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1.
Front Cell Infect Microbiol ; 14: 1419568, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38983115

RESUMEN

Background: Helicobacter pylori infection poses a significant health burden worldwide, and its virulence factor CagA plays a pivotal role in its pathogenesis. Methods: In this study, the interaction between H. pylori-infected AGS cells and silver nanoparticles (AgNPs) was investigated, with a focus on the modulation of CagA-mediated responses, investigated by western blotting. Both, the dose-dependent efficacy against H. pylori (growth curves, CFU assay) and the impact of the nanoparticles on AGS cells (MTT assay) were elucidated. Results: AGS cells infected with H. pylori displayed dramatic morphological changes, characterized by elongation and a migratory phenotype, attributed to CagA activity. Preincubation of H. pylori with AgNPs affected these morphological changes in a concentration-dependent manner, suggesting a correlation between AgNPs concentration and CagA function. Conclusion: Our study highlights the nuanced interplay between host-pathogen interactions and the therapeutic potential of AgNPs in combating H. pylori infection and offers valuable insights into the multifaceted dynamics of CagA mediated responses.


Asunto(s)
Antígenos Bacterianos , Proteínas Bacterianas , Infecciones por Helicobacter , Helicobacter pylori , Nanopartículas del Metal , Transducción de Señal , Plata , Helicobacter pylori/efectos de los fármacos , Proteínas Bacterianas/metabolismo , Antígenos Bacterianos/metabolismo , Plata/farmacología , Plata/metabolismo , Humanos , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/tratamiento farmacológico , Transducción de Señal/efectos de los fármacos , Interacciones Huésped-Patógeno , Células Epiteliales/microbiología , Factores de Virulencia/metabolismo , Línea Celular , Antibacterianos/farmacología , Línea Celular Tumoral
2.
Neoplasma ; 70(3): 390-401, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-37498075

RESUMEN

Silver nanoparticles (AgNPs) exhibit unique physicochemical properties, making these nanomaterials attractive for various medical applications. Among them, AgNPs have shown great potential in the treatment of cancer by inducing apoptosis in cancer cells, inhibiting tumor growth, and enhancing the efficacy of conventional cancer treatments such as chemotherapy and radiation therapy. Despite the promising therapeutical advantage of AgNPs, there are several challenges that need to be addressed. One of the most important is AgNPs' toxicity, which in case of treatment might be extended to non-cancerous cells and tissues. In our study, we therefore investigated the effects of spherical AgNPs with the silver core size of 10, 30, and 45 nm coated with polyacrylic acid (PAA-AgNPs) in an in vitro model using cancer (A549) and non-cancer (HEL299) cells. We estimated the impact of these nanoparticles on cell viability, cell proliferation, and cell actin cytoskeleton remodeling. Moreover, changes in the expression of TNFA, IL-10, FN1, and SOD1 mRNA induced by PAA-AgNPs were determined. Our results suggest that the smallest (10 nm) PAA-AgNPs are the most effective in apoptosis induction, however, they are also the most toxic from the three AgNPs types to both, cancer and non-cancer cells, while bigger (30 and 45 nm) PAA-AgNPs showed fewer undesirable effects in these pulmonary cells.


Asunto(s)
Neoplasias Pulmonares , Nanopartículas del Metal , Humanos , Plata/farmacología , Plata/química , Nanopartículas del Metal/química , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/metabolismo , Apoptosis , Pulmón/metabolismo
3.
Small ; 19(51): e2207216, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36703534

