Cure and care at the cradle of innovation

“Illness is the night-side of life” tying one’s up in its own body and weaknesses leading either to curative or care spaces that instead of bringing hope bring to mind loneliness and death. Even if the tendency is to believe in the efficiency of medical processes, the collective memory of healthcare buildings is related to discomfort. Ill bodies enter a machine where they are homogenized, losing autonomy and privacy. Intimacy is exposed in a public domain. In healthcare buildings the focus is on medical procedure and not on the prostrate body, which is the real origin and dimensional parameter of these space

An Intangible Heritage in Use. Portuguese Institute of Oncology

The Portuguese Institute of Oncology (IPO) built in modern Lisbon, between 1927 and 1948, and added to until 1996, is the result of the Francisco Gentil effort to study and treat cancer. It is part of the Portuguese modern healthcare network and a reference concerning social, urban and architecture innovations, where the architects Cristino da Silva (1896–1936), Carlos Ramos (1897–1969), Raul Lino (1879–1974), Ernest Koop (1890–1962), Walter Diestel (1904–) and Raul Rodrigues de Lima (1909–1980) took part. By highlighting its cultural value this essay aims to stress the importance of achieving public and institutional awareness, in dealing with its everyday intensive use and transformation, towards a sustainable future

Physicochemical characteristics of organic honey samples of africanized honeybees from Paraná River islands.

This research was carried out to evaluate the physicochemical composition of organic honey in Paraná River islands, in Porto Brasílio, State of Paraná. Honey was harvested directly from super of the colonies in three apiaries spread in the Floresta and Laranjeira Islands, from August 2005 to August 2006. Twenty-four samples of organic honey produced by Africanized honeybees were evaluated. The following parameters were analyzed: pH, acidity, formol index, hydroxymethylfurfural, ashes, color, electric conductivity and moisture. Three replications per sample were performed for laboratorial analysis, giving the means and standard deviation. Most honey samples were in conformity with the Normative Instruction 11 from October 20, 2000. However, 4.17% were not in accordance with the moisture standards, 8.33% showed high concentrations of hydroxymethylfurfural, thus, totalizing 12.50% of non-complying samples. Nevertheless, 87.50% of the analyzed honey samples are within the standards, being characterized as an organic product of excellent quality, with good commercialization perspectives in the market

Genetic testing for germline variants in homologous recombination repair genes, other than BRCA1 and BRCA2, in patients with suspected hereditary cancer syndromes

Homologous recombination repair (HRR) is the cellular mechanism for error-free repair of DNA double-strand breaks. Pathogenic germline variants in BRCA1 and BRCA2 lead to HRR deficiency associated with breast, ovarian, prostate, pancreatic cancers and are sensitive to PARP inhibitors (PARPi). Defects in HRR genes beyond BRCA1/2 could also result in HRR deficiency and sensitize the tumor to PARPi, thus expanding the subset of patients that can benefit from these targeted therapy cancer drugs. We studied 56 DNA samples obtained from patients with personal and family history of cancer. Genes involved in HRR (ATM, BAP1, BLM, BRIP1, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, NBN, PALB2, RAD51C, RAD51D) were analysed by NGS using TruSight® Hereditary Cancer. Sequence alignment and annotation included DRAGEN Enrichment and Variant Interpreter - Illumina®. Variant classification, according to ACMG-AMP 1, was based on VEP, HSF, Alamut, VarSome and several databases (ex. HGMD, gnomAD, dbSNP). Variants of uncertain significance (VUS) were also classified with the stepwise ABC system2. All pathogenic/likely pathogenic SNVs and CNVs were confirmed by Sanger sequencing or MLPA. We identified 156 SNVs and one CNV, of these 125 were benign/likely benign. Seven clinically actionable variants were found in 10.7% of the patients: 3 pathogenic variants in FANCA, FANCD2 and FANCI give rise to premature stop codons and one pathogenic CNV in FANCA (deletion of exons 38 and 39); 2 likely pathogenic variants in BLM and FANCI affecting splicing and one frameshift in FANCG. Classification of 18 VUS with the ABC system resulted in: 8 class 0 (normal finding), 7 class E (potential interest) and 3 class D (low penetrance) variants. In addition, 7 SNVs were classified as hypomorphic alleles. This study confirmed: i) the importance of extending the molecular study beyond BRCA1/2 to other genes involved in HRR, ii) some variants require functional/family studies to establish their pathogenicity, and iii) these genes could potentially be considered for specific and clinical studies involving PARPi therapy.Funding: FCT/MCTES, Projects - ToxOmics and Human Health (UIDB/00009/2020) and GenomePT (POCI-01-0145-FEDER-022184).info:eu-repo/semantics/publishedVersio

RIBEIRA | Entre duas escarpas, uma linha de água

info:eu-repo/semantics/publishedVersio

The Adaptive Reuse of the Lisbon Riverside Area: the MMC Case Study: International docomomo Workshop

Lisbon, Portugal, 201

Search for alterations in genes involved in DNA repair by homologous recombination, excluding BRCA1 and BRCA2, associated with hereditary cancer.

