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Amphibian secretions have been extensively investigated for the production of bioactive molecules. Salamandrin-I is an antioxidant peptide, isolated from the skin secretion of the fire salamander, that has induced no toxicity in microglia or erythrocytes. Importantly, the administration of antioxidants may constitute an adequate therapeutic approach to cancer treatment. Here, with the purpose of better characterizing the therapeutic potential of salamandrin-I, we investigated whether this antioxidant peptide also exerts anticancer activity, using the human leukemia cell line HL-60 as a cancer model. Salamandrin-I treatment induced a significant reduction in HL-60 proliferation, which was accompanied by cell cycle arrest. Furthermore, the peptide-induced cell death showed a significant increase in the LDH release in HL-60 cells. The cellular toxicity exerted by salamandrin-I is possibly related to pyroptosis, since the HL-60 cells showed loss of mitochondrial membrane potential and hyperexpression of inflammasome components following the peptide treatment. This is the first demonstration of the anticancer potential of the salamandrin-I peptide. Such results are important, as they offer relevant insights into the field of cancer therapy and allow the design of future bioactive molecules using salamandrin-I as a template.

Lycopene is a carotenoid with potential use in the treatment of chronic illnesses. Here, different formulations of lycopene were studied: lycopene-rich extract from red guava (LEG), purified lycopene from red guava (LPG) and a self-emulsifying drug delivery system loaded with LPG (nanoLPG). The effects of administering orally various doses of LEG to hypercholesterolemic hamsters were evaluated regarding the liver function of the animals. The cytotoxicity of LPG in Vero cells was analyzed by a crystal violet assay and by fluorescence microscopy. In addition, nanoLPG was employed in stability tests. LPG and nanoLPG were tested for their cytotoxic effect on human keratinocytes and antioxidant capacity on cells in an endothelial dysfunction model in an isolated rat aorta. Finally, the effect of different nanoLPG concentrations on the expression of immune-related genes (IL-10, TNF-α, COX-2 and IFN-γ) from peripheral blood mononuclear cells (PBMC) using real-time PCR was also analyzed. Results suggest that LEG, despite not being able to improve blood markers indicative of liver function in hypercholesterolemic hamsters, reduced hepatic degenerative changes. Additionally, LPG did not show cytotoxicity in Vero cells. In relation to nanoLPG, the effects produced by heat stress evaluated by Dynamics Light Scattering (DLS) and visually were loss of color, texture change and phase separation after 15 days without interfering with the droplet size, so the formulation proved to be efficient in stabilizing the encapsulated lycopene. Although LPG and nanoLPG showed moderate toxicity to keratinocytes, which may be related to cell lineage characteristics, both revealed potent antioxidant activity. LPG and nanoLPG showed vasoprotective effects in aortic preparations. The gene expression assay indicates that, although no significant differences were observed in the expression of IL-10 and TNF-α, the PBMCs treated with nanoLPG showed a reduction in transcriptional levels of IFN-γ and an increased expression of COX-2. Thus, the work adds evidence to the safety of the use of lycopene by humans and shows that tested formulations, mainly nanoLPG due to its stability, stand out as promising and biosafe products for the treatment of diseases that have oxidative stress and inflammation in their etiopathology.

© 2022 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited, Tryptophyllins constitute a heterogeneous group of peptides that are one of the first classes of peptides identified from amphibian's skin secretions. Here, we report the structural characterization and antioxidant properties of a novel tryptophyllin-like peptide, named PpT-2, isolated from the Iberian green frog Pelophylax perezi. The skin secretion of P. perezi was obtained by electrical stimulation and fractionated using RP-HPLC. De novo peptide sequencing was conducted using MALDI MS/MS. The primary structure of PpT-2 (FPWLLS-NH2 ) was confirmed by Edman degradation and subsequently investigated using in silico tools. PpT-2 shared physicochemical properties with other well-known antioxidants. To test PpT-2 for antioxidant activity in vitro, the peptide was synthesized by solid phase and assessed in the chemical-based ABTS and DPPH scavenging assays. Then, a flow cytometry experiment was conducted to assess PpT-2 antioxidant activity in oxidatively challenged murine microglial cells. As predicted by the in silico analyses, PpT-2 scavenged free radicals in vitro and suppressed the generation of reactive species in PMA-stimulated BV-2 microglia cells. We further explored possible bioactivities of PpT-2 against prostate cancer cells and bacteria, against which the peptide exerted a moderate antiproliferative effect and negligible antimicrobial activity. The biocompatibility of PpT-2 was evaluated in cytotoxicity assays and in vivo toxicity with Galleria mellonella. No toxicity was detected in cells treated with up to 512 µg/ml and in G. mellonella treated with up to 40 mg/kg PpT-2. This novel peptide, PpT-2, stands as a promising peptide with potential therapeutic and biotechnological applications, mainly for the treatment/prevention of neurodegenerative disorders., This work was financed by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operacional Programme for Competitiveness and Internationalization (POCI), and by Portuguese funds through FCT - Fundação para a Ciência e a Tecnologia in the framework of the project POCI-01-0145-FEDER-031158 – PTDC/BII-BIO/31158/2017. The authors would like to thank the participation and scientific support of the Unit projects UIDB/50006/2020 | UIDP/50006/2020, and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Universal Faixa ‘B’ (grant number 32103/2018-0). A.P. is a recipient of a post-doctoral grant from the project PTDC/BII-BIO/31158/2017. The authors would like to thank the researcher Roberto Resendes (CiBio, University of the Azores, Ponta Delgada, São Miguel, Azores, Portugal) for the logistical support in the collection of samples. C.P.A acknowledges FCT-MCTES fellowship PD/BD/136860/2018. A.B.-N. and F.C.D.A.L. acknowledge CNPq (grants 420449/2018-3 and 428211/2018-6) for financial support.

Background Self-emulsifying drug delivery systems (SEDDSs) have attracted attention because of their effects on solubility and bioavailability of lipophilic compounds. Herein, a SEDDS loaded with lycopene purified from red guava (nanoLPG) was produced. The nanoemulsion was characterized using dynamic light scattering (DLS), zeta potential measurement, nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR), lycopene content quantification, radical scavenging activity and colloidal stability in cell culture medium. Then, in vivo toxicity and tissue distribution in orally treated mice and cytotoxicity on human prostate carcinoma cells (DU-145) and human peripheral blood mononuclear cells (PBMC) were evaluated. Results NanoLPG exhibited physicochemical properties with a size around 200 nm, negative zeta-potential, and spherical morphology. The size, polydispersity index, and zeta potential parameters suffered insignificant alterations during the 12 month storage at 5 °C, which were associated with lycopene stability at 5 °C for 10 months. The nanoemulsion showed partial aggregation in cell culture medium at 37 °C after 24 h. NanoLPG at 0.10 mg/mL exhibited radical scavenging activity equivalent to 0.043 ± 0.002 mg Trolox/mL. The in vivo studies did not reveal any significant changes in clinical, behavioral, hematological, biochemical, and histopathological parameters in mice orally treated with nanoLPG at 10 mg/kg for 28 days. In addition, nanoLPG successfully delivered lycopene to the liver, kidney and prostate in mice, improved its cytotoxicity against DU-145 prostate cancer cells—probably by pathway independent on classical necrosis and apoptosis—and did not affect PBMC viability. Conclusions Thus, nanoLPG stands as a promising and biosafe lycopene delivery system for further development of nanotechnology-based health products. Graphical Abstract

