RESUMO
In this study, we isolated a novel lectin from the marine sponge Aiolochroia crassa, named AcrL. The lectin showed a preference for glycans containing sialic acid terminal residues, as indicated by the strongest inhibition with fetuin and bovine submaxillary mucin. Primary structure determination by mass spectrometry revealed that AcrL is a galectin with conserved amino acid residues typically involved in carbohydrate binding. Structural modeling indicated that AcrL adopts a typical galectin ß-sandwich motif, featuring two anti-parallel ß-sheets with five strands each. Docking calculations revealed a carbohydrate-binding site composed of a main site, capable of hosting galactopyranosides, and an extended site, facilitating the binding of complex carbohydrates. AcrL inhibited significant biofilm formation against Staphylococcus aureus, S. epidermidis, and Escherichia coli with concentrations ranging from 500 to 15.6 µg.mL-1 for S. aureus, 7.8 µg.mL-1 for S. epidermidis, and 500 µg.mL-1 for E. coli. Furthermore, when combined with different antibiotics, AcrL potentiated their effect against pathogenic bacteria. The antimicrobial mechanism of AcrL was investigated using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The analysis indicates that AcrL induces damage to the bacterial membrane. These findings underscore the discovery of a novel galectin in a basal organism and the comprehensive biochemical characterization conducted in this research, highlighting the potential of AcrL as a novel antibacterial agent and emphasizing its importance in combating bacterial infections.
Assuntos
Galectinas , Poríferos , Animais , Galectinas/química , Galectinas/farmacologia , Galectinas/metabolismo , Galectinas/isolamento & purificação , Poríferos/química , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Antibacterianos/farmacologia , Antibacterianos/química , Escherichia coli/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Sequência de Aminoácidos , Sinergismo Farmacológico , Simulação de Acoplamento MolecularRESUMO
This data descriptor presents a curated dataset for pathogen detection and identification (Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans) directly from whole-blood samples. The dataset was created using differential cell lysis combined with rapid extraction, digestion, and mass spectrometry-based proteomics. Our method offers a rapid diagnostic alternative to traditional culture, enabling timely disease management, such as sepsis. Highlighting our dataset's uniqueness, it features a three-tier structure: Spectral Libraries of Pathogens for identifying peptide peaks for putative biomarkers; Spiked pathogen in blood MS data for biomarker panel optimization through varied concentration samples; and Parallel Reaction Monitoring (PRM) data from sepsis patients for validating our biomarker panel, achieving 83.3% sensitivity within seven hours without microbial enrichment culture. This dataset serves as a comprehensive reference for bioinformatic tool development and biomarker panel proposals, advancing microbial detection, antimicrobial resistance, and epidemiological studies.
Assuntos
Biomarcadores , Candida albicans , Proteômica , Pseudomonas aeruginosa , Sepse , Staphylococcus aureus , Humanos , Proteômica/métodos , Biomarcadores/sangue , Sepse/sangue , Sepse/diagnóstico , Sepse/microbiologia , Espectrometria de MassasRESUMO
The present study aimed to evaluate the anti-staphylococcal, antibiofilm, cytotoxicity and trypanocidal activity, mechanisms of parasite death and immunomodulatory effect of CrataBL encapsulated into liposomes (CrataBL-Lipo). CrataBL-Lipo were prepared by the freeze-thaw technique and characterized. Anti-staphylococcal and antibiofilm activities of CrataBL and CrataBL-Lipo were evaluated against standard and clinical strains of Staphylococcus aureus susceptible and resistant. Thus, broth microdilution method was performed to determine the Minimum Inhibitory Concentration (MIC). Antibiofilm activity at subinhibitory concentrations was evaluated using the crystal violet staining method. Cytotoxicity of CrataBL-Lipo was verified in L929 fibroblasts and J774A.1 macrophages by determining the inhibitory concentration necessary to kill 50 % of cells (IC50). Trypanocidal activities of CrataBL-Lipo was evaluated in Trypanosoma cruzi and the efficacy was expressed as the concentration necessary to kill 50 % of parasites (EC50). The mechanisms of parasite death and immunomodulatory effect of CrataBL-Lipo were evaluated using flow cytometry analysis. CrataBL-Lipo presented Ø of 101.9 ± 1.3 nm (PDI = 0.245), ζ of +33.8 ± 1.3 mV and %EE = 80 ± 0.84 %. CrataBL-Lipo presented anti-staphylococcal activity (MIC = 0.56 mg/mL to 0.72 mg/mL). CrataBL-Lipo inhibited 45.4 %-75.6 % of biofilm formation. No cytotoxicity of CrataBL-Lipo was found (IC50 > 100 mg/L). CrataBL-Lipo presented EC50 of 1.1 mg/L, presenting autophagy, apoptosis and necrosis as death profile. In addition, CrataBL-Lipo reduced the production of IL-10 and TNF-α levels, causing an immunomodulatory effect. CrataBL-Lipo has a therapeutic potential for the treatment of staphylococcal infections and Chagas disease exhibiting a high degree of selectivity for the microorganism, and immunomodulatory properties.
