Polymorphism in ficolin-1 (FCN1) gene is associated with an earlier onset of type 1 diabetes mellitus in children and adolescents from northeast Brazil

10siFicolins are innate immune proteins able to activate the complement system by the lectin pathway. Single nucleotide polymorphisms (SNPs) of FCN1 and FCN2 genes, encoding for ficolin 1 and 2, have been related to the susceptibility to infectious and autoimmune disease. This study aims at investigating the association of SNPs at FCN1 and FCN2 gene with the development of type 1 diabetes mellitus (T1D). Two SNPs at FCN1: rs2989727 and rs1071583 and three at FCN2: rs17514136, rs3124954 and rs7851696 were studied in 204 children diagnosed with T1D and 193 healthy individuals. No direct associations were found with the T1D onset or with the insurgence of T1D related celiac disease (CD) and autoimmune thyroiditis (AIDT). However, the genotype T/T (rs1071583) of FCN1 was associated with an early age at T1D diagnosis compared with C/C or C/T genotypes (p= 0.05), around two years of difference. Thus, our results suggest that the T/T genotype (rs1071583) is not directly involved in the initial steps of T1D onset, but, after the trigger that induces T1D, individual carrying this genotype could increase/accelerate the pancreatic autoimmune response.openopenAnjosa, Zilma Pereira Dos; Santos, Manuella Maria Silva; Rodrigues, Natassia Javorski; Lacerda, Glaucia Alyne Nunes De; Araujo, Jaqueline; Silva, Jaqueline De Azevêdo; Tavares, Nathália De Alencar Cunha; Guimarães, Rafael Lima; Crovella, Sergio; Brandão, Lucas André CavalcantiAnjosa, Zilma Pereira Dos; Santos, Manuella Maria Silva; Rodrigues, Natassia Javorski; Lacerda, Glaucia Alyne Nunes De; Araujo, Jaqueline; Silva, Jaqueline De Azevêdo; Tavares, Nathália De Alencar Cunha; Guimarães, Rafael Lima; Crovella, Sergio; Brandão, Lucas André Cavalcant

Identification of Eschweilenol C in derivative of Terminalia fagifolia Mart. and green synthesis of bioactive and biocompatible silver nanoparticles

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 IMMinfo:eu-repo/semantics/publishedVersio

Antibacterial and antifungal activity of curcumin and methylene blue associated with laser on bacterial and fungal strains

Objective: To analyze the effect of methylene blue and 10% curcumin in fungi and bacteria through an in vitro study using photodynamic therapy (PDT). Methods: Curcumin and methylene blue were photosensitized by a Photon Lase III laser applied for 90 s in a dark environment within a laminar flow chamber. Enterococcus faecalis and Candida albicans strains were cultured and standardized. Then, a minimum inhibitory concentration (MIC) assay was conducted for these photosensitizers, with concentration variations and incubation to evaluate their antimicrobial activity. Results: With PDT, Curcumin had significant antibacterial activity against E. faecalis (MIC = 250 µg/mL). In contrast, methylene blue had antibacterial activity against E. faecalis (MIC < 12.5 µg/mL with PDT) and antifungal activity against C. albicans (MIC <12.5 µg/mL with or without PDT). Both agents showed greater efficacy in the presence of the laser. The results suggest that curcumin and methylene blue associated with laser may effectively treat microbial infections. Conclusion: Further research is needed to evaluate the efficacy and safety of using these agents in animal and human models and their effectiveness against different bacterial and fungal strains.

Evaluation of the Antibacterial Activity of New Dermaseptin Derivatives against <i>Acinetobacter baumannii</i>

Nosocomial infections represent one of the biggest health problems nowadays. Acinetobacter baumannii is known as an opportunistic pathogen in humans, affecting people with compromised immune systems, and is becoming increasingly important as a hospital-derived infection. It is known that in recent years, more and more bacteria have become multidrug-resistant (MDR) and, for this reason, the development of new drugs is a priority. However, these products must not affect the human body, and therefore, cytotoxicity studies are mandatory. In this context, antimicrobial peptides with potential antibacterial proprieties could be an alternative. In this research, we describe the synthesis and the bioactivity of dermaseptins and their derivatives against Acinetobacter baumannii. The cytotoxicity of these compounds was investigated on the HEp-2 cell line by MTT cell viability assay. Thereafter, we studied the morphological alterations caused by the action of one of the active peptides on the bacterial membrane using atomic force microscopy (AFM). The cytotoxicity of dermaseptins was concentration-dependent at microgram concentrations. It was observed that all tested analogs exhibited antibacterial activity with Minimum Inhibitory Concentrations (MICs) ranging from 3.125 to 12.5 μg/mL and Minimum Bactericidal Concentrations (MBCs) ranging from 6.25 to 25 μg/mL. Microscopic images obtained by AFM revealed morphological changes on the surface of the treated bacteria caused by K4S4(1-16), as well as significant surface alterations. Overall, these findings demonstrate that dermaseptins might constitute new lead structures for the development of potent antibacterial agents against Acinetobacter baumannii infections

