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The demand for products based on natural ingredients is increasing among cosmetic and textile consumers. Cork extracts contain components of interest with special properties, including antioxidant, anti-inflammatory, and antibacterial activities, that might improve the effectiveness of cosmetic formulations currently on the market and may impart new characteristics to textiles. The main goal of this work was to investigate the effect of the incorporation of three cork extracts into two commercial cosmetic formulations (formulation A and B) and evaluate their role as textile dyeing agents. The extracts (E1, E2, and E3) were obtained from cork powder using natural deep eutectic solvents (NADES) (E1-NADES 1: lactic acid:glycerol, E2-NADES 2: lactic acid:glycine, and E3-NADES 3: lactic acid:sodium citrate) and applied in combination with the solvent. The impact of the extracts on the cosmetic formulations’ properties was evaluated in terms of pH, viscosity, antioxidant activity, transdermal permeation capacity, cytotoxicity, and organoleptic characteristics (odor, color, and appearance). The results demonstrated that the cork extracts improved the antioxidant performance of the formulations (90% reduction in DPPH (1,1-difenil-2-picril-hidrazil)). Moreover, low concentrations (5 mg/mL and 10 mg/mL) of extract did not present a cytotoxic effect on keratinocytes. Cotton fabrics were efficiently dyed with the NADES-based cork extracts which conferred to these substrates antioxidant (78% in DPPH reduction) and antibacterial abilities (inhibition halos: 12–15 mm). The application of cork extracts as ingredients in cosmetics or as dyeing/coloration agents for textile coloration is revealed to be a promising and green route to replace harmful ingredients normally used in industry., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit. D.S.F. also thanks the Portuguese Foundation for Science and Technology (FCT) for funding (SFRH/BD/147190/2019). A.R. thanks to Portuguese Foundation for Science and Technology (FCT) for is contract under the CEEC-Individual—4th Edition with the reference 2021.02803.CEECIND., info:eu-repo/semantics/publishedVersion

The poor mechanical performance and difficulty of using wound dressings without discomfort to the patient motivated the development of textile composites, incorporating peptide hydrogels in textile matrices. Thus, 70% viscose and 30% polyester non-woven and 100% cotton bandages were functionalized by impregnation with a hydrogel resulting from physical crosslinking of Z-Tyr-Phe-OH dipeptide. The healing properties of these structures resulted from the incorporation of curcumin into the polymer. The textile composites were characterized by Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), Thermogravimetric Analysis (TGA), and colour strength (K/S) with differences being observed after functionalization with the hydrogel (higher surface smoothness and thermal stability). The breathability and moisture management properties of the new composites were similar to those of commercial dressings, and the pH value of the aqueous extract resembled skin. The swelling of the composites increased significantly due to the contribution of the hydrogel. Furthermore, the textile composites released the curcumin for a long time with an initial burst release in the first 2 h and showed good antibacterial and antifungal properties, without displaying significant cytotoxicity on fibroblasts. Overall, the composites here were developed to meet the main requirements of dressings for the management and treatment of skin wounds., This study was supported by FCT – Foundation for Science and Technology within the scope of the project PTDC/QUI-QOR/29015/2017 and by FCT/OE, PIDDAC funds within the scope of the projects UID/CTM/00264/2021, UIDB/04469/2020 and UIDB/04423/2020., info:eu-repo/semantics/publishedVersion

Ohmic heating is a thermal processing method based on the application of electric fields directly into a semi-conductive medium. In this study, we explored for the first time the use of ohmic heating to obtain keratin films. The properties of the films prepared by ohmic heating and conventional heating were evaluated and compared under similar thermal profiles. A lower increase in free thiols' concentration was obtained for the keratin solutions and keratin films submitted to ohmic heating (16% increase for the keratin solution extracted from virgin hair, pHâ¯9, submitted to ohmic heating and 23% when submitted to conventional heating). Significant differences in the swelling results were observed for the films prepared with keratin extracted from virgin hair, with a swelling decrease in about 55% for the films prepared by ohmic heating. Generally, the keratin films obtained by ohmic heating showed distinct properties comparatively to the films produced by conventional methods. The application of a fusion protein on the keratin films demonstrated their capacity to be used as substitutes to hair fibers when evaluating the potential of new cosmetic products. This work suggests that ohmic heating show potential to tailor keratin films properties depending on an intended application or functionality., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Ana Tinoco and Rui M. Rodrigues thanks FCT for funding the scholarships with the references SFRH/BD/114035/2015 and SFRH/BD/110723/2015, respectively., info:eu-repo/semantics/publishedVersion

Eutectic-based formulations were developed aiming to modify the shape and improve the mechanical properties of Asian hair. These formulations comprised a eutectic mixture (choline chloride and urea) as a solvent, a natural reducing agent (l-cysteine), and a decapeptide. The treatment of Asian hair with the eutectic-based formulations increased the resistance of hair to mechanical wear by about 10\\% when compared with hair treated with a styling commercial formulation (chemical treatment). For these samples, the change in hair shape is associated with the presence of cysteine and the decapeptide, which participates in the disruption and in the rearrangement of new disulfide bonds with the keratin fibers. The eutectic-based formulations here presented show great potential to be used as a new class of hair styling agents in haircare research. These new formulations will avoid/reduce the application of harsh chemicals, which induce irreversible damage in hair structure and have a negative impact on the environment., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of the UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020., info:eu-repo/semantics/publishedVersion

Tomato bacterial spot (Bs), caused by Xanthomonas spp., including X. euvesicatoria (Xeu) remains a major threat for tomato production. The emergence of copper resistance strains of Xeu calls urgently for eco-friendly phytosanitary treatments as sustainable green alternatives for disease control. Satureja spp. essential oil (EO) has antimicrobial activity against xanthomonads and combined with zein nanoparticles (ZNPs), might offer a viable option for field applications. This study aims to evaluate the effects of S. montana EO, of ZNPs, and their combination in a nanoformulation, on Xeu quantity, and how these compounds modulate molecular and physiological changes in the pathosystem. Uninfected and infected tomato plants (var. Oxheart) were treated with EO; ZNPs and nanoformulation (EO + ZNPs). Treatments reduced Xeu amount by a minimum of 1.6-fold (EO) and a maximum of 202-fold (ZNPs) and improved plants health. Nanoformulation and ZNPs increased plants phenolic content. ZNPs significantly increased GPX activity and reduced CAT activity. Overall treatments upregulated transcripts of the phenylpropanoid pathway in infected plants, while ZNPs and nanoformulation upregulated those transcripts in uninfected plants. Both sod and aao transcripts were downregulated by treatments in infected plants. These findings demonstrate that S. montana EO, ZNPs and their nanoformulation are suitable to integrate tomato bacterial spot management strategies, mainly due to their antimicrobial activity on Xeu, however further field studies clarifying the long-term action of these products are required. These results also support the prophylactic potential of ZNPs on tomato bacterial spot., The authors would like to thank other funding sources that supported the researchers involved in this work, such as, the European Investment Funds through the COMPETE 2020—Operational Programme for Competitiveness and Internationalization (POCI) and National Funds by the Portuguese Foundation for Science and Technology (FCT) (02/SAICT/2017, Ref. PTDC/BAAAGR/31131/2017, Acronym EOIS-CropProt). National Funds through FCT (Portugal) also funded this work within the scope of GreenUPorto, FCUP research unit: UIDB/05748/2020 and UIDP/05748/2020 projects; CITAB research unit, UTAD: project UIDB/04033/2020). Furthermore, N.M-P. was funded by a PhD grant SFRH/BD/138187/2018. This study was also supported by the European Union’s Horizon 2020 Research and Innovation Program (Grant Agreement Num ber 857251)., info:eu-repo/semantics/publishedVersion

This work describes the development of a capacitive-type sensor created from nanoporous anodic aluminium oxide (NP-AAO) prepared by the one-step anodization method conducted in potentiostatic mode and performed in a low-cost homemade system. A series of samples were prepared via an anodization campaign carried out on different acid electrolytes, in which the anodization parameters were adjusted to investigate the effect of pore size and porosity on the capacitive sensing performance. Two sensor test cases are investigated. The first case explores the use of highly uniform NP-AAO structures for humidity sensing applications while the second analyses the use of NP-AAO as a capacitive touch sensor for biological applications, namely, to detect the presence of small objects such as bacterial colonies of Escherichia Coli. A mathematical model based on equivalent electrical circuits was developed to evaluate the effect of humidity condensation (inside the pores) on the sensor capacitance and also to estimate the capacitance change of the sensor due to pore blocking by the presence of a certain number of bacterial microorganisms. Regarding the humidity sensing test cases, it was found that the sensitivity of the sensor fabricated in a phosphoric acid solution reaches up to 39 (pF/RH%), which is almost three times higher than the sensor fabricated in oxalic acid and about eight times higher than the sensor fabricated in sulfuric acid. Its improved sensitivity is explained in terms of the pore size effect on the mean free path and the loss of Brownian energy of the water vapour molecules. Concerning the touch sensing test case, it is demonstrated that the NP-AAO structures can be used as capacitive touch sensors because the magnitude of the capacitance change directly depends on the number of bacteria that cover the nanopores; the fraction of the electrode area activated by bacterial pore blocking is about 4.4% and 30.2% for B1 (E. Coli OD600nm=0.1) and B2 (E. Coli OD600nm=1) sensors, respectively., This research was funded by: the Portuguese Foundation for Science and Technology (FCT) under the strategic funding grants UIDB/04029/2020, UIDB/04650/2020 and UIDB/04469/20 units; and, the European Regional Development Fund under the scope of Norte2020 program grant NORTE-01-0145-FEDER-000004, BioTecNorte., info:eu-repo/semantics/publishedVersion

