The role of the neuropilins in tumour angiogenesis and tumour progression

Neuropilins (NRPs) are multifunctional receptors for class 3 semaphorins, which are responsible for axon guidance during the development of the nervous system in vertebrates, and for vascular endothelial growth factors (VEGFs), essential for vascular development and angiogenesis in disease. There is now a large body of evidence that NRPs also mediate tumour angiogenesis and progression, and they have also emerged as novel therapeutic targets in cancer. Many neoplastic cell types express NRPs, and NRP1 and NRP2 upregulation is positively correlated with tumour progression and poor patient prognosis in several cancer types (Pellet-Many et al. Biochem J 411:211–226, 2008). Recently, NRPs have been shown to play novel roles in the tumour stem cell niche and in regulation of tumour immunity. This chapter focuses on the role of NRPs in tumour angiogenesis and tumour progression, focusing on the role of the NRPs as modulators of VEGF function and highlighting approaches to therapeutic targeting of NRPs in cancer

Demethylation of the Coding Region Triggers the Activation of the Human Testis-Specific PDHA2 Gene in Somatic Tissues

Human PDHA2 is a testis-specific gene that codes for the E1α subunit of Pyruvate Dehydrogenase Complex (PDC), a crucial enzyme system in cell energy metabolism. Since activation of the PDHA2 gene in somatic cells could be a new therapeutic approach for PDC deficiency, we aimed to identify the regulatory mechanisms underlying the human PDHA2 gene expression. Functional deletion studies revealed that the −122 to −6 promoter region is indispensable for basal expression of this TATA-less promoter, and suggested a role of an epigenetic program in the control of PDHA2 gene expression. Indeed, treatment of SH-SY5Y cells with the hypomethylating agent 5-Aza-2′-deoxycytidine (DAC) promoted the reactivation of the PDHA2 gene, by inducing the recruitment of the RNA polymerase II to the proximal promoter region and the consequent increase in PDHA2 mRNA levels. Bisulfite sequencing analysis revealed that DAC treatment induced a significant demethylation of the CpG island II (nucleotides +197 to +460) in PDHA2 coding region, while the promoter region remained highly methylated. Taken together with our previous results that show an in vivo correlation between PDHA2 expression and the demethylation of the CpG island II in testis germ cells, the present results show that internal methylation of the PDHA2 gene plays a part in its repression in somatic cells. In conclusion, our data support the novel finding that methylation of the PDHA2 coding region can inhibit gene transcription. This represents a key mechanism for absence of PDHA2 expression in somatic cells and a target for PDC therapy

Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli

Triglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils. In this work we describe TAG accumulation in the industrially appropriate microorganism Escherichia coli expressing the heterologous enzyme tDGAT, a wax ester synthase/triacylglycerol:acylCoA acyltranferase (WS/DGAT). With this purpose, we introduce a codon-optimized gene from the thermophilic actinomycete Thermomonospora curvata coding for a WS/DGAT into different E. coli strains, describe the metabolic effects associated to the expression of this protein and evaluate neutral lipid accumulation. We observe a direct relation between the expression of this WS/DGAT and TAG production within a wide range of culture conditions. More than 30% TAGs were detected within the bacterial neutral lipids in 90 minutes after induction. TAGs were observed to be associated with the hydrophobic enzyme while forming round intracytoplasmic bodies, which could represent a bottleneck for lipid accumulation in E. coli. We detected an increase of almost 3- fold in the monounsaturated fatty acids (MUFA) occurring in the recombinant strains. These MUFA were predominant in the accumulated TAGs achieving 46% of the TAG fatty acids. These results set the basis for further research on the achievement of a suitable method towards the sustainable production of these neutral lipids

Small molecule neuropilin-1 antagonists combine anti-angiogenic and anti-tumour activity with immune modulation through reduction of transforming growth factor beta (TGFβ) production in regulatory T-cells

