipsPublication . Andaluz, Stephanie; Shen, Ren; Brito, Miguel; Liu, Tzu-Ming; Jia, YanweiSickle Cell Disease (SCD) refers to a genetic blood disorder that affects the haemoglobin protein found within red blood cells (RBCs). This condition is caused by a point mutation in the gene that encodes the β-globin chain (HBB), leading to a structural variant of the normal adult haemoglobin tetramer, Haemoglobin A (HbA), into Haemoglobin S (HbS) or Haemoglobin C (HbC), leading the RBC to change shape. While high-income countries have achieved significant progress in managing SCD, low-income countries face challenges in providing accessible and effective interventions, where more than 80% of individuals born with SCD remain undiagnosed, and less than half of them survive beyond the age of 5. This paper introduces a novel PCR-based genetic diagnostic testing technology for SCD, designed for implementation on a disposable microfluidic chip. NOX technology is based on the kinetic hairpin oligonucleotide blockers model proposed by Jia et al. 2014. By adjusting the stem length and temperature conditions of hairpin oligonucleotide blockers, we tailored the binding process to target sequences effectively; by optimizing the blocker binding temperature within specific steps of the PCR process, we enhanced its preference for hybridizing with the target template (Fig. 1). Because with this technology we “block” our targets, avoiding the primers from binding and amplifying them, a positive result is read as the absence of fluorescence; therefore, we called this novel approach NOX. Our preliminary results, depicted in Figure 2, demonstrate that the incorporation of the hairpin blocker effectively retards the amplification process of 1,000 to 10,000 copies of the target sequence. These results indicate that the hairpin blocker effectively inhibits the amplification of its matched target up to 100 copies per reaction. To increase the potential of this technology, we designed a three-layer disposable microfluidic chip (Figure 3A-B). The 3x3 cm chip, consisting of a single inlet, eight reaction chambers, and outlets, was designed in a snowflake shape to aid the loading of the DNA samples by centrifugal force. The fabrication materials were chosen following the work of Bae et al. (2018) for a disposable multi-chamber film-based PCR chip. The top and bottom layers are PET film, while the middle layer holding the chambers and channels is PVC double-adhesive film. Inlets, outlets, reaction chambers, and channels are obtained by cutting with a plotting cutter, and fabrication is done by sequentially stacking the films. A 3D view of the chip can be seen in Figure 3B. The implementation of this microfluidic chip by the hand of our new technology promises to bring affordable genetic testing, contributing to better management of SCD in underserved regions.
- Chemotherapy-associated cardiotoxicity in women with breast cancer: assessment of left atrial strain by transthoracic echocardiographyPublication . Sousa, R. M.; Fonseca, Virgínia; Henriques, M.; Cavaco, S.; Silva, J.; Clérigo, Anália MatosBackground: Transthoracic echocardiography (TTE) is considered the gold-standard method for assessing cardiac function, with left ventricular ejection fraction (LVEF) and global longitudinal strain (GLS) being the main parameters used for early detection of cardiotoxicity. Left atrial (LA) strain represents the myocardial deformation of this structure and is assessed based on three main parameters, among which peak atrial longitudinal strain (PALS) stands out for best reflecting global LA function. Purpose: To evaluate left atrial strain by TTE as a predictor of cardiotoxicity at different stages of treatment with anthracyclines and trastuzumab in women with breast cancer.
- Chemotherapy-associated cardiotoxicity in women with breast cancer: assessment of myocardial work index by transthoracic echocardiographyPublication . Sousa, R. M.; Fonseca, Virgínia; Azevedo, C.; Simão, M.; Cobas, V.; Clérigo, Anália MatosBackground: Breast cancer is one of the most common malignancies worldwide. Despite advances in treatment, these therapies can increase the risk of cardiotoxicity, making cardiac monitoring crucial in this population. The Myocardial Work Index (MWI) has emerged as a tool to predict the risk of cardiotoxicity. Using the left ventricular pressure-strain loop technique, MWI evaluates myocardial work and calculates global work index (GWI), global wasted work (GWW), global constructive work (GCW), and global work efficiency (GWE). Purpose: To determine the added value of myocardial work indices compared with global longitudinal strain (GLS) assessed by transthoracic echocardiography (TTE) in the diagnosis of chemotherapy-associated cardiotoxicity in women with breast cancer.