Resveratrol (RSV) and omega 3 (3), because of their biological favorable properties, have become subjects of interest for researchers in dermocosmetic and pharmaceutical industries; however, these bioactives present technological limitations that hinder their effective delivery to the target skin layer. To overcome the stability and skin permeation limitations of free bioactives, this work proposes a combined strategy involving two different lipid nanosystems (liposomes and lipid nanoparticles) that include 3 in their lipid matrix. Additionaly, RSV is only encapsulated in liposomes that provid an adequate amphiphilic environment. Each formulation is thoroughly characterized regarding their physicalchemical properties. Subsequently, the therapeutic performance of the lipid nanosystems is evaluated based on their protective roles against lipid peroxidation, as well as inhibition of cicloxygenase (COX) and nitric oxid (NO) production in the RWA264.7 cell line. Finally, the lipid nanosystems are incorporated in hydrogel to allow their topical administration, then rheology, occlusion, and RSV releasediffusion assays are performed. Lipid nanoparticles provide occlusive effects at the skin surface. Liposomes provide sustained RSV release and their flexibility conferred by edge activator components enhances RSV diffusion, which is required to reach NO production cells and COX cell membrane enzymes. Overall, the inclusion of both lipid nanosystems in the same semisolid base constitutes a promising strategy for autoimmune, inflammatory, and cancerous skin diseases., This research was funded by FCT/MCTES—Foundation for Science and Technology I.P. from the Minister of Science, Technology, and Higher Education (PIDDAC) and European Regional Development Fund (ERDF) by the COMPETE—Programa Operacional Factores de Competitividade (POFC) through the project CONCERT (POCI-01-0145-FEDER-032651 and PTDC/NAN-MAT/326512017) and the Strategic Funding UID/Multi/04546/2019, UIDP/04423/2020 (Group of Natural Products and Medicinal Chemistry CIIMAR), and “Contrato-Programa” UIDB/04469/2020 (CF-UM-UP) and UIDB/04050/2020 (CBMA), and UIDB/04469/2020 (CEB), as well as the Research Center of the Portuguese Oncology Institute of Porto (project no. PI86-CI-IPOP-66-2019), and BioTecNorte operation (NORTE-01-0145-FEDER- 000004) funded by the European Regional Development Fund under the scope of Norte 2020 - Programa Operacional Regional do Norte. Marlene Lúcio thanks FCT and ERDF for her doctoral position (CTTI-150/18-CF (1) within the scope of the CONCERT project. Raul Machado acknowledges FCT I.P. for funding within the Scientific Employment Stimulus project (CEECIND/00526/2018)., info:eu-repo/semantics/publishedVersion

Cosmetics procedures and products combined with environmental insults and daily routines induce irreversible changes in hair. As result of damage, the hair loses some of its properties like strength, elasticity, and smoothness. Recent studies revealed the positive effects of protein-based cosmetics in providing protection to hair. Additionally, these cosmetic products have also shown a great ability to modify hair fibers. We review the effect of protein-based cosmetic formulations on hair properties like color, scent, strength, shape, and volume, highlighting the potential of keratin-based particles and keratin-fusion proteins. In the future, incorporating multifunctional proteins and peptides in the development of alternative hair formulations will result in advanced, sustainable, ecofriendly cosmetic products with a great impact on the cosmetic industry., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of the UIDB/04469/2020 unit. A.T. and M.M. have work contracts supported by the European Social Fund under the scope of Norte2020 – Programa Operacional Regional do Norte (NORTE-06-3559-FSE-000141)., info:eu-repo/semantics/publishedVersion

The perming of hair is a common styling procedure with negative impact on the overall properties of the hair fibers. Usually, this process uses harsh chemicals to promote the disruption of disulfide bonds and the formation of new bonds to change the shape of hair. Here, we explored bovine serum albumin (BSA), silk fibroin (SF), keratin and two fusion recombinant proteins (KP-UM and KP-Cryst) as new perming agents. A phosphate buffer prepared at different pH values (5, 7 and 9) was used to apply the proteins to virgin Asian hair, and a hot BaByliss was used to curl the hair fibers. To assess the potential of the protein formulations for hair styling, the perming efficiency and the perming resistance to wash were measured. Furthermore, the fiber water content was evaluated to assess if the proteins protected the hair during the styling process. Despite all of the proteins being able to assist in the curling of Asian hair, the best perming efficiency and perming resistance to wash results were observed for BSA and keratin. These proteins showed perming efficiency values close to that measured for a commercial perming product (chemical method), particularly at pH 5 and 9. The increase in the hairs internal and external water contents revealed a protective effect provided by the proteins during the application of heat in the styling procedure. This study shows the potential of proteins to be used in the development of new eco-friendly hair styling products., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit., info:eu-repo/semantics/publishedVersion

The olfactory system of mammals allows the detection and discrimination of thousands of odors from the environment. In mammals, odorant-binding proteins (OBPs) are considered responsible to carry odorant molecules across the aqueous nasal mucus to the olfactory receptors (ORs). The three-dimensional structure of these proteins presents eight antiparallel ?-sheets and a short ?-helical segment close to the C terminus, typical of the lipocalins family. The great ability of OBPs to bind differentiated ligand molecules has driven the research to understand the mechanisms underlying the OBP function in nature and the development of advanced biotechnological applications. This review describes the role of mammalian OBPs in the olfactory perception, highlighting the influence of several key parameters (amino acids, temperature, ionic strength, and pH) in the formation of the OBP/ligand complex. The information from the literature regarding OBP structure, affinity, the strength of binding, and stability inspiring the development of several applications herein detailed., This study was financially supported by the Portuguese Foundation for Science and Technology (Fundac¸~ao para a Ciencia e a Tecnologia) under the scope of the strategic funding of UID/BIO/04469/2019 unit and BioTecNorte operation [NORTE-01-0145-FEDER-000004] funded by European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. F.G. thanks Fundac~ao para a Ci^ encia e a Tecnologia for their funding [SFRH/BD/114684/2016]., info:eu-repo/semantics/publishedVersion

Hair perfumes are becoming a trend all over the world and new cosmetic formulations have been developed to address this recent need. In the present study, we developed a system for the controlled delivery of perfumes on hair, based on zein, a protein derived from maize, and on keratin. The Keratin:Zein particles, obtained using different strategies and proportions of zein and keratin, presented a high stability along storage, related with the presence of keratin on the particles formulation. When applied on hair, the particles formed a film-like structure over the fibers. Fragrance diffusion from the hair-coated particles was dependent on the temperature, the method of particles preparation and the fragrances physicochemical properties (melting point and vapor pressure). The particles formulations showed also the capacity to improve hairs mechanical properties and hydration degree. The new system based on Keratin:Zein particles revealed high potential for the development of personalized hair cosmetic products allowing adjustments depending on the personal sensory preferences., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER- 000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Ana Tinoco, Filipa Gonçalves and David S. Freitas thanks FCT for funding the scholarships with the references SFRH/BD/114035/2015, SFRH/BD/ 114684/2016 and SFRH/BD/147190/2019, respectively, info:eu-repo/semantics/publishedVersion

Ohmic heating (OH) is recognised as an emerging processing technology which recently is gaining increasing attention due to its ability to induce and control protein functionality. In this study, OH was used for the first time in the production of scaffolds for tissue engineering. BSA/casein solutions were processed by OH, promoting protein denaturation and aggregation, followed by cold-gelation through the addition of Ca2+. The formation of stable scaffolds was mostly dependent on the temperature and treatment time during OH processing. The variations of the electric field (EF) induced changes in the functional properties of both gel forming solutions and final scaffolds (contact angle, swelling, porosity, compressive modulus and degradation rate). The scaffolds biological performance was evaluated regarding their ability to support the adhesion and proliferation of human fibroblast cells. The production process resulted in a non-cytotoxic material and the changes imposed by the presence of the EF during the scaffolds production improved cellular proliferation and metabolic activity. Protein functionalization assisted by OH presents a promising new alternative for the production of improved and tuneable protein-based scaffolds for tissue engineering., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte., info:eu-repo/semantics/publishedVersion

The antimicrobial functionality of composites constituted by chitosan with silver-doped zeolites was developed and characterized. A composite with chitosan particles and silver-doped zeolites was synthesized using an ionic gelation process with sodium tripolyphosphate. The chitosan silver-doped zeolites composite obtained presented sizes up to 5 mu m, while the silver-doped zeolites had an average size between 0.5 mu m and 3.3 mu m. The synthesized chitosan silver-doped zeolites composites, as well as the silver-doped zeolites and the chitosan dissolution, were characterized through X-ray diffraction, Fourier Transform Infrared spectroscopy and scanning electron microscopy. The electro kinetic behaviour of chitosan, silver-doped zeolites and chitosan silver-doped zeolites composite was evaluated under different pH conditions. The antimicrobial activity of the composites was evaluated in terms of minimum inhibitory concentrations and minimum lethal concentrations and the results suggest that the chitosan silver-doped zeolites composites show antimicrobial activity against gram-negative and gram-positive bacteria, Escherichia coli and Staphylococcus aureus, respectively and against Candida albicans. The results here presented support the potential application of the composite of chitosan with silver-doped zeolites in the functionalization of textiles with antimicrobial properties., Fabio A. P. Scacchetti thanks to the CNPq-Brazil (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico-Brasil/National Council of Scientific and Technological Development-Brazil) for the doctoral scholarship (233550/2014-3). Artur Ribeiro and Artur Cavaco-Paulo thanks to the Portuguese Foundation for Science and Technology (FCT) for the strategic funding of UID/BIO/04469/2019 unit and BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by European Regional Development Fund under the scope of Norte2020-Programa Operacional Regional do Norte. This work is also financed by Project UID/CTM/00264/2019 of 2C2T-Centro de Ciencia e Tecnologia Textil, funded by National Founds through FCT/MCTES.