Assuntos
Antibacterianos , Biofilmes , Lipossomos , Testes de Sensibilidade Microbiana , Staphylococcus aureus , Tripanossomicidas , Trypanosoma cruzi , Biofilmes/efeitos dos fármacos , Trypanosoma cruzi/efeitos dos fármacos , Animais , Camundongos , Staphylococcus aureus/efeitos dos fármacos , Linhagem Celular , Antibacterianos/farmacologia , Tripanossomicidas/farmacologia , Macrófagos/efeitos dos fármacos , Lectinas/farmacologia , Fibroblastos/efeitos dos fármacos , Concentração Inibidora 50 , Sobrevivência Celular/efeitos dos fármacosRESUMO
AIMS: This study aimed to assess the antimicrobial potential of Bp1-AdE, produced by Bacillus pumilus 64-1, and to investigate its mode of action against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). METHODS AND RESULTS: Bp-1AdE, derived from sponge-associated B. pumilus, exhibited bactericidal activity at 1 550 µg ml-1 against S. aureus ATCC29213 and MRSA strains. Light and fluorescence microscopy revealed drastic cell lysis of S. aureus treated with Bp-1AdE. Scanning and transmission electron microscopy suggested that Bp-1AdE disrupts the cytoplasmic membrane. Toxicity assays showed that Bp-1AdE was non-toxic to Tenebrio molitor larvae. Liquid chromatography-mass spectrometry and Global Natural Product Social spectral libraries identified four substances within Bp-1AdE, including aliphatic alcohols [3,4-dipentylhexane-2,5-diol and 1,1'-(4,5-dibutyl-3,6-dimethylcyclohexane-1,2-diyl)bis(ethan-1-one)] and terpenoids (cholic acid and canrenone). CONCLUSIONS: Bp-1AdE demonstrated selective toxicity and bactericidal activity, highlighting its potential for controlling infections caused by multidrug-resistant S. aureus strains.
Assuntos
Antibacterianos , Bacillus pumilus , Staphylococcus aureus Resistente à Meticilina , Testes de Sensibilidade Microbiana , Staphylococcus aureus , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Bacillus pumilus/efeitos dos fármacos , Antibacterianos/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Animais , Farmacorresistência Bacteriana Múltipla , Poríferos/microbiologiaRESUMO
Drug-resistant bacteria such as Escherichia coli and Staphylococcus aureus represent a global health problem that requires priority attention. Due to the current situation, there is an urgent need to develop new, more effective and safe antimicrobial agents. Biotechnological approaches can provide a possible alternative control through the production of new generation antimicrobial agents, such as silver nanoparticles (AgNPs) and bacteriocins. AgNPs stand out for their antimicrobial potential by employing several mechanisms of action that can act simultaneously on the target cell such as the production of reactive oxygen species and cell wall rupture. On the other hand, bacteriocins are natural peptides synthesized ribosomally that have antimicrobial activity and are produced, among others, by lactic acid bacteria (LAB), whose main mechanism of action is to produce pores at the level of the cell membrane of bacterial cells. However, these agents have disadvantages. Nanoparticles also have limitations such as the tendency to form aggregates, which decreases their antibacterial activity and possible cytotoxic effects, and bacteriocins have a narrow spectrum of action, require high doses to be effective, and can be degraded by proteases. Given these limitations, nanoconjugates of these two agents have been developed that can act synergistically in the control of pathogenic bacteria resistant to antibiotics. This review focuses on knowing relevant aspects of the antibiotic resistance of E. coli and S. aureus, the characteristics of these new generation antibacterial agents, and their effect alone or forming nanoconjugates that are more effective against the multiresistant mentioned bacteria.