RESUMEN

Tackling the current problem of antimicrobial resistance (AMR) requires fast, inexpensive, and effective methods for controlling and detecting antibiotics in diverse samples at the point of interest. Cost-effective, disposable, point-of-care electrochemical biosensors are a particularly attractive option. However, there is a need for conductive and versatile carbon-based materials and inks that enable effective bioconjugation under mild conditions for the development of robust, sensitive, and selective devices. This work describes a simple and fast methodology to construct an aptasensor based on a novel graphene derivative equipped with alkyne groups prepared via fluorographene chemistry. Using click chemistry, an aptamer is immobilized and used as a successful platform for the selective determination of ampicillin in real samples in the presence of interfering molecules. The electrochemical aptasensor displayed a detection limit of 1.36 nM, high selectivity among other antibiotics, the storage stability of 4 weeks, and is effective in real samples. Additionally, structural and docking simulations of the aptamer shed light on the ampicillin binding mechanism. The versatility of this platform opens up wide possibilities for constructing a new class of aptasensor based on disposable screen-printed carbon electrodes usable in point-of-care devices.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Grafito , Grafito/química , Química Clic , Alquinos , Aptámeros de Nucleótidos/química , Técnicas Electroquímicas/métodos , Carbono/química , Técnicas Biosensibles/métodos , Electrodos , Oro/química , Ampicilina , Antibacterianos , Límite de Detección
4.
Sci Rep ; 12(1): 5222, 2022 03 25.
Artículo en Inglés | MEDLINE | ID: mdl-35338239

RESUMEN

The number of antibiotic-resistant bacterial strains is increasing due to the excessive and inappropriate use of antibiotics, which are therefore becoming ineffective. Here, we report an effective way of enhancing and restoring the antibacterial activity of inactive antibiotics by applying them together with a cyanographene/Ag nanohybrid, a nanomaterial that is applied for the first time for restoring the antibacterial activity of antibiotics. The cyanographene/Ag nanohybrid was synthesized by chemical reduction of a precursor material in which silver cations are coordinated on a cyanographene sheet. The antibacterial efficiency of the combined treatment was evaluated by determining fractional inhibitory concentrations (FIC) for antibiotics with different modes of action (gentamicin, ceftazidime, ciprofloxacin, and colistin) against the strains Escherichia coli, Pseudomonas aeruginosa, and Enterobacter kobei with different resistance mechanisms. Synergistic and partial synergistic effects against multiresistant strains were demonstrated for all of these antibiotics except ciprofloxacin, which exhibited an additive effect. The lowest average FICs equal to 0.29 and 0.39 were obtained for colistin against E. kobei and for gentamicin against E. coli, respectively. More importantly, we have experimentally confirmed for the first time, that interaction between the antibiotic's mode of action and the mechanism of bacterial resistance strongly influenced the combined treatment's efficacy.


Asunto(s)
Antibacterianos , Colistina , Antibacterianos/química , Antibacterianos/farmacología , Ciprofloxacina/farmacología , Colistina/farmacología , Sinergismo Farmacológico , Escherichia coli , Gentamicinas/farmacología , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa
5.
Adv Sci (Weinh) ; 8(12): 2003090, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-34194925

RESUMEN

The ability of bacteria to develop resistance to antibiotics is threatening one of the pillars of modern medicine. It was recently understood that bacteria can develop resistance even to silver nanoparticles by starting to produce flagellin, a protein which induces their aggregation and deactivation. This study shows that silver covalently bound to cyanographene (GCN/Ag) kills silver-nanoparticle-resistant bacteria at concentrations 30 times lower than silver nanoparticles, a challenge which has been so far unmet. Tested also against multidrug resistant strains, the antibacterial activity of GCN/Ag is systematically found as potent as that of free ionic silver or 10 nm colloidal silver nanoparticles. Owing to the strong and multiple dative bonds between the nitrile groups of cyanographene and silver, as theory and experiments confirm, there is marginal silver ion leaching, even after six months of storage, and thus very high cytocompatibility to human cells. Molecular dynamics simulations suggest strong interaction of GCN/Ag with the bacterial membrane, and as corroborated by experiments, the antibacterial activity does not rely on the release of silver nanoparticles or ions. Endowed with these properties, GCN/Ag shows that rigid supports selectively and densely functionalized with potent silver-binding ligands, such as cyanographene, may open new avenues against microbial resistance.


