ESS - APCT - Posters apresentados em eventos científicos
URI permanente para esta coleção:
Navegar
Entradas recentes
- Psychostimulants and neuroinflammation: finding critical players in the crostalk between glial cells and neuronsPublication . Bravo, Joana; Andrade, Elva Bonifácio; Vieira, Renato; Lorga, Inês; Azevedo, M.; Rodrigues, J.; Magalhães, Ana; Relvas, João B.; Summavielle, Teresa; Summavielle, Teresa; Bonifácio Andrade, Elva; Bravo, JoanaExposure to psychostimulants has been classically associated with damage to neuronal terminals. However, it is now accepted that interaction between neuronal and glial cells also contributes to the addictive behavior. We have recently shown that acute methamphetamine (Meth), a powerful psychostimulant, causes microgliosis and increases microglia activation through astrocytic-TNF release. We are now interested in clarifying the progression of neuroinflammation under chronic drug exposure and how different brain and immune cells contribute to this inflammatory process.To explore this, firstly, we performed a proteomic analysis, in different phases of the addictive process, in mice exposed to an escalating dosing of Meth for ten days (Meth10d). To validate the conditioning power of our model,mice were tested in a condition place preference (CPP) at 10d of Meth, and 2 or 10 days of withdrawal (WD). At all these time points, mice were seen to be strongly conditioned by Meth. Next, we conducted a proteomic analysis to compare the different time points (using the hippocampus, where we previously found robust microgliosis underMeth). We found a proteome profile that varied substantially with exposure (Meth10d) and after a short- (WD2d)and long-term withdrawal (WD10d) periods. Interestingly, the most altered pathways were neuro transmitter-related. However, we also identified significant differences in Wnt signaling, which was previously linked to regulation of microglia reactivity. As such, we evaluated the microglia profile after chronic Meth exposure and at withdrawal. In the hippocampus, the number of microglia cells was significantly increased at Meth10d and remained also increased at WD2d. Microglia presented a more ameboid-like shape at Meth10d, but its ramified morphology was recovered atWD2d. Importantly, our proteomic data also revealed that during Meth withdrawal, several microglial receptors were down regulated, suggesting that microglia was in a “primed” state. In addition, as the crosstalk between neurons and microglia seems to be relevant for the behavioral expression of Meth, we are dissecting the modulation of microglia by neurons under Meth exposure, to evaluate neuroimmune regulatory ligand-receptor pairs that seem to impact onthe neuron-microglia interaction. Of note, some these ligand-receptor pairs seem to be down regulated by chronic Meth and during abstinence, which may be associated with reduced neuronal ability to down regulate microglia reactivity, and lead to increased neuronal damage.We fore see that these receptors may prove to be interesting therapeutic targets for the treatment of addiction, and therefore we will manipulate them to confirm their value in reducing relapse rates and improve addiction treatments.
- Blockingmethamphetamine-induced microglia reactivity by targeting glutamate receptorsPublication . Summavielle, Teresa; Canedo, Teresa; Silva, Ana Isabel; Andrade, Elva Bonifácio; Almeida, Tiago O.; Bravo, Joana; Terceiro, Ana Filipa; Canedo, Teresa; Silva, Ana Isabel; Magalhães, Ana; Relvas, João B.; Bonifácio Andrade, Elva; Bravo, JoanaExposure to psychostimulants has been classically associated with damage to neuronal terminals. However, it is now accepted that interaction between neuronal and glial cells also contributes to the addictive behavior. We have recently shown that acute methamphetamine (Meth), a powerful psychostimulant, causes microgliosis and increases microglia activation through astrocytic-TNF release1. We are now interested in clarifying the progression of neuroinflammation under chronic drug exposure and how different brain and immune cells contribute to this inflammatory process.To explore this, firstly, we performed a proteomic analysis, in different