Absence of light exposure increases pathogenicity of Pseudomonas aeruginosa pneumonia-associated clinical isolates

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/publishedVersio

Surface charge mediated cell-surface interaction on piezoelectric materials

Cell–material interactions play an essential role in the development of scaffold-based tissue engineering strategies. Cell therapies are still limited in treating injuries when severe damage causes irreversible loss of muscle cells. Electroactive biomaterials and, in particular, piezoelectric materials offer new opportunities for skeletal muscle tissue engineering since these materials have demonstrated suitable electroactive microenvironments for tissue development. In this study, the influence of the surface charge of piezoelectric poly(vinylidene fluoride) (PVDF) on cell adhesion was investigated. The cytoskeletal organization of C2C12 myoblast cells grown on different PVDF samples was studied by immunofluorescence staining, and the interactions between single live cells and PVDF were analyzed using an atomic force microscopy (AFM) technique termed single-cell force spectroscopy. It was demonstrated that C2C12 myoblast cells seeded on samples with net surface charge present a more elongated morphology, this effect being dependent on the surface charge but independent of the poling direction (negative or positive surface charge). It was further shown that the cell deadhesion forces of individual C2C12 cells were higher on PVDF samples with an overall negative surface charge (8.92 ± 0.45 nN) compared to those on nonpoled substrates (zero overall surface charge) (4.06 ± 0.20 nN). These findings explicitly demonstrate that the polarization/surface charge is an important parameter to determine cell fate as it affects C2C12 cell adhesion, which in turn will influence cell behavior, namely, cell proliferation and differentiationPortuguese Foundation for Science and Technology (FCT) in the framework of the Strategic Funding UID/FIS/04650/2019, UID/BIA/04050/2013, UID/BIO/04469, project POCI-01-0145-FEDER-028237 and under BioTecNorte operation (NORTE-01-0145-FEDER-000004). The authors also thank the FCT for the SFRH/BD/111478/2015 (S.R.) and SFRH/BPD/90870/2012 (C.R.) grants. Funds provided by FCT in the framework of EuroNanoMed 2016 call, Project LungChek ENMed/0049/2016 are also gratefully acknowledged. The authors acknowledge funding by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-3-R (AEI/FEDER, UE) and from the Basque Government Industry and Education Department under the ELKARTEK, HAZITEK and PIBA (PIBA-2018-06

Satisfação dos utentes com o atendimento prestado nas farmácias comunitárias

A satisfação dos utentes é considerada uma medida importante com efeitos nos resultados de cuidados de saúde. Objetivos: Determinar o grau de satisfação dos utentes com o atendimento nas farmácias comunitárias, e verificar se o grau de satisfação é igual, independentemente, da idade e do género do utente. Material e Métodos: Este estudo de caracter transversal, observacional e correlacional teve como base uma amostra não probabilística acidental, constituída por 525 inquiridos. O questionário foi aplicado no período de Outubro de 2012 a Janeiro de 2013 em quatro localidades seleccionadas por conveniência na zona norte de Portugal, foi utilizada a escala “Pharmacy Services Questionnaire” (FSQ) validada para a língua portuguesa, desenvolvida por Larson et al. (2002). Em relação ao género dos inquiridos, verificou-se que 60,62% pertenciam ao sexo feminino e 39,38% eram do sexo masculino. Com idades compreendidas entre os 18 e os 83 anos, com uma média de 41,81 anos (D.P.±16,262, o que permite concluir que a amostra não é homogénea relativamente à idade). Em relação à escolaridade dos inquiridos, 2,9% não possuíam estudos; 16,8% tinham entre 1 a 4 anos de escolaridade; 9,5% possuíam o 2º ciclo; 12,4% possuíam o 3º ciclo; 31,6% concluíram o ensino secundário e 23,2% possuíam habilitações superiores. Resultados: Os indicadores de validade da análise fatorial mostraram um KMO=0,972. Foram identificados duas dimensões, nomeadamente, o atendimento e acompanhamento farmacoterapêutico, que explicavam no conjunto 64,230% da variância total. O Alfa Cronbach total foi de 0,962, o que indica uma fiabilidade da consistência interna muito boa. A contribuição das dimensões para a satisfação global dos utentes foi por ordem de importância, o atendimento (r=0,864; p<0,01) e acompanhamento farmacoterapêutico (r=0,848; p<0,01). O grau de satisfação dos utentes com os serviços das farmácias comunitárias foi o seguinte: reduzido para 5,3%, médio para 39%, bom para 46,1% e muito bom para 9%. Tendo em conta o fator atendimento por género, o feminino registou um grau de satisfação mais elevado, enquanto que, no fator acompanhamento farmacoterapêutico não se verificou qualquer diferença entre géneros. Relativamente à classe etária, verificou-se que são os utentes com idades mais avançadas, que registam um grau de satisfação mais elevado. Discussão e Conclusão: Concluiu-se que, de um modo geral, os utentes se encontram satisfeitos com o atendimento nas farmácias comunitárias, sendo o sexo feminino e os utentes com idades mais avançadas, os que apresentam um grau de satisfação superior. O aspeto a melhorar para o aumento da satisfação dos utentes, relativamente ao acompanhamento farmacoterapêutico, é “O modo como o profissional de farmácia lhe explica os possíveis efeitos secundários”. Em relação ao atendimento o aspeto a melhorar é “O modo como o profissional de farmácia responde às suas perguntas”