Assuntos
Antibacterianos , Bacteriocinas , Farmacorresistência Bacteriana Múltipla , Escherichia coli , Nanopartículas Metálicas , Nanocompostos , Prata , Staphylococcus aureus , Bacteriocinas/farmacologia , Bacteriocinas/química , Prata/farmacologia , Prata/química , Escherichia coli/efeitos dos fármacos , Nanopartículas Metálicas/química , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/farmacologia , Antibacterianos/química , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Nanocompostos/química , Testes de Sensibilidade Microbiana , Lactobacillales/metabolismo , Lactobacillales/efeitos dos fármacosRESUMO
OBJECTIVES: Photodynamic inactivation (PDI) is a powerful technique for eradicating microorganisms, and our group previously demonstrated its effectiveness against planktonic cultures of Staphylococcus aureus bacteria using 5,10,15,20-tetrakis[4-(3-N,N-dimethylaminopropoxy)phenyl]porphyrin (TAPP) and visible light irradiation. However, biofilms exhibit a lower sensitivity to PDI, mainly due to limited penetration of the photosensitizer (PS). In the context of emerging antibacterial strategies, near-infrared treatments (NIRTs) have shown promise, especially for combating resistant strains. NIRT can act either through photon absorption by water, causing a thermal effect on bacteria, or by specific chromophores without a significant temperature increase. Our objective was to enhance biofilm sensitivity to TAPP-PDI by pretreatment with NIRT. This combined approach aims to disrupt biofilms and increase the efficacy of TAPP-PDI against bacterial biofilms. MATERIALS AND METHODS: In vitro biofilm models of S. aureus RN6390 were utilized. NIRTs involved a 980 nm laser (continuous mode, 7.5 W/cm2, 30 s, totaling 225 J/cm2) post-TAPP exposure to enhance photosensitizer accumulation. Subsequent visible light irradiation at 180 J/cm2 was employed to perform PDI. Colony-forming unit counts evaluated the synergistic effect on bacterial viability. Scanning electron microscopy visualized the architectural changes in the biofilm structure. TAPP was extracted from bacteria to estimate the impact of NIRT on biofilm penetration. RESULTS: Using in vitro biofilm models, NIRT application following biofilm exposure to TAPP increased PS accumulation per bacteria. Under these conditions, NIRT induced a transient increase in the temperature of PBS to 46.0 ± 2.6°C (ΔT = 21.5°C). Following exposure to visible light, a synergistic effect emerged, yielding a substantial 4.4 ± 0.1-log CFU reduction. In contrast, the PDI and NIRT treatments individually caused a decrease in viability of 0.9 ± 0.1 and 0.8 ± 0.2-log respectively. Interestingly, preheating TAPP-PBS to 46°C had no significant impact on TAPP-PDI efficacy, suggesting the involvement of thermal and nonthermal effects of NIR action. In addition to the enhanced TAPP penetration, NIRT dispersed the biofilms and induced clefts in the biofilm matrix. CONCLUSION: Our findings suggest that NIR irradiation serves as a complementary treatment to PDI. This combined strategy reduces bacterial numbers at lower PS concentrations than standalone PDI treatment, highlighting its potential as an effective and resource-efficient antibacterial approach.
Assuntos
Biofilmes , Fotoquimioterapia , Fármacos Fotossensibilizantes , Staphylococcus aureus , Biofilmes/efeitos dos fármacos , Biofilmes/efeitos da radiação , Staphylococcus aureus/efeitos dos fármacos , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Raios Infravermelhos , Porfirinas/farmacologiaRESUMO
Liquid smoke, an alternative to traditional wood burning smoking, enhances product value by imparting desirable characteristics such as aroma, flavor, and color. Furthermore, it contains components with inherent antimicrobial and antioxidant properties. This study compares the effects of liquid smoke and conventional smoking methods in bacon processing. Over a 90-day storage period at 22°C, physical-chemical stability, sensory attributes, and microbiological characteristics of the bacon were evaluated. The antimicrobial and antioxidant properties of liquid smoke were assessed. Liquid smoke exhibited antioxidant activity, with an inhibitory concentration (IC50) value of 0.19 mg/mL, indicating the concentration of the extract needed to inhibit 50% of DPPH (2,2'-diphenyl-1-picrylhydrazyl) radicals. Moreover, it demonstrated antimicrobial effects against Escherichia coli, Salmonella choleraesius, Staphylococcus aureus, and Listeria monocytogenes, with a minimum bactericidal concentration ranging from 7.5% to 10%. Throughout the storage, bacon treated with liquid smoke showed no signs of rancid odor, supported by thiobarbituric acid reactive substances values below 0.85 mg MDA/kg (where MDA is malondialdehyde). The utilization of liquid smoke yielded visually attractive bacon with enhanced color attributes, including a distinct yellow and red hue, as well as increased luminosity and brightness, surpassing the effects of traditional smoke. Remarkably, liquid smoke application significantly reduced processing time from 30 h to approximately 5 h, leading to substantial cost savings in the processing phase.