Asunto(s)
Antibacterianos/uso terapéutico , Infecciones Bacterianas/tratamiento farmacológico , Farmacorresistencia Bacteriana/efectos de los fármacos , Nanopartículas del Metal/uso terapéutico , Plata/uso terapéutico , Nanopartículas del Metal/química , Plata/química
6.
Colloids Surf B Biointerfaces ; 202: 111680, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33714189

RESUMEN

The use of Ag-modified nanomaterials continues to attract attention in biological contamination control, their potential cytotoxicity is often overlooked. Herein, biocompatible carbon nitride is modified with 1 and 5 wt.% Ag and effects of different nanomaterial dose and Ag content on antimicrobial activity and cytotoxicity is studied. Pure Ag nanoparticles and AgNO3 is tested for comparison, together with ten bacterial strains including pan-resistant Pseudomonas aeruginosa. Cytotoxicity is then investigated in three adherent and two suspension human cell lines, and results confirm that cancer adherent cell lines are the most immune lines and human cervical adenocarcinoma cells (HeLa) are more resilient than human lung adenocarcinoma cells (A549). The HeLa remains over 90 % viable even after 24 -h treatment with the highest concentration of 5%Ag/g-C3N4 (300 mg L-1) while A549 sustained viability only up to 100 mg L-1. Higher concentrations then induce cytotoxicity and A549 cell viability decreases. Our results show the importance of complementary testing of cytotoxicity by LIVE/DEAD assay using flow cytometry with more different human cell lines, which might be less immune to tested nanomaterials than HeLa and A549. Combined controls of new antibacterial agent activity tests then provide increased knowledge of their biocompatibility.


Asunto(s)
Nanopartículas del Metal , Plata , Antibacterianos/farmacología , Humanos , Nitrilos
7.
Sci Rep ; 11(1): 6240, 2021 03 18.
Artículo en Inglés | MEDLINE | ID: mdl-33737512

RESUMEN

Targeted and effective therapy of diseases demands utilization of rapid methods of identification of the given markers. Surface enhanced Raman spectroscopy (SERS) in conjunction with streptavidin-biotin complex is a promising alternative to culture or PCR based methods used for such purposes. Many biotinylated antibodies are available on the market and so this system offers a powerful tool for many analytical applications. Here, we present a very fast and easy-to-use procedure for preparation of streptavidin coated magnetic polystyrene-Au (or Ag) nanocomposite particles as efficient substrate for surface SERS purposes. As a precursor for the preparation of SERS active and magnetically separable composite, commercially available streptavidin coated polystyrene (PS) microparticles with a magnetic core were utilized. These composites of PS particles with silver or gold nanoparticles were prepared by reducing Au(III) or Ag(I) ions using ascorbic acid or dopamine. The choice of the reducing agent influences the morphology and the size of the prepared Ag or Au particles (15-100 nm). The prepare composites were also characterized by HR-TEM images, mapping of elements and also magnetization measurements. The content of Au and Ag was determined by AAS analysis. The synthesized composites have a significantly lower density against magnetic composites based on iron oxides, which considerably decreases the tendency to sedimentation. The polystyrene shell on a magnetic iron oxide core also pronouncedly reduces the inclination to particle aggregation. Moreover, the preparation and purification of this SERS substrate takes only a few minutes. The PS composite with thorny Au particles with the size of approximately 100 nm prepared was utilized for specific and selective detection of Staphylococcus aureus infection in joint knee fluid (PJI) and tau protein (marker for Alzheimer disease).


Asunto(s)
Enfermedad de Alzheimer/diagnóstico , Oro/química , Nanopartículas Magnéticas de Óxido de Hierro/química , Plata/química , Espectrometría Raman/métodos , Infecciones Estafilocócicas/diagnóstico , Proteínas tau/análisis , Enfermedad de Alzheimer/sangre , Enfermedad de Alzheimer/genética , Ácido Ascórbico/química , Biomarcadores/análisis , Dopamina/química , Humanos , Nanopartículas Magnéticas de Óxido de Hierro/ultraestructura , Tamaño de la Partícula , Poliestirenos/química , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus aureus/patogenicidad , Estreptavidina/química , Líquido Sinovial/microbiología , Proteínas tau/sangre , Proteínas tau/genética
8.
Nat Commun ; 12(1): 713, 2021 01 29.
Artículo en Inglés | MEDLINE | ID: mdl-33514738

