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A promising method for additive manufacturing that makes it possible to produce intricate and personalized parts is selective laser melting (SLM). However, the mechanical properties of as-corroded SLM parts are still areas of concern. This research investigates the mechanical behavior of SLM parts that are exposed to a saline environment containing a 3.5% NaCl solution for varying lengths of time. The exposure times chosen for this study were 10 days, 20 days, and 30 days. The results reveal that the tensile strength of the parts is significantly affected by the duration of exposure. Additionally, the study also examined the influence of porosity on the corrosion behavior of the parts. The analysis included studying the mass loss of the parts over time, and a regression analysis was conducted to analyze the relationship between exposure time and mass loss. In addition, the utilization of scanning electron microscopy (SEM) and X-ray photo spectroscopy (XPS) techniques yielded valuable insights into the fundamental mechanisms accountable for the observed corrosion and mechanical behavior. It was found that the presence of corrosion products (i.e., oxide layer) and pitting contributed to the degradation of the SLM parts in the saline environment. This research emphasizes the importance of considering part thickness in the design of SLM components for corrosive environments and provides insights for enhancing their performance and durability.
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Concept of microorganisms has largely been perceived from their pathogenic view point. Nevertheless, it is being gradually revisited in terms of its significance to human health and now appears to be the most dominant force that shapes the immune system of the human body and also determines an individual's predisposition to diseases. Human inhabits bacterial diversity (which is predominant among all microbial communities in human body) occupying 0.3% of body mass, known as microbiota. On birth, a part of microbiota that child obtains is essentially a mother's legacy. So, the review was initiated with this critical topic of microbiotal inheritance. Since, each body site has distinct physiological specifications; therefore, they contain discrete microbiome composition that has been separately discussed along with dysbiosis-induced pathologies originating in different body organs. Factors affecting microbiome composition and may cause dysbiosis like antibiotics, delivery, feeding method etc. as well as the strategies that immune system adopts to prevent dysbiosis have been highlighted. We also tried to bring into attention the topic of dysbiosis induced biofilms, that enables cohort to survive stresses, evolve, disseminate and infection resurgence that is still in dormancy. Eventually, we put spotlight on microbiome significance in medical therapeutics. We didn't merely confine article to gut microbiota, that is being studied more extensively. Numerous community forms at diverse body sites are inter-related, and being exposed to awfully variable perturbations appear to be challenging to evaluate perturbation risks holistically. All aspects have been elaborately discussed to achieve a global depiction of human microbiota in order to meet urgent necessity for protocol standardisation. Demonstrates that environmental challenges (antibiotic use, alterations in diet, stress, smoking etc.) might cause dysbiosis i.e. transition of healthy microbiome composition to the one in which pathogenic microorganisms become more abundant, and eventually results in an infected state.
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Microbioma Gastrointestinal , Microbiota , Humanos , Bacterias/genética , Biopelículas , Disbiosis/microbiología , Microbiota/fisiología , Recién NacidoRESUMEN
Bacterial cells are surrounded by a peptidoglycan (PG) cell wall, which is essential for cell integrity and intrinsic biogenesis pathways; hence, the cell wall is a potential target for several antibiotics. Among several lytic transglycosylases (LTs), the mltG gene plays a crucial role in the synthesis of peripheral PG. It localises the re-modelled PGs for septum formation and cleavage across the bacterial cell wall during daughter cells separation. However, the role of mltG gene in bacterial virulence, particularly in Gram-positive bacteria during dentine biofilm and caries development, has remained unexplored. Hence, we exploited Gram-positive Streptococcus mutans cells for the very first time to construct a mltG knock-out bacterial strain, e.g., ΔmltG S. mutans. Systematic comparative investigations revealed that doubling time (Td), survival, enzymatic efficiencies, pH tolerance, bio-synthesise of lipid, proteins and DNA, biofilm formation and dentine lesions were significantly (p < 0.001) compromised in case of ΔmltG S. mutans than wild type strain. The qRT-PCR based gene expression profiling revealed that transcriptional expression of critically important genes involved in biofilm, metabolism, and stress response were dysregulated in the mutant. Besides, an incredible reduction in dentine caries development was found in the molar teeth of Wistar rats and also in human extracted teeth. Concisely, these trends obtained evidently advocated the fact that the deletion of mltG gene can be a potential target to impair the S. mutans virulence through severe growth retardation, thereby reducing the virulence potential of S. mutans.
