From genes to radioresistance in head and neck squamous cell carcinoma

Head and Neck Cancers (HNC) are a group of tumours located in the upper aero-digestive tract. Head and Neck Squamous Cell Carcinoma (HNSCC) represent about 90% of all HNC cases. It has been considered the sixth most malignant tumour worldwide and, despite clinical and technological advances, the five-year survival rate has not improved much in the last years. Nowadays, HNSCC is well established as a heterogeneous disease and that its development is due to accumulation of genetic events. Apart from the majority of the patients being diagnosed in an advanced stage, HNSCC is also a disease with poor therapeutic outcome. One of the therapeutic approaches is radiotherapy. However, this approach has different drawbacks like the radioresistance acquired by some tumour cells, leading to a worse prognosis. A major knowledge in radiation biology is imperative to improve this type of treatment and avoid late toxicities, maintaining patient quality of life in the subsequent years after treatment. Then, identification of genetic markers associated to radiotherapy response in patients and possible alterations in cells after radiotherapy are essential steps towards an improved diagnosis, higher survival rate and a better life quality. Not much is known about the radiation effects on cells, so, the principal aim of this study was to contribute to a more extensive knowledge about radiation treatment in HNSCC. For this, two commercial cell lines, HSC-3 and BICR-10, were used and characterized resorting to karyotyping, aCGH and MS-MLPA. These cell lines were submitted to different doses of irradiation and the resulting genetic and methylation alterations were evaluated. Our results showed a great difference in radiation response between the two cell lines, allowing the conclusion that HSC-3 was much more radiosensitive than BICR-10. Bearing this in mind, analysis of cell death, cell cycle and DNA damages was performed to try to elucidate the motifs behind this difference. The characterization of both cell lines allowed the confirmation that HSC-3 was derived from a metastatic tumour and the hypothesis that BICR-10 was derived from a dysplasia. Furthermore, this pilot study enabled the suggestion of some genetic and epigenetic alterations that cells suffer after radiation treatment. Additionally, it also allowed the association of some genetic characteristics that could be related to the differences in radiation response observable in this two cell lines. Taken together all of our results contribute to a better understanding of radiation effects on HNSCC allowing one further step towards the prediction of patients’ outcome, better choice of treatment approaches and ultimately a better quality of life.

Evaluation of circulating endothelial progenitor cells by multicolor flow cytometry in chronic kidney disease patients

Endothelial dysfunction and impaired endothelial regenerative capacity play a key role in the pathogenesis of cardiovascular disease, which is one of the major causes of mortality in chronic kidney disease (CKD) patients. Circulating endothelial cells (CEC) may be an indicator of vascular damage, while circulating endothelial progenitor cells (EPC) may be a biomarker for vascular repair. However, the simultaneously evaluation of CEC and EPC circulating levels and its relation were not previously examined in CKD population. A blood sample (18ml) of healthy subjects (n=10), early CKD (n=10) and advanced CKD patients (n=10) was used for the isolation of early and late EPCs, CECs, and hematopoietic cells, identified by flow cytometry (BD FACSCanto™ II system) using a combination of fluorochrome-conjugated primary antibodies: CD31-PE, CD45-APC Cy7, CD34-FITC, CD117-PerCp Cy5.5, CD133-APC, CD146-Pacific Blue, and CD309-PECy7. Exclusion of dead cells was done according to a fixable viability dye staining. This eightcolor staining flow cytometry optimized protocol allowed us to accurate simultaneously identify EPCs, CECs and hematopoietic cells. In addition, it was also possible to distinguish the two subpopulations of EPCs, early and late EPCs subpopulation, by CD45intCD31+CD34+CD117-CD133+CD309-CD146- and CD45intCD31+CD34+CD117-CD133-CD309+CD146- multiple labeling, respectively. Moreover, the identification of CECs and hematopoietic cells was performed by CD45-CD31+CD34-/lowCD117-CD133-CD309-CD146+ and CD34+CD117+, respectively. The levels of CECs were non-significantly increased in early CKD (312.06 ± 91.34) and advanced CKD patients (191.43±49.86) in comparison with control group (103.23±24.13). By contrast, the levels of circulating early EPCs were significantly reduced in advanced CKD population (17.03±3.23) in comparison with early CKD (32.31±4.97), p=0.04 and control group (36.25 ± 6.16), p=0.03. In addition the levels of late EPCs were significantly reduced in both advanced (6.60±1.89), p=0.01, and early CKD groups (8.42±2.58), p=0.01 compared with control group (91.54±29.06). These results were accompanied by a dramatically reduction in the recruitment, differentiation and regenerative capacity indexes in CKD population. Taken together, these results suggest an imbalance in the process of endothelial repairment in CKD population, and further propose that the indexes of recruitment, differentiation and regenerative capacity of EPCs, may help to select the patients to benefit from guiding intervention strategies to improve cardiovascular health by inducing vascular protection.