Assuntos
Antioxidantes , Listeria monocytogenes , Produtos da Carne , Fumaça , Staphylococcus aureus , Antioxidantes/farmacologia , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Produtos da Carne/microbiologia , Produtos da Carne/análise , Staphylococcus aureus/efeitos dos fármacos , Humanos , Escherichia coli/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Manipulação de Alimentos/métodos , Anti-Infecciosos/farmacologia , Antibacterianos/farmacologia , Paladar , Armazenamento de Alimentos/métodos , Conservação de Alimentos/métodos , CorRESUMO
The incorporation of bactericidal properties into textiles is a widely sought-after aspect, and silver nanoparticles (AgNPs) can be used for this. Here, we evaluate a strategy for incorporating AgNPs into a cotton fabric. For this purpose, a bactericidal textile coating based on a composite of AgNPs and kappa-carrageenan (k-CA) was proposed. The composite was obtained by heating the silver precursor (AgNO3) directly in k-CA solution for green synthesis and in situ AgNPs stabilization. Cotton substrates were added to the heated composite solution for surface impregnation and hydrogel film formation after cooling. Direct synthesis of AgNPs on a fabric was also tested. The results showed that the application of a coating based on k-CA/AgNPs composite can achieve more than twice the silver loading on the fabric surface compared to the textile subjected to direct AgNPs incorporation. Furthermore, silver release tests in water showed that higher Ag+ levels were reached for k-CA/AgNPs-coated cotton. Therefore, inoculation tests with the bacteria Staphylococcus aureus (SA) using the agar diffusion method showed that samples covered with the composite resulted in significantly larger inhibition halos. This indicated that the use of the composite as a coating for cotton fabric improved its bactericidal activity against SA.
Assuntos
Antibacterianos , Carragenina , Fibra de Algodão , Teste de Materiais , Nanopartículas Metálicas , Testes de Sensibilidade Microbiana , Tamanho da Partícula , Prata , Staphylococcus aureus , Prata/química , Prata/farmacologia , Carragenina/química , Carragenina/farmacologia , Nanopartículas Metálicas/química , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/síntese química , Têxteis , Propriedades de SuperfícieRESUMO
The New World screwworm, Cochliomyia hominivorax Coquerel (Diptera: Calliphoridae), was officially eliminated from Costa Rica in 2000, but it was reintroduced in 2023. A myiasis by C. hominivorax in a 71-year-old man with a 4-month history of foot hyperkeratosis and interdigital ulcers is reported. The myiasis was detected before sampling for bacterial culture. Approximately 160 first- and second-instar larvae were recovered and identified as C. hominivorax. Morphological identification was based mainly on characteristics of the cephalopharyngeal skeleton, spiracles, and pigmented dorsal tracheal trunks. Sequencing of a cytochrome c oxidase subunit I gene fragment confirmed the identity. The ulcers healed after extraction of the larvae and ciprofloxacin treatment of a concurrent Staphylococcus aureus and Pseudomonas aeruginosa infection. Given the reintroduction of C. hominivorax in Costa Rica and the risk of northward expansion, this report highlights its impact on public health and calls for awareness among clinicians and healthcare practitioners.
Assuntos
Calliphoridae , Humanos , Costa Rica/epidemiologia , Masculino , Animais , Idoso , Infecção por Mosca da Bicheira/epidemiologia , Larva , Miíase/diagnóstico , Miíase/parasitologia , Miíase/epidemiologia , Antibacterianos/uso terapêutico , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/epidemiologia , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/genética , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/epidemiologia , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/genética , Ciprofloxacina/uso terapêutico , DípterosRESUMO
Dressings should protect wounds, promote healing, absorb fluids, and maintain moisture. Bacterial cellulose is a biopolymer that stands out in biomaterials due to its high biocompatibility in several applications. In the area of dressings, it is already marketed as an alternative to traditional dressings. However, it lacks any intrinsic activity; among these, the need for antimicrobial activity in infected wounds stands out. We developed a cationic cellulose film by modifying cellulose with 1-(5-carboxypentyl)pyridin-1-ium bromide, enhancing its wettability (contact angle: 26.6°) and water retention capacity (2714.37 %). This modified film effectively retained oxacillin compared to the unmodified control. Liposomal encapsulation further prolonged oxacillin release up to 11 days. Both oxacillin-loaded films and liposomal formulations demonstrated antimicrobial activity against Staphylococcus aureus. Our findings demonstrate the potential of chemically modified cellulose as a platform for controlled anionic antibiotics and/or their formulations delivery in wound care.
Assuntos
Antibacterianos , Bandagens , Celulose , Preparações de Ação Retardada , Liberação Controlada de Fármacos , Lipossomos , Oxacilina , Staphylococcus aureus , Celulose/química , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Oxacilina/administração & dosagem , Oxacilina/farmacologia , Oxacilina/química , Cátions/química , Ânions/química , Cicatrização/efeitos dos fármacos , MolhabilidadeRESUMO
Herein, four different grafted chitosans were synthesized by covalent attachment of glycine, L-arginine, L-glutamic acid, or L-cysteine to the chitosan chains. All products were subsequently permethylated to obtain their corresponding quaternary ammonium salts to enhance the inherent antimicrobial properties of native chitosan. In all cases, transparent hydrogels with the following remarkable characteristics were obtained: i) high-water absorption capacity (32-44 g H2O per g of polymer), ii) viscoelastic behavior at low deformations, iii) flexibility when subjected to deformations and iv) stability over long time scales. All the permethylated derivatives successfully inhibited 100 % of the growth of S. aureus. They also exhibited higher antimicrobial activity against E. coli than native chitosan. The structure of the chemically crosslinked products was more stable under external perturbations than that of the physically crosslinked ones. Between the chemically crosslinked products, the permethylated glutamic acid-grafted chitosan exhibited a noteworthy higher water absorption capacity with respect to that modified with cysteine, which makes it the most promising material for various industrial applications, including biomedical and food industries. Regarding biomedical applications, this derivative met the required physicochemical criteria for wound dressings, which encourages the pursuit of biological studies necessary to ensure the safety of its use for this application.
