PurposeThe biomedical applications of silver nanoparticles (AgNPs) are heavily investigated due to their cytotoxic and antimicrobial properties. However, the scientific literature is lacking in data on the aggregation behavior of nanoparticles, especially regarding its impact on biological activity. Therefore, to assess the potential of AgNPs in therapeutic applications, two different AgNP samples were compared under biorelevant conditions.MethodsCitrate-capped nanosilver was produced by classical chemical reduction and stabilization with sodium citrate (AgNP@C), while green tea extract was used to produce silver nanoparticles in a green synthesis approach (AgNP@GTs). Particle size, morphology, and crystallinity were characterized using transmission electron microscopy. To observe the effects of the most important biorelevant conditions on AgNP colloidal stability, aggregation grade measurements were carried out using UV-Vis spectroscopy and dynamic light scatterig, while MTT assay and a microdilution method were performed to evaluate the effects of aggregation on cytotoxicity and antimicrobial activity in a time-dependent manner.ResultsThe aggregation behavior of AgNPs is mostly affected by pH and electrolyte concentration, while the presence of biomolecules can improve particle stability due to the biomolecular corona effect. We demonstrated that high aggregation grade in both AgNP samples attenuated their toxic effect toward living cells. However, AgNP@GT proved less prone to aggregation thus retained a degree of its toxicity.ConclusionTo our knowledge, this is the first systematic examination regarding AgNP aggregation behavior with simultaneous measurements of its effect on biological activity. We showed that nanoparticle behavior in complex systems can be estimated by simple compounds like sodium chloride and glutamine. Electrostatic stabilization might not be suitable for biomedical AgNP applications, while green synthesis approaches could offer new frontiers to preserve nanoparticle toxicity by enhancing colloidal stability. The importance of properly selected synthesis methods must be emphasized as they profoundly influence colloidal stability, and therefore biological activity.

Due to obvious disadvantages of the classical chemical methods, green synthesis of metallic nanoparticles has attracted tremendous attention in recent years. Numerous environmentally benign synthesis methods have been developed yielding nanoparticles via low-cost, eco-friendly, and simple approaches. In this study, our aim was to determine the suitability of coffee and green tea extracts in green synthesis of silver nanoparticles as well as to compare the performance of the obtained materials in different biological systems. We successfully produced silver nanoparticles (C-AgNP and GT-AgNP) using coffee and green tea extracts; moreover, based on our comprehensive screening, we delineated major differences in the biological activity of C-AgNPs and GT-AgNPs. Our results indicate that although GT-AgNPs exhibited excellent antimicrobial activity against all the examined microbial pathogens, these particles were also highly toxic to mammalian cells, which limits their potential applications. On the contrary, C-AgNPs manifested substantial inhibitory action on the tested microbes but were nontoxic to human and mouse cells, indicating an outstanding capacity to discriminate between potential pathogens and mammalian cells. These results clearly show that the various green materials used for stabilization and for reduction of metal ions have a defining role in determining and fine-tuning the biological activity of the obtained nanoparticles.

The nanomaterial industry generates gigantic quantities of metal-based nanomaterials for various technological and biomedical applications; however, concomitantly, it places a massive burden on the environment by utilizing toxic chemicals for the production process and leaving hazardous waste materials behind. Moreover, the employed, often unpleasant chemicals can affect the biocompatibility of the generated particles and severely restrict their application possibilities. On these grounds, green synthetic approaches have emerged, offering eco-friendly, sustainable, nature-derived alternative production methods, thus attenuating the ecological footprint of the nanomaterial industry. In the last decade, a plethora of biological materials has been tested to probe their suitability for nanomaterial synthesis. Although most of these approaches were successful, a large body of evidence indicates that the green material or entity used for the production would substantially define the physical and chemical properties and as a consequence, the biological activities of the obtained nanomaterials. The present review provides a comprehensive collection of the most recent green methodologies, surveys the major nanoparticle characterization techniques and screens the effects triggered by the obtained nanomaterials in various living systems to give an impression on the biomedical potential of green synthesized silver and gold nanoparticles.

Mitochondrial genome diversity in closely related species provides an excellent platform for investigation of chromosome architecture and its evolution by means of comparative genomics. In this study, we determined the complete mitochondrial DNA sequences of eight Candida species and analyzed their molecular architectures. Our survey revealed a puzzling variability of genome architecture, including circular- and linear-mapping and multipartite linear forms. We propose that the arrangement of large inverted repeats identified in these genomes plays a crucial role in alterations of their molecular architectures. In specific arrangements, the inverted repeats appear to function as resolution elements, allowing genome conversion among different topologies, eventually leading to genome fragmentation into multiple linear DNA molecules. We suggest that molecular transactions generating linear mitochondrial DNA molecules with defined telomeric structures may parallel the evolutionary emergence of linear chromosomes and multipartite genomes in general and may provide clues for the origin of telomeres and pathways implicated in their maintenance.

As a part of our initiative aimed at a large-scale comparative analysis of fungal mitochondrial genomes, we determined the complete DNA sequence of the mitochondrial genome of the yeast Candida subhashii and found that it exhibits a number of peculiar features. First, the mitochondrial genome is represented by linear dsDNA molecules of uniform length (29 795 bp), with an unusually high content of guanine and cytosine residues (52.7 %). Second, the coding sequences lack introns; thus, the genome has a relatively compact organization. Third, the termini of the linear molecules consist of long inverted repeats and seem to contain a protein covalently bound to terminal nucleotides at the 5′ ends. This architecture resembles the telomeres in a number of linear viral and plasmid DNA genomes classified as invertrons, in which the terminal proteins serve as specific primers for the initiation of DNA synthesis. Finally, although the mitochondrial genome of C. subhashii contains essentially the same set of genes as other closely related pathogenic Candida species, we identified additional ORFs encoding two homologues of the family B protein-priming DNA polymerases and an unknown protein. The terminal structures and the genes for DNA polymerases are reminiscent of linear mitochondrial plasmids, indicating that this genome architecture might have emerged from fortuitous recombination between an ancestral, presumably circular, mitochondrial genome and an invertron-like element.