Qualidade de vida em pacientes com a doença de Graves Basedow

Dissertação de Mestrado apresentada no Instiuto Superior de Psicologia Aplicada para obtenção do grau de Mestre na especialidade de Psicologia Clínica

Prevalência de úlceras de pressão em dois hospitais portugueses: Impacto da Escala de Braden associada a um protocolo de prevenção

As úlceras de pressão constituem um importante problema de saúde pública comum a diversos cenários de cuidados, em todo o mundo, e também em Portugal, nomeadamente para a população mais idosa. O presente estudo teve como objectivos determinar o impacto da escala de Braden associada a um protocolo de prevenção de úlceras de pressão na prevalência destas lesões, assim como, monitorizar o problema das úlceras de pressão em dois hospitais portugueses, em que o Hospital A utiliza estes instrumento e o Hospital 8 não usa esses instrumentos. Foi utilizado o Instrumento de Prevalência de Úlceras de Pressão validado para Portugal na recolha dos dados. Os dados foram obtidos através da observação directa dos doentes. Foram realizadas duas avaliações com um intervalo de três meses em cada hospital. Os resultados encontrados revelaram que as úlceras de pressão constituem um problema de saúde para os dois hospitais observados. No entanto, o Hospital A apresenta menor taxa de prevalência de úlceras de pressão, menor número de úlceras de pressão mais graves, menor possibilidade de um doente adquirir uma úlcera de pressão e maior utilização de medidas preventivas. Os valores encontrados foram estatisticamente significativos para p <0,05. Confirma-se que a utilização da escala de Braden associada a um protocolo de prevenção de úlceras de pressão é uma medida eficaz na redução do número de úlceras de pressão. Para uma maior eficácia deste instrumento será importante a sua utilização como ferramenta de controlo da adequação das medidas preventivas. ABSTRACT; Pressure ulcers are an important public health problem common to several scenarios of care in the world, and also in Portugal, particularly for the elderly. This study aims to determine the impact of the Braden scale associated with a protocol for prevention of pressure ulcers in prevalence of these lesions, as well as monitor the problem of pressure ulcers in two Portuguesa hospitais, the Hospital A uses these instrument and Hospital B doesn’t. It was used the instrument of Prevalence of pressure ulcers validated for Portugal in the data collection. Data were collected through direct observation of patients. There were two assessments with an interval of three months in each hospital. The results showed that the pressure ulcers are a health problem for both hospitals observed. However, the Hospital A provides a lower prevalence rate of pressure ulcers, lower number of pressure ulcers more serious, less possibility of a patient acquiring a pressure ulcer and increased use of preventive measures. The values were statistically significant for p<0.05. lt is confirmed that the use of the Braden scale associated with a protocol for prevention of pressure ulcers is an effective measure in reducing the number of pressure ulcers. For greater efficiency will be important to use it as a tool for monitoring the adequacy of preventive measures.