Assuntos
Bandagens , Quitosana , Hidrogéis , Quitosana/química , Quitosana/farmacologia , Hidrogéis/química , Hidrogéis/farmacologia , Hidrogéis/síntese química , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Antibacterianos/farmacologia , Antibacterianos/química , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Água/química , Cicatrização/efeitos dos fármacos , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologiaRESUMO
Importance: Individuals with atopic dermatitis are frequently colonized and infected with Staphylococcus aureus. Empirical antibiotic therapy for individuals with atopic dermatitis is common, but data about the antimicrobial susceptibility profiles of S aureus strains isolated from these individuals are scarce for those living in particular geographic areas. Objective: To determine the antimicrobial susceptibility of S aureus from individuals with atopic dermatitis and analyze differences according to the income level of the country of origin and the data collection period. Data Sources: A meta-analysis of the literature was performed from the inception of the included databases (MEDLINE, Embase, Web of Science, Scopus, and Cochrane) to June 20, 2023, using predetermined Medical Subject Headings. Study Selection: Studies were included if they reported antibiotic susceptibility profiles of 1 or more S aureus cutaneous isolates from individuals with atopic dermatitis. Articles written in English, Spanish, French, or German were included. Data Extraction and Synthesis: Working in pairs, 6 of the authors conducted the data extraction. The guidelines from the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) were followed. Main Outcomes and Measures: The outcome of interest was antimicrobial susceptibility. Results: A total of 61 studies reported 4091 S aureus isolates from individuals with atopic dermatitis. For 4 of the 11 commonly used antibiotics (36.4%), antimicrobial susceptibility was 85% or less, including for methicillin (binomial proportion, 0.85 [95% CI, 0.76-0.91]), erythromycin (binomial proportion, 0.73 [95% CI, 0.61-0.83]), fusidic acid (binomial proportion, 0.80 [95% CI, 0.62-0.91]), and clindamycin (binomial proportion, 0.79 [95% CI, 0.65-0.89]). Most studies (46; 75.4%) were conducted in high-income countries. Antimicrobial susceptibility to erythromycin, methicillin, and trimethoprim and sulfamethoxazole was significantly lower in lower middle-income countries and upper middle-income countries. Regarding the temporal trends, 33 studies (54.1%) reported data collected from 1998 to 2010. Antimicrobial susceptibility patterns have not changed over time. Conclusions and Relevance: In this systematic review and meta-analysis, antimicrobial susceptibility of S aureus to ß-lactams, erythromycin, clindamycin, and fusidic acid may be suboptimal for empirical use in individuals with atopic dermatitis. Significant differences in antimicrobial susceptibility patterns were found in high-income countries and in lower middle-income countries and upper middle-income countries for some antibiotics.
Assuntos
Antibacterianos , Dermatite Atópica , Infecções Cutâneas Estafilocócicas , Staphylococcus aureus , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Dermatite Atópica/microbiologia , Dermatite Atópica/tratamento farmacológico , Testes de Sensibilidade Microbiana , Infecções Cutâneas Estafilocócicas/microbiologia , Infecções Cutâneas Estafilocócicas/tratamento farmacológico , Infecções Cutâneas Estafilocócicas/epidemiologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificaçãoRESUMO
The cefazolin inoculum effect (CzIE) has been associated with poor clinical outcomes in patients with methicillin-susceptible Staphylococcus aureus (MSSA) infections. We aimed to investigate the point prevalence of the CzIE among nasal colonizing MSSA isolates from ICU patients in a multicenter study in Colombia (2019-2023). Patients underwent nasal swabs to assess for S. aureus colonization on admission to the ICU, and some individuals had follow-up swabs. We performed cefazolin MIC by broth microdilution using standard and high inoculum and developed a modified nitrocefin-based rapid test to detect the CzIE. Whole-genome sequencing was carried out to characterize BlaZ types and allotypes, phylogenomics, and Agr-typing. A total of 352 patients were included; 46/352 (13%) patients were colonized with S. aureus and 22% (10/46) and 78% (36/46) with MRSA and MSSA, respectively. Among 36 patients who contributed with 43 MSSA colonizing isolates, 21/36 (58%) had MSSA exhibiting the CzIE. BlaZ type A and BlaZ-2 were the predominant type and allotype in 56% and 52%, respectively. MSSA belonging to CC30 were highly associated with the CzIE, and single-nucleotide polymorphism (SNP) analyses supported possible transmission of MSSA exhibiting the CzIE among some patients of the same unit. The modified nitrocefin rapid test had 100%, 94.4%, and 97.7% sensitivity, specificity, and accuracy, respectively. We found a high point prevalence of the CzIE in MSSA colonizing the nares of critically ill patients in Colombia. A modified rapid test was highly accurate in detecting the CzIE in this patient population.
