Repositório RCAAP

A polysulfide material was synthesized by the direct reaction of sulfur and d-limonene, by-products of the petroleum and citrus industries, respectively. The resulting material was processed into functional coatings or molded into solid devices for the removal of palladium and mercury salts from water and soil. The binding of mercury(II) to the sulfur-limonene polysulfide resulted in a color change. These properties motivate application in next-generation environmental remediation and mercury sensing.

In drug discovery and drug development, it is estimated that around 40% of commercialized and 90% of under-study drugs have inadequate pharmaceutical properties, severely impairing its therapeutic efficacy. Thus, there is a strong demand to find strategies to enhance the delivery of such drugs. Ionic liquids are a novel class of liquids composed of a combination of organic salts that are of particular interest alone or in combination with drug delivery systems. This review is focused on the recent efforts using ionic liquids in drug solubility, formulation and drug delivery with specific emphasis on nanotechnology. The latest developments using hybrid delivery systems obtained upon the combination of drug delivery systems and ionic liquids will also be addressed.

UIS4 is a key protein component of the host-parasite interface in the liver stage of the rodent malaria parasite Plasmodium berghei and required for parasite survival after invasion. In the infectious sporozoite, UIS4 protein has variably been shown to be translated but also been reported to be translationally repressed. Here we show that uis4 mRNA translation is regulated by the P. berghei RNA binding protein Pumilio-2 (PbPuf2 or Puf2 from here on forward) in infectious salivary gland sporozoites in the mosquito vector. Using RNA immunoprecipitation we show that uis4 mRNA is bound by Puf2 in salivary gland sporozoites. In the absence of Puf2, uis4 mRNA translation is de-regulated and UIS4 protein expression upregulated in salivary gland sporozoites. Here, using RNA immunoprecipitation, we reveal the first Puf2-regulated mRNA in this parasite.

INTRODUÇÃO: Mais de 70% dos doentes com cancro apresenta dor. Os doentes oncológicos podem ter quadros dolorosos de complexidade variada. Na maioria dos casos é possível alcançar um bom controlo da dor, contudo, alguns doentes com síndromes álgicas complexas podem ser difíceis de tratar. A intensidade da dor é, sem dúvida, uma das dimensões clinicamente mais relevantes na experiência da dor. Várias características são bem conhecidas como podendo influenciar a resposta a diferentes tratamentos. OBJECTIVOS: Pretendeu-se demonstrar que a intensidade da dor na avaliação inicial (Di) era um factor preditivo de complexidade no tratamento da dor oncológica. A complexidade congregava: tempo para controlar a dor (Tm); prescrição de opiáceos (PO); dose final de opiáceos (DDEM); tolerância final aos opiáceos (índice de escalada de opiáceos, IEO>4% dia); prescrição de co-analgésicos e de adjuvantes (COAA); utilização de atendimentos profissionais não programados (ANP). Pretendeu-se também identificar outros factores inerentes a características individuais, clínicas, tumorais e dolorosas, que pudessem afectar a expressão da complexidade relacionada com a Di. MÉTODOS: Estudo quantitativo, experimental, prospectivo, não controlado; envolvendo doentes consecutivos com cancro e com dor oncológica, ambulatórios, seguidos numa Consulta de Dor. A Di foi medida utilizando uma escala numérica graduada de 0 a 10. Os participantes foram avaliados durante 70 dias através de 3 consultas (inicial; intermédia, às 5 semanas; e final) e 8 telefonemas semanais (4 após a consulta inicial; 4 após a consulta intermédia). Foram calculadas as associações entre Di e: Tm (Kaplan-Meier); PO, IEO e COAA (Wilcoxon); Tipo de opiáceos (TO), DDEM e ANP (Kruskal-Wallis). Foram também analisadas as associações entre Di e várias características individuais, clínicas, tumorais e relativas à dor dos participantes. RESULTADOS: 264 incluídos (Sobreviventes=184, Óbitos=65, Transferidos=15). Sucesso=88% (dor controlada). Complexidade) Tm=21dias, 96% PO, DDEM= 104.3mg, mais de 22% com IEO>4% dia, COAA [80% AINE, anti-inflamatórios não esteróides; 49% AD (antidepressivos); 8% MR (miorelaxantes)], 97% ANP (média 3.16+1.40). Individuais) 50% mulheres, idades 63.74±21.07 anos, 65% casados, 70% escolaridade básica, 72% reformados, 87% rendimento mensal<450 euros. Clínicos) 81% com comorbilidades (50% cardiovasculares), 88% com estado ansioso/depressivo, 59% dependentes de terceiros, ECOG=1.48 (média), 57% em cuidados paliativos. Tumorais) 90% carcinomas (30% Cabeça/Pescoço, 25% Intestino), 76% metástases (35% ósseas). Dor) Di=7.92 (média), 67% dor mista (neuropática e nociceptiva), 78% dor episódica. Estatística) Di associada a: Tm (p=0.002), PO (p=0.002), DDEM final (p=0.009), tolerância IEO>4% (p=0.009), AINE (p=0.015), AD (p=0.002), MR (p=0.03), Histologia tumoral (p=0.046). Tm associado a: PO (p<0.001), TO (p=0.001). ANP: Telefonema Dor associado a Consulta Dor (p=0.0004). Sem outras correlações detectadas na análise bivariada. CONCLUSÕES: A Di é um factor preditivo significativo de complexidade do tratamento da dor oncológica. Os doentes com dor insuportável necessitam, para controlar a dor, de: mais Tm (que depende da PO e do TO), opiáceos, AINE, RM e AD. Os doentes com dor insuportável fazem doses superiores de opiáceos e desenvolvem mais tolerância. Os doentes com carcinomas têm dores de pior prognóstico. Os doentes que mais utilizam o recurso telefónico são os que mais utilizam as Consultas de Dor não programadas por sintomatologia descontrolada.