Users satisfaction regarding the service provided in community pharmacies

During the last century, the pharmacy professionals have significantly being professionalized, the user is a high priority and not just limited to the sale of pharmaceuticals. As such, it is necessary to know the users, their needs and expectations, their opinions and preferences. This transversal study, observational, correlational, aimed to: determine the degree of satisfaction with the attendance in community pharmacies, if there is a relationship between factors of a personal nature (age and gender) and satisfaction, trying to identify which aspects improve in order to increase satisfaction levels. The sample consisted of 525 respondents. Data collection was conducted in the period from October 2012 to January 2013 in four locations in northern Portugal. It was used a questionnaire consisting of three parts: the first contained items of personal characterization, the second consisted of the scale "Pharmacy Services Questionnaire" (FSQ) validated for the Portuguese language, developed by Larson et al. (2002) and the last had questions about the respondents' opinion. The results showed that the degree of satisfaction with the services of community pharmacies was in majority an average to 39 % and good for 46,1% . Having in mind the factor by gender, females showed a higher degree of satisfaction. Regarding the age group, older users had the highest degree of satisfaction. It was concluded that, generally, community pharmacies bring a higher degree of satisfaction in the older and the female users. "How the pharmacy professionals explain possible side effects" and "how the pharmacy professional answers your questions.", are the two aspects that must be improved to achieve a higher degree of satisfaction. KEYWORDS Satisfaction, Users, Customer Service, Pharmacotherapy Monitoring, Community Pharmacie

Fusion proteins with chromogenic and keratin binding modules

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/publishedVersio

Development of therapeutic and cosmetic formulations based on sardine-based products

Sardine is one of the most common fish of the Portuguese coast and has important nutritional features. Sardine oil is also a source of nutrients with proven benefits for human health, being rich in 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 such as rheumatoid arthritis or asthma, mental disorders and prevention of various types of cancer [2]. The aim of this work was to characterize in a systematic way the potential protective role of sardine oil and derived PUFAs. To evaluate the antioxidant and antiinflammatory effect of sardine oil and PUFAs, human fibroblasts (BJ-5ta), human melanocytes (A375) and human keratinocytes (NCTC2544) were used. Cell viability was affected for concentrations higher than 8mg/ml for sardine oil and higher than 0.1mg/ml for PUFAs. However and regarding PUFAs, melanocytes revealed a higher susceptibility. With the lowest tested concentrations, sardine-based compounds promoted cell proliferation and protected cells from induced oxidative stress, with higher protection conferred by PUFAs. 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

Multifunctional platform based on electroactive polymers and silica nanoparticles for tissue engineering applications

Poly(vinylidene fluoride) nanocomposites processed with different morphologies, such as porous and non-porous films and fibres, have been prepared with silica nanoparticles (SiNPs) of varying diameter (17, 100, 160 and 300 nm), which in turn have encapsulated perylenediimide (PDI), a fluorescent molecule. The structural, morphological, optical, thermal, and mechanical properties of the nanocomposites, with SiNP filler concentration up to 16 wt %, were evaluated. Furthermore, cytotoxicity and cell proliferation studies were performed. All SiNPs are negatively charged independently of the pH and more stable from pH 5 upwards. The introduction of SiNPs within the polymer matrix increases the contact angle independently of the nanoparticle diameter. Moreover, the smallest ones (17 nm) also improve the PVDF Youngs modulus. The filler diameter, physico-chemical, thermal and mechanical properties of the polymer matrix were not significantly affected. Finally, the SiNPs inclusion does not induce cytotoxicity in murine myoblasts (C2C12) after 72 h of contact and proliferation studies reveal that the prepared composites represent a suitable platform for tissue engineering applications, as they allow us to combine the biocompatibility and piezoelectricity of the polymer with the possible functionalization and drug encapsulation and release of the SiNP.This work was supported by the Portuguese Foundation for Science and Technology (FCT) in the framework of the Strategic Funding UID/FIS/04650/2013 and UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) and project POCI-01-0145-FEDER-028237 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); and also under the scope of the strategic funding of UID/BIO/04469 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. The authors also thank the FCT for the SFRH/BD/111478/2015 (S.R.), SFRH/BPD/96707/2013 (T.R.), SFRH/BPD/90870/2012 (C.R.) and SFRH/BPD/121526/2016 (D.C) grants. The authors acknowledge funding from the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-3-R (AEI/FEDER, UE) and from the Basque Government Industry and Education Departments under the ELKARTEK, HAZITEK and PIBA (PIBA-2018-06) programs, respectively.info:eu-repo/semantics/publishedVersio