Influenza vaccine effectiveness in Portugal: season 2015/2016 report

This report was prepared as part of the Project “Monitoring Influenza vaccine effectiveness during influenza seasons and pandemics in the European Union” and describes the results obtained in Portugal under the Protocol Agreement celebrated between EpiConcept SARL, Paris and National Health Institute Dr. Ricardo Jorge, Lisbon. Data and activities related to the individuals 65 years and more were funded by European Union’s Horizon 2020 research and innovation programme under grant agreement no 634446.

FTIR, a potential tool to dementia diagnosis trough analysis of plasma

Nowadays it is still difficult to perform an early and accurate diagnosis of dementia, therefore many research focus on the finding of new dementia biomarkers that can aid in that purpose. So scientists try to find a noninvasive, rapid, and relatively inexpensive procedures for early diagnosis purpose. Several studies demonstrated that the utilization of spectroscopic techniques, such as Fourier Transform Infrared Spectroscopy (FTIR) and Raman spectroscopy could be an useful and accurate procedure to diagnose dementia. As several biochemical mechanisms related to neurodegeneration and dementia can lead to changes in plasma components and others peripheral body fluids, blood-based samples and spectroscopic analyses can be used as a more simple and less invasive technique. This work is intended to confirm some of the hypotheses of previous studies in which FTIR was used in the study of plasma samples of possible patient with AD and respective controls and verify the reproducibility of this spectroscopic technique in the analysis of such samples. Through the spectroscopic analysis combined with multivariate analysis it is possible to discriminate controls and demented samples and identify key spectroscopic differences between these two groups of samples which allows the identification of metabolites altered in this disease. It can be concluded that there are three spectral regions, 3500-2700 cm -1, 1800-1400 cm-1 and 1200-900 cm-1 where it can be extracted relevant spectroscopic information. In the first region, the main conclusion that is possible to take is that there is an unbalance between the content of saturated and unsaturated lipids. In the 1800-1400 cm-1 region it is possible to see the presence of protein aggregates and the change in protein conformation for highly stable parallel β-sheet. The last region showed the presence of products of lipid peroxidation related to impairment of membranes, and nucleic acids oxidative damage. FTIR technique and the information gathered in this work can be used in the construction of classification models that may be used for the diagnosis of cognitive dysfunction.

Fisioterapia em cuidados paliativos: da evidência à prática

Com início nos anos 50, através de um empenho invejável de Cicely Saunders, em Inglaterra, que avaliando as diferentes necessidades dos doentes em fim de vida, foi enfermeira, assistente social e médica, colmatando desta forma as diferentes e complexas necessidades destes doentes. Portugal começou muito mais tarde, em 1994, mas tem vindo a crescer o interesse de muitos profissionais de saúde implicados em deseuvolver os Cuidados Paliativos, na organização, no trabalho no terreno e na investigação, de modo a permitir que cada vez mais portugueses tenham acesso a este tipo de cuidados. Quando se aborda este tema, fala-se sobre o sofrimento que acompanha as doenças graves, progressivas e incuráveis. Este sofrimento pode ser de carácter físico, psicológico, espiritual e social. Os cuidados paliativos, tradicionalmente dirigidos para as doenças do foro oncológico, têm vindo a tomar uma importância significativa nas patologias não oncológicas: falências de órgãos, doenças neurológicas degenerativas, nomeadamente as demências, doentes vegetativos, etc. que, pelas alterações demográficas que se esperam acontecer, podem vir a predominar na necessidade de cuidados específicos que se encaixam na filosofia dos cuidados paliativos. Os cuidados paliativos caracterizam-se por uma filosofia que implica a intervenção de vários grupos profissionais. De acordo com Twycioss, uma equipa básica deve incluir: um médico, um enfermeiro e um assistente social. As situações em cuidados paliativos podem apresentai uma complexidade tão significativa que exigem intervenção de outros grupos profissionais. Twycross afirma, ainda, que uma equipa é tão mais rica, quanto mais grupos profissionais diferentes a compõem.