RESUMEN

Despite proteotoxic stress and heat shock being implicated in diverse pathologies, currently no methodology to inflict defined, subcellular thermal damage exists. Here, we present such a single-cell method compatible with laser-scanning microscopes, adopting the plasmon resonance principle. Dose-defined heat causes protein damage in subcellular compartments, rapid heat-shock chaperone recruitment, and ensuing engagement of the ubiquitin-proteasome system, providing unprecedented insights into the spatiotemporal response to thermal damage relevant for degenerative diseases, with broad applicability in biomedicine. Using this versatile method, we discover that HSP70 chaperone and its interactors are recruited to sites of thermally damaged proteins within seconds, and we report here mechanistically important determinants of such HSP70 recruitment. Finally, we demonstrate a so-far unsuspected involvement of p97(VCP) translocase in the processing of heat-damaged proteins. Overall, we report an approach to inflict targeted thermal protein damage and its application to elucidate cellular stress-response pathways that are emerging as promising therapeutic targets.


Asunto(s)
Proteínas HSP70 de Choque Térmico/metabolismo , Respuesta al Choque Térmico , Análisis de la Célula Individual/métodos , Proteína que Contiene Valosina/metabolismo , Línea Celular Tumoral , Calor/efectos adversos , Humanos , Nanopartículas del Metal/química , Complejo de la Endopetidasa Proteasomal/metabolismo , Plata/química , Resonancia por Plasmón de Superficie , Ubiquitina/metabolismo , Proteína que Contiene Valosina/genética
9.
Photodiagnosis Photodyn Ther ; 33: 102140, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33307229

RESUMEN

As resistance of bacterial strains to antibiotics is a major problem, there is a need to look for alternative treatments. One option is antimicrobial photodynamic inactivation (aPDI). The pathogenic cells are targeted by a nontoxic photosensitizer while the surrounding healthy tissue is relatively unaffected. The photosensitizer is activated by light of t appropriate wavelength resulting in the generation of reactive oxygen species that are cytotoxic for the pathogens. In this work, the photosensitizer TMPyP and silver nanoparticles (AgNPs) were investigated for their synergistic antibacterial effect. We tested these two substances on two bacterial strains, methicillin-resistant Staphylococcus aureus 4591 (MRSA) and extended-spectrum beta-lactamases-producing Klebsiella pneumoniae 2486 (ESBL-KP), to compare their effectiveness. The bacteria were first incubated with TMPyP for 45 min or 5 h, then irradiated with a LED source with the total fluence of 10 or 20 J/cm2 and then placed in a microbiological growth medium supplemented with AgNPs. To accomplish the synergistic effect, the optimal combination of TMPyP and AgNPs was estimated as 1.56-25 µM for TMPyP and 3.38 mg/l for AgNPs in case of MRSA and 1.56-50 µM for TMPyP and 3.38 mg/l for AgNPs in case of ESBL-KP at 45 min incubation with TMPyP and fluence of 10 J/cm2. Longer incubation and/or longer irradiation led to a decrease in the maximum values of the photosensitizer concentration to produce the synergistic effect. From this work it can be concluded that the combination of antimicrobial photodynamic inactivation with a treatment including silver nanoparticles could be a promising approach to treat bacterial infection.


Asunto(s)
Antiinfecciosos , Nanopartículas del Metal , Staphylococcus aureus Resistente a Meticilina , Fotoquimioterapia , Porfirinas , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Klebsiella pneumoniae , Resistencia a la Meticilina , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Porfirinas/farmacología , Plata/farmacología
10.
Chemosphere ; 269: 128759, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33153849

RESUMEN

Androgen receptor (AR) signalling is triggered by androgens that have lipophilic nature. Since it was indicated that graphene oxide (GO) might facilitate passive diffusion of lipophilic compounds probably via Trojan horse-like mechanism, we tested the hypothesis if this suggestion would apply for androgens as well. Thus, we investigated if GO affects dihydrotestosterone (DHT)-triggered signalling of AR in two prostate cancer-derived cell lines, 22Rv1 and LNCaP. These cell lines differ in number of AR variants, i.e. there are two variants in 22Rv1 cells (full length and truncated) but only one in LNCaP cells (full length). Graphene oxide had no effect on basal luciferase activity but significantly decreased DHT-inducible AR-dependent luciferase activity in stably transfected cells. In 22Rv1 cells, it induced concentration-dependent decrease of DHT-inducible KLK3 mRNA and PSA protein after 24 h. While there was no effect on UBE2C mRNA (regulated by truncated variant), there was synergistic effect of DHT and GO on UBE2C protein level. Translocation of full-length AR (AR-FL) was potentiated by GO in the presence of DHT in 22Rv1 cells but it was suppressed in LNCaP cells. DHT-stimulated enrichment of AR-FL on KLK3 promoter was not significantly affected by GO in any tested cell line neither was KLK3 mRNA at 4 h of incubation. In conclusion, GO affects DHT-triggered signalling in both types of cells in similar manner, but ligand-triggered redistribution of AR-FL is affected differently. One of the reasons may be the presence of truncated variant of androgen receptor.