Dissertação de Mestrado em Genética Clínica Laboratorial apresentada à Faculdade de MedicinaCancer occurs when, due to uncorrected DNA errors in our cells, there is a proliferation of this population of cells, which have the ability to invade other tissues. About 5-10% of cancers correspond to hereditary cancer, where the patient inherits a causative germline variant that, in the case of an autosomal dominant inheritance pattern, will be present in heterozygosity in all cells. When there is a 2nd event, this means, that a somatic pathogenic/pathogenic variant occurs in the second allele of the gene, according to the two-hit theory, the gene becomes non-functional making it difficult to repair the error originating in the founder cell, promoting the process of carcinogenesis. There are 5 distinct but interconnected repair pathways, one of them, repair by homologous recombination with several proteins involved, such as BRCA1/2. Currently, 4 inhibitors of PARP, enzymes involved in the base excision repair pathway, are approved in breast, ovarian, prostate, and pancreatic cancers if likely pathogenic/pathogenic variants in the BRCA1 and BRCA2 genes are identified due to their crucial role in the DNA repair by homologous recombination pathway. When likely pathogenic/pathogenic variants are present in the genes, involved in the homologous recombination repair pathway, there is no repair of errors by this pathway, and by inhibiting the activity of PARP enzymes both repair pathways are inhibited leading to accumulation of errors and subsequent cell death, termed selective lethality. Thus, the main objective of this work was to screen variants in genes involved in the repair pathway by homologous recombination, excluding BRCA1 and BRCA2, in 56 samples corresponding to patients with suspected hereditary cancer referred to the National Institute of Health Doutor Ricardo Jorge. The 18 genes analyzed were the following: ATM, BAP1, BLM, BRIP1, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, NBN, PALB2, RAD51C, RAD51D.To this end, a molecular analysis was performed using next-generation sequencing (NGS) methodology, in silico analysis using different predictors, software and databases of variants with allele frequencies below 5%; SNV's, SV's and CNV's were included in the analysis. In this analysis the ACMG-AMP guidelines were used for variant classification according to the established criteria. The identified probably pathogenic/pathogenic variants were confirmed using cycle Sanger sequencing methodology, or MLPA. For variants of uncertain significance (VUS), the stepwise ABC classification system was subsequently applied. A total of 156 SNVs were identified in this study, 3 probably pathogenic and 3 pathogenic, all confirmed by cycle Sanger sequencing methodology. Of these 6 causal variants identified, 4 of them most likely are targeted for NMD mechanism culminating in total allele loss. Of these 156 SNV's, 18 were classified as VUS, and with the subsequent application of the stepwise ABC classification system, 8 were classified as class 0, 7 as class E and 3 as class D. According to the studies published in the literature, other 7 of the identified variants can be classified as hypomorphic alleles.In silico analysis of CNV's/SV's suggested that some samples can have this type of variants, although only the deletion of exons 38 and 39 in the FANCA gene in sample HRR-47 was confirmed by MLPA. This deletion, based in silico results, can be related with a structural rearrangement involving exon 37. As it was not possible to confirm the mechanism of this rearrangement, two hypotheses were put forward, the first involving an initial duplication of exon 37 and subsequent deletion of the segment covering exons 37, 38 and 39; the second hypothesis involves the interstitial insertion of exon 37 in another region of the genome and the mentioned deletion in the FANCA locus. With this study, it was possible to assess the importance of functional studies, the confirmation of the variants identified by other methods, and evidence the importance of extending the molecular studies to other genes, as in 10.7% of the samples a causal variant was identified. Since the genes studied are involved in the same repair pathway as the BRCA1 and BRCA2, they may be considered as targets for iPARP therapy.O cancro ocorre quando, devido a erros (não corrigidos) no DNA das nossas células, ocorre a proliferação desregulada das células em causa, que adquirem a capacidade de invadir outros tecidos. Cerca de 5-10% dos cancros correspondem a cancro hereditário, onde o indivíduo herda uma variante germinativa causal que, no caso de um padrão de hereditariedade autossómico dominante, estará presente em heterozigotia em todas as células. Quando existe um 2º evento, ou seja, uma variante provavelmente patogénica/patogénica somática no segundo alelo do gene, segundo a teoria two-hit, este fica não funcional, perdendo a capacidade de reparar erros que ocorrem no DNA ou de regular a divisão celular, originando-se assim uma célula