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Owing to unique nano-scale properties, TiO2-NPs (T-NPs) are employed as food-quality enhancers in >900 processed food products. Whereas, epigallocatechin-3-gallate (EGCG), a green tea polyphenol is consumed in traditional brewed tea, globally. Taken together, we aimed to investigate whether human gastric-acid digested T-NPs and complex tea catechins yield ionic species (Ti4+, Ti3+ etc.) and active EGCG forms to meet favourable conditions for in vivo bio-genesis of EGCG-coronated TiO2-NPs (ET-NPs) in human gut. Secondly, compared to bare-surface micro and nano-scale TiO2, i.e., T-MPs and T-NPs, respectively, how EGCG coronation on ET-NPs in the gut facilitates the modulation of intrinsic propensity of internalization of TiO2 species into bacteria, body-organs, and gut-microbiota (GM), and immune system. ET-NPs were synthesized in non-toxic aqueous solution at varied pH (3-10) and characterised by state-of-the-arts for crystallinity, surface-charge, EGCG-encapsulation, stability, size, composition and morphology. Besides, flow-cytometry (FCM), TEM, EDS, histopathology, RT-PCR, 16S-rRNA metagenomics and ELISA were also performed to assess the size and surface dependent activities of ET-NPs, T-NPs and T-MPs vis-a-vis planktonic bacteria, biofilm, GM bacterial communities and animal's organs. Electron-microscopic, NMR, FTIR, DLS, XRD and EDS confirmed the EGCG coronation, dispersity, size-stability of ET-NPs, crystallinity and elemental composition of ET-NPs-8 and T-NPs. Besides, FCM, RT-PCR, 16S-rRNA metagenomics, histopathology, SEM and EDS analyses exhibited that EGCG coronation in ET-NPs-8 enhanced the penetration into body organs (i.e., liver and kidney etc.) and metabolically active bacterial communities of GM.
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Té , Titanio , Animales , Humanos , Té/química , AlimentosRESUMEN
A simple, facile, moist-heating (e.g., autoclave), one-step procedure for EGCG-mediated biosynthesis of narrow-size magnetic iron oxide (α-Fe2O3) nanoparticles (EGCG-MINPs) was developed. The influence of pH of the reaction mixture over the size distribution of as-synthesized EGCG-MINPs was investigated systematically by employing UV-visible (UV-vis) spectroscopy and dynamic light scattering (DLS)-based hydrodynamic size, surface charge (zeta-potential), and polydispersity index (PDI). The FE-SEM, TEM, and XRD characterizations revealed that the EGCG-MINPs synthesized at pH 5.0 were in the size range of 6.20-16.7 nm and possess well-crystalline hexagonal shaped nanostructures of hematite (α-Fe2O3) crystal phase. The role of EGCG in Fe3+ ion reduction and EGCG-MINP formation was confirmed by FTIR analysis. The VSM analysis has revealed that EGCG-MINPs were highly magnetic nanostructures with the hysteretic feature of saturation magnetization (Ms), remanent magnetization (Mr), and coercivity (Hc) as 33.64 emu/g, 12.18 emu/g, and 0.33 Oe, respectively. Besides, significant (p < 0.001) dose-dependent (250-1000 µg/mL) antibacterial and antibiofilm activities against the NDM-1-producing Gram-negative Escherichia coli (AK-33), Klebsiella pneumoniae (AK-65), Pseudomonas aeruginosa (AK-66), and Shigella boydii (AK-67) bacterial isolates warranted the as-synthesized EGCG-MINPs as a promising alternative for clinical management of chronic bacterial infections in biomedical settings. In addition, molecular docking experiments revealed that compared to free Fe3+ and EGCG alone, the EGCG-MINPs or Fe-EGCG complex possess significantly high binding affinity toward HSA and hence can be considered as promising biocompatible nanodrug carriers in in vivo drug delivery systems.