During the treatment of acute lymphoblastic leukemia (ALL) with asparaginase (ASN) there is an accumulation of ammonia in the body as result of asparagine hydrolysis. This accumulation known as hyperammonemia is one of the main side-effects of this therapy. To avoid hyperammonemia is urgent to develop new strategies for ammonia retention. Herein is presented the immobilization of ASN into bovine serum albumin/poloxamer 407 (BSA/Pol407) nanoparticles. The ability of the developed nanoparticles to hydrolyze asparagine while retaining the forming ammonia is also explored. Different percentages of ASN were entrapped into BSA nanoparticles coated with Poloxamer 407 and were prepared by high-pressure homogenization. The nanoparticles were characterized regarding their physico-chemical properties, stability, capacity to retain ammonia and safety using zebrafish embryos as an in vivo model of toxicity. The BSA/ASN25%/Pol407 nanoparticles were selected as the best formulation to hydrolyze asparagine using the lowest nanoparticle concentration. These nanoparticles presented physical characteristics suitable for an intravenous application and were capable to retain the forming ammonia decreasing the negative effect of free ASN on zebrafish survival. These nanoparticles could potentially be used to prevent hyperammonemia during ALL treatment with ASN., This study was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01- 0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145- FEDER-000004) and Nanotechnology Based Functional Solutions (NORTE-01-0145-FEDER-000019) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. We also acknowledge the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through Fundação para a Ciência e a Tecnologia (FCT) and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI). Marisa P. Sárria was supported by Marie Curie COFUND funding from the European Union’s 7th Framework Programme for research, technological development and demonstration under grant agreement 600,375. Artur Ribeiro and Ana Tinoco thanks FCT for funding the scholarships with the references SFRH/BPD/98388/2013, SFRH/BD/ 114035/2015, respectively., info:eu-repo/semantics/publishedVersion

In this work, the anchorage of pig odorant binding protein (OBP-I) into liposomal membrane was promoted by the fusion of OBP-I with the anchor SP-DS3 peptide and with the (GQ)20 spacer. The presence of the (GQ)20 spacer in the construct confers flexibility to the protein and increases the distance between the OBP binding site and the liposomal surface. The engineered proteins, OBP::SP-DS3 and OBP::(GQ)20::SP-DS3, were produced in Escherichia coli BL21(DE3) and characterized by circular dichroism spectroscopy and MALDI-TOF. The functionalization of liposomes with the OBP proteins was performed through ethanol injection, and similar liposomal anchorage (9297%) was found for both OBP constructs. The effect of OBPs proximity to the liposomes membrane on 1-aminoanthracene (1-AMA, model ligand) transduction was evaluated by measuring the amount of 1-AMA transduced into liposomes by fluorescence spectroscopy. While protein flexibility, given by the presence of the (GQ)20 spacer, seems to influence the binding efficiency, 45% for OBP::(GQ)20::SP-DS3 and 29% for OBP::SP-DS3, the distance between the proteins binding site and the liposomal membrane determines their ability to transduce the 1-AMA into the liposomes (23% for OBP::SP-DS3 and 19% for OBP::(GQ)20::SP-DS3). The anchorage capacity and proximity effect were confirmed by an experimental control where the wild-type (wt) OBP was added to the liposomes, resulting in low 1-AMA transduction (3.5%) and low binding to OBPwt (9%). These findings evidence the effect of anchorage, carrier proteins flexibility, and proximity as key features for the entrapment of molecules into the liposomal membrane. The developed OBP-based devices are thus promising anchorage systems for the capture and storage of odors with potential applications in textile and cosmetic industries., This work was supported by the Portuguese Fundação para a Ciência e a Tecnologia (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). F.G. and A.R. thank FCT for their scholarships (SFRH/BD/114684/2016 and SFRH/BPD/98388/2013). C.S. is an investigator FCT (SFRH/IF/00186/2018). We thanks to the BioTecNorte operation (NORTE-01-0145- FEDER390 000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte., info:eu-repo/semantics/publishedVersion

Given the growing harmful effects of ultraviolet (UV) radiation there is an urgent need to develop effective strategies to prevent skin damages. Therefore, hydrogel or cotton and wool fabrics were impregnated with avobenzone (AVO)-loaded and omega-3 (3)-enriched lipid formulations (liposomes LIP, nanostructured lipid carriers NLC and solid lipid nanoparticles SLN) to obtain UV blocking sunscreen gel or functional textiles. To evaluate the physicochemical characteristics of AVO and the sun blocking performance of the formulations incorporated in hydrogels or textiles, in silico and in vitro studies were conducted. In silico studies indicated that AVO can permeate the epithelium, and lipid formulations could be a promising strategy for incorporating and retaining AVO. Furthermore, all lipid formulations showed an entrapment efficiency of AVO of ca. 100 % and a loading capacity between 712 %. The presence of AVO was confirmed by ATR-FTIR and SEM analysis. In vitro release profiles showed that the UV filter was almost totally retained within the lipid formulations (release of only 4.2 ± 2.5 % at 6 h) with similar results obtained for textiles. After assessing the cytotoxicity, cationic formulations loaded with AVO showed to be cytotoxic in a human keratinocyte cell line (NCTC 2544 cells) as opposed to AVO-loaded anionic formulations after 24 h. Photodegradation studies demonstrated that lipid formulations have photoprotective properties by stabilizing AVO but the photoprotective role could not be attributed to 3 despite its antioxidant effect evaluated by ATR-FTIR. Overall, it is possible to conclude that SLN and NLC were more effective as UV protectors than LIP, and that each formulation can have different applications. Negatively charged NLC and SLN can be used for UV protection in contact with the skin either in sunscreen gel or clothing, while positively charged NLC and SLN should be reserved as UV protectors of tissues not in contact with skin (e.g., umbrellas, curtains, etc.)., This work was supported by Fundação para a Ciência e Tecnologia (FCT I.P.) in the framework of the Strategic Funding [UID/FIS/04650/2019 and UIDB/04469/2020], “Contrato-Programa” UIDB/ and by the project CONCERT [POCI-01-0145-FEDER-032651 and PTDC/NAN-MAT/326512017], co-financed by the European Regional Development Fund (ERDF), through COMPETE 2020, under Portugal 2020, and FCT I.P. Marlene Lúcio thanks FCT and ERDF for doctoral position [CTTI-150/18-CF (1)] in the scope of the project CONCERT. Raul Machado acknowledges FCT I.P. for funding within the Scientific Employment Stimulus instrument (CEECIND/00526/2018) The authors also would like to express appreciation for the support of the sponsors [Programme - COMPETE and by national funds through FCT – Foundation for Science and Technology within the scope of the project POCI-01-0145-FEDER-007136]. Maria João Faria and Ana Rita Caldas thank for their grants (TSSiPRO—NORTE‐01‐0145‐ FEDER‐000015—supported by the regional operational program NORTE 2020, under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund), info:eu-repo/semantics/publishedVersion

Objective The capacity of hair to absorb water causes changes in its physical and cosmetic properties under different environmental conditions. Hence, the control of hair volume in variable relative humidity settings is an important topic in cosmetics. The behaviour of two types of hair, Caucasian and Asian, was studied regarding their volume change in different relative humidity conditions. The ability of a peptide as a hair volume treatment was evaluated in two climate control formulations. Methods Tresses of the two types of hair were tested in two relative humidity (RH) conditions: (A) variable relative humidity (2 hours 40% RH, followed by 2 hours 90% RH and 2 hours of 40% RH), and (B) continuous high relative humidity (90% RH for 6 hours). Changes in the hair tress volume were assessed throughout time. Hair treated with two climate control formulations, with and without a peptide (KP peptide), were tested under the two relative humidity conditions. Results Caucasian hair had a higher change in volume compared to the Asian hair in variable and high relative humidity conditions. The hair volume increase when subject to high air humidity, and it was lower with the incorporation of a peptide into climate control formulations. Conclusion Caucasian hair showed higher volume than Asian hair when submitted to both relative humidity conditions. The incorporation of the peptide into the climate control formulations, a base (mostly composed of water 94%) and an ethanolic, was found to reduce the volume of Caucasian hair tresses. The presence of the peptide improved the hair volume change more than 60% in high relative humidity conditions., We thank the Industry and Processes Laboratory (LIP) from Centre of Biological Engineering (CEB) for allowing the use of the climate chamber Binder KBF 115. This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI01-0145-FEDER-006684) and under the Project RECI/BBB-EBI/ 0179/2012 (FCOMP-01-0124-FEDER-027462). This study was also supported by BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by the European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. C elia F. Cruz and Artur Ribeiro thank FCT for SFRH/BD/ 100927/2014 and SFRH/BPD/98388/2013 grants, respectively., info:eu-repo/semantics/publishedVersion

In the present study, we coated cotton fabrics with protein-based nanoparticles containing vitamin E (-tocopherol) by the pad-cure method. Scanning electron microscopy, Fourier transform infra-red spectroscopy, and air permeability analysis of coated samples confirmed the fixation of the nanoparticles onto the fabrics surface. The antioxidant activity of the coated fabrics was evaluated by 2,2-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals reduction. Samples coated with nanoparticles containing the highest amount of encapsulated vitamin E (20% of the oil phase) showed the highest antioxidant activity. The protein-based coating was maintained for at least 10 washing cycles, demonstrating the reliability of the pad-cure method for the fixation of nanoparticles onto cotton surfaces. A methodology for nanoparticles release from the coated surfaces and their transfer to other substrates was demonstrated by the simple crock meter rubbing in the presence of sweat and protease. A high amount of material can be transferred and released to other substrates, such as textiles and skin, through the synergistic effect of sweat/protease and abrasion. An array of cosmetic and medical applications are possible with the developed coating and release methodology in which vitamin E would impart vital benefits as skin protection, anti-aging product, or skin moisturizer., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/ BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER006684) and BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by the European Regional Development Fund under the scope of Norte2020 − Programa Operacional Regional do Norte. Artur Ribeiro thanks FCT for the SFRH\BPD\98388\2013 grant. Fatemeh Shahmoradi Ghaheh thanks the Iran Ministry of Science, Research and Technology for the monetary support., info:eu-repo/semantics/publishedVersion