Dissertação de Mestrado apresentada no Ispa – Instituto Universitário para obtenção de grau de Mestre na especialidade de Psicologia Clínica A investigação tem mostrado que a visualização de pornografia se associa à qualidade da relação e à propensão para comportamentos extradiádicos. Contudo, não se analisou ainda se estas associações podem advir da discrepância entre os conteúdos pornográficos que as pessoas visualizam e as suas atividades sexuais na relação. Como tal, pretendemos analisar em que medida a visualização mais frequente de conteúdos e atividades que não são postas em prática na relação sexual (i.e., discrepância no sentido da pornografia) se associa à qualidade da relação e ao desejo por comportamentos extradiádicos. Pretendemos igualmente perceber se estas associações são explicadas pela necessidade de novidade sexual e pela satisfação sexual, bem como se diferem consoante o género. Num estudo correlacional com pessoas numa relação amorosa (N = 733), verificámos que a maior discrepância no sentido da pornografia se associou a menor novidade sexual e menor satisfação sexual. Contudo, apenas a satisfação sexual se associou a menor qualidade da relação e a maior desejo de comportamentos extradiádicos, tanto sexuais como emocionais. Ainda que mulheres reportassem maior satisfação sexual relacional e homens reportassem maior desejo de comportamentos extradiádicos, o género não emergiu como moderador dos resultados. Os resultados deste estudo sugerem que a pornografia pode servir como forma de combate às necessidades sexuais através das fantasias sexuais, mas também aumentando as frustrações sexuais não atendidas. ABSTRACT: Research has shown that viewing pornography is associated with relationship quality and propensity for extradyadic behavior. However, it has not yet been examined whether these associations may stem from the discrepancy between the pornographic content people view and their sexual activities in the relationship. As such, we want to examine to what extent more frequent viewing of content and activities that are not put into practice in the sexual relationship (i.e., pornography discrepancy) is associated with relationship quality and desire for extradadic behavior. We also want to understand whether these associations are explained by the need for sexual novelty and sexual satisfaction, and whether they differ by gender. In a correlational study of people in a romantic relationship (N = 733), we found that greater discrepancy in the direction of pornography was associated with less sexual novelty and less sexual satisfaction. However, only sexual satisfaction was associated with lower relationship quality and greater desire for extradyadic behaviors, both sexual and emotional. Even though women reported higher relational sexual satisfaction and men reported higher desire for extradyadic behaviors, gender did not emerge as a moderator of the results. The results of this study suggest that pornography may serve as a way to combat sexual needs through sexual fantasies, but also by increasing unmet sexual frustrations.

The skin glands of amphibian species hold a major component of their innate immunity, namely a unique set of antimicrobial peptides (AMPs). Although most of them have common characteristics, differences in AMP sequences allow a huge repertoire of biological activity with varying degrees of efficacy. We present the first study of the AMPs from Pleurodema somuncurence (Anura: Leptodactylidae: Leiuperinae). Among the 11 identified mature peptides, three presented antimicrobial activity. Somuncurin-1 (FIIWPLRYRK), somuncurin-2 (FILKRSYPQYY), and thaulin-3 (NLVGSLLGGILKK) inhibited Escherichia coli growth. Somuncurin-1 also showed antimicrobial activity against Staphylococcus aureus. Biophysical membrane model studies revealed that this peptide had a greater permeation effect in prokaryotic-like membranes and capacity to restructure liposomes, suggesting fusogenic activity, which could lead to cell aggregation and disruption of cell morphology. This study contributes to the characterization of peptides with new sequences to enrich the databases for the design of therapeutic agents. Furthermore, it highlights the importance of investing in nature conservation and the power of genetic description as a strategy to identify new compounds. Fil: Cancelarich, Natalia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina Fil: Wilke, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina Fil: Fanani, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina Fil: Moreira, Daniel C.. Universidade do Brasília; Brasil Fil: Perez, Luis Orlando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico de Ciencias Sociales y Humanas; Argentina Fil: Alves Barbosa, Eder. Universidade do Brasília; Brasil Fil: Plácido, Alexandra. Universidad de Porto; Portugal Fil: Socodato, Renato. Universidad de Porto; Portugal Fil: Portugal, Camila C.. Universidad de Porto; Portugal Fil: Relvas, João B.. Universidad de Porto; Portugal Fil: De la Torre, Beatriz G.. University of Kwazulu-Natal; Sudáfrica Fil: Albericio, Fernando. University of Kwazulu-Natal; Sudáfrica Fil: Basso, Nestor Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Diversidad y Evolución Austral; Argentina Fil: Leite, José. Universidade do Brasília; Brasil Fil: Marani, Mariela Mirta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina

Made available in DSpace on 2022-04-29T08:36:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 In addition to the morphophysiological changes experienced by amphibians during metamorphosis, they must also deal with a different set of environmental constraints when they shift from the water to the land. We found that Pithecopus azureus secretes a single peptide ([M + H]+ = 658.38 Da) at the developmental stage that precedes the onset of terrestrial behaviour. De novo peptide and cDNA sequencing revealed that the peptide, named PaT-2, is expressed in tandem and is a member of the tryptophyllins family. In silico studies allowed us to identify the position of reactive sites and infer possible antioxidant mechanisms of the compounds. Cell-based assays confirmed the predicted antioxidant activity in mammalian microglia and neuroblast cells. The potential neuroprotective effect of PaT-2 was further corroborated in FRET-based live cell imaging assays, where the peptide prevented lipopolysaccharide-induced ROS production and glutamate release in human microglia. In summary, PaT-2 is the first peptide expressed during the ontogeny of P. azureus, right before the metamorphosing froglet leaves the aquatic environment to occupy terrestrial habitats. The antioxidant activity of PaT-2, predicted by in silico analyses and confirmed by cell-based assays, might be relevant for the protection of the skin of P. azureus adults against increased O 2 levels and UV exposure on land compared with aquatic environments. Laboratório de Síntese e Análise de Biomoléculas Instituto de Química Universidade de Brasília Núcleo de Pesquisa em Morfologia e Imunologia Aplicada (NuPMIA) Faculdade de Medicina Universidade de Brasília Laboratório de Imunologia Celular NuPMIA Faculdade de Medicina Universidade de Brasília Programa de Pós-graduação em Medicina Tropical Núcleo de Medicina Tropical Faculdade de Medicina Universidade de Brasília Laboratório de Hematologia e Celulas-tronco Faculdade de Ciências da Saúde Universidade de Brasília Laboratório de Microscopia e Microanálise Instituto de Ciências Biológicas Universidade de Brasília Instituto de Ciências Biológicas Universidade de Brasília Laboratório de Espectrometria de Massa Embrapa Recursos Genéticos e Biotecnologia Bioprospectum Lda UPTEC LAQV/REQUIMTE Departamento de Química e Bioquímica Faculdade de Ciências Universidade Do Porto Glial Cell Biology Lab Instituto de Investigacao e Inovacao em Saude and Instituto de Biologia Molecular e Celular Universidade Do Porto Instituto Federal de Educação Ciência e Tecnologia de São Paulo São Paulo State University (Unesp) Campus of Itapeva The Bridge School of Chemistry Joseph Banks Laboratories University of Lincoln Departamento de Química Fundamental Instituto de Química Universidade de São Paulo, SP São Paulo State University (Unesp) Campus of Itapeva

© The Author(s) 2020. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Cre-ative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not per-mitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/., Cutaneous secretions of amphibians have bioactive compounds, such as peptides, with potential for biotechnological applications. Therefore, this study aimed to determine the primary structure and investigate peptides obtained from the cutaneous secretions of the amphibian, Leptodactylus vastus, as a source of bioactive molecules. The peptides obtained possessed the amino acid sequences, GVVDILKGAAKDLAGH and GVVDILKGAAKDLAGHLASKV, with monoisotopic masses of [M + H]± = 1563.8 Da and [M + H]± = 2062.4 Da, respectively. The molecules were characterized as peptides of the class of ocellatins and were named as Ocellatin-K1(1-16) and Ocellatin-K1(1-21). Functional analysis revealed that Ocellatin-K1(1-16) and Ocellatin-K1(1-21) showed weak antibacterial activity. However, treatment of mice with these ocellatins reduced the nitrite and malondialdehyde content. Moreover, superoxide dismutase enzymatic activity and glutathione concentration were increased in the hippocampus of mice. In addition, Ocellatin-K1(1-16) and Ocellatin-K1(1-21) were effective in impairing lipopolysaccharide (LPS)-induced reactive oxygen species (ROS) formation and NF-kB activation in living microglia. We incubated hippocampal neurons with microglial conditioned media treated with LPS and LPS in the presence of Ocellatin-K1(1-16) and Ocellatin-K1(1-21) and observed that both peptides reduced the oxidative stress in hippocampal neurons. Furthermore, these ocellatins demonstrated low cytotoxicity towards erythrocytes. These functional properties suggest possible to neuromodulatory therapeutic applications., Alexandra Plácido is a recipient of a post-doctoral grant from the project FCT (PTDC/BII-BIO/31158/2017). Renato Socodato and Camila Cabral Portugal hold postdoctoral fellowships from FCT (Refs: SFRH/BPD/91833/2012 and FRH/BPD/91962/2012, respectively). This work was funded through project UID/QUI/50006/2013-POCI/01/0145/FEDER/007265 (LAQV/REQUIMTE) with financial support from FCT/MEC through national funds and co-financed by FEDER, under the Partnership Agreement PT 2020