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Multi-drug resistant (MDR) bacterial cells embedded in biofilm matrices can lead to the development of chronic cariogenesis. Here, we isolated and identified three Gram-positive MDR oral cocci, (1) SJM-04, (2) SJM-38, and (3) SJM-65, and characterized them morphologically, biochemically, and by 16S rRNA gene-based phylogenetic analysis as Georgenia sp., Staphylococcus saprophyticus, and Rothia mucilaginosa, respectively. These three oral isolates exhibited antibiotic-resistance against nalidixic acid, tetracycline, cefuroxime, methicillin, and ceftazidime. Furthermore, these Gram positive MDR oral cocci showed significant (p < 0.05) variations in their biofilm forming ability under different physicochemical conditions, that is, at temperatures of 28, 30, and 42 °C, pH of 6.4, 7.4, and 8.4, and NaCl concentrations from 200 to 1000 µg/mL. Exposure of oral isolates to TiO2NPs (14.7 nm) significantly (p < 0.05) reduced planktonic cell viability and biofilm formation in a concentration-dependent manner, which was confirmed by observing biofilm architecture by scanning electron microscopy (SEM) and optical microscopy. Overall, these results have important implications for the use of tetragonal anatase phase TiO2NPs (size range 5-25 nm, crystalline size 13.7 nm, and spherical shape) as an oral antibiofilm agent against Gram positive cocci infections. We suggest that TiO2NPs pave the way for further applications in oral mouthwash formulations and antibiofilm dental coatings.
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Green synthesized nanoparticles (NPs) have attracted enormous attention for their clinical and non-clinical applications. A natural polyphenol, gallo-tannin (GT) was used to reduce and cap the Fe2O3-NPs. GT-Fe2O3-NPs were synthesized following co-precipitation of FeCl3 and FeSO4·7H2O with GT. Fe2O3-NPs absorbed light at 380 nm. Physicochemically, Fe2O3-NPs were spherical with slight aggregation and average diameter of 12.85 nm. X-ray diffraction confirmed crystallinity and EDX revealed the elemental percentage of iron and oxygen as 21.7% and 42.11%, respectively. FT-IR data confirmed the adsorption of gallo-tannin functional groups. Multiple drug-resistant (MDR) Escherichia coli (ESßL), Pseudomonas aeruginosa (ESßL), and Staphylococcus aureus were found susceptible to 500-1000 µg GT-Fe2O3-NPs per ml. In synergy, Fe2O3-NPs enhanced the efficiency of some antibiotics. GT-Fe2O3 NPs showed significant (P ≤ 0.05) inhibition of growth and biofilm against MDR E. coli, P. aeruginosa, and S. aureus causing morphological and biofilm destruction. Violacein production (quorum sensing mediated) by C. violaceum was inhibited by GT-Fe2O3-NPs in a concentration-dependent manner with a maximum decrease of 3.1-fold. A decrease of 11-fold and 2.32-fold in fungal mycelial growth and human breast cancer (MCF-7) cell viability, respectively was evident. This study suggests a plausible role of gallo-tannin capped Fe2O3-NPs as an alternative antibacterial, antiquorum sensing, antibiofilm, antifungal, and anti-proliferative agent.
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The unregulated discharge of nanoparticles (NPs) from various nanotechnology industries into the environment is expected to alter the composition and physiological functions of soil microbiota. Considering this knowledge gap, the impact of five NPs (Ag, ZnO, CuO, Al2O3, and TiO2) differing in size and morphology on growth behavior and physiological activity of Azotobacter chroococcum, Bacillus thuringiensis, Pseudomonas mosselii, and Sinorhizobium meliloti were investigated. Various biochemical and microscopic approaches were adopted. Interestingly, all bacterial strains were found sensitive to Ag-NPs and ZnO-NPs but showed tolerance toward CuO, Al2O3, and TiO2-NPs. The loss of cellular respiration due to NPs was coupled with a reduction in population size. ZnO-NPs at 387.5 µg mL-1 had a maximum inhibitory impact on A. chroococcum and reduced its population by 72%. Under Ag-NP stress, the reduction in IAA secretion by bacterial strains followed the order S. meliloti (74%) > P. mosselii (63%) > A. chroococcum (49%). The surface of bacterial cells had small- or large-sized aggregates of NPs. Also, numerous gaps, pits, fragmented, and disorganized cell envelopes were visible. Additionally, a treated cell surface appeared corrugated with depressions and alteration in cell length and a strong heterogeneity was noticed under atomic force microscopy (AFM). For instance, NPs induced cell roughness for P. mosselii followed the order 12.6 nm (control) > 58 nm (Ag-NPs) > 41 nm (ZnO-NPs). TEM analysis showed aberrant morphology, cracking, and disruption of the cell envelope with extracellular electron-dense materials. Increased permeability of the inner cell membrane caused cell death and lowered EPS production. Ag-NPs and ZnO-NPs also disrupted the surface adhering ability of bacteria, which varied with time and concentration of NPs. Conclusively, a plausible mechanism of NP toxicity to bacteria has been proposed to understand the mechanistic basis of ecological interaction between NPs and resourceful bacteria. These results also emphasize to develop strategies for the safe disposal of NPs.