Iron is an essential micronutrient but is also highly toxic. In yeast and plant cells, a key detoxifying mechanism involves iron sequestration into intracellular storage compartments, mediated by members of the vacuolar iron-transporter (VIT) family of proteins. Here we study the VIT homologue from the malaria parasites Plasmodium falciparum (PfVIT) and Plasmodium berghei (PbVIT). PfVIT-mediated iron transport in a yeast heterologous expression system is saturable (Km ∼ 14.7 μM), and selective for Fe(2+) over other divalent cations. PbVIT-deficient P. berghei lines (Pbvit(-)) show a reduction in parasite load in both liver and blood stages of infection in mice. Moreover, Pbvit(-) parasites have higher levels of labile iron in blood stages and are more sensitive to increased iron levels in liver stages, when compared with wild-type parasites. Our data are consistent with Plasmodium VITs playing a major role in iron detoxification and, thus, normal development of malaria parasites in their mammalian host.

Mutations in the gene encoding the mitochondrial kinase PINK1 cause early-onset familial Parkinson's disease. To understand the biological function of PINK1 and its role in the pathogenesis of Parkinson's disease, it is useful to study its kinase activity towards substrates both in vivo and in vitro. For in vitro kinase assays, a purified Triboleum castaneum PINK1 insect orthologue is often employed, because it displays higher levels of activity when compared to human PINK1. We show, however, that the activity requirements, and more importantly the substrate specificity, differ between both orthologues. While Triboleum castaneum PINK1 readily phosphorylates the PINKtide peptide and Histone H1 in vitro, neither of these non-physiological substrates is phosphorylated by human PINK1. Nonetheless, both Tc and human PINK1 phosphorylate Parkin and Ubiquitin, two physiological substrates of PINK1. Our results show that the substrate selectivity differs among PINK1 orthologues, an important consideration that should be taken into account when extrapolating findings back to human PINK1.

Globally, thyroid cancer accounts for 2 % of all cancer diagnoses, and can be classified as well-differentiated or undifferentiated. Currently, differentiated thyroid carcinomas have good prognoses, and can be treated with a combination of therapies, including surgical thyroidectomy, radioactive iodine therapy and hormone-based therapy. On the other hand, anaplastic thyroid carcinoma, a subtype of undifferentiated thyroid carcinoma characterized by the loss of thyroid-like phenotype and function, does not respond to either radioactive iodine or hormone therapies. In most cases, anaplastic thyroid carcinomas are diagnosed in later stages of the disease, deeming them inoperable, and showing poor response rates to systemic chemotherapy. Recently, treatment courses using multiple-target agents are being explored and clinical trials have shown very promising results, such as overall survival rates, progression-free survival and tumor shrinkage. This review is focused on thyroid carcinomas, with particular focus on anaplastic thyroid carcinoma, exploring its undifferentiated nature. Special interest will be given to the treatment approaches currently available and respective obstacles or drawbacks. Our purpose is to contribute to understand why this malignancy presents low responsiveness to current treatments, while overviewing novel therapies and clinical trials.

