Vedran Milosavljevic

Selenium nanoparticles as a nutritional supplement.

Selenium is an essential trace element in the diet, required for maintenance of health and growth; however, its toxicity could cause serious damage depending on dose and chemical form. Selenium nanoparticles (SeNPs) represent what we believe to be a novel prospect for nutritional supplementation because of their lower toxicity and ability to gradually release selenium after ingestion. In this review, we discuss various forms and types of SeNPs, as well as the way they are synthesized. We also discuss absorption and bioavailability of nanoparticles within the organism. SeNPs demonstrate anticancer and antimicrobial properties that may contribute to human health, not only as dietary supplements, but also as therapeutic agents.

3D-printed chip for detection of methicillin-resistant Staphylococcus aureus labeled with gold nanoparticles

Methicillin‐resistant Staphylococcus aureus (MRSA) is a dangerous pathogen occurring not only in hospitals but also in foodstuff. Currently, discussions on the issue of the increasing resistance, and timely and rapid diagnostic of resistance strains have become more frequent and sought. Therefore, the aim of this study was to design an effective platform for DNA isolation from different species of microorganisms as well as the amplification of mecA gene that encodes the resistance to β‐lactam antibiotic formation and is contained in MRSA. For this purpose, we fabricated 3D‐printed chip that was suitable for bacterial cultivation, DNA isolation, PCR, and detection of amplified gene using gold nanoparticle (AuNP) probes as an indicator of MRSA. Confirmation of the MRSA presence in the samples was based on a specific interaction between mecA gene with the AuNP probes and a colorimetric detection, which utilized the noncross‐linking aggregation phenomenon of DNA‐functionalized AuNPs. To test the whole system, we analyzed several real refractive indexes, in which two of them were positively scanned to find the presence of mecA gene. The aggregation of AuNP probes were reflected by 75% decrease of absorbance (λ = 530 nm) and change in AuNPs size from 3 ± 0.05 to 4 ± 0.05 nm (n = 5). We provide the one‐step identification of mecA gene using the unique platform that employs the rapid, low‐cost, and easy‐to‐use colorimetric method for MRSA detection in various samples.

Synthesis of carbon quantum dots for DNA labeling and its electrochemical, fluorescent and electrophoretic characterization

Nanoparticles as a progressively developing branch offer a tool for studying the interaction of carbon quantum dots (CQDs) with DNA. In this study, fluorescent CQDs were synthesized using citric acid covered with polyethylene glycol (PEG) as the source of carbon precursors. Furthermore, interactions between CQDs and DNA (double-stranded DNA and single-stranded DNA) were investigated by spectral methods, gel electrophoresis, and electrochemical analysis. Primarily, the fluorescent behavior of CQDs in the presence of DNA was monitored and major differences in the interaction of CQDs with tested single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) were observed at different amounts of CQDs (µg mL−1: 25, 50, 100, 250, 500). It was found that the interaction of ssDNA with CQDs had no significant influence on the CQDs fluorescence intensity measured at the excitation wavelengths of 280 nm, 350 nm, and 400 nm. However, in the presence of dsDNA, the fluorescence intensity of CQDs was significantly increased. Our results provide basic understanding of the interaction between CQDs and DNA. Such fabricated CQDs-DNA might be of great benefit for the emerging nanomaterials based biosensing methods.

Comparative study on toxicity of extracellularly biosynthesized and laboratory synthesized CdTe quantum dots

Nanobiosynthesis belongs to the most recent methods for synthesis of nanoparticles. This type of synthesis provides many advantages including the uniformity in particle shape and size. The biosynthesis has also a significant advantage regarding chemical properties of the obtained particles. In this study, we characterized the basic properties and composition of quantum dots (QDs), obtained by the extracellular biosynthesis by Escherichia coli. Furthermore, the toxicity of the biosynthesized QDs was compared to QDs prepared by microwave synthesis. The obtained results revealed the presence of cyan CdTe QDs after removal of substantial amounts of organic compounds, which stabilized the nanoparticle surface. QDs toxicity was evaluated using three cell lines Human Foreskin Fibroblast (HFF), Human Prostate Cancer cells (PC-3) and Breast Cancer cells (MCF-7) and the MTT assay. The test revealed differences in the toxicity between variants of QDs, varying about 10% in the HFF and 30% in the MCF-7 cell lines. The toxicity of the biosynthesized QDs to the PC-3 cell lines was about 35% lower in comparison with the QDs prepared by microwave synthesis.

Application of CdTe/ZnSe quantum dots in in vitro imaging of chicken tissue and embryo.

The present work is aimed to synthesize CdTe/ZnSe core/shell quantum dots (QDs) in an easy way and to explore the possibilities of its application in in vitro imaging of chicken tissue and embryo. The QDs were prepared using microwave irradiation with different temperatures, which is a very easy and less time‐consuming method. Subsequently, these QDs were characterized by spectrofluorimetry, Transmission Electron Microscopy, X‐ray fluorescence analysis and Dynamic Light Scattering measurement. A blueshifting of the emission was found when ZnSe was deposited on CdTe QDs. The QDs showed its fluorescence emission quantum yields up to 25%. They were applied into chicken embryos and breast muscle tissues to study their efficiency in in vitro imaging. All the QDs of different color were able to visualize in in vitro imaging. The highest fluorescence intensity was detected in the case of red QDs prepared at 100°C. The green and red QDs were possible to detect up to the depth of 3 and 4 mm of the tissue, respectively.

