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The insufficient UV photoprotection of approved sunscreens and the negative impact of their compositions on ecosystems and human health makes the utility of sunscreen questionable. Therefore, discovering UV filters with significant antioxidant activity and improved topical performance and photostability is crucial. The use of nanosized natural molecules incorporated in sunscreens has been a scientific hot topic, since they provide a synergistic effect with synthetic UV filters, improving overall SPF and antioxidant activity, higher epidermis retention, and less toxicity. Pedro Fonte from iBB and colleagues reviewed about formulations containing nanosized flavonoids with improved safety and UVA photoprotection becoming promising sunscreens.

A Wednesday afternoon spent learning about cutting-edge science while enjoying a beer with friends in a "Hygge" setting is undoubtedly appealing. This is the value proposition from the “PubHD” initiative. In this week’s edition, the IBB/IST will be represented by Rui Oliveira-Silva.

To enhance the performance of (bio)sensors and obtain information about a given system at a specific time, Rui concentrated on the development of nanomaterials during his doctoral studies. The availability of these (bio)sensors, enable early detection, process control of pharmaceuticals, or assess the effectiveness therapies and to follow-up patients. These (bio)sensors render information that was previously impossible, and are increasingly crucial in contemporary precision medicine.

During past years, lipid nanoparticles (LNPs) have emerged as promising carriers for RNA delivery, with several clinical trials focusing on both infectious diseases and cancer. More recently, the success of messenger RNA (mRNA) vaccines for the treatment of severe diseases, such as acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is partially justified by the development of LNPs encapsulating mRNA for efficient cytosolic delivery. In a collaborative work with researchers from Hovione Farmaciência SA and INESC-MN, Pedro Fonte from iBB published an invited review paper in Expert Opinion on Drug Delivery addressing the production and formulation of LNPs by using microfluidic devices, the status of mRNA-loaded LNPs therapeutics and explored spray drying process, as a promising dehydration process to enhance LNP stability and provide alternative administration routes. It is foreseen this work may contribute to advances in the field in the upcoming years.

Chitosan is a biodegradable and biocompatible natural polymer approved by FDA as considered GRAS (Generally Recognised as Safe) for biomedical applications. In a joint Portugal-India collaboration led by Pedro Fonte, and collaboration of iBB Master student Ana Patrício, it was published in Materials journal an overview on chitosan‐based nanoparticles, focusing on their versatile features and different applications in biomedical sciences and engineering, including their potential for drug delivery and tissue engineering. In this work, several methods that have been developed to produce chitosan-based nanoparticles are fully disclosed. More importantly, the characterization and application of such delivery systems in different pathologies are addressed. It is foreseen that this contribution could be a guidance, on developing chitosan-based nanocarrier systems for different applications.

It is usually accepted that only 0.1-1% of marine bacteria may be cultivated under laboratory conditions. This resulted mainly from the observation that only a small fraction of the total bacteria determined by microscopic counts could be enumerated by culture plate techniques, and has been a justification for the widely implementation of cultivation-independent molecular techniques. In a recent published paper, Carlos J.C. Rodrigues and Carla C.C.R. de Carvalho (BERG-iBB) promoted the growth of bacteria from a marine rock pond by culture dependent techniques and compared the results with culture independent methods. The highest efficiency of cultivation was 45% attained in marine agar. The study shows that it is possible to grow more than the traditionally considered 1% of bacteria from a seawater sample using standard agar plate techniques and laboratorial conditions, and to find bacteria not detected by molecular approaches.

DNA-origami biomanufacturing relies in many cases on the use of asymmetric PCR (aPCR) to generate 500-3500 base, single-stranded DNA (ssDNA) scaffolds that are then purified by agarose gel extraction, a technique that is laborious, not scalable and presents low recovery yields. In a paper just published in Separation and Purification Technology, Ana Rita Santos and Miguel Prazeres from BERG-iBB, in collaboration with Pedro Paulo from CQE, present alternative methods based on anion-exchange or multimodal chromatography that yield purified scaffolds from aPCR mixtures. The recovered scaffolds were further used to assemble DNA-origami nanostructures. The work indicates that chromatography could play an important role in scaling-up the purification of ssDNA scaffolds for the biomanufacturing DNA-origami nanostructures.

Carla CCR de Carvalho (iBB-BERG) presented the work entitled “Marine bioprocesses with bacteria from the Portuguese coast” at the “One Sustainable Ocean”, a side event of the UN Ocean conference (UNOC) held in Lisbon. The UN Ocean Conference, co-hosted by the Governments of Kenya and Portugal, aimed at starting a new chapter of global ocean action.

