A new jatrophane diterpene, pubescenol (1), known ent-abietane lactones, helioscopinolide A (2) a... more A new jatrophane diterpene, pubescenol (1), known ent-abietane lactones, helioscopinolide A (2) and B (3), and taraxerone, 24-methylenecycloartanol, and vanillin have been isolated from Euphorbia pubescens. Diterpenes 1-3 and previously described pubescene D (4) were shown to be moderate inhibitors of the growth of MCF-7, NCI-H460, and SF-268 human tumor cell lines, whereas compounds 2 and 3 also exhibited significant antibacterial activity against Staphylococcus aureus.
As the personal care industry evolves, formulators are seeking innovative solutions for their for... more As the personal care industry evolves, formulators are seeking innovative solutions for their formulations' needs, and for ingredients that can offer multiple functions within formulations. Considering that essential oils (EOs) may present a wide spectrum of biological activities, the composition, antimicrobial and antioxidant activity of Portuguese Thymbra capitata, Thymus caespititius and Myrtus communis EOs were assessed in order to evaluate them as preservatives and antioxidants in topical emulsions. The in vivo safety application of some emulsions was also tested. T. capitata EO was mainly constituted by carvacrol (73%), whereas α-terpineol (27%), p-cymene (14%) and carvacrol (10%) dominated Th. caespititius EO, and 1,8-cineole (37%) was dominant in M. communis EO. The minimum inhibitory concentration of T. capitata, Th. caespititius and M. communis EOs was 0.4 μg/mL against C. albicans and ranged between 0.4 and 30.7 μg/mL against A. brasiliensis. Gram-positive bacteria were more susceptible to each EO than Gram-negative bacteria. T. capitata EO showed significantly higher antioxidant activity than Th. caespititius and M. communis EOs. EOs incorporated in emulsions showed preservative activity against all microorganisms tested and T. capitata EO emulsions showed powerful reactive oxygen species (ROS) scavenging effects. A safety evaluation study was performed with 0.10% and 0.01% T. capitata EO emulsions according to the Human Repeat Insult Patch Test (HRIPT). All emulsions were considered safe for topical application. T. capitata, Th. caespititius and M. communis significantly improved the microbiological quality of the prepared emulsions and may constitute a powerful alternative to the current preservatives and antioxidants for use in topical formulations.
Infections caused by multiresistant bacterial pathogens are a significant problem worldwide, turn... more Infections caused by multiresistant bacterial pathogens are a significant problem worldwide, turning the search for natural compounds to act as alternatives to antibiotics of major importance. The aim of the present study was to investigate the in vitro antimicrobial activity of 7α-acetoxy-6β-hydroxyroyleanone (1), isolated from Plectranthus grandidentatus (Lamiaceae), and 11 additional royleanone abietane derivatives of 1 (2-12) against important Gram-positive human bacterial pathogens. Results showed that the aromatic and alkylic esters 2, 3 and 5 are more active than 1 against Enterococcus and Staphylococcus (minimum inhibitory concentration (MIC) values ranging from 0.98 to 62.50 µg/mL). Moreover, 7α-acetoxy-6β-hydroxy-12-O-(4-chloro)benzoylroyleanone (2) gave rise to a new antibacterial-prototype (MIC values of 3.91-15.63 µg/mL against Staphylococcus and of 0.98-3.91 µg/mL against Enterococcus). The results showed that the compounds under analysis also present antimicrobial activity against resistant bacteria. The hydrophobic extra-interactions with bacterial targets seem to play an important role on the activity of royleanones derivatives.
Pseudomonas aeruginosa is a Gram-negative opportunistic bacterium that presents resistance to sev... more Pseudomonas aeruginosa is a Gram-negative opportunistic bacterium that presents resistance to several antibiotics, thus, representing a major threat to human and animal health. Phage-derived products, namely lysins, or peptidoglycan-hydrolyzing enzymes, can be an effective weapon against antibiotic-resistant bacteria. Whereas in Gram-positive bacteria, lysis from without is facilitated by the exposed peptidoglycan layer, this is not possible in the outer membrane-protected peptidoglycan of Gram-negative bacteria. Here, we suggest the encapsulation of lysins in liposomes as a delivery system against Gram-negative bacteria, using the model of P. aeruginosa. Bioinformatic analysis allowed for the identification of 38 distinct complete prophages within 66 P. aeruginosa genomes (16 of which newly sequenced) and led to the identification of 19 lysins of diverse sequence and function, 5 of which proceeded to wet lab analysis. The four purifiable lysins showed hydrolytic activity against Gr...