Use of cashew gum for development of LBL films with antibacterial actibity for applicantion against foodborne pathogens

Escherichia coli are one ofthe most common etiological agents of diarrhea in developing countries. The appearance of resistant E. coliprevents these infections treatment, thus antimicrobial peptides and their biotechnological applications can make good alternatives to treat and prevent intestinal infections by these bacteria. The aim of this study was to evaluate the antibacterial activity of PcL342-354C peptide, derived from Cry1Ab16 Toxinfrom Bacillus thuringiensis, againsts trains of E. coli, as well as the synthesis, characterization and evaluation of the antibacterial activity of layer-by-layer(LbL) films containing PcL342-354C. The results showed that the PcL342-354Cpeptide inhibited the growth of E. coliATCC 25922 (Minimal Inhibitory Concentration -MIC-: 31.25 μg/mL), E. coliML1 (MIC: 31.25 μg/mL) and E. coli ATCC 35218 (MIC: 15.62 μg/mL). The Minimal Bactericidal Concentration was250 μg/mL for all strains used,proving a potential antibacterial activity against these Gram-negative bacteria. Morphological effect on E. coli,produced by Cry1Ab16 Toxin derived peptide at MIC, was significant. The morphology of the ITO/Cashew gum/PcL342-354C film was analysed using Atomic Force Microscopy (AFM) which showed an increase of roughness due to the increase in the number of layers. LbL film has antibacterial activity against E. coli NCTC 9001 in both tested conditions (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.Fil: Placido, Alexandra. Universidad de Porto; PortugalFil: Bragança, Idalina. Universidad de Porto; PortugalFil: 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; ArgentinaFil: Rodrigues de Araujo, Alyne. Universidade Federal do Piaui; BrasilFil: Vasconcelos, Adreanne G.. Universidade Federal do Piaui; BrasilFil: Batziou, Krystallenia. Universidad de Porto; PortugalFil: Domingues, Valentina. Universidad de Porto; PortugalFil: Eaton, Peter. Universidad de Porto; PortugalFil: de Souza de Almedia Leite, José Roberto. Universidade Federal do Piaui; BrasilFil: Delerue Matos, Cristina. Universidad de Porto; PortugalXIII Encontro de Química dos AlimentosPortoPortugalUniversidade do PortoSociedade Portuguesa de Químic

Photodynamic Therapy With Propolis: Antibacterial Effects on Staphylococcus aureus, Streptococcus mutans and Escherichia coli Analysed by Atomic Force Microscopy: Photodynamic Therapy With Propolis

Introduction: Photodynamic therapy (PDT) is a process that uses a light source (e.g. laser), oxygen molecules, and a photosensitizing agent. PDT aims to act against pathogens, including those resistant to antimicrobials. The association of PDT with natural drugs, such as Propolis, has not been widely studied.Methods: Therefore, this study aimed to evaluate the antimicrobial effect of PDT in vitro by using Propolis as a photosensitizing agent. For this purpose, the dry Propolis extract was used as a photosensitizer and a low-power laser (Photon Laser III model) was irradiated onto the microwells for 90 seconds. Gram-positive and Gram-negative bacterial strains were used in the tests at a concentration of 5 × 105 CFU/mL. Initially, the antibacterial activity of the photosensitizers without laser action was determined by using a serial microdilution method before the experiment with a laser. After the incubation of the plates in a bacteriological oven, resazurin (0.1%) was added and the minimum inhibitory concentration (MIC) was determined. Alterations in the morphology of the bacteria were analyzed using atomic force microscopy (AFM).Results: Bacteria were sensitive to Propolis with MICs ranging from 13.75 to 0.85 mg/mL, but no susceptibility was observed for methylene blue without laser application. A change was observed for MIC values of Propolis against Staphylococcus aureus after irradiation, which decreased from 1.71 mg/mL to 0.85 mg/mL. However, this behavior was not observed in Escherichia coli, the only gram-negative strain used. In addition, AFM images revealed alterations in the size of one of the bacteria tested.Conclusion: The Propolis is more active against gram-positive bacteria and PDT improved its activity against one of the strains tested

Characterization and Biological Activities of Ocellatin Peptides from the Skin Secretion of the Frog Leptodactylus pustulatus