phases of the addictive process, in mice exposed to an escalating dosing of Meth for ten days (Meth10d). To validate the conditioning power of our model, mice were tested in a condition place preference (CPP) at 10d of Meth, and 2 or 10 days of withdrawal (WD). At all these time points, mice were seen to be strongly conditioned by Meth. Next, we conducted a proteomic analysis to compare the different time points (using the hippocampus, where we previously found robust microgliosis underMeth1). We found a proteome profile that varied substantially with exposure (Meth10d) and after a short- (WD2d)and long-term withdrawal (WD10d) periods. Interestingly, the most altered pathways were neuro transmitter-related.However, we also identified significant differences in Wnt signaling, which was previously linked to regulation of microglia reactivity. As such, we evaluated the microglia profile after chronic Meth exposure and at withdrawal. In the hippocampus, the number of microglia cells was significantly increased at Meth10d and remained also increased at WD2d. Microglia presented a more ameboid-like shape at Meth10d, but its ramified morphology was recovered at WD2d. Importantly, our proteomic data also revealed that during Meth withdrawal, several microglial receptors were down regulated, suggesting that microglia was in a “primed” state. In addition, as the crosstalk between neurons and microglia seems to be relevant for the behavioral expression of Meth, we are dissecting the modulation of microgliaby neurons under Meth exposure, to evaluate neuroimmune regulatory ligand-receptor pairs that seem to impact on the neuron-microglia interaction. Of note, some these ligand-receptor pairs seem to be down regulated by chronic Meth and during abstinence, which may be associated with reduced neuronal ability to down regulate microglia reactivity, and lead to increased neuronal damage. We fore see that these receptors may prove to be interesting therapeutic targets for the treatment of addiction, and therefore we will manipulate them to confirm their value in reducing relapse rates and improve addiction treatments.
- Uso de filtros no processamento de Citologia de base líquida: Importante discriminar?Publication . Monteiro, Tiago; Fernandes, Sílvia; Silva, Regina; Silva, ReginaRegularmente em todo o mundo, sendo a filtração com recurso a filtros descartáveis a metodologia mais utilizada. Apesar de existirem filtros específicos para diversos tipos de processamento, pouco se sabe acerca das suas características, da forma como são empregues e do impacto da sua utilização no processamento de citologia de base líquida e consequente diagnóstico citológico. O estudo teve como principal objetivo avaliar e registar as características dos filtros usados em citologia de base líquida, e avaliar o impacto do seu uso indiscriminado, em Portugal. Para tal, estudou-se microscopicamente os poros de uma área de 7500µm2 de membrana de policarbonato de filtros de citologia de base líquida, indicados para processamento de amostras ginecológicas (Gyn) e não ginecológicas (Não Gyn), de três marcas diferentes. Procedeu-se à sua caracterização quanto ao número, diâmetro e distribuição dos poros. Foi ainda administrado um questionário a laboratórios de citologia em Portugal, e os dados recolhidos foram analisados com o intuito de caracterizar o uso das metodologias, os resultados obtidos permitem concluir que existem diferenças entre os filtros destinados aos vários tipos de processamento e entre filtros de diferentes marcas. Os filtros destinados ao processamento de amostras Gyn, para além de poros de maiores dimensões (7.2-7.5 µm), apresentam um menor número de poros (92-98) e respetiva sobreposição em 55 a 58% da área em análise. Por sua vez, os filtros recomendados para amostras Não Gyn apresentam menor diâmetro (5.7-6.1 µm), maior número de poros (107-122) e respetiva sobreposição numa maior área (60 a 83%).Considerados globalmente, os resultados revelam que as diferenças verificadas entre os filtros podem resultar na alteração ao nível da representação de estruturas celulares e de microrganismos, e o seu uso indiscriminado a nível laboratorial pode comprometer a avaliação citopatológica.