Bombesin mediates several biological activities in the gastrointestinal (GI) tract and central nervous system in mammals, including smooth muscle contraction, secretion of GI hormones and regulation of homeostatic mechanisms. Here, we report a novel bombesin-like peptide isolated from Boana raniceps. Its amino acid sequence, GGNQWAIGHFM-NH

Copper nanoparticles stabilized with cashew (CG-CuNPs) were synthesized by reduction reaction using ascorbic acid and sodium borohydride, using the cashew gum (CG) as a natural polymer stabilizer. Dynamic light scattering, atomic force microscopy, Fourier-transform infrared spectroscopy, UV-Vis spectrophotometry, and x-ray diffraction were used to characterize the nanoparticles (CG-CuNPs), and copper was quantified by electrochemical measurement. The UV-vis spectra of the CG-CuNPs confirmed the formation of nanoparticles by appearance of a surface plasmon band at 580 nm after 24 h of reaction. The Fourier-transform infrared spectrum of CG-CuNPs showed the peak at 1704 cm−1 from cashew gum, confirming the presence of the gum in the nanoparticles. The average size of CG-CuNPs by dynamic light scattering and atomic force microscopy was around 10 nm, indicating small, approximately spherical particles. Antimicrobial assays showed that CG-CuNPs had activity against Staphylococcus aureus ATCC 29213 with a minimal inhibitory concentration of 0.64 mM. The cytotoxicity assay on BALB/c murine macrophages showed lower cytotoxic effects for CG-CuNPs than CuSO4·5H2O. Viability cell assays for CG-CuNPs at (0.250 mM) inhibited by 70% the growth of 4T1 LUC (4T1 mouse mammary tumor cell line) and NIH 3T3 cells (murine fibroblast cells) over a 24-h period. Therefore, CG-CuNPs can be used as an antimicrobial agent with lower cytotoxic effects than the CuSO4·5H2O precursor., The author would like to thank at UCM for performingDPV, USP by X-ray diffraction experiment, REQUIMTE/LAQV for FTIR, UnB and UFPI for the cytotoxicityassays, as well as at UFPI for help with DLS, UV-Vis,AFM, and microbiological experiments. This work was supported by Project 400398/2014-1—Desenvolvimento de Nanopartículas Estabilizadas com Goma de Cajueiro para Aplicações Biotecnologicas, financed by CNPq. AlexandraPlácido is grateful to FCT by her grant SFRH/BD/97995/2013, financed by POPH-QREN-Tipologia 4.1-Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior.

Copyright © 2021 American Chemical Society and American Society of Pharmacognosy, Snake venoms are important sources of bioactive molecules, including those with antiparasitic activity. Cathelicidins form a class of such molecules, which are produced by a variety of organisms. Batroxicidin (BatxC) is a cathelicidin found in the venom of the common lancehead (Bothrops atrox). In the present work, BatxC and two synthetic analogues, BatxC(C-2.15Phe) and BatxC(C-2.14Phe)des-Phe1, were assessed for their microbicidal activity. All three peptides showed a broad-spectrum activity on Gram-positive and -negative bacteria, as well as promastigote and amastigote forms of Leishmania (Leishmania) amazonensis. Circular dichroism (CD) and nuclear magnetic resonance (NMR) data indicated that the three peptides changed their structure upon interaction with membranes. Biomimetic membrane model studies demonstrated that the peptides exert a permeabilization effect in prokaryotic membranes, leading to cell morphology distortion, which was confirmed by atomic force microscopy (AFM). The molecules considered in this work exhibited bactericidal and leishmanicidal activity at low concentrations, with the AFM data suggesting membrane pore formation as their mechanism of action. These peptides stand as valuable prototype drugs to be further investigated and eventually used to treat bacterial and protozoal infections., This work was supported by Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF, Brazil, concession number 00193.00001937/2018-83). A.D. thanks the Brazilian National Council for Scientific and Technological Development (CNPq) for the financial support (142099/2018-9). This work was supported by FCT (PTDC/BII-BIO/31158/2017). A.P. is a recipient of a postdoctoral grant from the same project. A.C.M. and J.M. are grateful to Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grant numbers 2016/22488-3 and 2019/25905-2; 2018/24077-6), Brazil. C.P.A. acknowledges FCT-MCTES fellowship PD/BD/136860/2018. We also acknowledge Project Norte-01-0145-FEDER-000012 [Structured program on bioengineered therapies for infectious diseases and tissue regeneration, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF)]. Bioprospectrum, Lda (UPTEC, Porto, Portugal) is acknowledged for the project management and logistical support in receiving the sample in Portugal. The authors thank Wanessa Feliz Cabral (NuPMIA/UnB/Brazil) for their technical support in the RP-HPLC analyses that make up the Supporting Information of this article. A.C.M and J.M are grateful to Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP grant numbers 2016/22488-3 and 2019/25905-2; 2018/24077-6) and HC-FMUSP, Brazil.

Made available in DSpace on 2021-06-25T10:44:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Universidade Estadual Paulista Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fingerprints have been used in forensic investigations as a powerful tool for human identification. However, it is desirable to find new sustainable sources and synthesis for fingerprint developers, in order to reduce the production costs, to improve the sensitivity and to reduce environment impact after the disposal. This report describes a study involving the production of acetylated cashew-gum-based silver nanoparticles (ACG-AgNPs), using three concentrations of ACG (0.5, 1.0 and 5.0 mg/mL). The nanoparticles were characterized by UV-VIS spectroscopy, AFM (atomic force microscopy) and TEM (transmission electron microscopy). In addition, computer simulations and evaluation of their effect as a potential fingerprint developer on paper were studied, using ninhydrin as a positive control. The results showed that samples were able to reveal the lines contained in the fingerprint residues on paper. Positive results were obtained for all the samples in tests performed 1, 6 and 24 h after application. Howeverthe best performance was observed with the sample containing ACG 0.5 mg/mL. ACG-AgNPs have potential for practical and rapid latent fingerprint development on porous surfaces, which allows the development of a product to be used even at the crime scene. Another advantage was the synthesis process: simple, fast, low-cost and using a biocompatible and biodegradable material as stabilizer, providing a non-toxic material. In addition, this material, is likely to cause less environmental damage than the reagents used by conventional techniques. Faculdade de Medicina do ABC FMABC Departamento de Polícia Técnico-Científica do Piauí DPTC-PI Núcleo de Pesquisa em Biodiversidade e Biotecnologia Biotec Universidade Federal do Delta do Parnaíba UFDPar LAQV-REQUIMTE Department of Chemistry and Biochemistry Faculdade de Ciências da Universidade do Porto Bioprospectum Lda UPTEC Identification Institute Civil Police of the Federal District Núcleo de Pesquisa em Morfologia e Imunologia Aplicada NuPMIA Area de Morfologia Faculdade de Medicina FM Universidade de Brasília UnB Instituto Federal de Educação Ciência e Tecnologia de São Paulo Universidade Estadual Paulista UNESP Campus Experimental de Itapeva Universidade Estadual Paulista UNESP Campus Experimental de Itapeva CNPq: 420449/2018-3 CNPq: 428211/2018-6

The skin glands of amphibian species hold a major component of their innate immunity, namely a unique set of antimicrobial peptides (AMPs). Although most of them have common characteristics, differences in AMP sequences allow a huge repertoire of biological activity with varying degrees of efficacy. We present the first study of the AMPs from

Amphibian skin is a multifunctional organ that plays key roles in defense, breathing, and water balance. In this study, skin secretion samples of the fire salamander (Salamandra salamandra) were separated using RP-HPLC and de novo sequenced using MALDI-TOF MS/MS. Next, we used an in silico platform to screen antioxidant molecules in the framework of density functional theory. One of the identified peptides, salamandrin-I, [M + H]+ = 1406.6 Da, was selected for solid-phase synthesis, it showed free radical scavenging activity against DPPH and ABTS radicals. Salamandrin-I did not show antimicrobial activity against Gram-positive and -negative bacteria. In vitro assays using human microglia and red blood cells showed that salamandrin-I has no cytotoxicity up to the concentration of 100 &micro, M. In addition, in vivo toxicity tests on Galleria mellonella larvae resulted in no mortality at 20 and 40 mg/kg. Antioxidant peptides derived from natural sources are increasingly attracting interest. Among several applications, these peptides, such as salamandrin-I, can be used as templates in the design of novel antioxidant molecules that may contribute to devising strategies for more effective control of neurological disease.