Herpetic infections caused by Herpes simplex virus (HSV) are among the most common human infections, affecting more than two quarters of the world's population. The standard treatment for orofacial herpes is the administration of antiviral drugs, mainly acyclovir (ACV). However, current products are mostly based on semisolid formulations that have limited ability to promote drug skin penetration and tend to leak from the application site, thus showing reduced ability to sustain local drug residence. This work reports on the production of poly (-caprolactone) (PCL) fibrous matrices with ACV and omega-3 fatty acids (3) for application as dressings to the topical treatment of orofacial herpes. PCL fibrous matrices with the co-incorporated bioactive compounds were obtained by electrospinning and characterized regarding their morphology, chemical, physical, and mechanical properties. The potential use of the developed polymeric fibrous matrices for topical applications was evaluated by: (i) the release kinetics of the bioactive compounds; (ii) the occlusive factor of the fibrous mat; (iii) ACV skin permeation capacity; and (iv) the cytotoxicity in a keratinocyte cell line. PCL fibrous matrices loaded with the bioactive compounds presented a smooth morphology and a good balance between flexibility and hardness essential to be durable for handling, while having a desirable texture to be used comfortably. The fibrous mat also provided a sustained release of ACV during 96 h and improved the skin permeability of this drug (Kp = 0.00928 ± 0.000867 cm/h) presenting also high porosity (74%) and a water vapor transmission rate (WVTR) of 881 ± 91 g/m2day, essential to maintain moist and oxygen for faster healing of herpes lesions. Furthermore, cytotoxicity studies suggest that the fibrous mat are safe for topical application. Overall, the PCL based electrospun fibrous matrices with ACV and 3 hereby described have the potential to be used as therapeutic bandage systems for the treatment of orofacial herpes., Financial support was provided by Fundação para a Ciência e Tecnologia (FCT) in the framework of the Strategic Funding UID/FIS/04650/2019 and in the ambit of the project CONCERT with reference POCI- 01-0145-FEDER-032651 and PTDC/NAN-MAT/326512017, co-financed by the European Regional Development Fund (ERDF), through COMPETE 2020, under Portugal 2020, and FCT I.P. This work was also supported by the strategic program UID/BIA/04050/2019 and project FunBioPlas with reference ERA-IB-2-6/0004/2014 funded by national Portuguese funds through FCT I.P. Funding was also provided by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-3-R (AEI/FEDER, UE)., info:eu-repo/semantics/publishedVersion

The present research relates to a fusion protein comprising a chromogenic blue ultramarine protein (UM) bound to a keratin-based peptide (KP). The KP-UM fusion protein explores UM chromogenic nature together with KP affinity towards hair. For the first time a fusion protein with a chromogenic nature is explored as a hair coloring agent. The KP-UM protein colored overbleached hair, being the color dependent on the formulation polarity. The protein was able to bind to the hair cuticle and even to penetrate throughout the hair fibre. Molecular dynamics studies demonstrated that the interaction between the KP-UM protein and the hair was mediated by the KP sequence. All the formulations recovered the mechanical properties of overbleached hair and KP-UM proved to be safe when tested in human keratinocytes. Although based on a chromogenic non-fluorescent protein, the KP-UM protein presented a photoswitch phenomenon, changing from chromogenic to fluorescent depending on the wavelength selected for excitation. KP-UM protein shows the potential to be incorporated in new eco-friendly cosmetic formulations for hair coloration, decreasing the use of traditional dyes and reducing its environmental impact., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2019 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. We also acknowledge the strategic program UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through Fundação para a Ciência e a Tecnologia (FCT) and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI). Ana Tinoco, Filipa Gonçalves and Carla Silva thanks FCT for funding the scholarships with the references SFRH/BD/114035/2015, FRH/BD/114684/2016 and IF/00186/2015, respectively., info:eu-repo/semantics/publishedVersion

The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fbioe. 2019.00298/full#supplementary-material, Styling hair with straightening irons is a popular daily hair routine that significantly damage the hair keratin fiber due to the high temperature applied. In this study, we investigate the effect of two fusion proteins based on the human eye D-crystallin conjugated with a keratin-based peptide (KP-Cryst Wt and KP-Cryst Mut) on hair exposed to thermal damage. The mutant form was designed to improve protein stability and promote interaction with the hair. Through the study, it was demonstrated the protection of Asian and Caucasian virgin hair's structure by the pretreatments with the KP-Cryst fusion proteins. After hair thermal exposure, a higher water content was quantified by TGA on the hair fibers pretreated with the fusion proteins (about 38% for the KP-Cryst Wt and 44% for the KP-Cryst Mut). Also, negligible alterations in hair fibers' stiffness were observed after iron application, demonstrating the proteins capacity to effectively prevent the conversion of keratin -helix structure into -sheets. The results proved the capacity of the fusion proteins to bind to hair and protect it against high temperatures', supporting the development of new formulations based on the KP-Cryst proteins., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2019 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. AT and CS thanks FCT for funding the scholarships with the references SFRH/BD/114035/2015 and IF/00186/2015, respectively. JG thanks to the fellowship UMINHO/BI/424/2018., info:eu-repo/semantics/publishedVersion

HIV/AIDS stands as a global burden, and vaginal microbicides constitute a promising strategy for topical pre-exposure prophylaxis. Preceding the development of a microbicide containing tenofovir disoproxil fumarate (TDF) and emtricitabine (FTC), in silico and in vitro studies were performed to evaluate the physicochemical characteristics of both drugs, and to study their biophysical impact in lipid model systems. Results from these pre-formulation studies defined hydrogels as adequate vehicles to incorporate TDF-loaded liposomes and FTC. After studying interactions with mucin, zwitterionic liposomes with a mean diameter of 134 &plusmn, 13 nm, an encapsulation TDF efficiency of approximately 84%, and a transition temperature of 41 &deg, C were selected. The chosen liposomal formulation was non-cytotoxic to HEC-1-A and CaSki cells, and was able to favor TDF permeation across polysulfone membranes (Jss = 9.9 &mu, g&middot, cm&minus, 2&middot, h&minus, 1). After the incorporation of TDF-loaded liposomes and FTC in carbomer hydrogels, the drug release profile was sustained over time, reaching around 60% for both drugs within 3&ndash, 6 h, and best fitting the Weibull model. Moreover, liposomal hydrogels featured pseudoplastic profiles that were deemed suitable for topical application. Overall, the proposed liposomal hydrogels may constitute a promising formulation for the vaginal co-delivery of TDF/FTC.

Perspiration as a response to daily activity and physical exercise results in unpleasant odors that cause social unrest and embarrassment. To tackle it, functional textiles incorporating fragrances could be an effective clothing deodorizing product. This work presents two strategies for the release of -citronellol from functionalized cotton with carbohydrate-binding module (CBM)-based complexes (OBP::GQ20::CBM/-citronellolapproach 1 and CBM::GQ20::SP-DS3-liposome/-citronellolapproach 2). CBM from Cellulomonas fimi was fused with the odorant-binding protein (OBP::GQ20::CBM) and with an anchor peptide with affinity to the liposome membrane (CBM::GQ20::SP-DS3). In approach 1, OBP fusion protein served as a fragrance container, whereas in approach 2, the fragrance was loaded into liposomes with a higher cargo capacity. The two strategies showed a differentiated -citronellol release profile triggered by an acidic sweat solution. OBP::GQ20::CBM complex revealed a fast release (31.9% and 25.8% of the initial amount, after 1.5 and 24 h of exposure with acidic sweat solution, respectively), while the CBM::GQ20::SP-DS3-liposome complex demonstrated a slower and controlled release (5.9% and 10.5% of the initial amount, after 1.5 and 24 h of exposure with acidic sweat solution, respectively). Both strategies revealed high potential for textile functionalization aimed at controlled release of fragrances. The OBP::GQ20::CBM/-citronellol complex is ideal for applications requiring fast release of a high amount of fragrance, whereas the CBM::GQ20::SP-DS3-liposome/-citronellol complex is more suitable for prolonged and controlled release of a lower amount of -citronellol., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2019 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. F.G. and C.S. thanks FCT for their funding (SFRH/BD/114684/2016; SFRH/IF/00186/2015)., info:eu-repo/semantics/publishedVersion

In this work we explored PEGylation as an efficient strategy to improve esterases catalytic performance. For this, we PEGylated three esterases, namely lipase from Candida antarctica B (CALB), lipase from Thermomyces lanuginosus (TL) and cutinase from Fusarium solani pisi (CUT) and evaluated their catalytic performance by using the biosynthesis of poly(ethylene glutarate) as model reaction. After PEGylation with a 5 kDa aldehydePEG, CALB and cutinase revealed an increase of activity against pnitrophenyl butyrate hydrolysis (2fold of increase for CALB and 4fold of increase for cutinase). Unmodified and PEGylated lipase TL displayed however similar activity results. The polymerase activity of native and PEGylated esterases was also assessed. The data revealed a higher polymerase activity for the lipase TL and cutinase PEGylated forms (88 % conversion for PEGlipase TL and 34 % for PEGcutinase). Molecular dynamics were used to evaluate the effect of PEG on the geometry of the active site of enzymes with lid domain (TL and CALB). These studies corroborate the experimental data revealing a more open active site cavity for the PEGylated catalysts facilitating the catalysis., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2019 unit and BioTecNorte operation (NORTE‐01‐0145‐FEDER‐000004) funded by European Regional Development Fund under the scope of Norte2020 ‐ Programa Operacional Regional do Norte. The authors thanks to FCT for funding their scholarship: Jennifer Noro (SFRH/BD/121673/2016), Filipa Gonçalves (SFRH/BD/114684/2016) and Artur Ribeiro (SFRH/BPD/98388/2013). Carla Silva is an investigator FCT (SFRH/IF/00186/2015). Tarsila Castro thanks the senior position funded by the European Union through the European Regional Development Fund (ERDF) under the Competitiveness Operational Program (BioCell‐NanoART=Novel Bio‐inspired Cellular Nano‐architectures, POC‐A1.1.4‐E‐2015 nr. 30/01. 09. 2016). Access to computing resources funded by the Project “Search‐ON2: Revitalization of HPC infrastructure of UMinho” (NORTE‐07‐0162‐FEDER‐000086), cofounded by the North Portugal Regional Operational Programme (ON.2‐O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF), is also gratefully acknowledged., info:eu-repo/semantics/publishedVersion