Assuntos
Antibacterianos , Cefazolina , Unidades de Terapia Intensiva , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas , Humanos , Colômbia/epidemiologia , Cefazolina/farmacologia , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/tratamento farmacológico , Antibacterianos/farmacologia , Feminino , Masculino , Pessoa de Meia-Idade , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Prevalência , Nariz/microbiologia , Idoso , Sequenciamento Completo do Genoma , AdultoRESUMO
OBJECTIVES: Investigate whether and which synoviocytes would acquire trained immunity characteristics that could exacerbate joint inflammation following a secondary Staphylococcus aureus infection. METHODS: Lipopolysaccharide (LPS) and S. aureus were separately or double injected (21 days of interval) into the tibiofemoral joint cavity of male C57BL/6 mice. At different time points after these stimulations, mechanical nociception was analyzed followed by the analysis of signs of inflammation and damage in the affected joints. The trained immunity markers, including the glycolytic and mTOR pathway, were analyzed in whole tissue or isolated synoviocytes. A group of mice was treated with Rapamycin, an mTOR inhibitor before LPS or S. aureus stimulation. RESULTS: The double LPS - S. aureus hit promoted intense joint inflammation and damage compared to single joint stimulation, including markers in synoviocyte activation, production of proinflammatory cytokines, persistent nociception, and bone damage, despite not reducing the bacterial clearance. The double LPS - S. aureus hit joints increased the synovial macrophage population expressing CX3CR1 alongside triggering established epigenetic modifications associated with trained immunity events in these cells, such as the upregulation of the mTOR signaling pathway (p-mTOR and HIF1α) and the trimethylation of histone H3. Mice treated with Rapamycin presented reduced CX3CR1+ macrophage activation, joint inflammation, and bone damage. CONCLUSIONS: There is a trained immunity phenotype in CX3CR1+ synovial macrophages that contributes to the exacerbation of joint inflammation and damage during septic arthritis caused by S. aureus.
Assuntos
Lipopolissacarídeos , Macrófagos , Camundongos Endogâmicos C57BL , Infecções Estafilocócicas , Staphylococcus aureus , Serina-Treonina Quinases TOR , Animais , Masculino , Infecções Estafilocócicas/imunologia , Macrófagos/imunologia , Serina-Treonina Quinases TOR/imunologia , Staphylococcus aureus/imunologia , Receptor 1 de Quimiocina CX3C/metabolismo , Membrana Sinovial/imunologia , Membrana Sinovial/patologia , Artrite Infecciosa/imunologia , Artrite Infecciosa/microbiologia , Camundongos , Citocinas/imunologia , Citocinas/metabolismo , Sirolimo/farmacologia , Inflamação/imunologia , Sinoviócitos/imunologia , Sinoviócitos/efeitos dos fármacos , Nociceptividade/efeitos dos fármacos , Imunidade TreinadaRESUMO
The aim of this study was to evaluate the activation pathway(s) triggered by Minthostachys verticillata essential oil (EO) in bovine mammary epithelial cells (MAC-T) challenged with a strain of bovine Staphylococcus aureus. MAC-T cells were stimulated with EO, S. aureus or pre-treated with EO and then challenged with S. aureus. Cytokine's release was measured by ELISA. The mRNA for TLR2, TLR4, NOD2, MyD88 and NFκB was quantified by RT-qPCR. S. aureus adherence and internalization was also evaluated. MAC-T cells stimulated with S. aureus synthesized high levels of IL-1ß and IL-6 were kept up to 48 h, while IL-4 levels were not altered. Cells pre-treated with EO for 2 and 6 h and then challenged with S. aureus showed a significant increase of IL-1ß and IL-6. However, in these cells, a decrease in IL-1ß and IL-6 levels and an increase of IL-4 values was observed from 24 h. No significant increase in the expression levels of TLR2 or NOD2 was detected in all stimulated cells. However, the expression of TLR4, MyD88 and NFκB was increased in cells stimulated with S. aureus at 2 and 6 h as well as in cells pre-treated with EO between 2 and 6 h and then challenged with S. aureus. The NFκB expression levels was similar to control at 24 h in all stimulated cells, although pro-inflammatory cytokine levels and TLR4 and MyD88 expression levels remained high in cells stimulated with S. aureus. This results suggested the activation of other pathways independent of MyD88 by the pathogen that involucrated the activation of others transcription factors. Pre-treatment with EO during 2, 6 and 24 h did not affect S. aureus adherence but decreased its internalization. In conclusion, pre-treatment with EO increased the IL-1ß and IL-6 synthesis during the first hours post-challenged with S. aureus up-regulating TLR4/MyD88/NFκB pathway. Furthermore, EO increased the IL-4 levels from 6 to 24 h down-regulating the NFκB and possibly other transcription factors activated by the pathogen, which decreased its internalization into MAC-T cells.
