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One of the officially approved medications for the treatment of the pandemic disease COVID-19, caused by the SARS-CoV-2 virus, is remdesivir. This antiviral molecule is a prodrug that is metabolized into its active form (an ATP analogue). Because of its hepatotoxicity and nephrotoxicity, it is necessary to monitor the serum concentrations of remdesivir. For the therapeutic drug monitoring of remdesivir, a method using liquid chromatography with tandem mass spectrometry (LC-MS/MS) in positive electrospray ionization mode was developed. Mass detection was done via triple quadrupole in the Multiple reaction monitoring mode. Separation was done on Zorbax C18 column at 35 °C in mobile phase gradient and flow 0.4 mL min–1 (A – 0.1% formic acid in water, B – 0.1% formic acid in 95% acetonitrile). Time of analysis was 4 minutes. LC-MS/MS method was successfully validated. Calibration was done in blood serum and plasma and it was linear in the range of tested concentrations (0–1000 ng mL–1). Samples were prepared by protein precipitation. The method was used to measure remdesivir concentration in a patient with SARS-CoV-2 infection. The measured concentration 60 minutes after remdesivir application was 175±15 ng mL–1.

Polysaccharide nanoparticles are promising materials in the wide range of disciplines such as medicine, nutrition, food production, agriculture, material science and others. They excel not only in their non-toxicity and biodegradability but also in their easy preparation. As well as inorganic particles, a protein corona (PC) around polysaccharide nanoparticles is formed in biofluids. Moreover, it has been considered that the overall response of the organism to nanoparticles presence depends on the PC. This review summarises scientific publications about the structural chemistry of polysaccharide nanoparticles and their impact on theranostic applications. Three strategies of implementation of the PC in theranostics have been discussed: I) Utilisation of the PC in therapy; II) How the composition of the PC is analysed for specific disease markers; III) How the formed PC can interact with the immune system and enhances the immunomodulation or immunoelimination. Thus, the findings from this review can contribute to improve the design of drug delivery systems. However, it is still necessary to elucidate the mechanisms of nano-bio interactions and discover new connections in nanoscale research.

According to the United States Centers for Disease Control and Prevention (CDC), as of July 11, 2016, the reported average incidence of children diagnosed with an autism spectrum disorder (ASD) was 1 in 68 (1.46%) among 8-year-old children born in 2004 and living within the 11 monitoring sites' surveillance areas in the United States of America (USA) in 2012. ASD is a multifaceted neurodevelopmental disorder that is also considered a hidden disability, as, for the most part; there are no apparent morphological differences between children with ASD and typically developing children. ASD is diagnosed based upon a triad of features including impairment in socialization, impairment in language, and repetitive and stereotypic behaviors. The increasing incidence of ASD in the pediatric population and the lack of successful curative therapies make ASD one of the most challenging disorders for medicine. ASD neurobiology is thought to be associated with oxidative stress, as shown by increased levels of reactive oxygen species and increased lipid peroxidation, as well as an increase in other indicators of oxidative stress. Children with ASD diagnosis are considered more vulnerable to oxidative stress because of their imbalance in intracellular and extracellular glutathione levels and decreased glutathione reserve capacity. Several studies have suggested that the redox imbalance and oxidative stress are integral parts of ASD pathophysiology. As such, early assessment and treatment of antioxidant status may result in a better prognosis as it could decrease the oxidative stress in the brain before it can induce more irreversible brain damage. In this review, many aspects of the role of oxidative stress in ASD are discussed, taking into account that the process of oxidative stress may be a target for therapeutic interventions.

Nanotechnology is one of the most promising future technologies for the food industry. Some of its applications have already been introduced in analytical techniques and food packaging technologies. This review summarizes existing knowledge about the implementation of nanotechnology in wine laboratory procedures. The focus is mainly on recent advancements in the design and development of nanomaterial-based sensors for wine compounds analysis and assessing wine safety. Nanotechnological approaches could be useful in the wine production process, to simplify wine analysis methods, and to improve the quality and safety of the final product.

Silver nanoparticles are the most important nanoparticles in connection with the antimicrobial effect. Nowadays, the green synthesis of various types of nanoparticles is rapid, effective and produce less toxic nanoparticles often with specific properties. In our experiment we have developed and described in details various types of silver nanoparticles synthesized chemically or by the green synthesis. Nine different silver nanoparticles were synthesized, three by citrate method at different pHs (8; 9; 10), four using gallic acid at alkaline pHs (10; 11), and two by green synthesis using green tea and coffee extracts, both at pH 9. Characterisation of silver nanoparticles was performed using dynamic light scattering, scanning electron microscopy, and ultraviolet-visible absorption spectroscopy. Silver nanoparticles prepared by green synthesis showed the highest antioxidant activity and also ability for quenching of free radicals. Antibacterial activity of silver nanoparticles was determined on bacterial cultures such as Staphylococcus aureus and Escherichia coli. Silver nanoparticles synthesized using green tea and coffee extracts showed the highest antibacterial activity for both bacterial strains. Minimal inhibition concentration for both strains was found to be 65 μM at each silver nanoparticle synthesized using green synthesis.