The pandemic brought migrant farm workers into the limelight once again, as has happened repeatedly in the last three decades, in Italy as in many other parts of the world. Here I examine how intersecting and sometimes conflicting discourses and interventions, that have this biopolitically conceived population as their object, decide upon these subjects’ worthiness of attention, care, and sympathy through criminalizing, victimizing, and humanitarian registers. I reflect on some of the affective dynamics that sustain both the governmental operations through which these populations were (sought to be) managed and reactions against them from a situated perspective, as an accomplice to many of the forms of struggle in which migrant farm workers have engaged in the last decade in Italy. The stage for many such occurrences is what I have elsewhere defined as the “encampment archipelago” that many such workers, and particularly those who migrate from across West Africa, inhabit—labor or asylum-seeker camps, but also slums or isolated, derelict buildings, and various hybrid, in-between spaces among which people circulate.

Currently, insulin can only be administered through the subcutaneous route. Due to the flaws associated with this route, it is of interest to orally deliver this drug. However, insulin delivered orally has several barriers to overcome as it is degraded by the stomach’s low pH, enzymatic content, and poor absorption in the gastrointestinal tract. Polymers with marine source like chitosan are commonly used in nanotechnology and drug delivery due to their biocompatibility and special features. This work focuses on the preparation and characterization of mucoadhesive insulin-loaded polymeric nanoparticles. Results showed a suitable mean size for oral administration (<600 nm by dynamic laser scattering), spherical shape, encapsulation efficiency (59.8%), and high recovery yield (80.6%). Circular dichroism spectroscopy demonstrated that protein retained its secondary structure after encapsulation. Moreover, the mucoadhesive potential of the nanoparticles was assessed in silico and the results, corroborated with ex-vivo experiments, showed that using chitosan strongly increases mucoadhesion. Besides, in vitro and in vivo safety assessment of the final formulation were performed, showing no toxicity. Lastly, the insulin-loaded nanoparticles were effective in reducing diabetic rats’ glycemia. Overall, the coating of insulin-loaded nanoparticles with chitosan represents a potentially safe and promising approach to protect insulin and enhance peroral delivery.

The Angolan village of Cusseque, established during thecountry’s civil war, lost its military purpose when the conflictended in 2002. The village’s neighbors assumed that Cusseque’sremaining residents would leave; most stayed. They have sincefought to legitimize their presence. Fieldwork with Cussequeresidents helps illuminate why they assert their merging withthe land—a merging that I callbodyland—through the agencyof honeybees. These men and women exert a compelling bodypolitics, one that is subject to the more-than-human agenciesthat dissolve the contours of the human. Moreover, Cusseque’sresidents contribute to anthropological discussions abouthuman-land relations by blending the question of the humanwith that of the land. They defy pervasive humanist regimes inwhich the legitimacy of human presence is dissociated from theinterdependence between body and land. [human-landrelationship,legitimacy,transcorporeality,honey,honeybees,postwar,body politics,Angola]

Synucleins belong to a family of intrinsically unstructured proteins that includes alpha-synuclein (aSyn), beta-synuclein (bSyn) and gamma-synuclein (gSyn). aSyn is the most studied member of the synuclein family due to its central role in genetic and sporadic forms of Parkinson's disease and other neurodegenerative disorders known as synucleionopathies. In contrast, bSyn and gSyn have been less studied, but recent reports also suggest that, unexpectedly, these proteins may also cause neurotoxicity. Here, we explored the yeast toolbox to investigate the cellular effects of bSyn and gSyn. We found that bSyn is toxic and forms cytosolic inclusions that are similar to those formed by aSyn. Moreover, we found that bSyn shares similar toxicity mechanisms with aSyn, including vesicular trafficking impairment and induction of oxidative stress. We demonstrate that co-expression of aSyn and bSyn exacerbates cytotoxicity, due to increased dosage of toxic synuclein forms, and that they are able to form heterodimers in both yeast and in human cells. In contrast, gSyn is not toxic and does not form inclusions in yeast cells. Altogether, our findings shed light into the question of whether bSyn can exert toxic effects and confirms the occurrence of aSyn/bSyn heterodimers, opening novel perspectives for the development of novel strategies for therapeutic intervention in synucleinopathies.