Supplementary data to this article can be found online at https:// doi.org/10.1016/j.reactfunctpolym.2018.11.015., Textile fabrics made up of cotton, wool, and polyethylene terephthalate (PET) were coated with poly(catechol) and poly(p-phenylenediamine) through the in situ enzymatic polymerization of catechol and p-phenylenediamine, assisted by laccase under high-pressure homogenization. All coated fabrics showed antimicrobial activity against both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) strains and revealed antioxidant character, measured in terms of (ABTSâ¢+)-scavenging activity. The coating of PET and cotton fabrics with both polymers did not affect the viability of foreskin fibroblasts. The methodology proposed for the in situ coating of textile materials assisted by laccase showed to be a promising approach to produce colored antimicrobial textiles with vast potential applications, namely, clothing and medical devices, among others., This study was supported by the Chinese Government Scholarship under China Scholarship Council [grant no. 201606790036], the Chinese Foundation Key Projects of Governmental Cooperation in International Scientific and Technological Innovation [grant no. 2016 YFE0115700], and the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/ 04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. The work was also supported by the FCT – Fundação para a Ciência e a Tecnologia [grant nº SFRH/BD/121673/2016, SFRH/BPD/98388/2013, and IF/00186/ 2015]., info:eu-repo/semantics/publishedVersion

Pseudomonas aeruginosa can alter its lifestyle in response to changes in environmental conditions. The switch to a pathogenic host-associated lifestyle can be triggered by the luminosity settings, resorting to at least one photoreceptor which senses light and regulates cellular processes. This study aimed to address how light exposure affects the dynamic and adaptability of two P. aeruginosa pneumonia-associated isolates, HB13 and HB15. A phenotypic characterization of two opposing growth conditions, constant illumination and intensity of full-spectrum light and total absence of light, was performed. Given the nature of P. aeruginosa pathogenicity, distinct fractions were characterized, and its inherent pathogenic potential screened by comparing induced morphological alterations and cytotoxicity against human pulmonary epithelial cells (A549 cell line). Growth in the dark promoted some virulence-associated traits (e.g., pigment production, LasA proteolytic activity), which, together with higher cytotoxicity of secreted fractions, supported an increased pathogenic potential in conditions that better mimic the lung microenvironment of P. aeruginosa. These preliminary findings evidenced that light exposure settings may influence the P. aeruginosa pathogenic potential, likely owing to differential production of virulence factors. Thus, this study raised awareness towards the importance in controlling light conditions during bacterial pathogenicity evaluation approaches, to more accurately interpret bacterial responses., FCT -Fundação para a Ciência e a Tecnologia(SFRH/BD/98558/2013), info:eu-repo/semantics/publishedVersion

Hair mechanical properties are primary indicators of hair integrity. Here we describe a methodology based on hair's resistance to mechanical wear to evaluate cosmetic treatments. Using a vibrational vortex cycle, we developed a protocol that relies on the plastic deformation and degradation of hair, due to frictional forces between hardened steel balls and hair fragments. We demonstrate that the balls when in contact with the hair fragments promoted extended hair degradation. The degree of degradation was related to hair's resistance to mechanical wear. Lower degradation rates reflect high resistance to mechanical wear. We found differences according to hair ethnicity and type of treatment. This methodology shows great potential for the screening and evaluation of the effect of cosmetic treatments on hair., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by the European Regional Development Fund under the scope of Norte 2020 - Programa Operacional Regional do Norte. AT thanks FTC for funding the scholarship with the reference SFRH/BD/114035/2015., info:eu-repo/semantics/publishedVersion

Dispersions in transcutol/isopropyl myristate make C60 fullerene molecules suitable for transdermal delivery. We found that C60 can successfully permeate the skin using pig skin in Franz diffusion cells. Molecular dynamics simulations and transmission electron microscopy confirmed these observations. Basic cosmetic formulations with transcutol/ isopropyl myristate without harsh organic solvents show a high potential for delivery of C60 for biopharmaceutical and cosmetics applications., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI01-0145-FEDER-006684).). Artur Ribeiro thanks FCT for the SFRH\BPD\98388\2013 grant., info:eu-repo/semantics/publishedVersion

Chemical straightening of curly human hair fibres involves the use of strong reducing agents at alkaline pH. Human hair is made of keratin, and the fixation of fibre shape involves the reduction and reformation of new disulphide bonds between keratin molecules. Here, we propose an alternative and green methodology using keratin peptide sequences (10-13 residues) derived from the human genome. In a previous study, we analysed 1235 cysteine-containing peptides encoded by all human genes of hair keratin and keratin-associated proteins. These peptide fragments have been designed by nature to interact with keratin. Here we tested eight peptides, which were select based on their affinity for human hair keratin solution as shown by Matrix-Assisted Laser-Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF/TOF) and by molecular dynamics simulation. The peptides were characterized in detail regarding their ability to act as hair straightening modulators and to improve the tensile strength and elasticity of hair. Of the eight tested peptides, PepE, PepG and KP showed the highest ability to interact with a keratin peptide model, and to improve hair mechanical properties and straightening efficiency. The proposed solutions presented here will replace harsh reducing agents at alkaline pH by peptide formulations acting at neutral pH to change hair shape through the re-conformation of disulphide bonds. Here, we provide experimental evidence which explains at a molecular level how keratin decapeptides can interact with large keratin molecules in human hair, opening an innovative green approach to changing the shape of hair fibre., This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of the UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and under the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462). This study was also supported by BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. Célia F. Cruz and Artur Ribeiro thank FCT for SFRH/BD/100927/2014 and SFRH/BPD/98388/2013 grants, respectively., info:eu-repo/semantics/publishedVersion

Supplementary data to this article can be found online at https://doi.org/10.1016/j.reactfunctpolym.2020.104664., In this work, zein, a by-product of the agricultural industry of corn, was used to impart hydrophobicity and antimicrobial activity to textiles. The cotton textiles were functionalized with zein in two different forms, free or as particles. The optimized coating conditions which rendered the highest hydrophobic character to the textiles were found to be 50g/L zein in the free form prepared with 70% ethanol. To enhance the antimicrobial activity of the coated textiles, ellagic acid was encapsulated into the zein particles. These presented stable diameters, between 300 and 450nm, and positive surface charge revealing high encapsulation efficiency. After textiles coating, the controlled release of ellagic acid was tested against different mimetic biologic solutions. The release profile was dependent on the ethanol concentration, on the sweat solution pH and on the external media used during dialysis. The functionalized textiles revealed differentiated antimicrobial activity against Escherichia coli (Gram-negative) and Staphylococcus aureus (Gram-positive). For E. coli the activity was mainly related with the action of zein while for S. aureus a combined effect between the protein and the ellagic acid was observed. The results herein presented pave the way for the development of new zein-based products for a wide range of applications., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. The authors also thanks to FCT for funding: Jennifer Noro (SFRH/BD/121673/2016),Filipa Gonçalves (SFRH/BD/114684/2016) and Carla Silva (SFRH/IF/00186/2015)., info:eu-repo/semantics/publishedVersion

This work reports the synthesis of nanocomposite photocatalytic materials based on nitrogen-doped TiO2nano, SiO2 and different percentages of HY zeolite (0, 12, 25 and 50%). These materials were characterized by using Fourier transformed infrared spectroscopy, X-ray diffraction, N2 adsorptiondesorption, UV-vis diffuse reflectance spectroscopy and scanning electron microscopy. The nanocomposites, which presented an energy band gap of about 3.03 eV, were immobilized on cotton fabric and their self-cleaning properties were investigated by decolourization of rhodamine B (RhB) dye in aqueous solution under simulated solar irradiation. The fabrics coated with the photocatalysts, containing and not containing zeolites, showed the same RhB decolourization (about 95%) after 5 h, excluding the situation where a large amount of HY (50%) was used in the nanocomposites. However, results obtained from high performance liquid chromatography analysis depicted that in the presence of the HY zeolite a more effective RhB degradation was achieved. In fact, even after the use of five consecutive cycles, the RhB decolourization remained high (about 85%). Generally, the photodegradation of RhB solution in the presence of cotton fabrics functionalized with TiO2nano, TiO2nano-SiO2 and TiO2nano-SiO2-0.25 HY resulted in the formation of products that exhibited a similar cytotoxic effect when compared to the untreated RhB solution and subjected to the same tested concentrations and incubation times., Salmon Landi Jr. thanks the Capes from Brazil for supports his Doctoral Fellowship (Proc. no. 13346/13-0). The authors are also grateful to the financial support of the project “AIProcMat@N2020 – Advanced Industrial Processes and Materials for a Sustainable Northern Region of Portugal 2020”, with the reference NORTE-01-0145-FEDER-000006 and the project BioTecNorte (operation NORTE-01-0145-FEDER-000004) supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work also has been funded by ERDF through COMPETE2020 – Programa Operacional Competitividade e Internacionalizac¸ão (POCI), Project POCI-01-0145-FEDER006984 – Associate Laboratory LSRE-LCM and by national funds through FCT – Fundac¸ão para a Ciência e a Tecnologia for project PTDC/AAGTEC/5269/2014 and Centre of Chemistry (UID/QUI/00686/2013 and UID/QUI/0686/2016). Artur Ribeiro thanks FCT for the SFRH\BPD\98388\2013 grant., info:eu-repo/semantics/publishedVersion