Nicotinamide phosphoribosyltransferase (NAMPT) is located in both the nucleus and cytoplasm and has multiple biological functions including catalyzing the rate-limiting step in NAD synthesis. Moreover, up-regulated NAMPT expression has been observed in many cancers. However, the determinants and regulation of NAMPT's nuclear transport are not known. Here, we constructed a GFP–NAMPT fusion protein to study NAMPT's subcellular trafficking. We observed that in unsynchronized 3T3-L1 preadipocytes, 25% of cells had higher GFP–NAMPT fluorescence in the cytoplasm, and 62% had higher GFP–NAMPT fluorescence in the nucleus. In HepG2 hepatocytes, 6% of cells had higher GFP–NAMPT fluorescence in the cytoplasm, and 84% had higher GFP–NAMPT fluorescence in the nucleus. In both 3T3-L1 and HepG2 cells, GFP–NAMPT was excluded from the nucleus immediately after mitosis and migrated back into it as the cell cycle progressed. In HepG2 cells, endogenous, untagged NAMPT displayed similar changes with the cell cycle, and in nonmitotic cells, GFP–NAMPT accumulated in the nucleus. Similarly, genotoxic, oxidative, or dicarbonyl stress also caused nuclear NAMPT localization. These interventions also increased poly(ADP-ribosyl) polymerase and sirtuin activity, suggesting an increased cellular demand for NAD. We identified a nuclear localization signal in NAMPT and amino acid substitution in this sequence ((424)RSKK to ASGA), which did not affect its enzymatic activity, blocked nuclear NAMPT transport, slowed cell growth, and increased histone H(3) acetylation. These results suggest that NAMPT is transported into the nucleus where it presumably increases NAD synthesis required for cell proliferation. We conclude that specific inhibition of NAMPT transport into the nucleus might be a potential avenue for managing cancer.

Modern anticancer therapy aims to increase the effectiveness of tumor treatment. The aim of this work was to propose a new nanotransporter for targeted delivery of anthracycline antibiotics, which is characterized by its bioavailability, increased uptake of the drug from the bloodstream at the site of tumor tissue and as well as low toxicity to non-target tissue. Chitosan nanoparticles have attracted great attention in the field of drug delivery due to their stability, low toxicity and easy preparation. Deacetylated chitosan skeleton is composed of glucosamine units and has a high density of charged amino groups which allow strong electrostatic interactions with biomolecules, transition metals (Zn, Se) and peptides. We obtained the encapsulation effectiveness of chitosan 20%. Electrochemical detection of the bounded Zn2+ ions into the chitosan structure showed shift from -0.99 to -0.93 V. This result proved the formation of a chitosan-zinc complex. The ability of metallothione in to quench the 2,2-diphenyl-1-picrylhydrazylradicalin the presence of 50 {aelig}M doxorubicin was confirmed by the change of relative absorbance in the range of 50 to 60%.

Quantum dots (QDs) have a broad range of applications in cell biolabeling, cancer treatment, metastasis imaging, and therapeutic drug monitoring. Despite their wide use, relatively little is known about their influence on other molecules. Interactions between QDs and proteins can influence the properties of both nanoparticles and proteins. The effect of mercaptosuccinic acid-capped CdTe QDs on intercellular copper–zinc superoxide dismutase (SOD1)—one of the main enzymatic antioxidants—was investigated. Incubation of SOD1 with QDs caused an increase in SOD1 activity, unlike in the case of CdCl2, which inhibited SOD1. Moreover, this effect on SOD1 increased with the size and potential of QDs, although the effect became clearly visible in higher concentrations of QDs. The intensity of QD-SOD1 fluorescence, analyzed with the use of capillary electrophoresis with laser-induced fluorescence detection, was dependent on SOD1 concentration. In the case of green QDs, the fluorescence signal decreased with increasing SOD1 concentration. In contrast, the signal strength for Y-QD complexes was not dependent on SOD1 dilutions. The migration time of QDs and their complexes with SOD1 varied depending on the type of QD used. The migration time of G-QD complexes with SOD1 differed slightly. However, in the case of Y-QD complexes with SOD1, the differences in the migration time were not dependent on SOD concentration. This research shows that QDs interact with SOD1 and the influence of QDs on SOD activity is size-dependent. With this knowledge, one might be able to control the activation/inhibition of specific enzymes, such as SOD1.