- Computacional pathology: What’s newPublication . Coelho, Daniel; Assunção, Teresa; Borrecho, Gonçalo; Geraldes, Mariana; Vinagre, Tiago; Ferreira, Inês; Ferreira, Ana; Fernandes, Ana Isabel; França, Amélia; Vale, João; Curado, Mónica; Mendes, Fernando; Martins, DianaThe term computacional pathology (CPath) has become a buzz-word among the digital pathology community. Adances in scanning systems, imaging technologies and storage devices are generating an ever-increasing volume of whole-slide images (WSI) acquired in clinical settings, which can be computacionally analyzed using artificial intelligence (AI), such as deep learning technologies, in a new área of development called CPath. The purpose of the review is to disseminate the latest news and futures perspectives by CPath. Deep learning in the context of CPath has methodological contributions that can be distinguished into approaches based on the final purpose of the analysis: predicting clinical endpoints such as cancer subtype, patient survival or genetic mutations from WSI and AI-based assistive tools, such as segmentation methods for WSI or virtual staining. The emergence of multipex imaging, spatially resolver genomic assays and 3D pathology, among other methodologies, will accelerate this trend, providing new opportunities for multimodal integration and discovering new biomarkers. Additionally, these developments will help automating labor-intensive manual work and reducing inter-observer variability diagnosis between pathologists, contributing to a better patient care. CPath will underpin the development of the next generation of cancer therapies and diagnostics, changing the clinical research and ultimately leading towards new cures or improved patient outcomes.
- Caraterização de tumores sólidos numa amostra pediátrica do Instituto Português de Oncologia do Porto Francisco Gentil, EPEPublication . Santos, Ana; Serra, Carla; Mota, Marlene; Sousa, Manuela; Mendes, CarlosCaracterizar uma amostra pediátrica com TS quanto à sua frequência, óbito, sexo e idade ao diagnóstico. Estudar o possível envolvimento da MO ao diagnóstico, recorrendo a dados qualitativos de mielogramas. Estudo observacional descritivo transversal, dos registos dos dados de pacientes pediátricos com TS (n=148), diagnosticados entre 2000-2008. O TS mais frequente foi o Neuroblastoma (NB=37,2%), e os com maior registo de óbitos foram Rabdomiossarcoma (RMS=30,4%) e Família Tumores Ewing (FTE=28,0%). Nas raparigas os Tumor Wilms (TW=68,0%) e Osteossarcoma (58,3%) foram mais frequentes, enquanto nos rapazes foram os Retinoblastoma (75,0%), RMS (65,2%) e NB (61,8%). O NB manifestou-se, em 30,9% dos casos, até aos 12 meses de idade, o TW, em 32,0% dos casos, até aos 2 anos, e o RMS, em 69,6% dos casos, entre os 1-10 anos. A maioria dos pacientes com FTE (80,0%), NB (74,6%) e RMS (73,9%) realizaram mielograma ao diagnóstico. Observando os resultados qualitativos do mielograma, os tumores onde poderá ter ocorrido invasão da MO foram o NB e RMS. Na frequência dos TS verificou-se que seguem o descrito na literatura, excepto a FTE que surge em segundo lugar conjuntamente com o TW, sendo o NB mais frequente. Nos óbitos, o tumor com maior registo, RMS, não corresponde ao com maior gravidade (NB) descrito na investigação. Para estes resultados pode ter contribuído o número reduzido da amostra. Relativamente ao sexo, o NB e o RMS são mais frequentes nos rapazes, enquanto o TW é mais frequente nas raparigas, como descrito na literatura. Doentes com NB, TW e RMS apresentaram idades ao diagnóstico que indicam um melhor prognóstico. O NB, RMS e FTE, em alguns casos, disseminam-se para a MO, podendo ser esta a explicação para os resultados obtidos quanto à realização ou não de mielograma. Dos resultados obtidos nos mielogramas, verifica-se que poderá ter ocorrido invasão medular nos NB e RMS. Propõe-se continuar este estudo com amostra, por TS, mais representativa e com dados dos hemogramas e biópsias ósseas.