The aims of this study were to produce poly-ɛ-caprolactone lipid-core nanocapsules containing lycopene-rich extract from red guava (LEG), to characterize those nanoparticles and to evaluate their cytotoxic effects on human breast cancer cells. Lipid-core nanocapsules containing the extract (nanoLEG) were produced by the method of interfacial deposition of the preformed polymer. The nanoparticles were characterized by Dynamic Light Scattering (DLS), Polydispersity Index, Zeta Potential, pH, Encapsulation Efficiency, Nanoparticle Tracking Analysis (NTA), Atomic Force Microscopy (AFM) and Transmission Electron Microscopy (TEM). Cell viability was evaluated by the MTT dye reduction method in the human breast cancer MCF-7 cell line and inhibition of ROS and NF-κB was assayed in living human microglial cell line (HMC3) by time-lapse images microscopy. A hemolytic activity assay was carried out with sheep blood. Data showed that nanoparticles average size was around 200 nm, nanoparticles concentration/mL was around 0.1 µM, negative zeta potential, pH 5.0 and spherical shape, with low variation during a long storage period (7 months) at 5 °C, indicating stability of the system and protection against lycopene degradation. The percentage of encapsulation varied from 95% to 98%. The nanoLEG particles significantly reduced the viability of the MCF-7 cells after 24 h (61.47%) and 72 h (55.96%) of exposure, even at the lowest concentration tested (6.25-200 μg/ml) and improved on the cytotoxicity of free LEG to MCF-7. NanoLEG inhibited LPS-induced NF-kB activation and ROS production in microglial cells. The particles did not affect the membrane integrity of sheep blood erythrocytes at the concentrations tested (6.25-200 μg/mL). Thus, the formulation of lipid-core nanocapsules with a polysorbate 80-coated poly-ɛ-caprolactone wall was efficiently applied to stabilize the lycopene-rich extract from red guava, generating a product with satisfactory physico-chemical and biological properties for application as health-promoting nanotechnology-based nutraceutical, emphasizing its potential to be used as a cancer treatment.

Soybean (Glycine max L.) is the most important genetically modified (GM) crop, from which the glyphosate tolerant soybean, also known as Roundup Ready (RR), is one of the most representative. The present work intends to assess the presence of two RR soybean lines, namely GTS 40-3-2 and MON89788, in a wide range of processed foods commercialised in Portugal. For this purpose, two quantitative real-time PCR approaches were proposed using available and newly designed primers. The results showed that, from 90 food samples, 73 confirmed the presence of soybean, while 15 others suggest their incompliance with labelled soybean. RR soybean was detected in 9 samples, from which 5 were identified as being the GTS-40-3-2 event, 3 the MON89788 event and one contained both events. The developed real-time PCR methods were successfully validated through the participation in an interlaboratory proficiency program and applied to processed foods. The estimated RR soybean contents of 8 samples ranged between 0.01 and 0.39%, but the sample containing both events accounted for 23.9% of GM ingredients. Such high level above the threshold for labelling regulations suggests the need of more strict control of GMO in foods.

The cultivation of genetically modified organisms (GMO) continues to expand worldwide. Still, many consumers express concerns about the use of GMO in food or feed, and many countries have legislated on labelling systems to indicate the presence of GMO in commercial products. To deal with the increased number of GMO events and to address related regulations, alternative detection methods for GMO inspection are required. In this work, a genosensor based on Surface Plasmon Resonance under continuous flow was developed for the detection and quantification of a genetically modified soybean (event GTS 40-3-2). In a single chip, the simultaneous detection of the event-specific and the taxon-specific samples were achieved, whose detection limits were 20 pM and 16 pM, respectively. The reproducibility was 1.4%, which supports the use of the chip as a reliable and cost-effective alternative to other DNA-based techniques. The results indicate that the proposed method is a versatile tool for GMO quantification in food and feed samples. This work received financial support from the European Union (FEDER funds through COMPETE), National Funds (FCT, Fundação para a Ciência e a Tecnologia) through project UID/QUI/ 50006/2013 and cofinanced by FEDER funds. Alexandra Plácido is grateful for the FCT grant (SFRH/BD/97995/2013) financed by POPH-QREN (subsidized by FSE and MCTES).

A green synthetic route was developed to prepare silver nanoparticles (AgNPs) in aqueous solution for biological applications. Eschweilenol C, a compound derivative ellagic acid was identified as the main constituent of the aqueous fraction of the ethanolic extract of Terminalia fagifolia Mart. by NMR analysis. In the green synthesis, the ethanolic extract of T. fagifolia and its aqueous fraction were used to promote silver reduction and nanoparticle stabilization. The synthesized AgNPs presented a spherical or polygonal morphology shape by TEM analysis and AgNPs showed high levels of antioxidant and considerable antibacterial and antifungal activities. Synthesized nanoparticles presented significant antioxidant activity by sequestration of DPPH and ABTS radicals, in addition to iron reduction (FRAP assay) and measurement of antioxidant capacity in ORAC units, in addition, AgNP synthesized with the aqueous fraction also demonstrated antioxidant potential in microglial cells. Gram-positive and Gram-negative bacteria were susceptible to growth inhibition by the nanoparticles, among which the AgNPs formed by the ethanolic extract was the most effective. The data obtained by AFM images suggested that AgNPs could lead to the lysis of bacteria and subsequent death. The antifungal assays showed high efficiency against yeasts and dermatophytes. This work represents the first description of antifungal activity by AgNPs against Fonsecaea pedrosoi, the etiologic agent of chromoblastomycosis. In relation to biocompatibility, the AgNPs induced lower haemolysis than AgNO3., We thank Herbert Kogler and Reinhard Wimmer for the identification of Eschweilenol C. The NMR laboratory at Aalborg University is supported by the Obel Family, SparNord and Carlsberg foundations.The authors are grateful to Carla Eiras (LIMAV/CT/UFPI) and to FCT and EU for financial support through project UID/QUI/50006/2013– POCI-01-0145-FEDER-007265 from COMPETE and projectNORTE-01-0145-FEDER-000011 from COMPETE. Thanks to Andreia Pinto for help with the TEM measurements at Instituto de Medicina Molecular (IMM). This work was supported by the Histology and Comparative Pathology Laboratory of the IMM

The natural alkaloid epiisopiloturine has recently become the focus of study for various medicinal properties, particularly for its anti-inflammatory and antischistosomal effect. The incorporation of active molecules in natural polymeric matrices has garnered increasing interest during recent decades. A new derivative of cashew gum successfully obtained by gum acetylation has shown great potential as a carrier in controlled drug release systems. In this work, epiisopiloturine was encapsulated in acetylated cashew gum nanoparticles in order to increase solubility and allow slow release, whereas the morphology results were supported by computer simulations. The particles were produced under a variety of conditions, and thoroughly characterized using light scattering and microscopic techniques. The particles were spherical and highly stable in solution, and showed drug incorporation at high levels, up to 55% efficiency. Using a dialysis-based in vitro assay, these particles were shown to release the drug via a Fickian diffusion mechanism, leading to gradual drug release over approximately 6 h. These nanoparticles show potential for the use as drug delivery system, while studies on their potential anti-inflammatory action, as well as toxicity and efficacy assays would need to be performed in the future to confirm their suitability as drug delivery candidates., This work was conducted in partnership with the Polymer Laboratory of the Federal University of Ceará for polymer modification. The authors thanks Foundation for Science and Technology (FCT) for the fellowships SFRH/BD/97995/2013 (AP) and SFRH/BD/95983/2013 (MPA), in the context of the POCH program. The work at UCIBIO/REQUIMTE was supported by FCT through project UID/MULTI/04378/2013 – POCI/01/0145/FEDER/007728 with financial support from FCT/MCTES through national funds and co-financed by FEDER, under the Partnership Agreement PT2020. The work at REQUIMTE/LAQV received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT) through project UID/QUI/50006/2013. The computational time was provided by GRID-Unesp, SICC/IFSP and CENAPAD/SP. The authors also acknowledge CNPq and CAPES for a scholarship and financial aid.

