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  1. Article ; Online: Virulence Factors in Coagulase-Negative Staphylococci

    Angela França / Vânia Gaio / Nathalie Lopes / Luís D. R. Melo

    Pathogens, Vol 10, Iss 2, p

    2021  Volume 170

    Abstract: Coagulase-negative staphylococci (CoNS) have emerged as major pathogens in healthcare-associated facilities, being S. epidermidis , S. haemolyticus and, more recently, S. lugdunensis , the most clinically relevant species. Despite being less virulent ... ...

    Abstract Coagulase-negative staphylococci (CoNS) have emerged as major pathogens in healthcare-associated facilities, being S. epidermidis , S. haemolyticus and, more recently, S. lugdunensis , the most clinically relevant species. Despite being less virulent than the well-studied pathogen S. aureus , the number of CoNS strains sequenced is constantly increasing and, with that, the number of virulence factors identified in those strains. In this regard, biofilm formation is considered the most important. Besides virulence factors, the presence of several antibiotic-resistance genes identified in CoNS is worrisome and makes treatment very challenging. In this review, we analyzed the different aspects involved in CoNS virulence and their impact on health and food.
    Keywords coagulase-negative staphylococci ; biofilms ; virulence factors ; Medicine ; R
    Language English
    Publishing date 2021-02-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Author Correction

    Luís D. R. Melo / R. Ferreira / Ana R. Costa / H. Oliveira / J. Azeredo

    Scientific Reports, Vol 11, Iss 1, Pp 1-

    Efficacy and safety assessment of two enterococci phages in an in vitro biofilm wound model

    2021  Volume 1

    Abstract: An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...

    Abstract An amendment to this paper has been published and can be accessed via a link at the top of the paper.
    Keywords Medicine ; R ; Science ; Q
    Language English
    Publishing date 2021-05-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Efficacy and safety assessment of two enterococci phages in an in vitro biofilm wound model

    Luís D. R. Melo / R. Ferreira / Ana R. Costa / H. Oliveira / J. Azeredo

    Scientific Reports, Vol 9, Iss 1, Pp 1-

    2019  Volume 12

    Abstract: Abstract Chronic wounds affect thousands of people worldwide, causing pain and discomfort to patients and represent significant economical burdens to health care systems. The treatment of chronic wounds is very difficult and complex, particularly when ... ...

    Abstract Abstract Chronic wounds affect thousands of people worldwide, causing pain and discomfort to patients and represent significant economical burdens to health care systems. The treatment of chronic wounds is very difficult and complex, particularly when wounds are colonized by bacterial biofilms which are highly tolerant to antibiotics. Enterococcus faecium and Enterococcus faecalis are within the most frequent bacteria present in chronic wounds. Bacteriophages (phages) have been proposed as an efficient and alternative against antibiotic-resistant infections, as those found in chronic wounds. We have isolated and characterized two novel enterococci phages, the siphovirus vB_EfaS-Zip (Zip) and the podovirus vB_EfaP-Max (Max) to be applied during wound treatment. Both phages demonstrated lytic behavior against E. faecalis and E. faecium. Genome analysis of both phages suggests the absence of genes associated with lysogeny. A phage cocktail containing both phages was tested against biofilms formed in wound simulated conditions at a multiplicity of infection of 1.0 and a 2.5 log CFU.mL−1 reduction in the bacterial load after at 3 h of treatment was observed. Phages were also tested in epithelial cells colonized by these bacterial species and a 3 log CFU.mL−1 reduction was observed using both phages. The high efficacy of these new isolated phages against multi-species biofilms, their stability at different temperatures and pH ranges, short latent periods and non-cytotoxicity to epithelial cells suggest their therapeutic use to control infectious biofilms present in chronic wounds.
    Keywords Medicine ; R ; Science ; Q
    Subject code 572
    Language English
    Publishing date 2019-04-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Identification of the first endolysin Cell Binding Domain (CBD) targeting Paenibacillus larvae

    Sílvio B. Santos / Ana Oliveira / Luís D. R. Melo / Joana Azeredo

    Scientific Reports, Vol 9, Iss 1, Pp 1-

    2019  Volume 9

    Abstract: Abstract Bacteriophage endolysins present enormous biotechnological potentials and have been successfully used to control and detect bacterial pathogens. Endolysins targeting Gram-positive bacteria are modular, displaying a cell binding (CBD) and an ... ...