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Chemically synthesized copper oxide nanoparticles (CuONPs) involve the generation of toxic products, which narrowed its biological application. Hence, we have developed a one-pot, green method for CuONP production employing the leaf extract of Cymbopogon citratus (CLE). Gas chromatography-mass spectrometry (GC-MS) analysis confirmed the capping of CuONPs by CLE esters (CLE-CuONPs). Fourier-transform infrared (FTIR) showed phenolics, sugars, and proteins mediated nucleation and stability of CLE-CuONPs. X-ray diffraction (XRD) and transmission electron microscopy (TEM) revealed CLE-CuONPs between 11.4 to 14.5 nm. Staphylococcus aureus-1 (MRSA-1), Staphylococcus aureus-2 (MSSA-2) exposed to CLE-CuONPs (1500 µg/mL) showed 51.4%, 32.41% survival, while Escherichia coli-336 (E. coli-336) exposed to 1000 µg/mL CLE-CuONPs showed 45.27% survival. Scanning electron microscopy (SEM) of CLE-CuONPs treated E. coli-336, MSSA-2 and MRSA-1 showed morphological deformations. The biofilm production by E. coli-336 and MRSA-1 also declined to 33.0 ± 3.2% and 49.0 ± 3.1% at 2000 µg/mL of CLE-CuONPs. Atomic absorption spectroscopy (AAS) showed 22.80 ± 2.6%, 19.2 ± 4.2%, and 16.2 ± 3.6% accumulation of Cu2+ in E. coli-336, MSSA-2, and MRSA-1. Overall, the data exhibited excellent antibacterial and antibiofilm efficacies of esters functionalized CLE-CuONPs, indicating its putative application as a novel nano-antibiotic against multi drug resistance (MDR) pathogenic clinical isolates.
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Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Cobre/química , Cymbopogon/metabolismo , Nanopartículas del Metal/química , Extractos Vegetales/farmacología , Escherichia coli/efectos de los fármacos , Cromatografía de Gases y Espectrometría de Masas , Tecnología Química Verde , Concentración de Iones de Hidrógeno , Pruebas de Sensibilidad Microbiana , Microscopía Electrónica de Rastreo , Nanotecnología , Hojas de la Planta/metabolismo , Polvos , Espectrofotometría Atómica , Espectroscopía Infrarroja por Transformada de Fourier , Staphylococcus aureus/efectos de los fármacos , Difracción de Rayos XRESUMEN
We provide a novel one-step/one-pot bio-inspired method of synthesis for Myristica fragrans leaf ester (MFLE) cappedzinc oxide nanoparticles (MFLE-ZnONPs). Antibacterial and antbiofilm efficacies of MFLE-ZnONPs were tested against the multi-drug resistant (MDR) Escherichia coli (E. coli-336), methicillin-resistant Staphylococcus aureus (MRSA-1) and methicillin-sensitive (MSSA-2) clinical isolates. Antibacterial screening using well diffusion assay revealed the cytotoxicity of MFLE-ZnONPs in the range of 500-2000⯵g/ml. MFLE-ZnONPs significantly increased the zone of growth inhibition of E. coli-336 (17.0⯱â¯0.5 to 19.25⯱â¯1.0â¯mm), MSSA-2 (16.75⯱â¯0.8 to 19.0⯱â¯0.7â¯mm) and MRSA-1 (16.25⯱â¯1.0 to 18.25⯱â¯0.5â¯mm), respectively. The minimum inhibitory concentration (MIC) and minimum bactericidal concentrations (MBC) against E. coli-336, MRSA-1 and MSSA-2 were found to be 1500, 1000 and 500⯵g/ml, and 2500, 2000 and 1500⯵g/ml, respectively. A time and dose dependent reduction in the cell proliferation were also found at the respective MICs of tested strains. Scanning electron microscopy (SEM) of MFLE-ZnONPs-treated strains exhibited cellular damage via loss of native rod and coccoid shapes because of the formation of pits and cavities. E. coli-336 and MRSA-1 strains at their MICs (1500 and 1000⯵g/ml) sharply reduced the biofilm production to 51% and 24%. The physico-chemical characterization via x-ray diffraction (XRD) ascertained the crystallinity and an average size of MFLE-ZnONPs as 48.32⯱â¯2.5â¯nm. Gas chromatography-mass spectroscopy (GC-MS) analysis of MFLE-ZnONPs unravelled the involvement of two bio-active esters (1) butyl 3-oxobut-2-yl ester and (2) α-monoolein) as surface capping/stabilizing agents. Fourier transform infrared (FTIR) analysis of MFLE and MFLE-ZnONPs showed the association of amines, alkanes, aldehydes, amides, carbonyl and amines functional groups in the corona formation. Overall, our data provide novel insights on the rapid development of eco-friendly, cost-effective bio-synthesis of MFLE-ZnONPs, showing their putative application as nano-antibiotics against MDR clinical isolates.