Asunto(s)
Neoplasias de la Próstata , Receptores Androgénicos , Andrógenos , Línea Celular Tumoral , Dihidrotestosterona/farmacología , Grafito , Humanos , Masculino , Receptores Androgénicos/genética
11.
Chemosphere ; 253: 126753, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32464781

RESUMEN

Graphene oxide (GO) is an engineered nanomaterial which was demonstrated to have outstanding capacity for adsorption of organic pollutants such as polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), the ligands and activators of the aryl hydrocarbon receptor (AhR). Due to the partially overlapping ligand capacity of AhR and pregnane X receptor (PXR), we tested the impact of GO particles on their signalling. While reporter gene assay revealed potentiating effect of GO on ligand-activated AhR-dependent luciferase activity, there was no effect for PXR. However, inducible target genes for AhR (CYP1A1) or PXR (ABCB1) were decreased at mRNA as well as protein levels by the presence of GO in HepG2 (for AhR), LS180 (for PXR) or primary human hepatocytes (both receptors). Moreover, the presence of GO diminished PXR and AhR protein levels in primary cultures of human hepatocytes. This was partially reversed by proteasome inhibitor MG132 for AhR but not for PXR. In conclusion, GO decreases ligand-stimulated activities of AhR and PXR in human cells.


Asunto(s)
Grafito/química , Xenobióticos/metabolismo , Biotransformación , Citocromo P-450 CYP1A1/metabolismo , Genes Reporteros , Hepatocitos/efectos de los fármacos , Humanos , Receptor X de Pregnano , ARN Mensajero/metabolismo , Receptores de Hidrocarburo de Aril/metabolismo , Receptores de Esteroides/genética , Transducción de Señal , Xenobióticos/química
12.
Neuro Endocrinol Lett ; 39(4): 299-304, 2018 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-30531708

RESUMEN

OBJECTIVES: The aim of this study was to establish and evaluate the mortality rate, hatching rate and observe the presence of sublethal changes in zebrafish embryos after exposure to silver ions and nanoparticles. METHODS: Tests were conducted on newly fertilized zebrafish embryos, according to the modified OECD guideline 236, using a semistatic method and 96 hour incubation time. Silver nitrate and two different silver nanoparticles, stabilized with 0.01% solution of maltose and gelatine in the first case, and stabilized with polyvinylpyrrolidone, in the latter, were tested. RESULTS: Significant differences in toxicity of tested substances were recorded. The value of 96hLC50 for silver nitrate was 58.44 µg/L. The value of 96hLC50, calculated for silver nanoparticles stabilized with 0.01% solution of maltose and gelatine, was nearly 100 times higher, 4.31 mg/L. The value 96hLC50 for silver nanoparticles stabilized with polyvinylpyrrolidone exceeded 100mg/L, occurrence of sublethal effects caused by silver nanoparticles stabilized with polyvinylpyrrolidone was insignificant in most of the exposition groups, but only in this substance caused decreased hatching rate. CONCLUSION: Properties of different silver nanoparticles play an important role in levels of their toxicity and predominant mechanisms of action. In general, silver nanoparticles are less toxic for Danio rerio embryos than silver ions.