    Abstract Abstract Bacteriophage endolysins present enormous biotechnological potentials and have been successfully used to control and detect bacterial pathogens. Endolysins targeting Gram-positive bacteria are modular, displaying a cell binding (CBD) and an enzymatically active domain. The CBD of phage endolysins are recognized by their high specificity and host affinity, characteristics that make them promising diagnostic tools. No CBD able to bind Paenibacillus larvae has been identified so far. P. larvae is a Gram-positive spore forming bacteria that causes the American Foulbrood. This highly contagious infection leads to honeybee larvae sepsis and death, resulting in an adverse impact on pollination and on the beekeeping industry. In this work, the first CBD targeting P. larvae was identified and its core binding sequence was investigated. Moreover, it was shown that the domain is highly specific, targeting exclusively P. larvae cells from all ERIC genotypes. The identification of such a domain represents a step forward for the development of effective methods to detect and control this pathogen.
    Keywords Medicine ; R ; Science ; Q
    Language English
    Publishing date 2019-02-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: A novel flow cytometry assay based on bacteriophage-derived proteins for Staphylococcus detection in blood

    Susana P. Costa / Nicolina M. Dias / Luís D. R. Melo / Joana Azeredo / Sílvio B. Santos / Carla M. Carvalho

    Scientific Reports, Vol 10, Iss 1, Pp 1-

    2020  Volume 13

    Abstract: Abstract Bloodstream infections (BSIs) are considered a major cause of death worldwide. Staphylococcus spp. are one of the most BSIs prevalent bacteria, classified as high priority due to the increasing multidrug resistant strains. Thus, a fast, specific ...

    Abstract Abstract Bloodstream infections (BSIs) are considered a major cause of death worldwide. Staphylococcus spp. are one of the most BSIs prevalent bacteria, classified as high priority due to the increasing multidrug resistant strains. Thus, a fast, specific and sensitive method for detection of these pathogens is of extreme importance. In this study, we have designed a novel assay for detection of Staphylococcus in blood culture samples, which combines the advantages of a phage endolysin cell wall binding domain (CBD) as a specific probe with the accuracy and high-throughput of flow cytometry techniques. In order to select the biorecognition molecule, three different truncations of the C-terminus of Staphylococcus phage endolysin E-LM12, namely the amidase (AMI), SH3 and amidase+SH3 (AMI_SH3) were cloned fused with a green fluorescent protein. From these, a higher binding efficiency to Staphylococcus cells was observed for AMI_SH3, indicating that the amidase domain possibly contributes to a more efficient binding of the SH3 domain. The novel phage endolysin-based flow cytometry assay provided highly reliable and specific detection of 1–5 CFU of Staphylococcus in 10 mL of spiked blood, after 16 hours of enrichment culture. Overall, the method developed herein presents advantages over the standard BSIs diagnostic methods, potentially contributing to an early and effective treatment of BSIs.
    Keywords Medicine ; R ; Science ; Q
    Subject code 500
    Language English
    Publishing date 2020-04-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: Characterization of a new podovirus infecting Paenibacillus larvae

    Henrique G. Ribeiro / Luís D. R. Melo / Hugo Oliveira / Maarten Boon / Rob Lavigne / Jean-Paul Noben / Joana Azeredo / Ana Oliveira

    Scientific Reports, Vol 9, Iss 1, Pp 1-

    2019  Volume 12

    Abstract: Abstract The Paenibacillus larvae infecting phage API480 (vB_PlaP_API480) is the first reported podovirus for this bacterial species, with an 58 nm icosahedral capsid and a 12 × 8 nm short, non-contractile tail. API480 encodes 77 coding sequences (CDSs) ... ...