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Ésteres/farmacología , Nanopartículas del Metal/química , Myristica/metabolismo , Extractos Vegetales/farmacología , Óxido de Zinc/farmacología , Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Hojas de la Planta/metabolismoRESUMEN
This study demonstrates a simple one-pot green method for biosynthesis of terpenoids encapsulated copper oxide nanoparticles (CuONPs) using aqueous leaf extract of Eucalyptus globulus (ELE), as reducing, dispersing, and stabilizing agent. Indeed, the greater attachment and internalization of ELE-CuONPs in Gram-positive and -negative biofilm producing clinical bacterial isolates validated the hypothesis that terpenoids encapsulated CuONPs are more stable and effective antibacterial and antibiofilm agent vis-à-vis commercially available nano and micro sized analogues. Gas chromatography-mass spectroscopy (GC-MS) analysis of pristine ELE identified 17 types of terpenoids based on their mass-to-charge (m/z) ratios. Amongst them four bioactive terpenoids viz. terpineols, 2,6-octadienal-3,7-dimethyl, benzamidophenyl-4-benzoate and ß-eudesmol were found associated with the CuONPs as ELE-cap, and most likely involved in the nucleation and stabilization of ELE-CuONPs. Further, the Fourier transformed infrared (FTIR) analysis of ELE-CuONPs also implicated other functional biomolecules like proteins, sugars, alkenes, etc. with ELE terpenoids corona. Flow cytometric (FCM) data exhibited significantly enhanced intracellular uptake propensity of terpenoids encapsulated ELE-CuONPs and accumulation of intracellular reactive oxygen species (ROS), which ensued killing of planktonic cells of extended spectrum ß-lactamases (ESßL) producing Escherichia coli-336 (E. coli-336), Pseudomonas aeruginosa-621 (P. aeruginosa-621) and methicillin-resistant Staphylococcus aureus-1 (MRSA-1) clinical isolates compared to the bare surface commercial nano-CuO and bulk sized CuO. The study for the first-time demonstrated the (i) differential bio-nano interface activities due to ELE surface and varied cell wall composition of test bacterial isolates, (ii) antibacterial effect and biofilm inhibition due to disruption of proteins involved in adhesion and biofilm formation triggered by CuONPs induced intracellular oxidative stress, and (iii) indigenous terpenoids-capped bio-inspired CuONPs are more stable and effective antibacterial and antibiofilm agent as compared with commercially available nano-CuO and bulk-CuO.