Asunto(s)
Embrión no Mamífero/efectos de los fármacos , Nanopartículas , Nitrato de Plata/toxicidad , Plata/toxicidad , Animales , Gelatina , Dosificación Letal Mediana , Maltosa , Povidona , Pez Cebra
13.
Sci Rep ; 8(1): 4589, 2018 03 15.
Artículo en Inglés | MEDLINE | ID: mdl-29545580

RESUMEN

The controlled preparation of Au nanoparticles (NPs) in the size range of 6 to 22 nm is explored in this study. The Au NPs were prepared by the reduction of tetrachloroauric acid using maltose in the presence of nonionic surfactant Tween 80 at various concentrations to control the size of the resulting Au NPs. With increasing concentration of Tween 80 a decrease in the size of produced Au NPs was observed, along with a significant decrease in their size distribution. The size-dependent catalytic activity of the synthesized Au NPs was tested in the reduction of 4-nitrophenol with sodium borohydride, resulting in increasing catalytic activity with decreasing size of the prepared nanoparticles. Eley-Rideal catalytic mechanism emerges as the more probable, in contrary to the Langmuir-Hinshelwood mechanism reported for other noble metal nanocatalysts.

14.
Ecotoxicol Environ Saf ; 147: 110-116, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-28841525

RESUMEN

Concerns regarding the potential toxic effects of zinc oxide nanoparticles (ZnO NPs) on aquatic organisms are growing due to the fact that NPs may be released into aquatic ecosystems. This study aimed to investigate the effects of dietary exposure to ZnO NPs on juvenile common carp (Cyprinus carpio). Fish were fed a spiked diets at doses 50 and 500mg of ZnO NPs per kg of feed for 6 weeks followed by a 2-week recovery period. Fish were sampled every 2 weeks for haematology trends, blood biochemistry measures, histology analyses, and determination of the accumulation of zinc in tissues. At the end of the exposure and post-exposure periods, fish were sampled for an assessment of lipid peroxidation levels. Dietborne ZnO NPs had no effects on haematology, blood biochemistry, and lipid peroxidation levels during the exposure period. After the recovery period, aspartate aminotransferase activity significantly (p < 0.05) increased and alanine transferase activity significantly (p < 0.05) decreased in the higher exposure group. The level of lipid peroxidation significantly (p < 0.05) decreased in liver of treated fish after 2 weeks post-exposure period. A histological examination revealed mild histopathological changes in kidneys during exposure. Our results did not show a significant increase of zinc content at the end of experiment in any of tested organs. However, chronic dietary exposure to ZnO NPs might affect kidney and liver function.


Asunto(s)
Carpas/fisiología , Monitoreo del Ambiente/métodos , Nanopartículas del Metal/toxicidad , Estrés Oxidativo/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Óxido de Zinc/toxicidad , Animales , Carpas/metabolismo , Dieta , Relación Dosis-Respuesta a Droga , Riñón/efectos de los fármacos , Riñón/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Factores de Tiempo , Distribución Tisular , Contaminantes Químicos del Agua/metabolismo , Óxido de Zinc/metabolismo
15.
Nat Nanotechnol ; 13(1): 65-71, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-29203912

RESUMEN

Silver nanoparticles have already been successfully applied in various biomedical and antimicrobial technologies and products used in everyday life. Although bacterial resistance to antibiotics has been extensively discussed in the literature, the possible development of resistance to silver nanoparticles has not been fully explored. We report that the Gram-negative bacteria Escherichia coli 013, Pseudomonas aeruginosa CCM 3955 and E. coli CCM 3954 can develop resistance to silver nanoparticles after repeated exposure. The resistance stems from the production of the adhesive flagellum protein flagellin, which triggers the aggregation of the nanoparticles. This resistance evolves without any genetic changes; only phenotypic change is needed to reduce the nanoparticles' colloidal stability and thus eliminate their antibacterial activity. The resistance mechanism cannot be overcome by additional stabilization of silver nanoparticles using surfactants or polymers. It is, however, strongly suppressed by inhibiting flagellin production with pomegranate rind extract.