- Molecular profiling of prognostic biomarkers in MelanomaPublication . Jesus, Marta; Correia, Beatriz; Costa, Catarina; João, Alexandre; Lopes, José Manuel; Barata, Catarina; Soares, Paula; Pópulo, HelenaCutaneous melanoma (CM) is the most aggressive and deadliest form of skin cancer (1). Its development is influenced by the genetic background, genetic mutations and environmental factors. The three most frequent mutations of CM appear in BRAF gene, which leads to uncontrolled cell growth, in NRAS gene, that disrupts normal cell signaling, and in TERT promoter, that increases telomerase activity and play a crucial role in tumor progression, influencing melanoma aggressiveness and patient outcome (2, 3). Despite advances in treatment, the prognosis of patients with advanced-stage remains poor (4). This work aims to improve prognosis determination by correlating identified molecular alterations with clinicopathological data. The analysis is being conducted on primary tumors collected at Hospital dos Capuchos. The study currently includes 29 samples collected between 2023 and 2024, 18 from male subjects and 11 from female subjects, with an average age of 69. Most of the melanomas were located on the trunk, and the most common diagnoses were nodular melanoma and superficial spreading melanoma. The mean Breslow thickness of the series is 3.78 mm, and 46% of samples have a mitotic index greater than 1. The samples were subjected to histopathological evaluation, followed by PCR and sequencing techniques, to identify the three mutations. Subsequent statistical analysis will be performed to assess the correlation between these genetic changes and clinicopathological features, with a focus on determining their impact on patient outcomes. The work is still ongoing, but the results will provide further information about the three mutations and their corresponding aggressiveness, as well as how the coexistence of different mutations influences patient prognosis. This study underlines the significance of molecular profiling in the prognosis assessment of melanoma patients, and it will provide a foundation for further research on molecular-based prognosis models.
- CytoPath®Easy: screening of cervical cancerPublication . Fernandes, Sílvia; Vilarinho, Ana Sofia; Silva, ReginaRecently, CytoPath®Easy kit was created by DiaPath S.p.A. and started to be commercially available for the screening of cervical cancer. Using this methodology, epithelial cells are immersed in a preservative liquid, and a thin-layer of cells in the slide is obtained through gravity sedimentation and filtration. The main objectives of this study are to evaluate the efficacy for the processing of cervical samples, for the detection of pre-neoplastic lesions and for the nucleic preservation and extraction by the kit. For this purpose, 215 cervical samples obtained by self-sampling were used: 174 were collected and processed by CytoPath® Easy and, as a control, 41 were collected and processed by the Thinprep® method. The samples were processed, stained by the Papanicolaou method, and independently evaluated microscopically for various morphological parameters; nucleic acids were isolated and evaluated for purity and integrity by spectrophotometry. Results obtained showed that both methods have a good performance, allowing the morphological evaluation of the cervical epithelium. However, the statistical analysis reveals that the methods are different from each other, with overall lower results being obtained in the method under study (p<0.001). In turn, both methods allow the extraction of good quality and quantity of DNA. Although some differences were found regarding morphology of the cells fixed and processed by the CytoPath®Easy method, this new methodology reveals efficient for the preservation of nucleic acids. Thus, its use in cervical cancer screening is recommended.
- Optimisation of a molecular methodology for the detection of virulence factors of enterotoxigenic Escherichia coli for the diagnosis of swine colibacillosisPublication . Campos, Ana; Oliveira, Ricardo; Almeida, Carina; Vieira, Filipa Quintela; Silva, Regina AugustaThe most common bacterial pathogen causing enteric infections in pigs is enterotoxigenic Escherichia coli (ETEC). Since pigs represent the largest livestock category in the European Union, ETEC-associated diseases, better known as swine colibacillosis leading to acute diarrhea and eventual death of the animal, result in significant costs to the pig industry. These diseases are traditionally prevented or treated with antibiotics, and this has had a huge impact on the emergence of resistant bacteria, correlating with the emergence of resistant infections in humans. Recognition of this problem has led the authorities to set ambitious goals for the reduction of this type of drug in animal husbandry, leading to the creation of a national project, APTAcoli, which aims to select aptamers (consisting of small single-stranded oligonucleotides capable of binding to target molecules with great affinity and specificity, due to the specific secondary and/ or tertiary structures they can acquire) as an alternative in the treatment of colibacillosis. The present experimental study, which is on the APTAcoli agenda, focused on the optimization of a molecular methodology - Multiplex PCR - for the detection of the main virulence factors of ETEC to be used in an epidemiological study to characterize fecal samples from pigs in Portuguese farms. After using different optimization techniques, the results were two multiplex PCR amplification sets, one for amplification of the main toxigenic factors of ETEC (STa, STb, LT and STx2e) and another for amplification of the main adhesion factors (F4, F5, F6, F18 and F41).