L-asparaginases have an acknowledge role in food and pharmaceutical industries. Accordingly, efforts to produce highly efficient, selective and stable L-asparaginases are currently underway. One of the strategies used to fulfill this goal involves the production of recombinant. In a recent paper published in Marine Biotechnology, Pedro Fernandes (iBB-BERG) and colleagues from the University of Hormozgan and Tarbiat Modares University in Iran report the production of a novel marine recombinant L‑asparaginase and provide its biochemical and structural characterization. The results obtained suggest the potential of the thermostable recombinant enzyme obtained for practical applications in food and pharmaceutical areas.

Lupanine is used as a building block in the synthesis of sparteine, a chiral selector in drug synthesis. This alkaloid is found in wastewaters derived from the debittering process that makes lupin beans edible. In a recently published work, carried out by researchers of the Faculty of Sciences and Faculty of Pharmacy from the University of Lisbon, and Teresa Esteves, Frederico Ferreira, Flávio Ferreira and Ana Mota from BERG-iBB, a computational chemistry approach was taken to design molecularly imprinted polymers (MIPs) selecting itaconic acid, a biobased building block, as a functional monomer that can provide higher affinities for lupanine. In this work, lupanine was concentrated from lupin bean wastewater by nanofiltration, extracted with ethyl acetate, and purified using the synthesized MIP, which was able to selectively recognize lupanine and improve its purity to 88%, with 82% recovery of the alkaloid, showing the potential application of this strategy to render the industrial process more sustainable.

When an idea in a coffee break in an international meeting turns into a paper, Pedro Fonte from BERG-iBB in collaboration with researchers from UFVJM, Brazil just published a work in JSFA Reports performing an optimization of the brewing parameters on coffee extraction using a central composite rotatable design. The effects of extraction time, particle size of ground coffee, extraction temperature, coffee-to-water ratio, stirring on caffeine yield, and soluble solids on caffeine concentrations were studied. Optimized parameters showed 45 min sufficed to perform a cold extraction at 4°C and 24°C. The parameters selected for validation were 24°C, 30% coffee-to-water ratio, a stirring of 400 rpm resulting in 3.98 mg/ml of extracted caffeine, 11.20 °Brix, and 93.9% of caffeine yield. The smaller particle size (595 μm) displayed the higher caffeine extraction of about 4 mg/ml. This study reveals the high efficiency of cold brew extraction and its potential at the industrial scale, decreasing costs with energy and extraction time, and producing a coffee rich in caffeine and soluble solids.

Extracellular vesicles (EVs) are membrane vesicles fundamental for cellular communication processes. Either by taking advantage of EVs innate properties, or by loading them with therapeutic agents, EVs have emerged as cell-free strategies with diverse clinical applications. Indeed, when compared to mesenchymal stromal cells (MSCs), similar or better therapeutic benefits have been reported for MSC-derived EVs, thus explaining the increased interest in EVs over the years. Despite appealing features, EVs implementation in the clinic is still limited by technical difficulties, like robust isolation methods capable of delivering a high-quality product with high yield. In this work, published in the journal Separation and Purification Technology, we described anion exchange chromatography as a robust and better alternative to already implemented techniques. Following this strategy, more than 85% of injected EVs could be recovered, while complying with the stringencies set by regulatory agencies. This study developed at iBB was led by Dr. Ana Fernandes-Platzgummer and Prof. Ana Azevedo and co-first-authored by PhD students Ricardo Silva and Sara Sousa Rosa.

Therapeutic proteins are bioactive compounds used for the treatment and prevention of several diseases, being usually well-tolerated and specific for their target. Nevertheless, they also present physicochemical instability, with susceptibility to suffer degradation. The use of lipid nanoparticles for therapeutic proteins encapsulation protects its structure, improving its bioavailability, and obtaining a sustained or controlled release. However, the loading of therapeutic proteins into lipid nanoparticles presents several challenges, so in a paper published in the  Journal of Drug Delivery Science and Technology, the researchers from iBB, Cláudia Viegas and Pedro Fonte addressed the challenges and advantages of loading therapeutic proteins into lipid nanoparticles. In the paper not only the different lipid structures, but also the protein encapsulation methods that do not compromise the structure and bioactivity of therapeutic proteins are discussed.

The increasing number of life-threatening infections observed in cancer patients has been ascribed to chemotherapy-induced neutropenia and to invasive medical procedures. In a recent published study, Carla C.C.R. de Carvalho (BERG-iBB) questioned if the infections could also be favored by an increased resistance of bacteria due to the adaptation to antineoplastic agents used in chemotherapy. After exposure to several antineoplastic agents, it was observed that cells of Staphylococcus aureus, Mycobacterium vaccae, Pseudomonas aeruginosa, and Escherichia coli changed the fatty acid profile of their cellular membranes, produced exopolymeric substances, and formed aggregates that adhered to surfaces. Additionally, when exposed to high concentrations of these compounds, a persister sub-population could be identified. After adaptation to antineoplastic agents, the minimum inhibitory concentration (MIC) of several antibiotics increased considerably in the tested strains. This should be a major concern during oncology treatments.