- MicroRNA-mediated post-transcriptional regulation of peripheral γδ T cell effector functionsPublication . Inácio, Daniel; Amado, Tiago; Sobral, Daniel; Enguita, Francisco; Cunha, Carolina; Silva-Santos, Bruno; Gomes, Anitaγδ T cells are unconventional lymphocytes able to rapidly produce large quantities of interleukin-17A (IL-17A) or interferon- γ (IFNγ), which often have major effects on the pathophysiology of diseases such as cancer or infections. Yet, we do not fully understand how the thymic-derived effector γδ T cell subsets making either IL-17A (γδ17 cells) or IFNγ (γδIFN cells) are activated and function in peripheral tissues. Here, we established an Il17a-GFP:Ifng-YFP double-reporter mouse strain to analyse at unprecedented depth the microRNA transcriptomes of pure γδ17 versus γδIFN cell populations from peripheral lymph nodes, and uncover new mechanisms of miRNA-mediated post-transcriptional regulation of γδ T cell effector functions. We found 103 differentially expressed microRNAs between γδ17 and γδIFN cells, from which we selected 10 candidates likely to target members of the transcriptional networks underlying IL-17A of IFN-γ production, to test their effects on γδ T cell differentiation or effector functions. Retroviral gain-of-function approaches in vitro highlighted a plethora of context-dependent microRNA-specific effects on the differentiation or expansion of γδ17 or γδIFN cells, and respective cytokine production. Furthermore, a detailed analysis of each candidate miRNA expression throughout ontogeny showed that in some cases it was imprinted in the thymus, whereas in others it was peripherally induced. This led to the observation that miR-181a-5p, highly expressed in early thymic γδ T cell subsets, favours thymic γδ17 cell commitment, while conversely promoting peripheral γδIFN activation and proliferation in response to TCR stimulation in vitro and upon malaria infection in vivo. On the contrary, we found miR-128-3p to act as a peripheral regulator of IFN-γ, preventing aberrant IFN-γ production upon TCR stimulation. These findings unravel a new layer of regulation of γδ T cell effector functions, which we are further characterizing with additional molecular assays.
- MicroRNA-mediated post-transcriptional regulation of peripheral γδ T cell effector functionsPublication . Inácio, Daniel; Amado, Tiago; Sobral, Daniel; Enguita, Francisco; Cunha, Carolina; Gomes, Anita; Silva-Santos, Brunoγδ T cells are unconventional lymphocytes able to rapidly produce large quantities of interleukin-17A (IL-17A) or interferon-γ (IFNγ), which often have major effects on the pathophysiology of diseases such as cancer or infections. Yet, we do not fully understand how the thymic-derived effector γδ T cell subsets making either IL-17A (γδ17 cells) or IFNγ (γδIFN cells) are activated and function in peripheral tissues. Here, we established an Il17a-GFP:Ifng-YFP double-reporter mouse strain to analyse at unprecedented depth the microRNA (miRNA) transcriptomes of pure γδ17 versus γδIFN cell populations from peripheral lymph nodes, and uncover new mechanisms of miRNA-mediated post-transcriptional regulation of γδ T cell effector functions. We found 103 differentially expressed miRNAs between γδ17 and γδIFN cells, from which we selected 10 candidates likely to target members of the transcriptional networks underlying IL-17A of IFN-γ production, to test their effects on γδ T cell differentiation or effector functions. Retroviral gain-of-function approaches in vitro highlighted a plethora of context-dependent miRNA-specific effects on the differentiation or expansion of γδ17 or γδIFN cells, and respective cytokine production. Furthermore, a detailed analysis of each candidate miRNA expression throughout ontogeny showed that in some cases it was imprinted in the thymus, whereas in others it was peripherally induced. This led to the observation that miR-181a-5p, highly expressed in early thymic γδ T cell subsets, favours thymic γδ17 cell commitment, while conversely promoting peripheral γδIFN activation and proliferation in response to TCR stimulation in vitro and upon malaria infection in vivo. On the contrary, we found miR-128-3p to act as a peripheral regulator of IFN-γ, preventing aberrant IFN-γ production upon TCR stimulation. These findings unravel a new layer of regulation of γδ T cell effector functions, which we are further characterizing with additional molecular assays.