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Cobre/química , Eucalyptus/química , Nanopartículas del Metal/química , Viabilidad Microbiana , Extractos Vegetales/farmacología , Especies Reactivas de Oxígeno/metabolismo , Biopelículas/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cristalización , Escherichia coli/efectos de los fármacos , Escherichia coli/ultraestructura , Humanos , Nanopartículas del Metal/ultraestructura , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Plancton/citología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/ultraestructura , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de SuperficieRESUMEN
Biogenic hematite (α-Fe2O3) nanoparticles (NPs) of average size <10â¯nm were synthesized using green approach with Aloe vera extract (ALE). The aim of the study was to assess the protective effect of extracellular polymeric substances (EPS) against antibacterial and antibiofilm activities of ALE-α-Fe2O3NPs in normal EPS producers (pristine) and experimentally modified (low-EPS) Pseudomonas aeruginosa (P. aeruginosa) cells and the mechanism of cell killing. Formation of ALE-α-Fe2O3NPs has been validated by X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM) and Fourier-transformed infrared spectroscopy (FTIR) analysis. The FTIR data suggested the possible role OH group bearing organic compounds of ALE in metal reduction and nucleation of NPs. Gas Chromatography-Mass spectroscopy (GC-MS) analysis revealed the presence of oxime-methoxy-phenyl, ethanone 1-phenyl, hexadecanoic acid, cyclohexanol 2,6-dimethyl, tetracontane, stigmast-5-en-3-ol, cyclohexanol 2,6-dimethyl, and cyclohexasiloxane dodecamethyl on the surface of ALE-α-Fe2O3NPs. Cell viability assay and SEM imaging revealed significantly greater bacteriostatic and/or bactericidal effect of ALE-α-Fe2O3NPs in low EPS cells compared to pristine cells or bare-α-Fe2O3NPs. This is attributed to thinner protective layer of EPS around the low EPS cells, and higher dispersibility and stability of ALE-α-Fe2O3NPs. Absorption of ALE-α-Fe2O3NPs and bare-α-Fe2O3NPs on EPS surface and within EPS matrix was ascertained by atomic absorption spectroscopy (AAS). The results suggest differential internalization of ALE-α-Fe2O3NPs and bare-α-Fe2O3NPs in P. aeruginosa cells. The flow cytometry (FCM) results exhibited increased intracellular granularity in low EPS (18.94%) as compared with pristine (10.94%) cells, which signifies the greater internalization of ALE-α-Fe2O3NPs. Moreover, the proportionate increase in intracellular ROS generation in low EPS (20.47%) via-a-vis pristine (7.56%) cells was observed. Overall, the results elucidate that ALE-α-Fe2O3NPs-bacterial interaction leads to attachment of NPs to EPS surface, migration within the EPS matrix and penetration into cell, which eventually results in growth inhibition due to intracellular ROS activity. Owing to significant antibacterial and antibiofilm activities, ALE-α-Fe2O3NPs may serve as a good candidate for clinical management of extended spectrum beta lactamases (ESBL) positive P. aeruginosa.
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Aloe/química , Compuestos Férricos/química , Nanopartículas del Metal/química , Polímeros/química , Pseudomonas aeruginosa/metabolismo , Aloe/metabolismo , Biopelículas/efectos de los fármacos , Cromatografía de Gases y Espectrometría de Masas , Tecnología Química Verde , Nanopartículas del Metal/toxicidad , Microscopía Electrónica de Rastreo , Tamaño de la Partícula , Extractos Vegetales/química , Polímeros/farmacología , Especies Reactivas de Oxígeno/metabolismo , Espectrofotometría Atómica , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos XRESUMEN
Titanium dioxide nanoparticles (TiO2-NPs) interaction with human serum albumin (HSA) and DNA was studied by UV-visible spectroscopy, spectrofluorescence, circular dichroism (CD), and transmission electron microscopy (TEM) to analyze the binding parameters and protein corona formation. TEM revealed protein corona formation on TiO2-NPs surface due to adsorption of HSA. Intrinsic fluorescence quenching data suggested significant binding of TiO2-NPs (avg. size 14.0 nm) with HSA. The Stern-Volmer constant (Ksv) was determined to be 7.6 × 102 M-1 (r2 = 0.98), whereas the binding constant (Ka) and number of binding sites (n) were assessed to be 5.82 × 102 M-1 and 0.97, respectively. Synchronous fluorescence revealed an apparent decrease in fluorescence intensity with a red shift of 2 nm at Δλ = 15 nm and Δλ = 60 nm. UV-visible analysis also provided the binding constant values for TiO2-NPs-HSA and TiO2-NPs-DNA complexes as 2.8 × 102 M-1 and 5.4 × 103 M-1. The CD data demonstrated loss in α-helicity of HSA and transformation into ß-sheet, suggesting structural alterations by TiO2-NPs. The docking analysis of TiO2-NPs with HSA revealed its preferential binding with aromatic and non-aromatic amino acids in subdomain IIA and IB hydrophobic cavity of HSA. Also, the TiO2-NPs docking revealed the selective binding with A-T bases in minor groove of DNA.