Assuntos
Citocinas , Fator 88 de Diferenciação Mieloide , NF-kappa B , Óleos Voláteis , Staphylococcus aureus , Receptor 4 Toll-Like , Animais , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Óleos Voláteis/farmacologia , Bovinos , Citocinas/metabolismo , Citocinas/genética , NF-kappa B/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Fator 88 de Diferenciação Mieloide/genética , Receptor 4 Toll-Like/metabolismo , Receptor 4 Toll-Like/genética , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Transdução de Sinais/efeitos dos fármacos , Feminino , Glândulas Mamárias Animais/efeitos dos fármacosRESUMO
Cotton fabrics with zinc oxide (ZnO) coating are of significant interest due to their excellent antibacterial performance. Thus, they are widely in demand in the textile industry due to their medical and hygienic properties. However, conventional techniques used to deposit ZnO on fabric require long processing times in deposition, complex and expensive equipment, and multiple steps for deposition, such as a separate process for nanoparticle synthesis and subsequent deposition on fabric. In this study, we proposed a new method for the deposition of ZnO on fabric, using cathodic cage plasma deposition (CCPD), which is commonly used for coating deposition on conductor materials and is not widely used for fabric due to the temperature sensitivity of the fabric. The effect of gas composition, including argon and a hydrogen-argon mixture, on the properties of ZnO deposition is investigated. The deposited samples are characterized by XRD, SEM, EDS, photocatalytic, and antibacterial performance against Staphylococcus aureus and Pseudomonas aeruginosa bacteria. It is observed that ZnO-deposited cotton fabric exhibits excellent photocatalytic degradation of methylene blue and antibacterial performance, specifically when a hydrogen-argon mixture is used in CCPD. The results demonstrate that CCPD can be used effectively for ZnO deposition on cotton fabric; this system is already used in industrial-scale applications and is thus expected to be of significant interest to garment manufacturers and hospitals.
Assuntos
Antibacterianos , Fibra de Algodão , Staphylococcus aureus , Óxido de Zinco , Óxido de Zinco/química , Óxido de Zinco/farmacologia , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/síntese química , Catálise , Staphylococcus aureus/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Eletrodos , Gases em Plasma/química , Gases em Plasma/farmacologia , Processos FotoquímicosRESUMO
The rapid resistance developed by pathogenic microorganisms against the current antimicrobial pool represents a serious global public health problem, leading to the search for new antibiotic agents. The scorpion Tityus stigmurus, an abundant species in Northeastern Brazil, presents a rich arsenal of bioactive molecules in its venom, with high potential for biotechnological applications. However, venom cytotoxicity constitutes a barrier to the therapeutic application of its different components. The objective of this study was to produce T. stigmurus-venom-loaded cross-linked chitosan nanoparticles (Tsv/CN) at concentrations of 0.5% and 1.0% to improve their biological antimicrobial activity. Polymeric nanoparticles were formed with a homogeneous particle size and spherical shape. Experimental formulation parameters were verified in relation to mean size (<180 nm), zeta potential, polydispersity index and encapsulation efficiency (>78%). Tsv/CN 1.0% demonstrated an ability to increase the antimicrobial venom effect against Staphylococcus aureus bacteria, exhibiting an MIC value of 44.6 µg/mL. It also inhibited different yeast species of the Candida genus, and Tsv/CN 0.5% and 1.0% led to a greater inhibitory effect of C. tropicalis and C. parapsilosis strains, presenting MIC values between 22.2 and 5.5 µg/mL, respectively. These data demonstrate the biotechnological potential of these nanosystems to obtain a new therapeutic agent with potential antimicrobial activity.
Assuntos
Quitosana , Testes de Sensibilidade Microbiana , Nanopartículas , Venenos de Escorpião , Escorpiões , Quitosana/química , Quitosana/farmacologia , Nanopartículas/química , Animais , Venenos de Escorpião/química , Venenos de Escorpião/farmacologia , Escorpiões/química , Staphylococcus aureus/efeitos dos fármacos , Anti-Infecciosos/farmacologia , Anti-Infecciosos/química , Candida/efeitos dos fármacos , Tamanho da Partícula , Antibacterianos/farmacologia , Antibacterianos/química , Animais PeçonhentosRESUMO
This study proposes an affordable plasma device that utilizes a parallel-plate dielectric barrier discharge geometry with a metallic mesh electrode, featuring a straightforward 3D-printed design. Powered by a high-voltage supply adapted from a cosmetic plasma device, it operates on atmospheric air, eliminating the need for gas flux. Surface modification of polyethylene treated with this device was characterized and showed that the elemental composition after 15 min of plasma treatment decreased the amount of C to ~80 at% due to the insertion of O (~15 at%). Tested against Candida albicans and Staphylococcus aureus, the device achieved a reduction of over 99% in microbial load with exposure times ranging from 1 to 10 min. Simultaneously, the Vero cell viability remained consistently high, namely between 91% and 96% across exposure times. These results highlight this device's potential for the surface modification of materials and various infection-related applications, boasting affordability and facilitating effective antimicrobial interventions.