We report the design, synthesis and comprehensive studybiological evaluation of a range ofsome potent small-molecule neuropilin-1 (NRP1) antagonists. NRP1 is implicated in the immune response to tumours, particularly in Treg cell fragility, required for PD1 checkpoint blockade. The design of these compounds was based on a previously identified compound EG00229, EG00229 which was used a starting point for optimisation. Through targeting of specific amino-acid residues additional H-bonding interactions were introduced, which led to increases in binding affinity and potency. The design of these molecules was informed and supported by X-ray crystal structures. Pharmacokinetic data was obtained for some of the most potent compounds, and cCompound 1 (EG01377) was identified as having properties suitable for further investigation. Compound 1 was then tested in several in vitro assays, and was shown to have anti-angiogenic, anti-migratory and anti-tumour effects. Remarkably, 1 was shown to be selective for NRP1 over the closely related protein NRP2. In purified Nrp1+, FoxP3+, CD25+ populations of Tregs from mice 1 was able to block a glioma conditioned medium induced increase in TGFβ production. This study therefore represents a comprehensive characterisation of a small-molecule NRP1 antagonist, and provides the basis for future in vivo studies

Política integrada e portugueses ciganos: o caso de educação de adultos de 2020 a 2022

Em Portugal a resposta das políticas públicas em matéria de educação de adultos tem passado por várias etapas, com avanços e recuos, até à atual configuração mais robusta e adaptada à realidade social do país. Contudo ainda persistem várias lacunas no que respeita ao alcance e à efetividade da mesma. As diferenças em termos de educação escolar entre os portugueses ciganos e a população em geral são claras devendo--se a um conjunto de fatores, entre os quais, desigualdades estruturais, diversidade sociológica e cultural e preconceito associado a uma imagem negativa “normalizada”. A situação dos/as portugueses/as ciganos/as é caracterizada por estas desigualdades múltiplas que reproduzem ciclos de exclusão e que tendem a condicionar o acesso aos bens e aos serviços públicos, dos quais são sujeitos de direito. Assim, o desenho e a implementação de políticas públicas ajustadas à realidade deste grupo social passam por um balanço entre uma abordagem mainstreaming/ transversal complementadas com medidas específicas de discriminação positiva, que contribuam para facilitar a efetivação dos direitos de cidadania. No presente capítulo analisamos a importância deste tipo de abordagem utilizando como foco a educação de adultos e a inovação política.info:eu-repo/semantics/publishedVersio

Computer Modeling Explains the Structural Reasons for the Difference in Reactivity of Amine Transaminases Regarding Prochiral Methylketones

Amine transaminases (ATAs) are pyridoxal-5′-phosphate (PLP)-dependent enzymes that catalyze the transfer of an amino group from an amino donor to an aldehyde and/or ketone. In the past decade, the enzymatic reductive amination of prochiral ketones catalyzed by ATAs has attracted the attention of researchers, and more traditional chemical routes were replaced by enzymatic ones in industrial manufacturing. In the present work, the influence of the presence of an α,β-unsaturated system in a methylketone model substrate was investigated, using a set of five wild-type ATAs, the (R)-selective from Aspergillus terreus (Atr-TA) and Mycobacterium vanbaalenii (Mva-TA), the (S)-selective from Chromobacterium violaceum (Cvi-TA), Ruegeria pomeroyi (Rpo-TA), V. fluvialis (Vfl-TA) and an engineered variant of V. fluvialis (ATA-256 from Codexis). The high conversion rate (80 to 99%) and optical purity (78 to 99% ee) of both (R)- and (S)-ATAs for the substrate 1-phenyl-3-butanone, using isopropylamine (IPA) as an amino donor, were observed. However, the double bond in the α,β-position of 4-phenylbut-3-en-2-one dramatically reduced wild-type ATA reactivity, leading to conversions of 99% ee. Computational docking simulations showed the differences in orientation and intermolecular interactions in the active sites, providing insights to rationalize the observed experimental results

Improving the geometry of a microstrip antenna for UWB signals through FDTD-CPML method and construction with fiberglass FR4

The Ultra Wideband (UWB) technology is a promising way to integrate data communications over short distances and at higher data rates. It is known that the design of the antenna is critical in the development of UWB systems. This paper proposes a new geometry for a microstrip UWB antenna. The antenna is analyzed using the Finite Difference Time Domain (FDTD) method, simulations using commercial software and experimental results. The analysis will be done by comparing the return loss simulated and measured of the antenna. The proposed antenna is constructed on a plate of FR4 fiber glass