    Abstract Abstract The Paenibacillus larvae infecting phage API480 (vB_PlaP_API480) is the first reported podovirus for this bacterial species, with an 58 nm icosahedral capsid and a 12 × 8 nm short, non-contractile tail. API480 encodes 77 coding sequences (CDSs) on its 45,026 bp dsDNA genome, of which 47 were confirmed using mass spectrometry. This phage has got very limited genomic and proteomic similarity to any other known ones registered in public databases, including P. larvae phages. Comparative genomics indicates API480 is a new species as it’s a singleton with 28 unique proteins. Interestingly, the lysis module is highly conserved among P. larvae phages, containing a predicted endolysin and two putative holins. The well kept overall genomic organisation (from the structural and morphogenetic modules to the host lysis, DNA replication and metabolism related proteins) confirms a common evolutionary ancestor among P. larvae infecting phages. API480 is able to infect 69% of the 61 field strains with an ERIC I genotype, as well as ERIC II strains. Furthermore, this phage is very stable when exposed to high glucose concentrations and to larval gastrointestinal conditions. This highly-specific phage, with its broad lytic activity and stability in hive conditions, might potentially be used in the biocontrol of American Foulbrood (AFB).
    Keywords Medicine ; R ; Science ; Q
    Subject code 500
    Language English
    Publishing date 2019-12-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article ; Online: Selection of aptamers against triple negative breast cancer cells using high throughput sequencing

    Débora Ferreira / Joaquim Barbosa / Diana A. Sousa / Cátia Silva / Luís D. R. Melo / Meltem Avci-Adali / Hans P. Wendel / Ligia R. Rodrigues

    Scientific Reports, Vol 11, Iss 1, Pp 1-

    2021  Volume 15

    Abstract: Abstract Triple-negative breast cancer is the most aggressive subtype of invasive breast cancer with a poor prognosis and no approved targeted therapy. Hence, the identification of new and specific ligands is essential to develop novel targeted therapies. ...

    Abstract Abstract Triple-negative breast cancer is the most aggressive subtype of invasive breast cancer with a poor prognosis and no approved targeted therapy. Hence, the identification of new and specific ligands is essential to develop novel targeted therapies. In this study, we aimed to identify new aptamers that bind to highly metastatic breast cancer MDA-MB-231 cells using the cell-SELEX technology aided by high throughput sequencing. After 8 cycles of selection, the aptamer pool was sequenced and the 25 most frequent sequences were aligned for homology within their variable core region, plotted according to their free energy and the key nucleotides possibly involved in the target binding site were analyzed. Two aptamer candidates, Apt1 and Apt2, binding specifically to the target cells with $$K_{d}$$ K d values of 44.3 ± 13.3 nM and 17.7 ± 2.7 nM, respectively, were further validated. The binding analysis clearly showed their specificity to MDA-MB-231 cells and suggested the targeting of cell surface receptors. Additionally, Apt2 revealed no toxicity in vitro and showed potential translational application due to its affinity to breast cancer tissue sections. Overall, the results suggest that Apt2 is a promising candidate to be used in triple-negative breast cancer treatment and/or diagnosis.
    Keywords Medicine ; R ; Science ; Q
    Subject code 616
    Language English
    Publishing date 2021-04-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  8. Article ; Online: Correction

    Ana Oliveira / Marta Leite / Leon D Kluskens / Sílvio B Santos / Luís D R Melo / Joana Azeredo

    PLoS ONE, Vol 10, Iss 8, p e

    The First Paenibacillus larvae Bacteriophage Endolysin (PlyPl23) with High Potential to Control American Foulbrood.

    2015  Volume 0136331

    Keywords Medicine ; R ; Science ; Q
    Language English
    Publishing date 2015-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  9. Article ; Online: The First Paenibacillus larvae Bacteriophage Endolysin (PlyPl23) with High Potential to Control American Foulbrood.

    Ana Oliveira / Marta Leite / Leon D Kluskens / Sílvio B Santos / Luís D R Melo / Joana Azeredo

    PLoS ONE, Vol 10, Iss 7, p e

    2015  Volume 0132095

    Abstract: Endolysins, which are peptidoglycan-degrading enzymes expressed during the terminal stage of the reproduction cycle of bacteriophages, have great potential to control Gram-positive pathogens. This work describes the characterization of a novel endolysin ( ...