Engineered odorant-binding proteins (OBPs) display tunable binding affinities triggered by temperature alterations. We designed and produced two engineered proteins based on OBP-I sequence: truncated OBP (tOBP) and OBP::GQ20::SP-DS3. The binding affinity of 1-aminoanthracene (1-AMA) to these proteins revealed that tOBP presents higher affinity at 25°C (kd=0.45M) than at 37°C (kd=1.72M). OBP::GQ20::SP-DS3 showed an opposite behavior, revealing higher affinity at 37°C (kd=0.58M) than at 25°C (kd=1.17M). We set-up a system containing both proteins to evaluate their temperature-dependent binding. Our data proved the 1-AMA differential and reversible affinity towards OBPs, triggered by temperature changes. The variations of the binding pocket size with temperature, confirmed by molecular modelling studies, were determinant for the differential binding of the engineered OBPs. Herein we described for the first time a competitive temperature-dependent mechanism for this class of proteins., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). A.R. and F.G. thank FCT for funding their scholarships with the references SFRH/BPD/98388/2013 and SFRH/BD/114684/2016, respectively. T.G.C. thanks senior position funded by the European Union through the European Regional Development Fund (ERDF) under the Competitiveness Operational Program (COP-A1-A1.1.4-E nr.30/01.09.2016). C.S. thanks to the BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Access to computing resources funded by the Project "Search-ON2: Revitalization of HPC infrastructure of UMinho" (NORTE-07-0162-FEDER-000086), co-funded by the North Portugal Regional Operational Programme (ON.2 - O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF), is also gratefully acknowledged., info:eu-repo/semantics/publishedVersion

Objective Human hair is an element with unquestionable relevance in society both for women and men. Therefore, it is of great importance to develop new cosmetic products for hair care capable to restore and improve hair's characteristics. Here we explore the potential of keratinbased particles in the protection and recovery of hair mechanical properties and thermal stability. Methods Keratinbased particles were obtained by high pressure homogenization (HPH) using keratin and silk fibroin. The particles were characterized regarding size, superficial charge and polydispersity index. Their safety to cells was assessed using human skin keratinocytes. Virgin and overbleached Asian hair were treated with eight keratinbased formulations. The effect of particles on hair's mechanical properties was evaluated in terms of stiffness and tensile strength. The impact of treatments in hair thermal performance was studied using differential scanning calorimetry (DSC). Results Keratinbased particles were capable to recover and/or improve the mechanical properties of virgin and overbleached hair. Virgin hair treated with K80SF20P particles presented an improvement of the mechanical properties around 40%. An increase in keratin helix denaturation enthalpy and in surface smoothness for both types of hair was also verified after treatment. These particles demonstrated stability over time and proved to be safe when tested in human keratinocytes. Conclusion The keratinbased particles here presented have the potential to be incorporated in the development of new and effective hair care cosmetic formulations., This study was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE2020 (POCI01-0145-FEDER-006684) and BioTecNorte operation (NORTE-010145-FEDER-000004) and Nanotechnology Based Functional Solutions (NORTE-01-0145- FEDER-000019) funded by the European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. We also acknowledge the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER007569) funded by national funds through Fundação para a Ciência e a Tecnologia (FCT) and by the ERDF through the COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI). Artur Ribeiro and Ana Tinoco thank FCT for funding the scholarships with the references SFRH/BPD/98388/2013 and SFRH/BD/ 114035/2015, respectively., info:eu-repo/semantics/publishedVersion

L-asparaginase (L-asparagine amino hydrolase, E.C.3.5.1.1) is an enzyme clinically accepted as an antitumor agent to treat acute lymphoblastic leukemia and lymphosarcoma. It catalyzes L-asparagine (Asn) hydrolysis to L-aspartate and ammonia, and Asn effective depletion results in cytotoxicity to leukemic cells. Microbial L-asparaginase (ASNase) production has attracted considerable attention owing to its cost effectiveness and eco-friendliness. The focus of this review is to provide a thorough review on microbial ASNase production, with special emphasis to microbial producers, conditions of enzyme production, protein engineering, downstream processes, biochemical characteristics, enzyme stability, bioavailability, toxicity and allergy potential. Some issues are also highlighted that will have to be addressed to achieve better therapeutic results and less side effects of ASNase use in cancer treatment: (a) search for new sources of this enzyme to increase its availability as a drug; (b) production of new ASNases with improved pharmacodynamics, pharmacokinetics and toxicological profiles, and (c) improvement of ASNase production by recombinant microorganisms. In this regard, rational protein engineering, directed mutagenesis, metabolic flux analysis and optimization of purification protocols are expected to play a paramount role in the near future., This work was supported by grants from Brazil, specifically those financed by CAPES (reference numbers: #2805/2010 and #2609/2013), CNPq (reference number: #552652/2011-3) and FAPESP (reference numbers: #2013/08617-7 and #2013/19486-0).

The synthesis of biodiesel from sunflower oil and methanol based on transesterification using the immobilized lipase from Thermomyces lanuginosus (Lipozyme TL-IM) has been investigated under silent conditions and under an ultrasound field. Ultrasound assisted process led to reduced processing time and requirement of lower enzyme dosage. We found for the first time that ratio 1:3 (oil to methanol) was favoured for the ultrasound assisted enzymatic process which is lower as that favoured for the silent process (ratio of 1.4). Our results indicate that intensification provided by ultrasound suppresses the need of the excess of the methanol reactant during the enzymatic biodiesel production. Ultrasound assisted enzymatic biodiesel production is therefore a faster and a cleaner processes., The authors acknowledge Brenntag India Pvt. Ltd. for kindly providing gift sample of lipase enzyme Lipozyme TL IM to carry out the research work. All authors acknowledge the funding of Department of Science and Technology and Portuguese Science Foundation under the Indo-Portuguese collaborative program.

This work is devoted to the study of the influence of different anodic aluminium oxide (AAO) morphologies on the sensitivity and performance of an AAO-based capacitive touch sensor. The AAO structures were fabricated in a cylindrical homemade anodization cell made from a solid polycarbonate billet via a lathe machining process. The AAO morphologies were obtained from the anodization of Al foil by using three different types of electrolyte (sulphuric acid, oxalic acid and phosphoric acid) and their morphologies are reported and compared using scanning electron microscopy (SEM) micrographs and currenttime characteristic curves. The sensors were fabricated by integrating the AAO structure with a nanotextured gold thin film deposited over the AAO layer by thermal evaporation, thus realizing a type of metal/insulator/metal parallel-plate capacitance sensor. It is demonstrated that AAO morphologies have influence on the performance of the AAO-based capacitive touch sensors. The variation of the capacitance of the sensors is investigated in this work for the AAO structures produced from anodization in an attempt to select anodizing conditions for a biological application aiming to detect small microorganisms such as bacterial colonies of Escherichia coli., This study was supported by the Portuguese Fundação para a Ciência e a Tecnologia (FCT) under the scope of the strategic funding of UID/FIS/04650/2013 and UID/BIO/04469/2013 units and COMPETE 2020 (POCI-01-0145-FEDER-006684). Artur Ribeiro thanks FCT for funding the scholarship with the reference SFRH/BPD/98388/2013., info:eu-repo/semantics/publishedVersion

The present work reports the coating of cotton fabrics with poly (ethylene glutarate) for the first time. Cotton fabric were soaked in diethyl glutarate and ethylene glycol diacetate without addition of extra solvent. The reactions were catalyzed by lipase from Thermomyces lanuginosus at 40 °C in water bath during 7 hours. The polyester coating was extracted and analyzed by MALDI-TOF and NMR. Monomers, dimers and trimers of poly (ethylene glutarate) were found. The modified cotton fabrics showed better hydrophobicity and an improvement in wrinkle recovery when compared with the untreated materials., This work was supported by the National Natural Science Foundation of China (Grant No. 31201134 and No. 31470509); the Industry-academic Joint Technological Prospective Fund Project of Jiangsu Province (Grant No.BY2013015-24) and the Opening Project of National Engineering Laboratory for Modern Silk (Grant No. KJS1312). This work was supported by FCT I.P through the strategic funding ID/BIO/04469/2013 and by Bioprocess and Bionanotechnology Research Group (BBRG) of Minho University and the Post Graduate Funding Program of Jiangnan University for Overseas Study. Artur Ribeiro thanks FCT for the SFRH\BPD\98388\2013 grant. All authors acknowledge the funding of Changjiang Scholars Program (Professor) in Chinese Ministry of Education., info:eu-repo/semantics/publishedVersion

Biofilm formation in urinary indwelling catheters is one of the most critical issues that patients face. Catheters were coated with poly(catechin)-antibiotic conjugates with enhanced antimicrobial properties. Catechin was conjugated with two antibiotics, namely trimethoprim (TMP) and sulfamethoxazole (SMZ) via activation with N,N′-disuccinimidyl carbonate (DSC) and subsequent coupling to molecules containing α-amine moieties. Silicone and polyurethane catheters were functionalized in situ through laccase oxidation of catechin-antibiotic conjugates. Four antimicrobial coatings were produced, namely with poly(catechin), poly(catechin)- TMP, poly(catechin)-SMZ and poly(catechin)-TMP-SMZ. The bacterial adhesion reduction was tested on the functionalized devices using gram-negative and gram-positive strains. The most significant reduction in adhesion was observed with poly(catechin)-TMP (gram-negative −85 % and grampositive −87 %) and with poly(catechin)-TMP-SMZ (gramnegative −85 % and gram-positive −91 %). The cytotoxicity to mammalian cells was tested by indirect contact for 5 days and revealed that all the tested coatings supported more than 90 % of viable cells. A promising approach for the increase of the indwelling catheter lifespan was developed aiming to reduce catheter-associated chronic infections., The authors would like to acknowledge Pronefro (Portugal) and Degania (Israel) for supplying polyurethane and silicone catheters, respectively. The author Idalina Goncalves would like to acknowledge the NOVO project (FP7-HEALTH-2011.2.3.1-5) for funding. This work was partly supported by FEDER through POFC-COMPETE and by Portuguese funds from Fundacao para a Ciencia e a Tecnologia (FCT) through the project PEst-OE/BIA/UI4050/2014. The authors Carla Silva and Teresa Matama would like to acknowledge FCT for their scholarships SFRH/BPD/46515/2008 and SFRH/BPD/47555/2008.