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ADN/química , ADN/metabolismo , Nanopartículas/química , Conformación de Ácido Nucleico , Albúmina Sérica Humana/química , Albúmina Sérica Humana/metabolismo , Titanio/química , Dicroismo Circular , Humanos , Simulación del Acoplamiento Molecular , Nanopartículas/ultraestructura , Unión Proteica , Dominios Proteicos , Espectrometría de Fluorescencia , Espectrofotometría Ultravioleta , Difracción de Rayos XRESUMEN
ZnO nanoparticles (ZnONPs) were synthesised through a simple and efficient biogenic synthesis approach, exploiting the reducing and capping potential of Aloe barbadensis Miller (A. vera) leaf extract (ALE). ALE-capped ZnO nanoparticles (ALE-ZnONPs) were characterized using UV-Vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and transmission electron microscopy (TEM) analyses. XRD analysis provided the average size of ZnONPs as 15 nm. FTIR spectral analysis suggested the role of phenolic compounds, terpenoids and proteins present in ALE, in nucleation and stability of ZnONPs. Flow cytometry and atomic absorption spectrophotometry (AAS) data analyses revealed the surface binding and internalization of ZnONPs in Gram +ve (Staphylococcus aureus) and Gram -ve (Escherichia coli) cells, respectively. Significant antibacterial activity of ALE-ZnONPs was observed against extended spectrum beta lactamases (ESBL) positive E. coli, Pseudomonas aeruginosa, and methicillin resistant S. aureus (MRSA) clinical isolates exhibiting the MIC and MBC values of 2200, 2400 µg/ml and 2300, 2700 µg/ml, respectively. Substantial inhibitory effects of ALE-ZnONPs on bacterial growth kinetics, exopolysaccharides and biofilm formation, unequivocally suggested the antibiotic and anti-biofilm potential. Overall, the results elucidated a rapid, environmentally benign, cost-effective, and convenient method for ALE-ZnONPs synthesis, for possible applications as nanoantibiotics or drug carriers.
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Antibacterianos/química , Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Nanopartículas/química , Staphylococcus aureus/efectos de los fármacos , Óxido de Zinc/química , Óxido de Zinc/farmacología , Aloe , Biopelículas/efectos de los fármacos , Escherichia coli/fisiología , Infecciones por Escherichia coli/tratamiento farmacológico , Tecnología Química Verde , Humanos , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/fisiología , Pruebas de Sensibilidad Microbiana , Oxidación-Reducción , Extractos Vegetales/química , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus aureus/fisiologíaRESUMEN
A simple and rapid microwave assisted method of green synthesis of silver nanoparticles (AgNPs) was developed using aqueous leaf extract of Eucalyptus globulus(ELE), and their antibacterial and antibiofilm potential investigated. With this aim, the aqueous solutions of ELE and AgNO3(1 mM) were mixed (1:4 v/v), and microwave irradiated at 2450 Mhz, for 30 sec. The instant color change of the ELE-AgNO3 mixture from pale yellow to dark brown indicated ELE-AgNPs synthesis. The intensity of peak at 428 nm in UV-Vis spectra, due to the surface plasmon resonance of AgNPs, varied with the amount of ELE, AgNO3 concentration, pH and time of incubation. The biosynthesized ELE-AgNPs were characterized by UV-visible spectroscopy, XRD, TEM, SEM-EDX, FTIR and TGA analyses. The size of ELE-AgNPs was determined to be in range of 1.9-4.3 nm and 5-25 nm, with and without microwave treatment, respectively. SEM exhibited the capping of AgNPs with the ELE constituents, and validated by FTIR analysis. The FTIR data revealed the presence of plant organic constituents and metabolites bound to ELE-AgNPs, which contributes for their stability. The antimicrobial activity of ELE-AgNPs was assessed by growth and biofilm inhibition of extended spectrum ß-lactamase (ESBL) producing Pseudomonas aeruginosa, Escherichia coli and methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) clinical bacterial isolates. The results demonstrated that S. aureus were more sensitive to ELE-AgNPs than E. coli and P. aeruginosa. MRSA exhibited higher sensitive than MSSA, whereas P. aeruginosa were more sensitive than E. coli to ELE-AgNPs treatment. Also, significant (83 ± 3% and 84 ± 5%) biofilm inhibition was observed in case of S. aureus and P. aeruginosa, respectively. The results elucidated environmentally friendly, economical and quick method for production of colloidal bio-functionalized ELE-AgNPs, for effectual clinical applications, as broad spectrum antibacterial agents and biofilm inhibitors.