Klebsiella pneumoniae (Kp) bacteria are an increasing threat to public health and represent one o... more Klebsiella pneumoniae (Kp) bacteria are an increasing threat to public health and represent one of the most concerning pathogens involved in life-threatening infections and antimicrobial resistance (AMR). To understand the epidemiology of AMR of Kp in Portugal, we analysed whole genome sequencing, susceptibility testing and other meta data on 509 isolates collected nationwide from 16 hospitals and environmental settings between years 1980 and 2019. Predominant sequence types (STs) included ST15 (n = 161, 32%), ST147 (n = 36, 7%), ST14 (n = 26, 5%) or ST13 (n = 26, 5%), while 31% of isolates belonged to STs with fewer than 10 isolates. AMR testing revealed widespread resistance to aminoglycosides, fluoroquinolones, cephalosporins and carbapenems. The most common carbapenemase gene was blaKPC-3. Whilst the distribution of AMR linked plasmids appears uncorrelated with ST, their frequency has changed over time. Before year 2010, the dominant plasmid group was associated with the extende...
The emergence of carbapenemase-producing Klebsiella pneumoniae strains has triggered the use of o... more The emergence of carbapenemase-producing Klebsiella pneumoniae strains has triggered the use of old antibiotics such as colistin which is consequently driving the emergence of colistin resistance in already multidrug resistant strains that can underlie life-threatening infections. This study analyses the mutational diversity of 22 genes associated with colistin resistance in 140 K. pneumoniae clinical isolates integrated in a high-resolution phylogenetic scenario. Colistin susceptibility was accessed by broth microdilution categorizing 98 isolates as susceptible and 16 as resistant, 10 of which were carbapenemase producers. Across the 22 genes examined, 171 non-synonymous mutations and 9 mutations associated with promoter regions were found. Eighty-five isolates had a truncation and/or deletion in at least one of the 22 genes. However, only seven mutations, the complete deletion of mgrB or insertion sequence mediated disruption were exclusively observed in resistant isolates, 4 of these (mgrBIle13fs, pmrBGly207Asp, phoQHis339Asp and ramAIle28Met) comprising novel mutations potentially involved in colistin resistance. The ISEcp1-blaCTX-M-15::mgrB disruption is herein noteworthy as a mechanism underlying co-resistance to third-generation cephalosporins and colistin. Moreover, the high-resolution phylogenetic context shows that most of the mutational diversity spans across multiple phylogenetic clades and that most of the mutations previously associated with colistin resistance are clade associated and present in susceptible isolates, showing no correlation with colistin resistance. In conclusion, the present study contributes with relevant data towards the genetic background of genes involved with colistin resistance deeply rooted across monophyletic groups providing a better understanding of the genes and mutations involved in colistin resistance.
A new jatrophane diterpene, pubescenol (1), known ent-abietane lactones, helioscopinolide A (2) a... more A new jatrophane diterpene, pubescenol (1), known ent-abietane lactones, helioscopinolide A (2) and B (3), and taraxerone, 24-methylenecycloartanol, and vanillin have been isolated from Euphorbia pubescens. Diterpenes 1-3 and previously described pubescene D (4) were shown to be moderate inhibitors of the growth of MCF-7, NCI-H460, and SF-268 human tumor cell lines, whereas compounds 2 and 3 also exhibited significant antibacterial activity against Staphylococcus aureus.
As the personal care industry evolves, formulators are seeking innovative solutions for their for... more As the personal care industry evolves, formulators are seeking innovative solutions for their formulations' needs, and for ingredients that can offer multiple functions within formulations. Considering that essential oils (EOs) may present a wide spectrum of biological activities, the composition, antimicrobial and antioxidant activity of Portuguese Thymbra capitata, Thymus caespititius and Myrtus communis EOs were assessed in order to evaluate them as preservatives and antioxidants in topical emulsions. The in vivo safety application of some emulsions was also tested. T. capitata EO was mainly constituted by carvacrol (73%), whereas α-terpineol (27%), p-cymene (14%) and carvacrol (10%) dominated Th. caespititius EO, and 1,8-cineole (37%) was dominant in M. communis EO. The minimum inhibitory concentration of T. capitata, Th. caespititius and M. communis EOs was 0.4 μg/mL against C. albicans and ranged between 0.4 and 30.7 μg/mL against A. brasiliensis. Gram-positive bacteria were more susceptible to each EO than Gram-negative bacteria. T. capitata EO showed significantly higher antioxidant activity than Th. caespititius and M. communis EOs. EOs incorporated in emulsions showed preservative activity against all microorganisms tested and T. capitata EO emulsions showed powerful reactive oxygen species (ROS) scavenging effects. A safety evaluation study was performed with 0.10% and 0.01% T. capitata EO emulsions according to the Human Repeat Insult Patch Test (HRIPT). All emulsions were considered safe for topical application. T. capitata, Th. caespititius and M. communis significantly improved the microbiological quality of the prepared emulsions and may constitute a powerful alternative to the current preservatives and antioxidants for use in topical formulations.
