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Applied Molecular Biosciences Unit
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New non-toxic n-alkyl cholinium-based ionic liquids as excipients to improve the solubility of poorly water-soluble drugs
Publication . Jesus, Ana R.; Raposo, Luís R.; Soromenho, Mário R.C.; Agostinho, Daniela A. S.; Esperança, José M. S. S.; Baptista, Pedro V.; Fernandes, Alexandra R.; Reis, Patrícia M.; LAQV@REQUIMTE; DQ - Departamento de Química; UCIBIO - Applied Molecular Biosciences Unit; MDPI - Multidisciplinary Digital Publishing Institute
In this work, we prepared new biocompatible N-alkyl cholinium-based ionic liquids to be used as cosolvents to improve the solubility of poorly water-soluble drugs, namely, sodium diclo-fenac and paracetamol. In this set of ionic liquids, we intend to understand the effect of increasing the asymmetry of the ionic liquid cation/anion by growing the length of one of the alkyl chains attached to the nitrogen center/sulfonate center on the dissolution capacity of the ionic liquid. The addition of these new ionic liquids to water increased the dissolution capacity of the drugs up to four-times that in water, and improved the pharmacodynamic properties of these drugs, especially the case of sodium diclofenac. The intermolecular interactions between the drugs and ionic liquids were investigated by NMR. Two-dimensional1H/1H nuclear overhauser effect spectroscopy (NO-ESY) revealed an interaction between sodium diclofenac and the alaninate anion from the [C2Ch]2[SucAla]. In the case of paracetamol and [C4Ch][C2SO3], it was possible to observe two inter-molecular interactions between the hydroxyl group of paracetamol and two protons from the cation [C4Ch]+. Interestingly, the ionic liquid bearing a succinyl-DL-alaninate anion, [SucAla]2−, and a N-ethyl cholinium cation, [C2Ch]+, which presented the highest ability to dissolve sodium diclofenac, showed no cytotoxicity up to 500 mM. Therefore, this ionic liquid is a potential candidate for drug delivery applications.
Molecular Beacon for Detection miRNA-21 as a Biomarker of Lung Cancer
Publication . Alexandre, Daniela; Teixeira, Bernardo; Rico, André; Valente, Salete; Craveiro, Ana; Baptista, Pedro V.; Cruz, Carla; DCV - Departamento de Ciências da Vida; UCIBIO - Applied Molecular Biosciences Unit; MDPI - Multidisciplinary Digital Publishing Institute
Lung cancer (LC) is the leading cause of cancer-related death worldwide. Although the diagnosis and treatment of non-small cell lung cancer (NSCLC), which accounts for approximately 80% of LC cases, have greatly improved in the past decade, there is still an urgent need to find more sensitive and specific screening methods. Recently, new molecular biomarkers are emerging as potential non-invasive diagnostic agents to screen NSCLC, including multiple microRNAs (miRNAs) that show an unusual expression profile. Moreover, peripheral blood mononuclear cells’ (PBMCs) miRNA profile could be linked with NSCLC and used for diagnosis. We developed a molecular beacon (MB)-based miRNA detection strategy for NSCLC. Following PBMCs isolation and screening of the expression profile of a panel of miRNA by RT-qPCR, we designed a MB targeting of up-regulated miR-21-5p. This MB 21-5p was characterized by FRET-melting, CD, NMR and native PAGE, allowing the optimization of an in-situ approach involving miR-21-5p detection in PBMCs via MB. Data show the developed MB approach potential for miR-21-5p detection in PBMCs from clinical samples towards NSCLC.
Development of Innovative Mediterranean-Style Semi-Hard Goat’s Cheese Supplemented with Seaweeds (Palmaria palmata and Ulva sp.) and Its Characterization
Publication . Campos, Bruno M.; Moreira-Leite, Bruno S.; Salgado, Abigail; Ramalho, Edgar; Marmelo, Isa; Malfeito-Ferreira, Manuel; Sousa, Paulo H. M. de; Henriques, Adolfo; Noronha, João P.; Diniz, Mário S.; Mata, Paulina; DQ - Departamento de Química; UCIBIO - Applied Molecular Biosciences Unit; LAQV@REQUIMTE; MDPI - Multidisciplinary Digital Publishing Institute
The main objective of this study was the development of two semi-hard goat cheeses supplemented with Palmaria palmata and Ulva sp. with the aim of developing innovative food products, increasing the concentration of nutrients in these cheeses and familiarizing consumers with seaweed-containing foods. The impact of seaweed addition was evaluated through physicochemical, microbiological, and organoleptic properties of the semi-hard goat cheeses. Carbohydrate content was relatively low, whereas the total lipid content was relatively high (particularly in semi-hard goat cheese supplemented with seaweeds). Crude protein content presented higher values in semi-hard goat cheese supplemented with Ulva sp. The semi-hard goat cheese supplemented with Ulva sp. shows increased levels of Ca, Fe, Mn, and Zn. Instrumental color and the textural parameters of semi-hard goat’s cheese varied significantly with seaweed addition. Most of the microbiological load complies with the Portuguese (INSA) and the United Kingdom’s (HPA) guidelines for assessing the microbiological safety of ready-to-eat foods placed on the market. Additionally, the Flash Profile scores of semi-hard goat cheeses supplemented with seaweeds highlighted aroma and flavor complexity. Overall, this study confirms the potential of using seaweeds as a viable alternative to produce semi-hard goat cheeses with less pungency or goat milk flavor, making this product more pleasant and appealing to consumers sensitive to these sensory characteristics.