Asunto(s)
Antibacterianos , Farmacorresistencia Bacteriana , Escherichia coli/efectos de los fármacos , Nanopartículas del Metal/química , Pseudomonas aeruginosa/efectos de los fármacos , Plata , Antibacterianos/química , Antibacterianos/farmacología , Estabilidad de Medicamentos , Pruebas de Sensibilidad Microbiana , Plata/química , Plata/farmacología
16.
Klin Mikrobiol Infekc Lek ; 23(1): 17-20, 2017 Mar.
Artículo en Checo | MEDLINE | ID: mdl-28467592

RESUMEN

OBJECTIVE: The aim was to evaluate the antibacterial effect of silver nanoparticles on anaerobic bacteria. MATERIAL AND METHODS: The microdilution method was used to determine the minimum inhibitory concentrations (MICs) of 28 nm silver nanoparticles, both unstabilized and stabilized by casein, gelatin and polyacrylic acid. The following anaerobic bacteria were tested: Bacteroides fragilis, Bacteroides thetaiotaomicron, Eggerthella lenta, Propionibacterium acnes, Clostridium perfringens, Clostridium difficile and Fusobacterium varium. RESULTS: Unstabilized silver nanoparticles exhibited antibacterial activity at concentrations ranging from 13 to 34 mg/L. A more significant effect with MIC values between 1 and 13 mg/L was shown for silver nanoparticles stabilized by casein. CONCLUSION: Unstabilized silver nanoparticles are active against anaerobic bacteria at concentrations proved to be cytotoxic to eukaryotic cells of human fibroblasts and multicellular organisms. Silver nanoparticles stabilized by casein appear to be more suitable for further research.


Asunto(s)
Antibacterianos/farmacología , Bacterias Anaerobias/efectos de los fármacos , Nanopartículas del Metal , Plata/farmacología , Humanos
17.
Sci Total Environ ; 579: 1504-1511, 2017 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-27916310

RESUMEN

Zinc oxide (ZnO) nanoparticles (NPs) have been dramatically used in industry, biology, and medicine. Despite their interesting physico-chemical properties for application in various industrial, medical, and consumer products, safe use of ZnO NPs are under challenges due to the inadequate information related to their toxicological endpoints. Proteomics was applied to evaluate the sub-lethal effects of dietary exposure to ZnO NPs on serum proteome profile of juvenile common carp, (Cyprinus carpio). Therefore, ZnO NPs solution (500mgkg-1 of feed) was added to a commercial carp feed for six weeks. We compared the serum proteome profile from 7 controls and 7 treated fish. In addition, zinc accumulation were measured in intestine, liver, gill and brain. In total, we were able to identify 326 proteins from 6845 distinct peptides. As a result of the data analysis, the abundance levels of four proteins were significantly altered (fold change (fc) ≥2 and p<0.05) after dietary exposure to ZnO NPs. The protein levels of the complement component C4-2 (fc 2.5) and the uncharacterised protein encoded by kng1 (fc 5.8) were increased and major histocompatibility class I (fc 4.9) and the uncharacterised protein encoded by lum (fc 3.5) were decreased (fc 2.5). Molecular pathway analysis revealed four canonical pathways including acute-phase response signalling, liver and retinoid X receptors activation, and intrinsic and extrinsic prothrombin activation pathways as significantly regulated in the treated fish. No significant difference was observed for zinc accumulation in exposed fish compared to controls. In summary, despite no apparent accumulation, ZnO NPs exposure to common carp probably disturbs the fish homeostasis by affecting proteins of the haematological and the immune systems.


Asunto(s)
Nanopartículas del Metal/toxicidad , Pruebas de Toxicidad Crónica , Contaminantes Químicos del Agua/toxicidad , Óxido de Zinc/toxicidad , Zinc/toxicidad , Animales , Carpas , Exposición Dietética , Peroxidación de Lípido/efectos de los fármacos , Estrés Oxidativo
18.
Food Chem Toxicol ; 96: 50-61, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27456126

RESUMEN

Silver nanoparticles (AgNPs) are the most frequently applied nanomaterials. In our experiments, we tested AgNPs (size 27 nm) manufactured by the Tollens process. Physico-chemical methods (TEM, DLS, AFM and spectrophotometry) were used for characterization and imaging of AgNPs. The effects of AgNPs and Ag(+) were studied in two experimental models (plant and mammalian cells). Human keratinocytes (SVK14) and mouse fibroblasts (NIH3T3) cell lines were selected to evaluate the cytotoxicity and genotoxicity effect on mammalian cells. Higher sensitivity to AgNPs and Ag(+) was observed in NIH3T3 than in SVK14 cells. AgNPs accumulated in the nucleus of NIH3T3 cells, caused DNA damage and increased the number of apoptotic and necrotic cells. Three genotypes of Solanum spp. (S. lycopersicum cv. Amateur, S. chmielewskii, S. habrochaites) were selected to test the toxicity of AgNPs and Ag(+) on the plant cells. The highest values of peroxidase activity and lipid peroxidation were recorded after the treatment of S. habrochaites genotype with AgNPs. Increased ROS levels were likely the reason for observed damaged membranes in S. habrochaites. We found that the cytotoxic and genotoxic effects of AgNPs depend not only on the characteristics of nanoparticles, but also on the type of cells that are treated with AgNPs.