Silver nanoparticles (AgNPs) have recently become very attractive for the scientific community due to their broad spectrum of applications in the biomedical field. The main advantages of AgNPs include a simple method of synthesis, a simple way to change their morphology and high surface area to volume ratio. Much research has been carried out over the years to evaluate their possible effectivity against microbial organisms. The most important factors which influence the effectivity of AgNPs against microorganisms are the method of their preparation and the type of application. When incorporated into fabric wound dressings and other textiles, AgNPs have shown significant antibacterial activity against both Gram-positive and Gram-negative bacteria and inhibited biofilm formation. In this review, the different routes of synthesizing AgNPs with controlled size and geometry including chemical, green, irradiation and thermal synthesis, as well as the different types of application of AgNPs for wound dressings such as membrane immobilization, topical application, preparation of nanofibers and hydrogels, and the mechanism behind their antimicrobial activity, have been discussed elaborately.

The aim of this study was to synthesize cadmium telluride nanoparticles (CdTe NPs) modified apoferritin, and examine if apoferritin is able to accommodate CdTe NPs. Primarily, the thermostability of horse spleen apoferritin was tested and it's unfolding at 70 °C was observed. Cadmium telluride nanoparticles (CdTe NPs) were synthesized both within apoferritin protein cage and on its surface. The thermal treatment of apoferritin with CdTe NPs resulted in the aggregation of cores, which was indicated by changes in the absorption spectra and the shape of apoferritin tryptophan fluorescence. The apoferritin modified with CdTe NPs was additionally modified with gold nanoparticles and attached to magnetic particles via oligonucleotide using gold affinity to thiol group. This anchor system was used to separate the construct using external magnetic field and to analyse the molecules attached to apoferritin.

The role of glutathione in autism spectrum disorder (ASD) is emerging as a major topic, due to its role in the maintenance of the intracellular redox balance. Several studies have implicated glutathione redox imbalance as a leading factor in ASD, and both ASD and many other neurodevelopmental disorders involve low levels of reduced glutathione (GSH), high levels of oxidized glutathione (GSSG), and abnormalities in the expressions of glutathione-related enzymes in the blood or brain. Glutathione metabolism, through its impact on redox environment or redox-independent mechanisms, interferes with multiple mechanisms involved in ASD pathogenesis. Glutathione-mediated regulation of glutamate receptors [e.g., N-methyl- d -aspartate (NMDA) receptor], as well as the role of glutamate as a substrate for glutathione synthesis, may be involved in the regulation of glutamate excitotoxicity. However, the interaction between glutathione and glutamate in the pathogenesis of brain diseases may vary from synergism to antagonism. Modulation of glutathione is also associated with regulation of redox-sensitive transcription factors nuclear factor kappa B (NF-κB) and activator protein 1 (AP-1) and downstream signaling (proinflammatory cytokines and inducible enzymes), thus providing a significant impact on neuroinflammation. Mitochondrial dysfunction, as well as neuronal apoptosis, may also provide a significant link between glutathione metabolism and ASD. Furthermore, it has been recently highlighted that glutathione can affect and modulate DNA methylation and epigenetics. Review analysis including research studies meeting the required criteria for analysis showed statistically significant differences between the plasma GSH and GSSG levels as well as GSH:GSSG ratio in autistic patients compared with healthy individuals (P = 0.0145, P = 0.0150 and P = 0.0202, respectively). Therefore, the existing data provide a strong background on the role of the glutathione system in ASD pathogenesis. Future research is necessary to investigate the role of glutathione redox signaling in ASD, which could potentially also lead to promising therapeutics.