Nanoparticles of bovine serum albumin (BSA) and silk fibroin (SF) with entrapped α-tocopherol were produced via ultrasonic emulsification. Populations with particle size of 200300 nm and highly negatively charged were obtained for all the tested formulations. Entrapment efficiencies of around 99% revealed the effective encapsulation of α-tocopherol into the produced nanoformulations. Generally, these nanodevices did not induce significant cytotoxicity to human skin keratinocytes for all the concentrations tested. The developed formulations showed free radical scavenging of ABTS.+ ability resulting from the synergistic effect between proteins in formulation and the entrapped tocopherol. Overall, the results contribute for the establishment of BSA:VO and BSA:SF:VO as biodegradable and non-toxic nanoformulations for the functionalization of textile devices and controlled delivery of tocopherol into the skin., This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE2020 (POCI-01-0145FEDER-006684) and BioTecNorte operation (NORTE-01-0145FEDER-000004) funded by European Regional Development Fund under the scope of Norte 2020-Programa Operacional Regional do Norte. Artur Ribeiro thanks FCT for the SFRH\BPD\98388\2013 grant. Fatemeh Shahmoradi would like to acknowledge the Iran Ministry of Science, Research and Technology (MSRT) for the monetary support., info:eu-repo/semantics/publishedVersion

The objective of this work was to exploit the fibronectin type II (FNII) module from human matrix metalloproteinase-2 as a functional domain for the development of silk-based biopolymer blends that display enhanced cell adhesion properties. The DNA sequence of spider dragline silk protein (6mer) was genetically fused with the FNII coding sequence and expressed in Escherichia coli. The chimeric protein 6mer + FNII was purified by non-chromatographic methods. Films prepared from 6mer + FNII by solvent casting promoted only limited cell adhesion of human skin fibroblasts. However, the performance of the material in terms of cell adhesion was significantly improved when 6mer + FNII was combined with a silk-elastin-like protein in a concentration-dependent behavior. With this work we describe a novel class of biopolymer that promote cell adhesion and potentially useful as biomaterials for tissue engineering and regenerative medicine. Statement of Significance This work reports the development of biocompatible silk-based composites with enhanced cell adhesion properties suitable for biomedical applications in regenerative medicine. The biocomposites were produced by combining a genetically engineered silk-elastin-like protein with a genetically engineered spider-silk-based polypeptide carrying the three domains of the fibronectin type II module from human metalloproteinase-2. These composites were processed into free-standing films by solvent casting and characterized for their biological behavior. To our knowledge this is the first report of the exploitation of all three FNII domains as a functional domain for the development of bioinspired materials with improved biological performance. The present study highlights the potential of using genetically engineered protein-based composites as a platform for the development of new bioinspired biomaterials., This work was supported by Fundação para a Ciência e Tecnologia (FCT – Portugal) Funded Project “Chimera” (PTDC/EBB-EBI/109093/2008), by FCT/MEC through Portuguese funds (PIDDAC) – PEst-OE/BIA/UI4050/2014, by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI). TC is thankful to the FCT, ESF and POPH for its support through the Investigador FCT Programme (IF/01635/2014). ARibeiro thanks FCT for the SFRH\BPD\98388\2013 grant. AMPereira, RMachado and AdaCosta acknowledge FCT for PD/BD/113811/2015, SFRH-BPD/86470/2012 and SFRH/BD/75882/2011 grants, respectively.

"Published online: 17 June 2016", Asparaginase reduces the levels of asparagine in blood, which is an essential amino acid for the proliferation of lymphoblastic malign cells. Asparaginase converts asparagine into aspartic acid and ammonia. The accumulation of ammonia in the bloodstream leads to hyperammonemia, described as one of the most significant side effects of asparaginase therapy. Therefore, there is a need for asparaginase formulations with the potential to reduce hyperammonemia. We incorporated 2 % of therapeutic enzyme in albumin-based capsules. The presence of asparaginase in the interface of bovine serum albumin (BSA) capsules showed the ability to hydrolyze the asparagine and retain the forming ammonia at the surface of the capsules. The incorporation of Poloxamer 407 in the capsule formulation further increased the ratio aspartic acid/ammonia from 1.92 to 2.46 (and 1.10 from the free enzyme), decreasing the levels of free ammonia. This capacity to retain ammonia can be due to electrostatic interactions and retention of ammonia at the surface of the capsules. The developed BSA/asparaginase capsules did not cause significant cytotoxic effect on mouse leukemic macrophage cell line RAW 264.7. The new BSA/asparaginase capsules could potentially be used in the treatment of acute lymphoblastic leukemia preventing hyperammonemia associated with acute lymphoblastic leukemia (ALL) treatment with asparaginase., Fundação para a Ciência e a Tecnologia - SFRH\BPD\98388\2013 ; UID/BIO/04469/2013 ; UID/BIA/04050/2013, COMPETE 2020 (POCI-01-0145-FEDER-006684)

Genetic engineering was used to produce an elastin-like polymer (ELP) with precise amino acid composition, sequence and length, resulting in the absolute control of MW and stereochemistry. A synthetic monomer DNA sequence encoding for (VPAVG)20, was used to build a library of concatemer genes with precise control on sequence and size. The higher molecular weight polymer with 220 repeats of VPAVG was biologically produced in Escherichia coli and purified by hot and cold centrifugation cycles, based on the reversible inverse temperature transition property of ELPs. The use of low cost carbon sources like lactose and glycerol for bacteria cells culture media was explored using Central Composite Design approach allowing optimization of fermentation conditions. Due to its self-assembling behaviour near 33 oC stable spherical microparticles with a size ~ 1μm were obtained, redissolving when a strong undercooling is achieved. The polymer produced showed hysteresis behaviour with thermal absorbing/releasing components depending on the salt concentration of the polymer solution., This work was supported by the ElastM Project, POCI/CTM/57177/2004 from Fundação para a Ciência e Tecnologia (Portugal) and FEDER, “Junta de Castilla y Leon” (VA087A06, VA016B08 and VA030A08), MEC (MAT2007-66275-C02-01 and NAN2004-08538), and by the Marie Curie RTN Biopolysurf (MRTN-CN-2004-005516). Raul Machado received a PhD grant: SFRH/BD/36754/2007 from Fundação para a Ciência e Tecnologia (Portugal). The authors would like to thank Magda Graça (CBMA) for all the technical support in sequencing, Lígia Rodrigues for the CCD enlightenment and Dan W. Urry for the stimulating discussions and encouragement to pursue the studies on protein-based polymers.

The present work explores the best conditions for the enzymatic synthesis of poly (ethylene glutarate) for the first time. The start-up materials are the liquids; diethyl glutarate and ethylene glycol diacetate, without the need of addition of extra solvent. The reactions are catalyzed by lipase B from Candida antarctica immobilized on glycidyl methacrylate-ter-divinylbenzene-ter-ethylene glycol dimethacrylate at 40 °C during 18 h in water bath with mechanical stirring or 1 h in ultrasonic bath followed by 6 h in vacuum in both the cases for evaporation of ethyl acetate. The application of ultrasound significantly intensified the polyesterification reaction with reduction of the processing time from 24 to 7 h. The same degree of polymerization was obtained for the same enzyme loading in less time of reaction when using the ultrasound treatment. The degree of polymerization for long-term polyesterification was improved approximately 8-fold due to the presence of sonication during the reaction. The highest degree of polymerization achieved was 31, with a monomer conversion of 96.77%. The ultrasound treatment demonstrated to be an effective green approach to intensify the polyesterification reaction with enhanced initial kinetics and high degree of polymerization., This study was supported by Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). The work was also supported by Bioprocess and Bionanotechnology Research Group (BBRG) of Minho University and the Post Graduate Funding Program of Jiangnan University for Overseas Study. All authors also acknowledge the funding of Department of Science and Technology and Portuguese Science Foundation under the Indo-Portuguese collaborative program.

"Available online 6 May 2016", An effective tissue engineering approach requires adjustment according to the target tissue to be engineered. The possibility of obtaining a protein-based formulation for the development of multivalent tunable scaffolds that can be adapted for several types of cells and tissues is explored in this work. The incremental substitution of bovine serum albumin (BSA) by human serum albumin (HSA), changing the scaffolds hydrophilic/hydrophobic ratio, on a previously optimized scaffold formulation resulted in a set of uniform porous scaffolds with different physical properties and associated cell proliferation profile along time. There was a general trend towards an increase in hydrophilicity, swelling degree and in vitro degradation of the scaffolds with increasing replacement of BSA by HAS. The set of BSA/HSA scaffolds presented distinct values for the storage (elastic) modulus and loss factor which were similar to those described for different native tissues such as bone, cartilage, muscle, skin and neural tissue. The preferential adhesion and proliferation of skin fibroblasts on the BSA25%HSA75% and HSA100% scaffolds, as predicted by their viscoelastic properties, demonstrate that the BSA/HSA scaffold formulation is promising for the development of scaffolds that can be tuned according to the tissue to be repaired and restored., This work was supported by FCT I.P through the strategic funding UID/BIO/04469/2013 and the strategic funding UID/BIA/04050/2013. Artur Ribeiro thanks FCT for the SFRHBPD983882013 grant.