- Role of microRNAs in effector versus regulatory CD4+ T cell differentiation during (auto)immune responses in vivoPublication . Cunha, Carolina; Romero, Paula Vargas; Inácio, Daniel; Sobral, Daniel; Costa, Marina; Papotto, Pedro; Enguita, Francisco; Gomes, Anita; Silva-Santos, BrunoIntroduction: MicroRNAs are an abundant class of evolutionarily conserved small non-coding RNA species that control gene expression at the post-transcriptional level and are estimated to regulate most developmental and physiological processes. They are involved in many aspects of immunity, notably in the differentiation and function of CD4+ T cell subsets. CD4+ T cells play critical roles in the host defense against pathogens, but they are also responsible for the generation of immune-mediated diseases. This arises from an incorrect balance between the various CD4+ T cell subsets, namely pro-inflammatory effector T cells, including the IFN-g-producers T helper 1 (Th1) cells and the IL-17-producers Th17 cells, and anti-inflammatory regulatory T cells (FoxP3+ subset). Although various individual miRNAs have been implicated in CD4+ T cell biology, data are missing on how miRNA regulatory networks may control the balance between effector and regulatory T cells in pathophysiological conditions in vivo. Aim of the study: To dissect the role of miRNAs in the post-transcriptional regulation of effector and regulatory CD4+ T cell differentiation in vivo.
- Emergent pan-azole-resistant Aspergillus fumigatus in poultry farms: implications for One HealthPublication . Gomes, Bianca; Ramos, Miguel J. N.; Nunes, Mónica; Viegas, Carla; Dias, RicardoAspergillus fumigatus is the leading etiological agent of aspergillosis in humans and animals. Azole resistance is an emerging public health concern. Poultry bedding material may act as an environmental reservoir for azole-resistant A. fumigatus. Understanding environmental reservoirs is essential to mitigate potential risks across the human–animal–environment interface. Aim of the study: To assess the occurrence of azole-resistant Aspergillus fumigatus in Portuguese poultry farms.
- Assessment of miocrobiological contamination exposure in the plastic waste industry using passive sampling methodsPublication . Viegas, Carla; Pena, Pedro; Cervantes, Renata; Kosicki, Robert; Twaruzek, Magdalena; Viegas, SusanaThis study assessed occupational exposure to microbiological contamination in the plastic waste sorting industry using passive sampling methods and aimed to identify suitable analytical approaches for characterizing microbial contamination in this environment.
- Comparative assessment of mycotoxin contamination and cytotoxic potential in indoor environments: grocery stores and fitness centers in PortugalPublication . Monteiro, Beatriz; Figueiredo, Filipa; Dias, Marta; Cervantes, Renata; Pena, Pedro; Twarużek, Magdalena; Kosicki, Robert; Soszczyńska, Ewelina; Viegas, Susana; Viegas, CarlaThe presence of mycotoxins in indoor environments is an emerging public health concern, particularly in specific indoor and occupational environments. Cytotoxicity assessment provides relevant information on the biological effects of complex environmental contaminant mixtures. Grocery stores and fitness centers represent distinct indoor environments with different exposure dynamics and contamination sources, requiring integrated evaluation approaches. This study aimed to compare mycotoxin contamination and cytotoxic potential, assessed using the HepG2 and A549 cell lines, of environmental matrices from two projects conducted in Portugal: grocery stores in 2021 and fitness centers in 2024. In grocery stores, only EDC samples were analyzed, whereas in fitness centers, a wider sampling campaign, using more sampling methods, was employed (EDCs, filters, and cleaning mops). Regarding mycotoxins, fumonisins B1, B2, and B3 were identified in EDC samples from grocery stores. In fitness centers, fumonisin B2 was detected in all matrices analyzed. Cytotoxicity assessment revealed distinct responses between settings and cell lines. In grocery stores, cytotoxic effects were limited, with only one EDC sample showing a positive response in the HepG2 cell line and one EDC sample in the A549 cell line. In fitness centers, the HepG2 cell line showed positive cytotoxic responses in filter and EDC samples, while mop samples did not induce cytotoxicity. The A549 cell line exhibited positive responses across all analyzed matrices. Overall, mycotoxins were detected in greater variety in grocery stores, while fitness centers showed higher cytotoxic potential, highlighting the need for integrated monitoring of contamination and biological effects in indoor environments.
- Fungal resistance and public health implications: the use of wastewater treatment plant (WWTP) sludge in agriculturePublication . Pimenta, Márcia; Cervantes, Renata; Pena, Pedro; Sabino, Raquel; Viegas, CarlaThis study aimed to assess microbial and fungal risks of commercial fertilizers and substrates associated with waste-derived materials in Portugal, integrating antifungal resistance screening under a One Health framework.