fundadora que promoverá o processo de carcinogénese. Existem 5 vias de reparação distintas, mas interligadas, sendo uma delas a reparação por recombinação homóloga onde estão envolvidas diversas proteínas tais como BRCA1/2. Atualmente encontram-se aprovados 4 inibidores da PARP, enzimas envolvidas na via de reparação base excision repair, em situações de cancro da mama, ovário, próstata, pâncreas em que sejam identificadas variantes provavelmente patogénicas/patogénicas nos genes BRCA1 e BRCA2 devido ao seu papel crucial na via de reparação do DNA por recombinação homóloga. Existindo variantes provavelmente patogénicas/patogénicas nos genes envolvidos na via de reparação por recombinação homóloga não existe reparação dos erros por esta via, e inibindo-se a atividade das enzimas PARP, ambas as vias de reparação são inibidas levando à acumulação de erros e posterior morte celular, denominada de letalidade seletiva. Desta forma, o principal objetivo deste trabalho, foi a pesquisa de variantes em genes envolvidos na via de reparação por recombinação homóloga, excluindo BRCA1 e BRCA2, em 56 amostras correspondentes a doentes com suspeita de cancro hereditário referenciados ao Instituto Nacional de Saúde Doutor Ricardo Jorge. Os 18 genes analisados foram os seguintes: ATM, BAP1, BLM, BRIP1, FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCL, FANCM, NBN, PALB2, RAD51C, RAD51D.Para tal foi feita a análise molecular pela metodologia de sequenciação de nova geração (NGS), análise in silico recorrendo a vários preditores, softwares e bases de dados das variantes com frequência alélica abaixo dos 5%, na análise foram incluídos SNV’s, SV’s e CNV’s. Nesta análise utilizou-se a classificação da ACMG-AMP para classificação de variantes, de acordo com os respetivos critérios definidos. As variantes provavelmente patogénicas/patogénicas identificadas foram confirmadas através da metodologia de sequenciação cíclica de Sanger, ou MLPA. Já para as variantes de significado incerto (VUS) foi aplicado posteriormente o sistema de classificação stepwise ABC. Neste estudo foram identificadas um total de 156 SNV’s, 3 provavelmente patogénicas e 3 patogénicas, todas confirmadas pela metodologia de sequenciação cíclica de Sanger. Destas 6 variantes causais identificadas, em 4 o mais provável será serem alvo de mecanismo NMD culminando na perda total do alelo. Destas 156 SNV’s, 18 foram classificadas como VUS, e com a posterior aplicação do sistema de classificação stepwise ABC, 8 foram classificadas como classe 0, 7 como classe E e 3 como classe D. De acordo com a análise da literatura científica, com base nos estudos funcionais realizados, 7 variantes poderão corresponder a alelos hipomórficos.A análise in silico de CNV’s/SV’s sugeriu a existência de variantes deste tipo em algumas amostras, contudo apenas a deleção do exão 38 e 39 no gene FANCA na amostra HRR-47 foi confirmada por MLPA. Esta deleção, de acordo com aos alertas obtidos na análise in silico, terá compreendido um rearranjo estrutural envolvendo o exão 37. Não tendo sido possível confirmar o mecanismo do referido rearranjo foram consideradas 2 hipóteses, a primeira envolvendo uma duplicação inicial do exão 37 e posterior deleção do segmento envolvendo os exões 37, 38 e 39; já a segunda hipótese envolve a inserção intersticial do exão 37 em outra região do genoma e a deleção nos exões em causa no respetivo locus do FANCA. Com o desenvolvimento deste trabalho foi possível aferir a importância dos estudos funcionais, da confirmação das variantes identificadas por outras metodologias e ainda evidenciar a importância do alargamento do estudo molecular a outros genes, dado que em 10,7% das amostras foi identificada uma variante causal. Estando os genes estudados envolvidos na mesma via de reparação que os genes BRCA1 e BRCA2 poderão ser tidos em conta como alvos de terapêutica com iPARP.Outro - Financiado pelo Instituto Nacional de Saúde Doutor Ricardo Jorge

Decreased viability and neurite length in neural cells treated with chitosan-dextran sulfate nanocomplexes.

CXCL12 is a chemokine known to regulate migration, proliferation, and differentiation of neural stem cells (NSCs) and to play a neuroprotective role in ischemic stroke. Chitosan-dextran sulfate nanocomplexes (Ch/DS NC) are known nanoparticulated systems used to efficiently deliver heparin-binding factors. Here we evaluate Ch/DS NC as carriers for CXCL12 in a mouse model of stroke. Free CXCL12 reduced the size of the ischemic brain lesion. However, when Ch/DS NC were administrated, the stroke volume increased. Neurotoxic screening revealed that Ch/DS NC reduced neuronal viability, decreased the extension of neurites and impaired NSC migration in vitro. To the best of our knowledge, neurotoxicity of Ch/DS NC has not been reported and further screenings will be needed in order to evaluate the biological safety of these nanocomposites. Our results add new data on nanoparticle neurotoxicity and may help us to better understand the complex interactions of the nanostructures with biological components