Functional and genomic characterization of Komagataeibacter uvaceti FXV3, a multiple stress resistant bacterium producing increased levels of cellulose
Publication . Nascimento, Francisco X.; Torres, Cristiana A. V.; Freitas, Filomena; Reis, Maria A. M.; Crespo, Maria T. B.; UCIBIO - Applied Molecular Biosciences Unit; DQ - Departamento de Química; Instituto de Tecnologia Química e Biológica António Xavier (ITQB); Elsevier BV
Bacterial cellulose is one of the most promising biomaterials for the development of a wide array of novel biotechnological solutions. Nevertheless, the commercial production of bacterial cellulose is still a challenge and obtaining novel strains presenting increased cellulose biosynthesis and stress resistance properties is of extreme importance. This work demonstrates the increased stress resistance, cellulose production abilities, and overall genomic properties of Komagataeibacter uvaceti FXV3, a novel cellulose-producing and stress resistant strain isolated from a fermented grape must. K. uvaceti FXV3 was able to grow under several stress conditions, including the presence of high concentrations of ethanol (up to 7.5 % v/v), a trait that is not observed in the model strain K. xylinus CECT 7351T. Moreover, K. uvaceti FXV3 produced increased concentrations of cellulose (4.31 mg/mL, 7 days after inoculation-DAI) when compared to K. xylinus CECT 7351T (1.42 mg/mL, 7 DAI). Moreover, the detailed analysis of strain FXV3 genome revealed the presence of several genes involved in cellulose and acetan biosynthesis, quorum-sensing and quenching mechanisms, carbohydrate, amino acid, alcohol and aldehyde metabolism, as well as several other genes involved in stress resistance. Additionally, comparative genomic analysis revealed the increased prevalence of stress resistance genes in K. uvaceti FXV3 when compared to K. xylinus CECT 7351T. Ultimately, this study reveals the increased biotechnological potential of K. uvaceti FXV3 and brings new insights into the genetics behind Komagataeibacter stress resistance and cellulose production abilities.
Valorization of Reground Pasta By-Product through PHA Production with Phototrophic Purple Bacteria
Publication . Marchetti, Angela; Palhas, Miguel; Villano, Marianna; Fradinho, Joana; DQ - Departamento de Química; UCIBIO - Applied Molecular Biosciences Unit; MDPI - Multidisciplinary Digital Publishing Institute
Annually, the food industry generates large amounts of waste and by-products, causing serious problems in their management and final disposal. In particular, by-products are mainly recovered as livestock feed. A most appealing strategy to valorize them has herein been investigated, through polyhydroxyalkanoate (PHA) production. In this view, a stream rich in volatile fatty acids deriving from the acidogenic fermentation of reground pasta (RP), a farinaceous food-industry by-product, was used as a carbon source for PHA production with a phototrophic purple bacteria (PPB) consortium. PPB are very versatile organisms that present a unique metabolism allowing them to adapt to a variety of environmental conditions. The PPB-PHA enrichment phase was performed in a lab-scale semi-continuous photo-bioreactor under a permanent carbon feast regime, with organic loading rate (OLR) increments from 14 to 19 mmolC/Ld. The results showed that the fermented RP solution composition (with 23.4% of HV precursors on a COD basis) was suitable for the PHBHV copolymer production, with the PPB consortium being capable of reaching a very high content in the hydroxyvalerate (HV) monomer, with a maximum of 60% (gHV/gPHA). Regarding the PHA accumulation stage where the light intensity was increased up to 20.2 W/L, a further increase in the culture PHA content by 76% after 12 h was obtained. Overall, these results open the possibility of valorizing food-industry by-products through the development of a biocatalytic process for PHA production with PPB, thus making the overall approach more sustainable from a green perspective.
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Fundação para a Ciência e a Tecnologia
Programa de financiamento
6817 - DCRRNI ID
Número da atribuição
UIDB/04378/2020