Infections caused by multiresistant bacterial pathogens are a significant problem worldwide, turn... more Infections caused by multiresistant bacterial pathogens are a significant problem worldwide, turning the search for natural compounds to act as alternatives to antibiotics of major importance. The aim of the present study was to investigate the in vitro antimicrobial activity of 7α-acetoxy-6β-hydroxyroyleanone (1), isolated from Plectranthus grandidentatus (Lamiaceae), and 11 additional royleanone abietane derivatives of 1 (2-12) against important Gram-positive human bacterial pathogens. Results showed that the aromatic and alkylic esters 2, 3 and 5 are more active than 1 against Enterococcus and Staphylococcus (minimum inhibitory concentration (MIC) values ranging from 0.98 to 62.50 µg/mL). Moreover, 7α-acetoxy-6β-hydroxy-12-O-(4-chloro)benzoylroyleanone (2) gave rise to a new antibacterial-prototype (MIC values of 3.91-15.63 µg/mL against Staphylococcus and of 0.98-3.91 µg/mL against Enterococcus). The results showed that the compounds under analysis also present antimicrobial activity against resistant bacteria. The hydrophobic extra-interactions with bacterial targets seem to play an important role on the activity of royleanones derivatives.
Pseudomonas aeruginosa is a Gram-negative opportunistic bacterium that presents resistance to sev... more Pseudomonas aeruginosa is a Gram-negative opportunistic bacterium that presents resistance to several antibiotics, thus, representing a major threat to human and animal health. Phage-derived products, namely lysins, or peptidoglycan-hydrolyzing enzymes, can be an effective weapon against antibiotic-resistant bacteria. Whereas in Gram-positive bacteria, lysis from without is facilitated by the exposed peptidoglycan layer, this is not possible in the outer membrane-protected peptidoglycan of Gram-negative bacteria. Here, we suggest the encapsulation of lysins in liposomes as a delivery system against Gram-negative bacteria, using the model of P. aeruginosa. Bioinformatic analysis allowed for the identification of 38 distinct complete prophages within 66 P. aeruginosa genomes (16 of which newly sequenced) and led to the identification of 19 lysins of diverse sequence and function, 5 of which proceeded to wet lab analysis. The four purifiable lysins showed hydrolytic activity against Gr...
Klebsiella pneumoniae (Kp) bacteria are an increasing threat to public health and represent one o... more Klebsiella pneumoniae (Kp) bacteria are an increasing threat to public health and represent one of the most concerning pathogens involved in life-threatening infections and antimicrobial resistance (AMR). To understand the epidemiology of AMR of Kp in Portugal, we analysed whole genome sequencing, susceptibility testing and other meta data on 509 isolates collected nationwide from 16 hospitals and environmental settings between years 1980 and 2019. Predominant sequence types (STs) included ST15 (n = 161, 32%), ST147 (n = 36, 7%), ST14 (n = 26, 5%) or ST13 (n = 26, 5%), while 31% of isolates belonged to STs with fewer than 10 isolates. AMR testing revealed widespread resistance to aminoglycosides, fluoroquinolones, cephalosporins and carbapenems. The most common carbapenemase gene was blaKPC-3. Whilst the distribution of AMR linked plasmids appears uncorrelated with ST, their frequency has changed over time. Before year 2010, the dominant plasmid group was associated with the extende...
The emergence of carbapenemase-producing Klebsiella pneumoniae strains has triggered the use of o... more The emergence of carbapenemase-producing Klebsiella pneumoniae strains has triggered the use of old antibiotics such as colistin which is consequently driving the emergence of colistin resistance in already multidrug resistant strains that can underlie life-threatening infections. This study analyses the mutational diversity of 22 genes associated with colistin resistance in 140 K. pneumoniae clinical isolates integrated in a high-resolution phylogenetic scenario. Colistin susceptibility was accessed by broth microdilution categorizing 98 isolates as susceptible and 16 as resistant, 10 of which were carbapenemase producers. Across the 22 genes examined, 171 non-synonymous mutations and 9 mutations associated with promoter regions were found. Eighty-five isolates had a truncation and/or deletion in at least one of the 22 genes. However, only seven mutations, the complete deletion of mgrB or insertion sequence mediated disruption were exclusively observed in resistant isolates, 4 of these (mgrBIle13fs, pmrBGly207Asp, phoQHis339Asp and ramAIle28Met) comprising novel mutations potentially involved in colistin resistance. The ISEcp1-blaCTX-M-15::mgrB disruption is herein noteworthy as a mechanism underlying co-resistance to third-generation cephalosporins and colistin. Moreover, the high-resolution phylogenetic context shows that most of the mutational diversity spans across multiple phylogenetic clades and that most of the mutations previously associated with colistin resistance are clade associated and present in susceptible isolates, showing no correlation with colistin resistance. In conclusion, the present study contributes with relevant data towards the genetic background of genes involved with colistin resistance deeply rooted across monophyletic groups providing a better understanding of the genes and mutations involved in colistin resistance.
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