Fungal invasive infections are an increasing health problem. The intrinsic complexity of pathogenic fungi and the unmet clinical need for new and more effective treatments requires a detailed knowledge of the infection process. During infection, fungal pathogens are able to trigger a specific transcriptional program in their host cells. The detailed knowledge of this transcriptional program will allow for a better understanding of the infection process and consequently will help in the future design of more efficient therapeutic strategies. Simultaneous transcriptomic studies of pathogen and host by high-throughput sequencing (dual RNA-seq) is an unbiased protocol to understand the intricate regulatory networks underlying the infectious process. This protocol is starting to be applied to the study of the interactions between fungal pathogens and their hosts. To date, our knowledge of the molecular basis of infection for fungal pathogens is still very limited, and the putative role of regulatory players such as non-coding RNAs or epigenetic factors remains elusive. The wider application of high-throughput transcriptomics in the near future will help to understand the fungal mechanisms for colonization and survival, as well as to characterize the molecular responses of the host cell against a fungal infection.

Endothelial cells (ECs) are plastic cells that can switch between growth states with different bioenergetic and biosynthetic requirements. Although quiescent in most healthy tissues, ECs divide and migrate rapidly upon proangiogenic stimulation. Adjusting endothelial metabolism to the growth state is central to normal vessel growth and function, yet it is poorly understood at the molecular level. Here we report that the forkhead box O (FOXO) transcription factor FOXO1 is an essential regulator of vascular growth that couples metabolic and proliferative activities in ECs. Endothelial-restricted deletion of FOXO1 in mice induces a profound increase in EC proliferation that interferes with coordinated sprouting, thereby causing hyperplasia and vessel enlargement. Conversely, forced expression of FOXO1 restricts vascular expansion and leads to vessel thinning and hypobranching. We find that FOXO1 acts as a gatekeeper of endothelial quiescence, which decelerates metabolic activity by reducing glycolysis and mitochondrial respiration. Mechanistically, FOXO1 suppresses signalling by MYC (also known as c-MYC), a powerful driver of anabolic metabolism and growth. MYC ablation impairs glycolysis, mitochondrial function and proliferation of ECs while its EC-specific overexpression fuels these processes. Moreover, restoration of MYC signalling in FOXO1-overexpressing endothelium normalizes metabolic activity and branching behaviour. Our findings identify FOXO1 as a critical rheostat of vascular expansion and define the FOXO1-MYC transcriptional network as a novel metabolic checkpoint during endothelial growth and proliferation.

The syntheses and antiplasmodial activities of various substituted aminoquinolines coupled to an adamantane carrier are described. The compounds exhibited pronounced in vitro and in vivo activity against Plasmodium berghei in the Thompson test. Tethering a fluorine atom to the aminoquinoline C(3) position afforded fluoroaminoquinolines that act as intrahepatocytic parasite inhibitors, with compound 25 having an IC50 = 0.31 μM and reducing the liver load in mice by up to 92% at 80 mg/kg dose. Screening our peroxides as inhibitors of liver stage infection revealed that the tetraoxane pharmacophore itself is also an excellent liver stage P. berghei inhibitor (78: IC50 = 0.33 μM). Up to 91% reduction of the parasite liver load in mice was achieved at 100 mg/kg. Examination of tetraoxane 78 against the transgenic 3D7 strain expressing luciferase under a gametocyte-specific promoter revealed its activity against stage IV-V Plasmodium falciparum gametocytes (IC50 = 1.16 ± 0.37 μM). To the best of our knowledge, compounds 25 and 78 are the first examples of either an 4-aminoquinoline or a tetraoxane liver stage inhibitors.