In synthesis of silver nanoparticles (AgNPs), the composition of the stabilizer used can be closely related to the effectiveness of the synthesis and to the shape of the final nanoparticles. Recently, the use of collagen as an effective nanoparticle stabilization agent was reported. In this work, synthesis of silver nanoparticles using mixed capping agents is reported. The capping agents used were cashew gum-hydrolyzed collagen; kappa carrageenan-hydrolyzed collagen, and agar-hydrolyzed collagen. We evaluated antibacterial action against Gram-positive and Gram-negative bacteria, as well as antifungal activity and cytotoxicity. Homogenized mixtures of collagen and aqueous cashew gum, carrageenan or agar respectively were used to produce the nanoparticles AgNPcolCashew, AgNPcolCarr and AgNPcolAgar. AgNP characterization was performed using Uv-vis, XRD, TEM and DLS and the biological activities were assayed using MIC and MBC analyses for both antibacterial and antifungal application. Results showed that the AgNPcollcar sample showed the strongest bacterial inhibition with MIC values of 62.5 and 31.25 μM/mL Ag against E. coli and P. aeruginosa respectively. Interestingly, AgNPcollAgar also presented the lowest cytotoxicity when compared with other AgNPs and AgNO3.

This work received financial support from the European Union (FEDER funds through COMPETE), National Funds (FCT, Fundação para a Ciência e a Tecnologia) through project UID/QUI/50006/2013. A.P. and M.F.B. are grateful to FCT grants SFRH/BD/97995/2013 and SFRH/BPD/78845/2011, financed by POPH–QREN–Tipologia 4.1–Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior. C.P. thanks FCT for the FCT Investigator contract IF/01080/2015., Development of expeditious analytical methods for the detection of genetically modified organisms (GMOs) is increasingly necessary, not only to verify compliance with labelling, but also to help industry to efficiently control the reception of raw materials. On the basis of this, a disposable electrochemical magnetogenoassay is proposed for simultaneous detection of two Roundup Ready (RR) soybean lines GTS 40-3-2 and MON89788, using gold-coated magnetic nanoparticles (Fe3O4@Au) as nanosupport. To perform this magnetogenoassay, a sandwich-type hybridization assay was used with different enzymatic labelling systems (fluorescein isothiocyanate and digoxigenin) and dual screen-printed carbon electrodes (SPdCEs), which allowed the simultaneous readout of each target. A linear relationship ranging from 0.1 to 2.5 nM and from 0.1 to 1.0 nM was achieved for GTS 40-3-2 and MON89788 events, respectively, and both assays showed a similar detection limit of about 0.1 nM. Furthermore, a good performance in terms of precision and selectivity was achieved. The proposed approach is a step forward for event-specific multiplex detection.

The growing microbial resistance to antibiotics and the adverse effects of the frequent use of antimicrobials in oral medicine require products as natural therapeutic alternatives. Croton betaceus and C. lundianus can be viable for in-depth studies in search of new bioproducts. This study evaluated the antioxidant and antimicrobial potential of ethanol extract and leaf fractions of Croton spp. The disk diffusion assay, FRAP and DPPH, were used for antimicrobial and antioxidant analysis, respectively. In addition to phytochemical prospecting and toxicity testing. Secondary metabolites with biological activities were found. Researched extracts and fractions have the potential to inhibit the growth of oral pathogens (Gram-positive, Gram-negative and yeast) in different concentrations. Enterococcus faecalis is the most susceptible to the extract. All fractions showed antioxidant potential. The hexane fraction (BHX and LHX) of both species can inhibit the growth of pathogens in low concentrations, and has a high antioxidant power. The fractions of the extracts of C. betaceus and C. lundianus have important biological activities that can be applied in the development of new antimicrobials for industry and other biotechnological applications.

The peptide PcL342-354C was obtained from the Cry1Ab16 toxin present in Bacillus thuringiensis ("Computational Modeling Deduced Three Dimensional Structure of Cry1Ab16 Toxin from B. thuringiensis AC11" (Kashyap, 2012) [1]). In this data article, we report the synthesis and characterization of the PcL342-354C peptide by MALDI-TOF/TOF mass spectrometry. In addition, the preparation of layer-by-layer films is shown based on interspersion of this peptide with both polyethylenimine (PEI) and poly(sodium 4-styrenesulfonate) (PSS), self-assembled on ITO (indium tin oxide) electrodes. The morphology of the ITO/PEI/PSS/PcL342-354C film was analyzed using atomic force microscopy (AFM). We also evaluated the effect of the number of bilayers in ITO/PEI/(PSS/PcL342-354C)n on the morphology of the film using AFM amplitude images. Further details about this study were published elsewhere, "Layer-by-layer films containing peptides of the Cry1Ab16 toxin from B. thuringiensis for potential biotechnological applications," (Plácido et al., 2016) [2]. Fil: Plácido, Alexandra. Instituto Superior de Engenharia Do Porto; Portugal Fil: de Oliveira Farias, Emanuel Airton. Universidade Federal do Piauí; Brasil Fil: Marani, Mariela Mirta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina Fil: Gomes Vasconcelos, Andreanne. Universidade Federal do Piauí; Brasil Fil: Leite, José R. S. A.. Universidade Federal do Piauí; Brasil Fil: Delerue Matos, Cristina. Instituto Superior de Engenharia Do Porto; Portugal

© 2019 Published by Elsevier Ltd., This study presents a green synthesis route to silver nanoparticles (AgNPs) stabilized with cashew gum (CG) or carboxymethylated cashew gum (CCG) using microwave-assisted synthesis and evaluates their antibacterial activity. The antimicrobial activity was measured by determining the minimum inhibitory concentration (MIC) with Staphylococcus aureus and Escherichia coli. In both cases of the presence of CG and CCG, it was found that higher pH lead to more efficient conversion of silver nitrate to AgNPs with well dispersed, spherical and stable particles as well as low crystallinity. CCG-capped AgNPs were slightly smaller (137.0 and 96.3 nm) than those coated with non-modified gum (144.7 and 100.9 nm). The samples presented promising antibacterial activity, especially on Gram-negative bacteria, resulting in significant membrane damage on treated bacteria in comparison to the untreated control, observed by atomic force microscopy. Thus, a quick and efficient synthesis route was applied to produce CGAgNPs and CCGAgNPs with antimicrobial potential., The authors gratefully acknowledge financial support from CAPES and FAPEPI; ACM are grateful to FAPESP (2014/02282-6 AND 2016/18023-5). YPM is grateful to CAPES (AUX-PERM-705/2009). The authors also acknowledge the CNPq and Capes for scholarship and financial aid. The authors are grateful to Fundação para a Ciência e a Tecnologia (MCTES funds) and European Union (European Social Fund and European Regional Development Fund), for financial support through project UID/QUI/50006/2013–POCI-01-0145-FEDER-007265 (LAQV-REQUIMTE), in the context of the COMPETE program from QREN, project NORTE-01-0145-FEDER-000011, in the context of the NORTE 2020 program, and fellowships SFRH/BD/97995/2013 (AP) and SFRH/BD/95983/2013 (MPdA), in the context of the POCH program, both from Portugal 2020 partnership agreement.