Antibody-free detection of infectious bacteria using quantum dots-based barcode assay.

HIGHLIGHTSDetection of S. aureus, methicillin‐resistant S. aureus and K. pneumoniae.The method was able to detect as low concentrations of bacteria as 102 CFU/mL.Detection was enabled using the bacteria‐specific genes (fnbA, mecA and wcaG).The proposed method is employing quantum dots and magnetic particles. ABSTRACT Staphylococcus aureus, methicillin‐resistant Staphylococcus aureus and Klebsiella pneumoniae are the most representative bacteria causing infectious diseases. Due to the increased application of antibiotics, the bacterial resistance is growing causing severe complications. Therefore, a sensitive determination of these pathogens is crucial for effective treatment. The aim of this study was to design an effective method for multiplex detection of Staphylococcus aureus, methicillin‐resistant Staphylococcus aureus and Klebsiella pneumoniae taking advantage from properties of magnetic particles as well as fluorescent nanoparticles (quantum dots). The method was able to detect as low concentrations of bacteria as 102 CFU/mL using the bacteria‐specific genes (fnbA, mecA and wcaG).

Antiviral activity of fullerene C60 nanocrystals modified with derivatives of anionic antimicrobial peptide maximin H5

Many active antiviral substances come from natural sources. In this way, peptides, isolated from Asian toad Bombina maxima, called maximins, are very promising. Most of them have good antimicrobial activity; however, derivatives of anionic 20 amino acids-long maximin H5 show also promising antiviral activity. The effect can be enhanced by binding to suitable nanocarriers such as fullerenes. In the present study, six mutants of maximin H5 were designed where aspartic acid at position 11 was replaced by asparagine, histidine, tyrosine, alanine, glycine, or valine. The binding yield of each peptide to fullerene C60 nanocrystals was studied by derivatization with fluorescent reagent fluorescamine. The antiviral activity of these peptides and peptides bound to fullerene C60 nanocrystals was studied using bacteriophage λ as a model virus. All of the designed peptides had higher antiviral activity compared to maximin H5. The highest antiviral activity was observed in case of maximin variants H5N, H5V, or H5Y. Moreover, the antiviral activity was dependent on the amount of peptide bound on the surface of fullerene C60 nanocrystals, which was enhanced by trimesic acid (benzene-1,3,5-tricarboxylic acid) treated fullerene C60 nanocrystals.Graphical abstract

The Zinc-Schiff Base-Novicidin Complex as a Potential Prostate Cancer Therapy

Prostate cancer cells control energy metabolism by chelating intracellular zinc. Thus, zinc delivery has been a popular therapeutic approach for prostate cancer. Here, we propose the use of the membrane-penetrating peptide Novicidin connected to zinc-Schiff base as a carrier vehicle for the delivery of zinc to prostate cells. Mass spectrometry, electrochemistry and spectrophotometry confirmed the formation/stability of this complex and provided insight regarding the availability of zinc for complex interactions. This delivery system showed minor toxicity in normal PNT1A cells and high potency towards PC3 tumor cells. The complex preferentially penetrated PC3 tumor cells in contrast to confinement to the membranes of PNT1A. Furthermore, zinc uptake was confirmed in both cell lines. Molecular analysis was used to confirm the activation of zinc stress (e.g., ZnT-1) and apoptosis (e.g., CASP-1). Our results strongly suggest that the zinc-Schiff base-Novicidin complex has great potential as a novel anticancer drug.

Using CdTe/ZnSe core/shell quantum dots to detect DNA and damage to DNA

CdTe/ZnSe core/shell quantum dot (QD), one of the strongest and most highly luminescent nanoparticles, was directly synthesized in an aqueous medium to study its individual interactions with important nucleobases (adenine, guanine, cytosine, and thymine) in detail. The results obtained from the optical analyses indicated that the interactions of the QDs with different nucleobases were different, which reflected in different fluorescent emission maxima and intensities. The difference in the interaction was found due to the different chemical behavior and different sizes of the formed nanoconjugates. An electrochemical study also confirmed that the purines and pyrimidines show different interactions with the core/shell QDs. Based on these phenomena, a novel QD-based method is developed to detect the presence of the DNA, damage to DNA, and mutation. The QDs were successfully applied very easily to detect any change in the sequence (mutation) of DNA. The QDs also showed their ability to detect DNAs directly from the extracts of human cancer (PC3) and normal (PNT1A) cells (detection limit of 500 pM of DNA), which indicates the possibilities to use this easy assay technique to confirm the presence of living organisms in extreme environments.

Advanced nanotechnologies in avian influenza: current status and future trends – A review

Abstract In the last decade, the control of avian influenza virus has experienced many difficulties, which have caused major global agricultural problems that have also led to public health consequences. Conventional biochemical methods are not sufficient to detect and control agricultural pathogens in the field due to the growing demand for food and subsidiary products; thus, studies aiming to develop potent alternatives to conventional biochemical methods are urgently needed. In this review, emerging detection systems, their applicability to diagnostics, and their therapeutic possibilities in view of nanotechnology are discussed. Nanotechnology-based sensors are used for rapid, sensitive and cost-effective diagnostics of agricultural pathogens. The application of different nanomaterials promotes interactions between these materials and the virus, which enables researchers to construct portable electroanalytical biosensing analyser that should effectively detect the influenza virus. The present review will provide insights into the guidelines for future experiments to develop better techniques to detect and control influenza viruses.