Objective Hair follicles are widely recognized as the preferential target and site of accumulation for nanoparticles after topical application. This feature is of particular importance for hair cosmetics, having the potential to refine the treatment of several hair follicle-related disorders. The aim of this work was to improve the preparation of Poly (D,L-lactide) (PLA) nanoparticles for in vivo follicular target and drug delivery. Methods Envisaging a future industrial scale-up of the process, nanoprecipitation method was used to prepare PLA nanoparticles: the effect of several processing parameters on their properties was examined and the yield of nanoparticles formation determined. Encapsulation efficiencies and in vitro release profiles of lipophilic and hydrophilic model compounds were also assessed. In vitro cytotoxicity and ex vivo penetration studies were performed on a reference skin cell line (NCTC2455, human skin keratinocytes) and porcine skin, respectively. Results Using acetone : ethanol (50 : 50, v/v) as the solvent phase, 0.6% (w/w) of Pluronic® F68 as a surfactant agent and agitation to mix the solvent and non-solvent phases, a monodispersed population of non-cytotoxic spherical nanoparticles of approximately 150 nm was obtained. The yield of nanoparticles for this formulation was roughly 90%. After encapsulation of model compounds, no significant changes were found in the properties of particles and the entrapment efficiencies were above 80%. The release kinetics of dyes from PLA nanoparticles indicate an anomalous transport mechanism (diffusion and polymer degradation) for Nile Red (lipophilic) and a Fickian diffusion of first order for fluorescein 5(6)-isothiocyanate (hydrophilic). Ex vivo skin penetration studies confirmed the presence of nanoparticles along the entire follicular ducts. Conclusions The optimized method allows the preparation of ideal PLA nanoparticles-based formulations for hair follicle targeting. PLA nanoparticles can effectively transport and release lipophilic and hydrophilic compounds into the hair follicles, and the yields obtained are acceptable for industrial purposes., Objectif Les follicules pileux sont largement reconnus comme la cible préférentielle et le site de l'accumulation des nanoparticules après application topique. Cette caractéristique est particulièrement importante pour les produits cosmétiques pour les cheveux, ayant la possibilité d'affiner le traitement de plusieurs troubles des follicules de cheveux. Le but de ce travail était d'améliorer la préparation de nanoparticules poly (D,L-lactide) (PLA) pour une administration folliculaire in vivo ciblée de drogues. Méthodes En envisageant un avenir à l’échelle industrielle du procédé, une méthode de nanoprécipitation a été utilisé pour préparer des nanoparticules de PLA: l'effet de plusieurs paramètres de traitement sur leurs propriétés a été examiné et le rendement de la formation des nanoparticules a été déterminé. Les efficacités d'encapsulation et de profils de libération in vitro de composés modèles lipophiles et hydrophiles ont également été évaluées. La cytotoxicité in vitro et ex vivo des études de pénétration a été effectuée sur une lignée de cellules de peau de référence (NCTC2455, des kératinocytes de peau humaine) et la peau de porc, respectivement. Resultats En utilisant l'acétone : éthanol (50 : 50, v/v) comme phase solvant, 0,6% (p/p) de Pluronic® F68 à titre d'agent tensioactif et l'agitation pour mélanger les phases de solvant et de non-solvant, une population monodispersée des nanoparticules sphériques non cytotoxiques d'environ 150 nm a été obtenue. Le rendement de nanoparticules pour cette formulation était d'environ 90%. Après encapsulation de composés modèles, aucune modification significative n'a été observée dans les propriétés des particules et les efficacités de piégeage ont été supérieures à 80%. La cinétique de libération de colorants de nanoparticules de PLA indique un mécanisme de transport anormal (diffusion et dégradation de polymère) pour le rouge Nil (lipophile) et une diffusion selon Fick de premier ordre pour FITC (hydrophile). Les études de pénétration ex vivo de la peau ont confirmé la présence de nanoparticules sur tous les conduits folliculaires. Conclusions La méthode optimisée permet la préparation de formulations à base de nanoparticules de PLA, idéales pour ciblage du follicule pileux. Les nanoparticules de PLA peuvent effectivement transporter et libérer des composés lipophiles et hydrophiles dans les follicules pileux; les rendements obtenus sont acceptables à des fins industrielles., The authors thank to the FCT Strategic Project PEst-OE/EQB/LA0023/2013 and to the Project “BioHealth – Biotechnology and Bioengineering approaches to improve health quality”, Ref. NORTE- 07-0124-FEDER-000027, co-funded by the Programa Operacional Regional do Norte (ON.2 – O Novo Norte), QREN, FEDER. This work was partly supported by FEDER through POFC–582 COMPETE and by Portuguese funds from FCT– Fundacão Para a Ciência e a Tecnologia through the project PEst-OE/BIA/UI4050/2014. The authors thank C. Botelho for technical assistance.

In the present work we explored the ABP-CM4 peptide properties from Bombyx mori for the creation of biopolymers with broad antimicrobial activity. An antimicrobial recombinant protein-based polymer (rPBP) was designed by cloning the DNA sequence coding for ABP-CM4 in frame with the N-terminus of the elastin-like recombinamer consisting of 200 repetitions of the pentamer VPAVG, here named A200. The new rPBP, named CM4-A200, was purified via a simplified nonchromatographic method, making use of the thermoresponsive behavior of the A200 polymer. ABP-CM4 peptide was also purified through the incorporation of a formic acid cleavage site between the peptide and the A200 sequence. In soluble state the antimicrobial activity of both CM4-A200 polymer and ABP-CM4 peptide was poorly effective. However, when the CM4-A200 polymer was processed into free-standing films high antimicrobial activity against Gram-positive and Gram-negative bacteria, yeasts and filamentous fungi was observed. The antimicrobial activity of CM4-A200 was dependent on the physical contact of cells with the film surface. Furthermore, CM4-A200 films did not reveal a cytotoxic effect against both normal human skin fibroblasts and human keratinocytes. Finally, we have developed an optimized ex vivo assay with pig skin demonstrating the antimicrobial properties of the CM4-A200 cast films for skin applications., This work was supported by FEDER through POFC - COMPETE, by Portuguese funds from FCT through Project PEst-OE/BIA/UI4050/2014, and by the Spanish Minister of Economy and Competitiveness (MAT2012-38043-C02-01) and Junta de Castilla y Leon-JCyL (VA152A12-2 and VA155A12-2), Spain. The authors also thank Matadouro - Central Carnes de Entre Douro e Minho, Lda for their availability to provide pig skin. A.C. and R.M. acknowledge FCT for SFRH/BD/75882/2011 and SFRH-BPD/86470/2012 Grants, respectively. T.C. is thankful to the FCT for its support through Programa Ciencia 2008.

Sardine is one of the most common fish of the Portuguese coast with important nutritional features. Sardine oil is also a natural source of nutrients with proven benefits for human health, being rich in omega-3 fatty acids, particularly EPA and DHA (polyunsaturated fatty acids-PUFAs) [1]. Several studies show that there is a direct link between a diet enriched in omega-3 and the prevention of diseases such as cardiovascular disease, inflammatory conditions, mental disorders and prevention of various types of cancer [2]. The aim of this work was to characterize the antioxidant role of sardine oil for the development of topical applications. To evaluate the antioxidant effect of sardine oil on skin, human fibroblasts (BJ-5ta), human melanocytes (A375) and human keratinocytes (NCTC2544) were used. Concentrations of oil higher than 8 mg/ml affected significantly the cell viability while for lower concentrations the effect was reduced. The lowest concentrations, 0.5 and 4 mg/ml, were tested to evaluate the protective role of sardine oil in situation of induced oxidative stress. These two concentrations were able to protect cells from damage with a higher effect measured for the fibroblasts. Moreover the incubation of cells with the sardine oil was able to activate a transcription factor Nuclear factor-erythroid-2-related factor 2 (Nrf2) which plays a crucial role in the coordinated induction of genes encoding many stress-responsive and cytoprotective enzymes and related proteins [3]. These results open the opportunity to develop new therapeutic and cosmetic applications based on sardine-derived compounds. Their incorporation in topical creams may contribute to a better treatment of inflammation and in the prevention of skin aging.

The authors regret that Telma C. Bernardo was inadvertently omitted in the author line-up. The correct authorship order should be as follows: Ana Margarida Pereira, Raul Machado, André da Costa, Artur Ribeiro, Telma C. Bernardo, Tony Collins, Andreia C. Gomes, Isabel B. Leonor, David L. Kaplan, Rui L. Reis, Margarida Casal. Telma C. Bernardo participated in recombinant 6mer+FNII production and purification. The authors regret the error and would like to apologize for any inconvenience caused., (undefined)

The objective of this work was to develop a new strategy to physically ‘repair’ chemically damaged hair. Hence the human eye γD-crystallin, a protein from the superfamily characterized structurally by the Greek key motif, was studied. The human γD-crystallin was chosen based on the ability of proteins belonging to this superfamily to be involved in the coating of specific structures. Two crystallins were used on the study, the wild type (Protein Data Bank ID: 1HK0) and the mutant protein. The mutant form was intended to induce a strong and quick protein polymerization as well to have new possible points of anchorage to hair., L'objectif de ce travail était de développer une nouvelle stratégie pour physiquement “réparer” les cheveux chimiquement endommagés. La protéine humaine γD-cristalline, une protéine de la superfamille caractérisée structurellement par le motif clé grecque, a été étudiée. Le γD-cristallin humain a été choisi en fonction de la capacité des protéines appartenant à cette superfamille d'être impliquées dans le revêtement des structures spécifiques. Deux cristallins ont été utilisés dans l'étude, le type sauvage (Protein Data Bank ID: 1HK0) et la protéine mutante. La forme mutante était destinée à induire une polymérisation de la protéine, forte et rapide ainsi d'avoir de nouveaux points d'ancrage possibles aux cheveux., This work is supported by FEDER through POFC-COMPETE and by national funds from FCT through the project PEst-C/BIA/UI4050/2011.

Transdermal perfusion of a large protein is reported for the first time, using a nanoemulsion of bovine serum albumin (66 kDa) of 160 nm prepared by a solid-in-oil (S/O) process. Molecular dynamics simulations confirmed skin permeation by these formulations, with integration of the protein into the lipid bilayers. These results demonstrate the real possibility of delivering large proteins transdermally for a range of medical and cosmetic applications., We thank Matadouro - Central Carnes de Entre Douro e Minho, Lda for their support on pig samples. The histological studies were supported by the Department of Histology from Life and Health Sciences Research Institute (ICVS), University of Minho. The confocal 3D images were performed by Department of Chemistry and Kanbar Laboratory for Nanomaterials Bar-Ilan University Center for Advanced Materials and Nanotechnology. Carla Silva would like to thanks Fundacao para a Ciencia e a Tecnologia for their funding (SFRH/BPD/46515/2008).