The integration of nanomaterials in the field of (bio)sensors has allowed developing strategies with improved analytical performance. In this work, ultrasmall core-shell Fe3O4@Au magnetic nanoparticles (MNPs) were used as the platform for the immobilization of event-specific Roundup Ready (RR) soybean and taxon-specific DNA sequences. Firstly, monodisperse Fe3O4 MNPs were synthesized by thermal decomposition and subsequently coated with a gold shell through reduction of Au(III) precursor on the surface of the MNPs in the presence of an organic capping agent. This nanosupport exhibited high colloidal stability, average particle size of 10.2 ± 1.3 nm, and spherical shape. The covalent immobilization of ssDNA probe onto the Au shell of the Fe3O4@Au MNPs was achieved through a self-assembled monolayer (SAM) created from mixtures of alkane thiols (6-mercapto-1-hexanol and mercaptohexanoic acid). The influence of the thiols ratio on the electrochemical performance of the resulting electrochemical genoassays was studied, and remarkably, the best analytical performance was achieved for a pure mercaptohexanoic acid SAM. Two quantification assays were designed; one targeting an RR sequence and a second targeting a reference soybean gene, both with a sandwich format for hybridization, signaling probes labelled with fluorescein isothiocyanate (FITC), enzymatic amplification and chronoamperometric detection at screen-printed carbon electrodes (SPCE). The magnetogenoassays exhibited linear ranges from 0.1 to 10.0 nM and from 0.1 to 5.0 nM with similar detection limits of 0.02 nM and 0.05 nM for the event-specific (RR) and the taxon-specific (lectin) targets, respectively. The usefulness of the approach was demonstrated by its application to detect genetically modified organisms (GMOs) in feed and food., This work received financial support from the European Union (FEDER funds through COMPETE), National Funds (FCT, Fundação para a Ciência e a Tecnologia) through project UID/QUI/50006/2013 and Regional Funds (Principado de Asturias government through Project FC15-GRUPIN14-025), and cofinanced by FEDER funds. A.P. and M.F.B. are grateful to FCT grants SFRH/BD/97995/2013 and SFRH/BPD/78845/2011, financed by POPH–QREN–Tipologia 4.1–Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior. C.P. thanks FCT for the FCT Investigator contract IF/01080/2015.

The amphibian skin plays an important role protecting the organism from external harmful factors such as microorganisms or UV radiation. Based on biorational strategies, many studies have investigated the cutaneous secretion of anurans as a source of bioactive molecules. By a peptidomic approach, a novel antioxidant peptide (AOP) with in vitro free radical scavenging ability was isolated from Physalaemus nattereri. The AOP, named antioxidin-I, has a molecular weight [M+H]+ = 1543.69Da and a TWYFITPYIPDK primary amino acid sequence. The gene encoding the antioxidin-I precursor was expressed in the skin tissue of three other Tropical frog species: Phyllomedusa tarsius, P. distincta and Pithecopus rohdei. cDNA sequencing revealed highly homologous regions (signal peptide and acidic region). Mature antioxidin-I has a novel primary sequence with low similarity compared with previously described amphibian's AOPs. Antioxidin-I adopts a random structure even at high concentrations of hydrophobic solvent, it has poor antimicrobial activity and poor performance in free radical scavenging assays in vitro, with the exception of the ORAC assay. However, antioxidin-I presented a low cytotoxicity and suppressed menadione-induced redox imbalance when tested with fibroblast in culture. In addition, it had the capacity to substantially attenuate the hypoxia-induced production of reactive oxygen species when tested in hypoxia exposed living microglial cells, suggesting a potential neuroprotective role for this peptide., The authors thank of the B2Tech platform facilities (Biochemical And Biophysical Technologies, Instituto de Investigação e Inovação em Saúde (I3S) da Universidade do Porto, Portugal) for the use of circular dichroism (CD). AP is grateful for the Fundação para a Ciência e a Tecnologia (FCT, Portugal) grants (SFRH/BD/97995/2013) financed by POPH-QREN (subsidized by FSE and MCTES). NV thanks FCT and FEDER (European Union) for funding through UID/MULTI/04378/2013, project grant IF/00092/2014 and IF2014 position. NV thanks also Fundação Manuel António da Mota (FMAM, Portugal) by support to Nuno Vale Lab. PG thanks FCT for funding through UID/QUI/50006/2013. RS and CCP, hold postdoctoral fellowships from FCT (Refs: SFRH/BPD/91833/2012 and FRH/BPD/91962/2012, respectively). ACM is grateful to FAPESP (Grants 2014/02282-6 and 2016/18023-5). YPM is grateful to CNPq by the Senior Researcher Grant (PQ-Sr 306036/2016-9). EAB is grateful to PNPD/CAPES (Grant No:1603966) for its postdoctoral fellowship. M. M. Marani is a researcher at CONICET.

Schistosomiasis, caused by helminth flatworms of the genus Schistosoma, is an infectious disease mainly associated with poverty that affects millions of people worldwide. Since treatment for this disease relies only on the use of praziquantel, there is an urgent need to identify new antischistosomal drugs. Piplartine is an amide alkaloid found in several Piper species (Piperaceae) that exhibits antischistosomal properties. The aim of this study was to evaluate the structure&ndash, function relationship between piplartine and its five synthetic analogues (19A, 1G, 1M, 14B and 6B) against Schistosoma mansoni adult worms, as well as its cytotoxicity to mammalian cells using murine fibroblast (NIH-3T3) and BALB/cN macrophage (J774A.1) cell lines. In addition, density functional theory calculations and in silico analysis were used to predict physicochemical and toxicity parameters. Bioassays revealed that piplartine is active against S. mansoni at low concentrations (5&ndash, 10 &micro, M), but its analogues did not. In contrast, based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays, piplartine exhibited toxicity in mammalian cells at 785 &micro, M, while its analogues 19A and 6B did not reduce cell viability at the same concentrations. This study demonstrated that piplartine analogues showed less activity against S. mansoni but presented lower toxicity than piplartine.

This study investigated a lycopene-rich extract from red guava (LEG) for its chemical composition using spectrophotometry, mass spectrometry, attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR), and computational studies. The cytotoxic activity of LEG and the underlying mechanism was studied in human breast adenocarcinoma cells (MCF-7), murine fibroblast cells (NIH-3T3), BALB/c murine peritoneal macrophages, and sheep blood erythrocytes by evaluating the cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and flow cytometry. Spectrophotometry analysis showed that LEG contained 20% of lycopene per extract dry weight. Experimental and theoretical ATR-FTIR suggests the presence of lycopene, whereas MS/MS spectra obtained after fragmentation of the molecular ion [M]+• of 536.4364 show fragment ions at m/z 269.2259, 375.3034, 444.3788, and 467.3658, corroborating the presence of lycopene mostly related to all-trans configuration. Treatment with LEG (1600 to 6.25μg/mL) for 24 and 72h significantly affected the viability of MCF-7 cells (mean half maximal inhibitory concentration [IC50]=29.85 and 5.964μg/mL, respectively) but not NIH-3T3 cells (IC50=1579 and 911.5μg/mL, respectively). Furthermore LEG at concentrations from 800 to 6.25μg/mL presented low cytotoxicity against BALB/c peritoneal macrophages (IC50≥800μg/mL) and no hemolytic activity. LEG (400 and 800μg/mL) caused reduction in the cell proliferation and induced cell cycle arrest, DNA fragmentation, modifications in the mitochondrial membrane potential, and morphologic changes related to granularity and size in MCF-7 cells; however, it failed to cause any significant damage to the cell membrane or display necrosis or traditional apoptosis. In conclusion, LEG was able to induce cytostatic and cytotoxic effects on breast cancer cells probably via induction of an apoptotic-like pathway., The authors acknowledge the computational time provided by CENAPAD/SP on the project proj697. Alexandra Plácido is gratefully to FCT by her grant SFRH/BD/97995/2013, financed by POPH–QREN–Tipologia 4.1–Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior. The work at REQUIMTE/LAQV received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT) through project UID/QUI/50006/2013. Adriany das G. N. Amorim is grateful to CAPES by for the doctoral fellowship process no. 99999.004236/2014-09 in Federal University of Piauí (UFPI). Eder A. Barbosa is grateful to PNPD/CAPES for its post-doctoral fellowship.