Com a evolução da web 2.0, a maioria das empresas elabora negócios através da Internet usando aplicações web. Estas aplicações detêm dados importantes com requisitos cruciais como confidencialidade, integridade e disponibilidade. A perda destas propriedades influencia directamente o negócio colocando-o em risco. A percepção de risco providencia o necessário conhecimento de modo a agir para a sua mitigação. Nesta tese foi concretizada uma colecção de honeypots web de alta interacção utilizando diversas aplicações e sistemas operativos para analisar o comportamento do atacante. A utilização de ambientes de virtualização assim como ferramentas de monitorização de honeypots amplamente utilizadas providencia a informação forense necessária para ajudar a comunidade de investigação no estudo do modus operandi do atacante, armazenando os últimos exploits e ferramentas maliciosas, e a desenvolver as necessárias medidas de protecção que lidam com a maioria das técnicas de ataque. Utilizando a informação detalhada de ataque obtida com os honeypots web, o comportamento do atacante é classificado entre diferentes perfis de ataque para poderem ser analisadas as medidas de mitigação de risco que lidam com as perdas de negócio. Diferentes frameworks de segurança são analisadas para avaliar os benefícios que os conceitos básicos de segurança dos honeypots podem trazer na resposta aos requisitos de cada uma e a consequente mitigação de risco.

In recent years, the scientific community has seen increasing use of natural compounds instead of chemical compounds in medicines, food supplements, cosmetics, and dermatological products. Nanotechnology, mainly used in the cosmetic and pharmaceutical industries, has also been increasingly used. In this context the purpose of nanotechnology is to increase the stability of active compounds, to modulate their release, as well as to improve the solubility of poorly water-soluble compounds. In this chapter, we will discuss key examples of natural products and their biological activities, as well as the advantages of using nanotechnology, combining them in food supplements, cosmetics, and dermatological products, with a brief overview of current products on the market.

Breast cancer is one of the most common cancers worldwide, which makes it a very impactful malignancy in the society. Breast cancers can be classified through different systems based on the main tumor features and gene, protein, and cell receptors expression, which will determine the most advisable therapeutic course and expected outcomes. Multiple therapeutic options have already been proposed and implemented for breast cancer treatment. Nonetheless, their use and efficacy still greatly depend on the tumor classification, and treatments are commonly associated with invasiveness, pain, discomfort, severe side effects, and poor specificity. This has demanded an investment in the research of the mechanisms behind the disease progression, evolution, and associated risk factors, and on novel diagnostic and therapeutic techniques. However, advances in the understanding and assessment of breast cancer are dependent on the ability to mimic the properties and microenvironment of tumors in vivo, which can be achieved through experimentation on animal models. This review covers an overview of the main animal models used in breast cancer research, namely in vitro models, in vivo models, in silico models, and other models. For each model, the main characteristics, advantages, and challenges associated to their use are highlighted.

Este relatório tem como foco o desenvolvimento e criação de um corpus de provas tipográficas pertencentes ao espólio1 de José Leite de Vasconcelos conservado no Museu Nacional de Arqueologia. Os pacotes2, que compõem o Legado de Leite de Vasconcelos, utilizados neste projeto foram os respeitantes à matéria de Filologia3. Numa primeira fase, identificou-se e organizou-se os materiais por tema e criou-se um catálogo dos mesmos para facilitar a posterior pesquisa. Numa segunda fase, fez-se fotografia de todas as provas tipográficas encontradas e conseguinte atribuição de cotas e edição de imagens. Ulteriormente, identificou-se a que obras pertenciam as provas e elaborou-se com essa informação tabelas de «correspondência»4. Por fim, este trabalho terá uma parte mais interpretativa, onde o foco será o chamado «dialeto brasileiro». Este foi um tema pouco tratado por Leite de Vasconcelos e, no entanto, teve relevância bastante para estar presente nos seus apontamentos e investigações. Foram encontrados, em específico, dois envelopes, que continham informação recolhida pelo autor, sobre este tema. Daí enveredou-se pelo estudo do seu conteúdo, fazendo uma descrição detalhada dos suportes materiais lá contidos.

Esta tese colocará em evidência a importância do triângulo constituído pelo desejo, imaginação e prazer, em A La Recherche du Temps Perdu, e com isso apresentar uma teoria poustiana sobre aquilo que constitui a criação artística. Far-se-á também ver como é que o processo desencadeado por este triângulo ganha forma em vários passos da Recherche, e com isso perceber o modo como Marcel se relaciona com os seus três vértices. Por fim, procurar-se-á mostrar de que forma essa teoria sobre a arte entra em diálogo com aquilo que Descartes defende em Meditações Metafísicas.

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