Assuntos
Candida albicans , Gases em Plasma , Staphylococcus aureus , Propriedades de Superfície , Candida albicans/efeitos dos fármacos , Gases em Plasma/química , Gases em Plasma/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Animais , Células Vero , Chlorocebus aethiops , Viabilidade Microbiana/efeitos dos fármacos , Polímeros/químicaRESUMO
The discovery that the lung harbors a diverse microbiome, as revealed by next-generation sequencing, has significantly altered our understanding of respiratory health and disease. Despite the association between the lung microbiota and disease, the nature of their relationship remains poorly understood, and culture isolation of these microorganisms could help to determine their role in lung physiology. Current procedures for processing samples from the lower respiratory tract have been shown to affect the viability of microorganisms, so it is crucial to develop new methods to improve their survival. This study aimed to improve the isolation and characterization of lung microorganisms using a bead-beating homogenization method in a mouse model. Microsphere diameter and bead-beating time affected the survival of the microorganisms (E. coli, S. aureus and C. albicans). Using 2.3 mm diameter microspheres for 60 s of bead-beating promoted the survival of both bacteria and yeast strains. After intratracheal instillation of these microorganisms in mice, approximately 70% of the cells were recovered after the tissue homogenization. To assess the efficiency of the proposed method, the diversity of bacteria was compared between the homogenate and lung tissue samples. Ninety-one genera were detected in the lung tissue, and 63 in the homogenate. Bacterial genera detected in the homogenate represented 84% of the total abundance of the microbiota identified in the lung tissue. Taken together, these results demonstrate that the tissue homogenization process developed in this study recovered the majority of the microorganisms present in the lung. This study presents a bead-beating homogenization method for effective cultivation of lung tissue microorganisms, which may help to improve the understanding of host-microbe interactions in the lung.
Assuntos
Pulmão , Microbiota , Animais , Pulmão/microbiologia , Camundongos , Microesferas , Staphylococcus aureus , Candida albicans/isolamento & purificação , Escherichia coli/isolamento & purificação , Bactérias/isolamento & purificação , Bactérias/classificação , Bactérias/genéticaRESUMO
BACKGROUND: In the pediatric population, Staphylococcus aureus infections are responsible for increased morbidity and mortality, length of hospitalization and the cost of inpatient treatment. The aim of this study is to describe the antimicrobial resistance profile of S. aureus isolated in clinical specimens from pediatric patients admitted to a tertiary hospital in Rio de Janeiro, Brazil. METHODS: Culture reports and medical records of hospitalized patients under 18 years of age with S. aureus infections between January 2015 and December 2022 were retrospectively analyzed. Information was collected on recent antibiotic use, previous hospital admission, inpatient unit, clinical specimen, time of infection (community or nosocomial), classification according to susceptibility to methicillin (methicillin sensitive - MSSA or methicillin resistant - MRSA) and sensitivity to other antimicrobials. We analyzed the distribution of the sensitivity profile of S. aureus infections over the 7 years evaluated in the study. RESULTS: Were included 255 unique clinical episodes, among which the frequencies of MSSA and MRSA were 164 (64%) and 91 (36%), respectively. Over the 7 years evaluated, there was stability in the prevalence percentage, with a predominance of MSSA in the range of 60 to 73.3%, except in 2020, when there was a drop in the prevalence of MSSA (from 73.3% in 2019 to 52.5%) with an increase in MRSA (from 26.7% in 2019 to 47.5%). Ninety-seven (38%) infections were community-acquired and 158 (62%) were healthcare-associated. The main clinical specimens collected were blood cultures (35.2%) and wound secretions (17%). The MRSA isolates presented percentages of sensitivity to trimethoprim-sulfamethoxazole from 90.4 to 100%, and to clindamycin from 77 to 89.8% in MRSA healthcare associated and MRSA community respectively. CONCLUSION: There was a constant predominance in the prevalence of MSSA with percentage values ââmaintained from 2015 to 2022, except in 2020, in which there was a specific drop in the prevalence of MSSA with an increase in MRSA. MSSA infections are still predominant in the pediatric population, but MRSA rates also present significant values, including in community infections, and should be considered in initial empiric therapy.