Eight new peptides were isolated from the skin secretion of the frog Leptodactylus pustulatus and their amino acid sequences determined by de novo sequencing and by cDNA cloning. Structural similarities between them and other antimicrobial peptides from the skin secretion of Leptodactylus genus frogs were found. Ocellatins-PT1 to -PT5 (25 amino acid residues) are amidated at the C-terminus, while ocellatins-PT6 to -PT8 (32 amino acid residues) have free carboxylates. Antimicrobial activity, hemolytic tests, and cytotoxicity against a murine fibroblast cell line were investigated. All peptides, except for ocellatin-PT2, have antimicrobial activity against at least one Gram negative strain. Ocellatin-PT8 inhibited the growth of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Salmonella choleraesuis strains with MICs in the 60−240 μM range. No significant effect was observed in human erythrocytes and in a murine fibroblast cell line after exposure to the peptides at MICs. A comparison between sequences obtained by both direct HPLC-MS de novo sequencing and cDNA cloning demonstrates the secretion of mature peptides derived from a pre-pro-peptide structure

Thaulin-1: The first antimicrobial peptide isolated from the skin of a Patagonian frog Pleurodema thaul (Anura: Leptodactylidae: Leiuperinae) with activity against Escherichia coli

Patagonia's biodiversity has been explored from many points of view, however, skin secretions of native amphibians have not been evaluated for antimicrobial peptide research until now. In this sense, Pleurodema thaul is the first amphibian specie to be studied from this large region of South America. Analysis of cDNA-encoding peptide in skin samples allowed identification of four new antimicrobial peptides. The predicted mature peptides were synthesized and all of them showed weak or null antimicrobial activity against Klebsiella pneumoniae, Staphylococcus aureus and Escherichia coli with the exception of thaulin-1, a cationic 26-residue linear, amphipathic, Gly- and Leu-rich peptide with moderate antimicrobial activity against E. coli (MIC of 24.7 M). AFM and SPR studies suggested a preferential interaction between these peptides and bacterial membranes. Cytotoxicity assays showed that thaulin peptides had minimal effects at MIC concentrations towards human and animal cells. These are the first peptides described for amphibians of the Pleurodema genus. These findings highlight the potential of the Patagonian region's unexplored biodiversity as a source for new molecule discovery.This work was partially supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) (PIP N° 11220120100050CO), and the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) (PICT N° 1199). M.M.M., L.O.P., S.A.C. and N.G.B. are researchers at CONICET. Work in AMT laboratory was supported by Project “NORTE-07-0124-FEDER-000002-Host-Pathogen Interactions” cofunded by Programa Operacional Regional do Norte under QREN, through FEDER (NORTE-07-0124-FEDER-000002-Host-Pathogen Interactions) and FCT. (SFRH/BD/97995/2013, SFRH/BD/93766/2013 and UID/MULTI/04378/2013) A.P. and A.G.G.A. are gratefully to FCT by their grants SFRH/BD/97995/2013 and SFRH/BD/93766/2013 respectively, financed by POPH–QREN–Tipologia 4.1–Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior. The authors thank N.L. Olivera for logistic support and helpful discussions regarding the article. P.E. thanks CNPq for PVE grant no. 400398/2014-1, and is supported by FCTvia the project UID/MULTI/04378/2013

Acetylated cashew gum and fucan for incorporation of lycopene rich extract from red guava (Psidium guajava L.) in nanostructured systems: antioxidant and antitumor capacity

Industrial application of lycopene is limited due to its chemical instability and low bioavailability. This study proposes the development of fucan-coated acetylated cashew gum nanoparticles (NFGa) and acetylated cashew gum nanoparticles (NGa) for incorporation of the lycopene-rich extract from red guava (LEG). Size, polydispersity, zeta potential, nanoparticles concentration, encapsulation efficiency, transmission electron microscopy (TEM) and atomic force microscopy (AFM) were used to characterize nanoparticles. The antioxidant activity was determinated and cell viability was evaluated in the human breast cancer cells (MCF-7) and human keratinocytes (HaCaT) by MTT assay. The toxic effect was evaluated by hemolysis test and by Galleria mellonella model. NFGa showed higher stability than NGa, having a size of 162.10 ± 3.21 nm, polydispersity of 0.348 ± 0.019, zeta potential -30.70 ± 0.53 mV, concentration of 6.4 × 109 nanoparticles/mL and 60% LEG encapsulation. Microscopic analysis revealed a spherical and smooth shape of NFGa. NFGa showed antioxidant capacity by ABTS method and ORAC assay. The NFGa presented significant cytotoxicity against MCF-7 from the lowest concentration tested (6.25-200 μg/mL) and did not affect the cell viability of the HaCaT. NFGa showed non-toxic effect in the in vitro and in vivo models. Therefore, NFGa may have a promising application in LEG stabilization for antioxidant and antitumor purposes.info:eu-repo/semantics/acceptedVersio