Asunto(s)
Apoptosis/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Nanopartículas del Metal/administración & dosificación , Plata/farmacología , Solanum lycopersicum/citología , Animales , Western Blotting , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Ensayo Cometa , Humanos , Técnicas In Vitro , Solanum lycopersicum/efectos de los fármacos , Nanopartículas del Metal/química , Ratones , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismo , Plata/química , Espectrometría Raman
19.
Ultrason Sonochem ; 32: 165-172, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-27150757

RESUMEN

A fast method for preparing of silver particle layers on glass substrates with high application potential for using in surface enhanced Raman spectroscopy (SERS) is introduced. Silver particle layers deposited on glass cover slips were generated in one-step process by reduction of silver nitrate using several reducing agents (ethylene glycol, glycerol, maltose, lactose and glucose) under ultrasonic irradiation. This technique allows the formation of homogeneous layers of silver particles with sizes from 80nm up to several hundred nanometers depending on the nature of the used reducing agent. Additionally, the presented method is not susceptible to impurities on the substrate surface and it does not need any additives to capture or stabilize the silver particles on the glass surface. The characteristics of prepared silver layers on glass substrate by the above mentioned sonochemical approach was compared with chemically prepared ones. The prepared layers were tested as substrates for SERS using adenine as a model analyte. The factor of Raman signal enhancement reached up to 5·10(5). On the contrary, the chemically prepared silver layers does not exhibit almost any pronounced Raman signal enhancement. Presented sonochemical approach for preparation of silver particle layers is fast, simple, robust, and is better suited for reproducible fabrication functional SERS substrates than chemical one.

20.
IET Nanobiotechnol ; 10(2): 62-8, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27074855

RESUMEN

In this study, culture supernatnats of Bacillus subtilis T-1 growing on brewery effluents and molasses was used for silver nanoparticles (Ag-NPs) synthesis. The biosurfactant production of B. subtilis T-1 was confirmed by the detection of genes in the genome and by the identification of the product in the supernatants. The genes for synthesis of surfactin (sfp, srfAA) and iturin (ituC) were noted by PCR reactions. Also, in examined culture supernatants the presence of C13, C14 and C15 surfactin homologues with the sodiated molecules [M + Na](+) at m/z 1030, 1044 and 1058 was confirmed using LC/MS/MS analysis. The formation of NPs in the culture supernatants was confirmed by UV-vis spectroscopy. The dynamic light scattering measurements and transmission electron microscopy images showed the nanometric sizes of the biosynthesised Ag-NPs which ranged from several nm to several tens of nm depending on the used culture supernatant. Biological properties of Ag-NPs were evaluated by binding of Ag-NPs with DNA isolated from the Escherichia coli ATCC 25922 and B. subtilis ATCC 6633. Biogenic Ag-NPs were actively bound to DNA in increased concentration which could be the one important mode of antibacterial action of the Ag-NPs.


Asunto(s)
Bacillus subtilis/metabolismo , Residuos Industriales , Nanopartículas del Metal/química , Plata/química , Plata/metabolismo , Tensoactivos/metabolismo , Agricultura , Bacillus subtilis/genética , Bacillus subtilis/crecimiento & desarrollo , Lipopéptidos/genética , Lipopéptidos/aislamiento & purificación , Lipopéptidos/metabolismo , Microscopía Electrónica de Transmisión , Melaza/microbiología , Tamaño de la Partícula , Péptidos Cíclicos/genética , Péptidos Cíclicos/aislamiento & purificación , Péptidos Cíclicos/metabolismo , Tensoactivos/química , Tensoactivos/aislamiento & purificación , Espectrometría de Masas en Tándem
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