Novel Biogenic Silver Nanoparticle-Induced Reactive Oxygen Species Inhibit the Biofilm Formation and Virulence Activities of Methicillin-Resistant Staphylococcus aureus (MRSA) Strain.

Emerging antibiotic-resistant bacteria result in increased mortality and have negative economic impacts. It is necessary to discover new strategies to create alternative antibacterial agents that suppress the bacterial resistance mechanism and limit the spread of serious infectious bacterial diseases. Silver nanoparticles may represent a new medicinal agents as alternative antibiotics affect different bacterial mechanisms such as virulence and resistance. In addition to that of silver nitrate (AgNO3) and ampicillin, for the first time, the inhibitory effect of silver nanoparticles synthesized using Desertifilum sp. (D-SNPs) was evaluated against five pathogenic bacteria using the agar well diffusion method. Also, the influence of D-SNPs and AgNO3 on bacterial antioxidant and metabolic activities was studied. The antibacterial activity of D-SNPs and AgNO3 against methicillin-resistant Staphylococcus aureus (MRSA) strains was studied at the morphological and molecular level. D-SNPs and AgNO3 have the ability to inhibit the growth of the five bacterial strains and resulted in an imbalance in the CAT, GSH, GPx and ATPase levels. MRSA treated with D-SNPs and AgNO3 showed different morphological changes such as apoptotic bodies formation and cell wall damage. Moreover, both caused genotoxicity and denaturation of MRSA cellular proteins. Additionally, TEM micrographs showed the distribution of SNPs synthesized by MRSA. This result shows the ability of MRSA to reduce silver nitrate into silver nanoparticles. These data indicate that D-SNPs may be a significant alternative antibacterial agent against different bacteria, especially MDR bacteria, by targeting the virulence mechanism and biofilm formation, leading to bacterial death.

Synthesis of Silver Nanoparticles Using a Novel Cyanobacteria Desertifilum sp. extract: Their Antibacterial and Cytotoxicity Effects.

BACKGROUND: The emergence of multi drug-resistant (MDR) bacterial infections and cancer has necessitated the development and discovery of alternative eco-safe antibacterial and anticancer agents. Biogenic fabrication of metallic nanoparticles is an emerging discipline for production of nanoproducts that exert potent anticancer and antibacterial activity, and do not suffer from the limitations inherent in physiochemical synthesis methods. METHODOLOGY: In this study, we isolated, purified, and characterized a novel cyanobacteria extract (Desertifilum IPPAS B-1220) to utilize in biofabrication of silver nanoparticles (D-SNPs). D-SNPs were produced by adding Desertifilum extract to silver nitrate solution under controlled conditions. Biofabrication of D-SNPs was confirmed using a UV-Vis spectrophotometer. The resultant D-SNPs were characterized using XRD, FTIR, SEM, and TEM. The toxicity of D-SNPs against five pathogenic bacteria and three cancer cell lines (MCF-7, HepG2, and Caco-2) was evaluated. RESULTS: Formation of D-SNPs was indicated by a color change from pale yellow to dark brown. The peak of the surface plasmon resonance of the D-SNPs was at 421 nm. The XRD detected the crystallinity of D-SNPs. FTIR showed that polysaccharides and proteins may have contributed to the biofabrication of D-SNPs. Under SEM and TEM, the D-SNPs were spherical with diameter ranges from 4.5 to 26 nm. The D-SNPs significantly suppressed the growth of five pathogenic bacteria, and exerted cytotoxic effects against MCF-7, HepG2, and Caco-2 cancer cells with IC50 values of 58, 32, and 90 µg/mL, respectively. CONCLUSION: These findings showed for the first time the potentiality of novel cyanobacteria strain Desertifilum IPPAS B-1220 to fabricate small SNPs that acted as potent anticancer and antibacterial material against different cancer cell lines and pathogenic bacterial strains. These findings encourage the researchers to focus on cyanobacteria in general and especially Desertifilum sp. IPPAS B-1220 for synthesizing different NPs that opening the window for new applications.

Cytotoxic effect of green silver nanoparticles against ampicillin-resistant Klebsiella pneumoniae.

Considering the harmful effects and high spread of drug-resistant Klebsiella pneumoniae, many researchers have been trying to produce new antibacterial agents to combat the emergence of multidrug-resistant (MDR) strains of this bacterium. Recent progress in the nanomedicine field has provided opportunities for synthesizing unique nanoagents to battle MDR bacteria by targeting virulence and resistance signalling. The biocidal effects of 14.9 nm silver nanoparticles fabricated using Nostoc sp. Bahar M (N-SNPs) and AgNO3 were examined against drug-resistant K. pneumoniae using the agar well diffusion method. Transmission electron microscopy (TEM) was used to detect the ultrastructural changes caused by N-SNPs and AgNO3. To address the mode of action of N-SNPs and AgNO3, CAT, GPx, LDH and ATPase levels were assessed. The toxicity of N-SNPs and AgNO3 was evaluated against the mfD, flu, hly, 23S, hns, hcp-1, VgrG-1 and VgrG-3 genes as well as cellular proteins. N-SNPs showed the greatest inhibitory activity against K. pneumoniae, with MIC and MBC values of 0.9 and 1.2 mg mL-1, respectively. Furthermore, N-SNPs and AgNO3 induced apoptotic features, including cell shrinkage and cell atrophy. N-SNPs were more potent bactericidal compounds than AgNO3, causing increased leakage of LDH and GPx activities and depletion of ATPase and CAT activities, resulting in induced oxidative stress and metabolic toxicity. Compared to AgNO3, N-SNPs exhibited the highest toxicity towards the selected genes and the greatest damage to bacterial proteins. N-SNPs were the most potent agents that induced bacterial membrane damage, oxidative stress and disruption of biomolecules such as DNA and proteins. N-SNPs may be used as effective nanodrugs against MDR bacteria.

Developing Spindlin1 small-molecule inhibitors by using protein microarrays.

The discovery of inhibitors of methyl- and acetyl-binding domains has provided evidence for the 'druggability' of epigenetic effector molecules. The small-molecule probe UNC1215 prevents methyl-dependent protein-protein interactions by engaging the aromatic cage of MBT domains and, with lower affinity, Tudor domains. Using a library of tagged UNC1215 analogs, we screened a protein-domain microarray of human methyllysine effector molecules to rapidly detect compounds with new binding profiles with either increased or decreased specificity. Using this approach, we identified a compound (EML405) that acquired a novel interaction with the Tudor-domain-containing protein Spindlin1 (SPIN1). Structural studies facilitated the rational synthesis of SPIN1 inhibitors with increased selectivity (EML631-633), which engage SPIN1 in cells, block its ability to 'read' H3K4me3 marks and inhibit its transcriptional-coactivator activity. Protein microarrays can thus be used as a platform to 'target-hop' and identify small molecules that bind and compete with domain-motif interactions.