Helichrysum italicum is a halophyte shrub with bright yellow flowers with a strong curry-like aroma that have been used in the production of cosmetics and pharmaceuticals, for seasoning and flavoring of food, and as natural food preservatives. In a study  recently published in Food Research International, researchers of Universidade Nova de Lisboa, Polytechnic of Leiria, University of Coimbra, University of Lincoln, the Quadram Institute, and Carla C.C.R. de Carvalho (BERG-iBB), characterize the composition, bioactive compounds, and nutritive value of H. italicum flowers. The flowers were found to be a good source of minerals, omega-6 and omega-3 PUFAs, polyphenols, carotenoids, curcuminoids and other bioactive compounds with relevant antibacterial, antifungal and antioxidant properties.

Photo details: Helichrysum italicum, Bj.schoenmakers, CC0, via Wikimedia Commons.

The Norwegian Minister of Fisheries and Ocean Policy Bjørnar Selnes Skjæran, the Minister of International Development Anne Beathe Tvinnereim, and the Embassador in Lisbon Tove Bruvik Westberg, visited the Instituto Superior de Agronomia as promotor of projects funded by EEA grants. Anabela Raymundo and Cristiana Nunes (LEAF-ISA), Carla de Carvalho (iBB-IST) and António Pagarete (Pagarete Microalgae Solutions) presented the collaborations they have had with Norwegian research groups including the project Yum Algae. This projects aims at finding enzymes to improve the sensory quality of microalgae ingredients in foods and during the visit several products, such as bread, cheese and ice cream containing microalgae, were tasted. The visit also included members of the “Unidade Nacional de Gestão” of EEA Grants and of “Direção Geral de Política do Mar”.

The ocean is an excellent source for new biocatalysts due to the tremendous genetic diversity of marine microorganisms, and it may contribute to the development of sustainable industrial processes. In a recent paper published in Microorganisms, Carlos J.C. Rodrigues and Carla C.C.R. de Carvalho (iBB-BERG) used a marine bacterium for the conversion of benzaldehyde to benzyl alcohol, which is an important chemical employed as a precursor for producing esters for cosmetics and other industries. A stirred reactor, using a fed-batch approach, enabled a 1.5-fold increase in benzyl alcohol productivity when compared with batch mode. However, product accumulation in the reactor hindered the conversion. The use of a continuous flow reactor packed with immobilized cells enabled a 9.5-fold increase in productivity when compared with the fed-batch stirred reactor system.

Greener, cost-effective methodologies for food processing and production have been actively looked after because of increased demand due to growing population. Enzymes, as green and highly efficient catalysts, have been a mainstay of food technology. However, the advantageous features of enzymes are negatively impacted by their long-term instability, harsh industrial operational conditions and challenges for enzyme recovery and reuse. Immobilized enzyme formulations have been developed to address these limitations. In a recently published book chapter, Filipe Carvalho and Pedro Fernandes (iBB-BERG) present the latest finding and trends on the design and application of immobilized enzyme for food production and processing. The chapter was published in the book Value Addition in Food Products and Processing using Enzyme Technology, published by Academic Press.

Pathogenic Vibrio species are responsible for human and animal illness and one of the main causes of human infection is related to the ingestion of undercooked seafood. Due to their filter-feeding habit, marine invertebrates, such as clams, are known to be a natural reservoir of specific microbial communities. In a recently published study carried out by researchers of ESTM, Polytechnic of Leiria, and Carla C.C.R. de Carvalho (BERG-iBB), the profile of antimicrobial resistance and the presence of virulence genes in the bacterial isolates from clams were evaluated. The presence of the mobilisable colistin resistance gene mcr-1 in three Vibrio spp. isolates highlights a potential threat to public health.

A new strategy to modify cellulose with the short antimicrobial hexapeptide MP196 (RWRWRW-NH2) is proposed by BERG and BSIRG researchers that uses fusions of Cys-terminated derivatives of MP196 and a carbohydrate binding module (CBM). CBM3-MP196-modified cellulose hydrogels displayed antibacterial activity that was significantly higher when compared with controls. This versatile concept offers a toolbox for the functionalization of different cellulose materials with a broad choice in peptides. the paper was published in Acta Biomaterialia. The work was funded by project CBM-X.

Lactococcus lactis is a food-grade, and generally recognized as safe, bacterium, which making it ideal for producing plasmid DNA (pDNA) or recombinant proteins for industrial or pharmaceutical applications. A paper published in Microorganisms by Sofia Duarte and Gabriel Monteiro from BERG-iBB reviews the major findings from L. lactis transcriptome and proteome studies, with an overexpression of native or recombinant proteins. These studies provide important insights on how to engineer the plasmid vectors and/or the strains in order to achieve high pDNA or recombinant proteins yields, with high quality standards.