- CINtec PLUS cytology as a cervical cancer screening test at the IPO of PortoPublication . Alves, Ana; Lopes, Ana Paula; Granja, Sara; Quintela Vieira, Filipa; Silva, Regina AugustaCervical squamous cell carcinoma of the cervix is the fourth most common cancer between women and seventh globally. There are many women undergoing colposcopy without real need, so more effective screening tests are needed to decide who among HPV-positive women should receive additional diagnostic evaluation to avoid unnecessary colposcopies. So, this study is intended to evaluate CINtec PLUS cytology as a screening test for cervical cancer in the IPO-FG of the Port of women with hpv test positive other than 16/18, improving the referral for colposcopy. CINtec PLUS cytology is an immunocytochemistry kit that simultaneously detects p16 and Ki-67 proteins. The presence of these proteins in the same cell indicates a deregulation of the cell cycle, and may be a marker of the persistence of HPV infections, their greater probability of progression and the severity of lesions secondary to the infection. An experimental research was carried out, in which the immunostaining of 73 non-16/18 HPV cervico-vaginal hpv samples was performed from 73 women integrated in cervical cancer screening in the North region carried out at the IPO-FG of Porto. The results obtained in CINtec PLUS were compared with those of cytology. 2 samples present unsatisfactory cytology for evaluation, being excluded. Cintec Plus cytology would reduce by 17% the number of women sent for colposcopy (7 (about 10%) by Cintec Plus vs 19 (about 27%) by cytology). Of the 52 NILM samples, 3 (about 6%) were positive for Cintec Plus, so they should have been referred for colposcopy and were not. Of the 10 ASC-US samples 100% are negative for Cintec Plus, as well as 5 LSIL 4 (80%) are negative, of the 3 HSIL 2 (about 67%) are negative. So, 16 women (about 23%) made colposcopy unnecessarily. In conclusion, Cintec Plus cytology would be an interesting tool as a second screening test in non-16/18 hpv cases in place of cytology, decreasing the number of women sent for colposcopy.
- Uncovering the microglia response during neonatal Group B Streptococcus meningitisPublication . Soares, Joana; Lorga, Inês; Bravo, Joana; Summavielle, Teresa; Nova, Manuel Vila; Bonifácio Andrade, ElvaGroup B Streptococcus (GBS) remains the most common bacterial cause of meningitis in neonates. Microglia, the brain resident immune cells, have a critical role in the development of neural circuits. However, the role of GBS infection on microglia activation and neurological sequelae remains poorly characterised. Here, we aimed to evaluate whether GBS induces changes in microglia profile during the acute phase of infection, using a mouse model that mimics key steps of GBS pathophysiology in humans. Female C57BL/6 mice were intra-vaginally inoculated with GBS during gestation, and CFU analysis was performed on postnatal days (P) 1, 3 and 5. Bacterial colonisation was found at all ages, peaking at P3. When analysing the status of microglia by flow cytometry in the whole brain of male pups at P3, an overall activation was observed in the infected group. Mainly, we found a significant increase in microglia frequency, as well as the mean fluorescence intensities (MFIs) of CD45, CD11b and F4/80. Additionally, we also analysed some microglial receptors that are important neuro-immune regulators with relevant functions during development. We observed increased CX3CR1 expression in microglia, whereas Sirp and CD200r were not altered. Moreover, analysing the cortex and hippocampus, relevant regions for cognition, we found similar numbers in Iba1+ cells, a known microglia marker, in the hippocampus of infected pups. In contrast, a significant decrease was observed in the cortex, suggesting altered migration of these cells. Furthermore, microglia phagocytosis was increased in the cortex of infected pups but not in the hippocampus. Interestingly, quantification of neurons revealed a significant decrease in the hippocampus of infected pups while being increased in the cortex, compared with age-match controls. Altogether, our results show that GBS meningitis alters the neonatal microglia profile. Further studies will be necessary to better understand the microglia inflammatory state after GBS infection.
- «
- 1 (current)
- 2
- 3
- »