Aim:To test ocellatin peptides (ocellatins-PT2-PT6) for antibacterial and antibiofilm activities and synergy with antibiotics against Pseudomonas aeruginosa. Materials & methods: Normal- and checkerboard-broth microdilution methods were used. Biofilm studies included microtiter plate-based assays and microscopic analysis by confocal laser scanning microscopy and atomic force microscopy. Results: Ocellatins were more active against multidrug-resistant isolates of P. aeruginosa than against susceptible strains. Ocellatin-PT3 showed synergy with ciprofloxacin and ceftazidime against multidrug-resistant isolates and was capable of preventing the proliferation of 48-h mature biofilms at concentrations ranging from 4 to 8× the MIC. Treated biofilms had low viability and were slightly more disaggregated. Conclusion: Ocellatin-PT3 may be promising as a template for the development of novel antimicrobial peptides against P. aeruginosa., This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e Tecnologia), under the Partnership Agreement PT2020 through project UID/QUI/50006/2013-POCI/01/0145/FEDER/007265 (LAQV/REQUIMTE) and from Programa Operacional Regional do Norte (ON.2 – O Novo Norte),under the Quadro de Referencia Estrat ˆ egico Nacional (QREN) and funded by Fundo Europeu de Desenvolvimento Regional ´ (Feder) through projects: NORTE-01-0145-FEDER-000024, NORTE-01-0145-FEDER-000011 and NORTE-07-0161-FEDER-000111.A Placido is grateful to FCT for financial support through the grant SFRH ´ /BD/97995/2013, POPH-QREN-Tipologia 4.1-Formação Avançada and Fundo Social Europeu and Ministerio da Ciência, Tecnologia e Ensino Superior. The authors have no other relevant ˆ affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Silver nanoparticles have been shown to possess considerable antibacterial activity, but in vivo applications have been limited due to the inherent, but low, toxicity of silver. On the other hand, silver nanoparticles could provide cutaneous protection against infection, due to their ability to liberate silver ions via a slow release mechanism, and their broad-spectrum antimicrobial action. Thus, in this work, we describe the development of a carboxymethyl cellulose-based hydrogel containing silver nanoparticles. The nanoparticles were prepared in the hydrogel in situ, utilizing two variants of cashew gum as a capping agent, and sodium borohydride as the reducing agent. This gum is non-toxic and comes from a renewable natural source. The particles and gel were thoroughly characterized through using rheological measurements, UV-vis spectroscopy, nanoparticles tracking analysis, and transmission electron microscopy analysis (TEM). Antibacterial tests were carried out, confirming antimicrobial action of the silver nanoparticle-loaded gels. Furthermore, rat wound-healing models were used and demonstrated that the gels exhibited improved wound healing when compared to the base hydrogel as a control. Thus, these gels are proposed as excellent candidates for use as wound-healing treatments.

The vasoactive proline-rich oligopeptide termed BPP-BrachyNH

Silver nanoparticles have been studied as an alternative for treatment of microbial infections and leishmaniasis, without promoting induction of microbial or parasite resistance. In this study, chitosan-based silver nanoparticles were synthesized from silver nitrate (AgNO3), sodium borohydride as a reducing agent, and the biopolymer chitosan as a capping agent. The chitosan-based silver nanoparticles were characterized by ultraviolet–visible, Fourier transform infrared, dynamic light scattering, zeta potential, atomic force microscopy, and transmission electron microscope. The antibacterial assay was performed by determination of the minimum inhibitory concentration. The antileishmanial and the cytotoxic effects induced by AgNO3, chitosan, and chitosan-based silver nanoparticles were analyzed by resazurin and MTT colorimetric assays, respectively. AgNO3, chitosan, and chitosan-based silver nanoparticles induced a marked activity against all bacterial strains and promastigote forms of Leishmania amazonensis at minimum inhibitory concentrations ranging from 1.69 to 3.38 µg Ag/mL. Interestingly, the chitosan-based silver nanoparticles presented less cytotoxicity than the AgNO3 alone and were more active against L. amazonensis than solely chitosan. Furthermore, the cytotoxic concentrations (CC50) of both chitosan and chitosan-based silver nanoparticles against macrophages were significantly higher than the IC50 against promastigotes. Thus, the chitosan-based silver nanoparticles represent a promising alternative for the treatment of microbial infections and leishmaniasis.

The vasoactive proline-rich oligopeptide termed BPP-BrachyNH2 (H-WPPPKVSP-NH2) induces in vitro inhibitory activity of angiotensin I-converting enzyme (ACE) in rat blood serum. In the present study, the removal of N-terminal tryptophan or C-terminal proline from BPP-BrachyNH2 was investigated in order to predict which structural components are important or required for interaction with ACE. Furthermore, the toxicological profile was assessed by in silico prediction and in vitro MTT assay. Two BPP-BrachyNH2 analogues (des-Trp(1)-BPP-BrachyNH2 and des-Pro(8)-BPP-BrachyNH2) were synthesized, and in vitro and in silico ACE inhibitory activity and toxicological profile were assessed. The des-Trp(1)-BPP-BrachyNH2 and des-Pro(8)-BPP-BrachyNH2 were respectively 3.2- and 29.5-fold less active than the BPP-BrachyNH2-induced ACE inhibitory activity. Molecular Dynamic and Molecular Mechanics Poisson-Boltzmann Surface Area simulations (MM-PBSA) demonstrated that the ACE/BBP-BrachyNH2 complex showed lower binding and van der Wall energies than the ACE/des-Pro(8)-BPP-BrachyNH2 complex, therefore having better stability. The removal of the N-terminal tryptophan increased the in silico predicted toxicological effects and cytotoxicity when compared with BPP-BrachyNH2 or des-Pro(8)-BPP-BrachyNH2. Otherwise, des-Pro(8)-BPP-BrachyNH2 was 190-fold less cytotoxic than BPP-BrachyNH2. Thus, the removal of C-terminal proline residue was able to markedly decrease both the BPP-BrachyNH2-induced ACE inhibitory and cytotoxic effects assessed by in vitro and in silico approaches. In conclusion, the aminoacid sequence of BPP-BrachyNH2 is essential for its ACE inhibitory activity and associated with an acceptable toxicological profile. The perspective of the interactions of BPP-BrachyNH2 with ACE found in the present study can be used for development of drugs with differential therapeutic profile than current ACE inhibitors.

Transgenic maize produced by the insertion of the Cry transgene into its genome became the second most cultivated crop worldwide. Cry gene from Bacillus thuringiensis kurstaki expresses protein derivatives of crystalline endotoxins which confer insect resistance onto the maize crop. Mandatory labeling of processed food containing or made by genetically modified organisms is in force in many countries, so, it is very urgent to develop fast and practical methods for GMO identification, for example, biosensors. In the absence of an available empirical structure of Cry1A(b)16 protein, a theoretical model was effectively generated, in this work, by homology modeling and molecular dynamics simulations based on two available homologous protein structures. Molecular dynamics simulations were carried out to refine the selected model, and an analysis of its global structure was performed. The refined models of Cry1A(b)16 showed a standard fold and structural characteristics similar to those seen in Bacillus thuringiensis Cry1A(a) insecticidal toxin and Bacillus thuringiensis serovar kurstaki Cry1A(c) toxin. After in silico analysis of Cry1A(b)16, two immunoreactive candidate peptides were selected and specific polyclonal antibodies were produced resulting in antibody–peptide interaction. Biosensing devices are expected to be developed for detection of the Cry1A(b) protein as a marker of transgenic maize in food. Proteins 2017; 85:1248–1257. Fil: Plácido, Alexandra. Instituto Politécnico do Porto; Portugal Fil: Coelho, Andreia. Instituto Politécnico do Porto; Portugal Fil: Nascimento, Lucas Abreu. Universidade Federal do Piauí; Brasil Fil: Vasconcelos, Andreanne Gomes. Universidade Federal do Piauí; Brasil Fil: Barroso, María Fátima. Instituto Politécnico do Porto; Portugal Fil: Ramos Jesús, Joilson. Universidade Federal do Piauí; Brasil Fil: Costa, Vladimir. Instituto de Doenças Tropicais Natan Portela; Brasil Fil: Lima, Francisco das Chagas Alves. Universidade Estadual do Piauí; Brasil Fil: Delerue Matos, Cristina. Instituto Politécnico do Porto; Portugal Fil: Ramos, Ricardo Martins. Universidade Estadual do Piauí; Brasil. Instituto Federal do Piauí; Brasil Fil: Marani, Mariela Mirta. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto Patagónico para el Estudio de los Ecosistemas Continentales; Argentina Fil: Leite, José Roberto de Souza Almeida. Universidade de Brasília; Brasil