Incidence of Rotavirus and Adenovirus: detection by molecular and immunological methods in human faeces

Rotaviruses and Adenoviruses are reported worldwide among the main agents of gastroenteritis and, consequently, the development and validation of sensitive and cost effective methods of detection is necessary. In this study, two approaches for detection of Rotavirus A and Adenovirus in samples of human faeces were compared: the immunological kit VIKIA Rota-Adeno and the nested-polymerase chain reaction (nested-PCR for Adenovirus) and Reverse Transcriptase-PCR (RT-PCR for Rotavirus) molecular methods. From January 2006 to July 2009, 467 samples of faeces from individuals with gastroenteritis symptoms assisted at the Hospital Infant D. Pedro (Aveiro, Portugal) were analysed for Rotavirus and Adenovirus, using the VIKIA kit. From the 467 samples, 59 (12.6%) were positive for Rotavirus and 5 (1.1%) for Adenovirus. Between December 2008 to July 2009, 18 samples were analysed by both immunologic and molecular methods. From the 18 samples, 10 were positive for Rotavirus (55.5%) and 16 for Adenovirus (88.9%) when analysed by RT-PCR and nested-PCR, respectively With VIKIA kit, 11 samples were positive for Rotavirus (61.1%) and only one was positive for Adenovirus (5.5%). Sequencing of PCR products confirmed the presence of Rotavirus in 1 sample and Adenovirus in the 10 samples that were classified as negative with VIKIA Kit. The results of VIKIA kit suggest that from the both viruses studied Rotavirus are the most incident viruses in gastroenteritis, however, molecular analysis results suggest that Adenovirus could be the most responsible for the viral gastroenteritis studied.The authors would like to thank University of Aveiro and Centre for Environmental and Marine Studies (CESAM, project Pest-C/MAR/LA0017/2011) for funding the study and to Hospital Infant D. Pedro of Aveiro for providing the faeces amples and to Dr. Albert Bosch from Barcelona University for the donation of Rotavirus and Adenovirus suspension.publishe

BSA/ASN/Pol407 nanoparticles for acute lymphoblastic leukemia treatment

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/publishedVersio

Evaluation of Arenaria montana L. hydroethanolic extract as a chemopreventive food ingredient: A case study focusing a dairy product (yogurt)

Natural ingredients are valuable options to be exploited in the design of innovative food formulations with health benefits. Therefore, it was evaluated the potential use of Arenaria montana L. hydroethanolic extract (rich in apigenin derivatives) as a chemopreventive agent in functional foods. Apigenin is recognized as inhibiting VEGFR-2, which is the key receptor involved in angiogenesis. The obtained extract was also able to inhibit the VEGFR-2 phosphorylation through an enzymatic assay (IC 50 = ~63 µg/mL). Thereafter, free and microencapsulated forms were incorporated in yogurt. The obtained products maintained the nutritional value along the tested 3 days of storage, as also free sugars and fatty acids profiles, in comparison with the control samples. Nevertheless, the VEGFR-2 phosphorylation inhibition was not exhibited as intended. Even this behavior for the microencapsulated forms can be attributed to the protecting effect of the alginate matrix, further studies are required in order to better understand the shown performance.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER for CIMO (UID/AGR/00690/2013) financial support. To POCI-01-0145-FEDER-006984 (LA LSRE-LCM) funded by ERDF through POCI-COMPETE2020 and FCT. To NORTE-01-0145-FEDER-000006, funded by NORTE 2020, under PT2020 through ERDF. L. Barros, R.C. Calhelha and J.C.M. Barreira acknowledge the FCT for their post-doctoral grants (SFRH/BPD/107855/2015, SFRH/BPD/68344/2010 and SFRH/BPD/72802/2010, respectively). The authors also thank Ana Maria Carvalho for providing Arenaria montana L. samples.info:eu-repo/semantics/publishedVersio