    Abstract Endolysins, which are peptidoglycan-degrading enzymes expressed during the terminal stage of the reproduction cycle of bacteriophages, have great potential to control Gram-positive pathogens. This work describes the characterization of a novel endolysin (PlyPl23) encoded on the genome of Paenibacillus larvae phage phiIBB_Pl23 with high potential to control American foulbrood. This bacterial disease, caused by P. larvae, is widespread in North America and Europe and causes important economic losses in apiculture. The restriction to antibiotic residues in honey imposed by the EU legislation hinders its therapeutic use to combat American foulbrood and enforces the development of alternative antimicrobial methods. The new endolysin described herein has an N-acetylmuramoyl-L-alanine amidase catalytic domain and exhibits a broad-spectrum activity against common P. larvae genotypes. Moreover, the enzyme displays high antimicrobial activity in a range of pH that matches environmental conditions (pH between 5.0 and 7.0), showing its feasible application in the field. At pH 7.0, a concentration of 0.2 μM of enzyme was enough to lyse 104 CFU.mL-1 of P. larvae in no more than 2 h. The presence of sucrose and of the substances present in the larvae gut content did not affect the enzyme activity. Interestingly, an increase of activity was observed when PlyPl23 was previously incubated in royal jelly. Furthermore, in vivo safety evaluation assays demonstrated that this enzyme is not toxic to the bee larvae. The present work describes for the first time an endolysin encoded in a P. larvae phage that presents high potential to integrate a commercial product to control the problematic American foulbrood.
    Keywords Medicine ; R ; Science ; Q
    Subject code 572
    Language English
    Publishing date 2015-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article: Susceptibility testing of Candida albicans and Candida glabrata to Glycyrrhiza glabra L

    Martins, Natália / Isabel C.F.R. Ferreira / Joana Azeredo / Luís D.R. Melo / Mariana Henriques / Sofia Costa-Oliveira / Sónia Silva

    Industrial crops and products. 2017 Dec. 01, v. 108

    2017  

    Abstract: Medicinal plants and their botanical formulations have gained a pivotal attention among scientific researchers mainly due to its action as health promoters. Licorice (Glycyrrhiza glabra L.) constitutes a great example, with an increasingly evidenced ... ...

    Abstract Medicinal plants and their botanical formulations have gained a pivotal attention among scientific researchers mainly due to its action as health promoters. Licorice (Glycyrrhiza glabra L.) constitutes a great example, with an increasingly evidenced antimicrobial potential. Opportunistic yeast infections constitute an alarming public health problem, highly exacerbated by the inefficacy of antifungal drugs and the increase of drug-resistant microorganisms, being Candida species one of the most common invaders. The present work aims to assess Candida glabrata and Candida albicans susceptibility to G. glabra methanol: water extract by using flow cytometry and transmission electron microscopy techniques. After 5min, licorice extract (1.5mg/mL) altered Candida membrane potential. Within an hour, it induced primary damages on Candida species cells, causing cell cytoplasm disorganization with high evidence of cell membrane invaginations, making cells turgid. Thus, based on the current findings, licorice extract seems to be a promising anti-Candida agent, without presenting any toxic potential at the effective concentrations used.
    Keywords antifungal agents ; Candida albicans ; Candida glabrata ; cell membranes ; cytoplasm ; drug resistance ; flow cytometry ; Glycyrrhiza glabra ; licorice ; medicinal plants ; membrane potential ; methanol ; public health ; researchers ; toxicity ; transmission electron microscopy ; yeasts
    Language English
    Dates of publication 2017-1201
    Size p. 480-484.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 1132158-1
    ISSN 1872-633X ; 0926-6690
    ISSN (online) 1872-633X
    ISSN 0926-6690
    DOI 10.1016/j.indcrop.2017.07.008
    Database NAL-Catalogue (AGRICOLA)

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