Background Cachexia affects 70-80% patients with pancreatic adenocarcinoma (PA). Systemic inflammation is a key feature of this multifactorial syndrome of unintended weight loss (WL). Despite being often described, cancer-associated cachexia (CAC) remains poorly understood and rarely recognized in clinical settings, therefore an unmet need. This study aims to acknowledge the effect of CAC in PA patients and to study the impact of WL, body mass index (BMI) and other available biomarkers (BM) on survival. Methods Retrospective analysis of PA patients treated in our institution between 2013 - 2018. Patients were categorized at baseline as having CAC if 1 of 2 criteria were met: WL > 5% from previous stable weight; BMI 2%. Serum hemoglobin, c-reactive protein (CRP), albumin, inflammation-based score (Glasgow Prognostic Score mGPS), neutrophil/lymphocyte ratio (NLR), platelet/lymphocyte ratio (PLR) and cancer specific antigen CA 19-9 were assessed at baseline and at 3-month. Data analysis with descriptive statistics, t-test, chi-square test, cox regression and log-rank test were performed with StataIC. Results 95 eligible patients (median age 72 [46;98], 60% female) were reviewed. At baseline, cachectic patients (CP) (74%), when compared to non-CP, had higher CA 19-9 (p = 0,047), more advanced disease (p = 0,001) lower BMI (p = 0,016), worse performance status (p = 0,016) and mean WL of 13,8 ± 6,7 (%) (p = 0,001). Also, NLR>3,5 (p = 0,001), PLR>150 (p = 0,018) and hypoalbuminemia (p = 0,004) were all statistically associated to CP. Median overall survival (OS) of non-CP was 19,13 months versus 5,03 months in CP (HR 2,69 95% CI 1,58-4,59, p = 0,001). In the latter, at baseline, higher CA 19.9 (HR 1,0 95% CI 1,01-1,021, p = 0,015), higher CRP (HR 1,21 95% CI 1,06-1,39, p = 0,006) and PLR>150 (HR 2,45; 95% CI 1,09-5,54, p = 0,031) were independent predictors of worst OS. In non-CP, normal baseline albumin (HR 0,04 95 CI 0,02-0,72, p = 0,029) was an independent predictor of better OS. Conclusions A better understanding of CAC in PA might improve outcomes for PA patients. Our study suggests that a prompt identification of prognostic BM may lead to a better standardized management of CAC and that nutritional parameters can be optimized, with impact on prognosis. Legal entity responsible for the study The authors. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

Made available in DSpace on 2019-10-06T16:30:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-08-10 European Regional Development Fund Ethnopharmacological Relevance: Folk knowledge transmitted between generations allows traditional populations to maintain the use of medicinal plants for the treatment of several diseases. In this context, the species Terminalia fagifolia Mart., native to Brazil, is used for the treatment of chronic and infectious diseases. Plants rich in secondary metabolites, such as this species and their derivatives, may represent therapeutic alternatives for the treatment of diseases that reduce the quality of life of people. Aim of the study: The aim of this study was to evaluate the antifungal and anti-inflammatory potential of aqueous fraction from ethanolic extract of T. fagifolia, with in silico study of the major compound of the fraction. Material and methods: The phytochemical study of the aqueous fraction was performed by HPLC, LC/MS and NMR. The antifungal activity was evaluated against yeasts, by determination of the minimum inhibitory concentration and minimum fungicidal concentration. The effect on Candida albicans was analyzed by AFM. The antibiofilm potential against biofilms of C. albicans was also tested. The anti-inflammatory potential of the aqueous fraction was evaluated in vivo by the carrageenan-induced paw edema and peritonitis. A microglial model of LPS-induced neuroinflammation was also studied. Further insights on the activation mechanism were studied using quantum chemistry computer simulations. Toxicity was evaluated in the Galleria mellonella and human erythrocytes models. Results: Eschweilenol C was identified as the major constituent of the aqueous fraction of the ethanolic extract of T. fagifolia. The aqueous fraction was active against all Candida strains used (sensitive and resistant to Fluconazole) with MICs ranging from 1000 to 0.4 μg/mL. By AFM it was possible to observe morphological alterations in treated Candida cells. The fraction significantly (p < 0.05) inhibited paw edema and decreased levels of malondialdehyde induced by carrageenan. In a microglial cell model, aqueous fraction demonstrated the ability to inhibit NF-κB after induction with lipopolysaccharide. The theoretical studies showed structural similarity between eschweilenol C and indomethacin and an excellent antioxidant potential. The aqueous fraction did not present toxicity in the studied models. Conclusion: The results indicate that the aqueous fraction of T. fagifolia has potential for biomedical applications with low toxicity. This finding can be attributed to the predominance of eschweilenol C in the aqueous fraction. The Northeast Biotechnology Network RENORBIO Federal University of Piaui Biotechnology and Biodiversity Center Research Biotec Federal University of Piaui Laboratory of Molecular Biology of Dimorphic and Pathogenic Fungi Institute of Biological Sciences University of Brasilia Glial Cell Biology Laboratory Institute for Research and Innovation in Health i3S University of Porto Bioprospectum Lda UPTEC Federal Institute of Education Science and Technology of São Paulo Campus Matão São Paulo State Universit UNESP Campus of Itapeva Medicinal Plants Research Center NPPM Federal University of Piaui LAQV/REQUIMTE Department of Chemistry and Biochemistry Faculty of Sciences of the University of Porto Center for Research in Applied Morphology and Immunology NuPMIA University of Brasilia São Paulo State Universit UNESP Campus of Itapeva European Regional Development Fund: PT 2020

The present work describes the development of an analytical method for the determination of methiocarb and its degradation products (methiocarb sulfoxide and methiocarb sulfone) in banana samples, using the QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure followed by liquid chromatography coupled to photodiode array detector (LC-PAD). Calibration curves were linear in the range of 0.5-10 mg L⁻¹ for all compounds studied. The average recoveries, measured at 0.1 mg kg⁻¹ wet weight, were 92.0 (RSD = 1.8%, n = 3), 84.0 (RSD = 3.9%, n = 3), and 95.2% (RSD = 1.9%, n = 3) for methiocarb sulfoxide, methiocarb sulfone, and methiocarb, respectively. Banana samples treated with methiocarb were collected from an experimental field. The developed method was applied to the analysis of 24 samples (peel and pulp) and to 5 banana pulp samples. Generally, the highest levels were found for methiocarb sulfoxide and methiocarb. Methiocarb sulfone levels were below the limit of quantification, except in one sample (not detected).

Escherichia coli is one of the most common etiological agents of diarrhea in developing countries. The appearance of resistant E. coli prevents treatment of these infections. Biotechnological products incorporating antimicrobial peptides are currently being considered in applications to prevent intestinal infections by these bacteria. The aim of this study was to evaluate the antibacterial activity of the peptide PcL342-354C, which is derived from the toxin Cry1Ab16 from Bacillus thuringiensis, against E. coli strains. We also report the preparation, characterization and evaluation of the antibacterial activity of LbL films containing PcL342-354C. The results showed that the PcL342-354C peptide inhibited the growth of different strains of E. coli with minimal inhibitory concentration ranging from 15.62–31.25 μg/mL and minimal bactericidal concentration was 250 μg/mL, indicating a potential antibacterial activity. The morphology of an ITO/Cashew gum/PcL342-354C film was analysed using atomic force microscopy which showed an increase of roughness due to the increase in the number of layers. The LbL films showed significant antibacterial activity against E. coli NCTC 9001 in both conditions tested (10 and 20 bilayers). Our results indicate that the peptide exhibits an antibacterial potential that can be tapped to develop biomaterials with antibacterial activity for use against foodborne pathogens.

BPP-BrachyNH2 is a proline-rich oligopeptide (PRO) firstly identified in skin secretion of the frog Brachycephalus ephippium, which possess in vitro inhibitory activity of angiotensin-I converting enzyme (ACE) and endothelium-dependent vasorelaxant activity. Considering its potential application in the treatment of cardiovascular diseases, the present work assessed the toxicological profile of the BPP-BrachyNH2. The in silico toxicity prediction was performed from the best model obtained through the optimization of the FASTA query peptide. This prediction study revealed that BPP-BrachyNH2 induced high predicted LD50 values for both humans and rats, and then is well-tolerated in the recommended range. The MTT assay was applied for the in vitro cytotoxic evaluation in murine macrophages. In this assay, a decrease of cell viability was not observed. The in vivo acute toxicological study was performed after the intraperitoneal administration of BPP-BrachyNH2 at doses of 5 and 50 mg/kg. After intraperitoneal administration, no death, alterations in behavioral parameters or weight gain curve was observed, as well as none in the serum biochemical parameters, and gross pathological and histopathological analyses. These observations demonstrates an acceptable safety profile for BPP-BrachyNH2, leading towards further studies focused on investigation of pharmacological and therapeutical applications for this peptide.