Breast cancer is the most common neoplasm, comprising 16% of all women's cancers worldwide. Research of Copper (Cu) concentrations in various body specimens have suggested an association between Cu levels and breast cancer risks. This systematic review and meta-analysis summarize the results of published studies and examine this association. We searched the databases PubMed, Scopus, Web of Science, and Google Scholar and the reference lists of relevant publications. The Standardized Mean Differences (SMDs) between Cu levels in cancer cases and controls and corresponding Confidence Intervals (CIs), as well as I2 statistics, were calculated to examine heterogeneity. Following the specimens used in the original studies, the Cu concentrations were examined in three subgroups: serum or plasma, breast tissue, and scalp hair. We identified 1711 relevant studies published from 1984 to 2017. There was no statistically significant difference between breast cancer cases and controls for Cu levels assayed in any studied specimen; the SMD (95% CI) was -0.01 (-1.06 - 1.03; P = 0.98) for blood or serum, 0.51 (-0.70 - 1.73; P = 0.41) for breast tissue, and -0.88 (-3.42 - 1.65; P = 0.50) for hair samples. However, the heterogeneity between studies was very high (P < 0.001) in all subgroups. We did not find evidence for publication bias (P = 0.91). The results of this meta-analysis do not support an association between Cu levels and breast cancer. However, due to high heterogeneity in the results of original studies, this conclusion needs to be confirmed by well-designed prospective studies.

Virus-induced haemorrhagic fevers often show a rapid, dramatic course. Infectivity and mortality are very high. Such viruses are known to attack primates as well as other animal species. Fast, sensitive, selective diagnostics can be very beneficial for early anti-epidemic measures right in the outbreak. Quantum dots (QDs) have excellent features that make them useful as detection labels. Their use increases the sensitivity of the analytical assay. The use of biologically active components in QDs synthesis brings new properties. Quantum dots are nanocrystals that due to their high stability, ease of preparation and biocompatibility are suitable for labeling biomolecules. CdTe QDs modified with mercaptosuccinic acid (MSA), reduced glutathione (GSH), and plant extracts were prepared. Green synthesized CdTe QDs showed emission depending on the amount of extract applied. Thus, QDs were prepared in full-color scale from blue to red. The modified biomolecules of QDs are applicable for the labeling of nucleic acids. Their long-term stability, including their application onto paper, was tested. CdTe QDs exhibit very good electrochemical detection with the limit of detection (LOD) in nanomolar concentration.

African Swine Fever Virus (ASFV) is a DNA virus of the Asfivirus genus of the Asfarviridae family that is found in blood, body fluids, and internal organs. ASFV was described more than 40 years ago. This virus spreads pandemically and the mortality rate of the virus-related disease ranges from 90 to 100 %. The aim of this study was to propose the detection of specific nucleic acid of ASFV using electrochemical hybridization biosensor. Determination of DNA was conducted by AdTSV DPV a CV (potential 0 V, end potential -1.8 V, step potential 5 mV, modulation amplitude 25 mV, 0.2 M acetate buffer pH 5.0). The volume of analysed sample was 10 mu L. CV signals CA (log -0.0373x, r 0.99, Ep -1.30 V) and P peak (log -0.0801x, r 0.99, Ep -1.52 V) were observed. To increase the sensitivity, a modification of ODN with CdTe was proposed. The CdTe signal was observed around the potential of -0.56 V and for modified ODNs the signal was -0.58 V. SPION were prepared to capture DNA. The interaction of DNA (PCR fragment, 280 bp) with SPION was very fast within 30 s. The technique will be further used for a microfluidic system.

Drugs are important xenobiotics in the environment. Their use increases with the growth of the human population, but also in agricultural primary production. Paracetamol (PAR) is a widely used analgesic and antipyretic and its production is still growing. Commonly available drug production technologies are being developed very intensively with nanotechnological modifications for their gradual and targeted release. Nanoparticles (ST/PAR) from starch were prepared: PAR (0, 1, 2, 3, 5 and 10 mg/L) was mixed with citric acid ester in a 1:8 v/v ratio for 30 min at 25 °C. By the centrifugation (16.000 g, 30 min) ST/PAR were obtained in the pellet. The effect of PAR was studied on Daphnia magna Straus (Cladocera, Crustacea). Adult females (70-400 mg) were used for self-evaluation. The EC50 was 3.749 mg/L after 48 h of PAR treatment. Total protein values determined by Lowry method were between 0.5-2.2 mg/mL and by Bradford method between 190-676 mg/L. Antioxidant activity values determined by CUPRAC method were between 4-15 μg/mL GAE and by ABTS method ranged between 40-103 μg/mL GAE. PAR values were between 9-40 μM. Subsequently, the biological activity of the prepared nanoparticles was tested.

11th International Conference on Nanomaterials - Research and Application, NANOCON 2019 -- 16 October 2019 through 18 October 2019 -- -- 159862 2-s2.0-85097160523 Some types of nanoparticles such as gold and silver show interesting chemical properties. Pseudoperoxidase activity has been found in iron- or gold-based materials, but some silver nanoparticles (AgNPs) also exhibit such properties. Pseudoperoxidase activity of AgNPs prepared by green synthesis using sage leaves was studied. The AgNPs (prepared by green synthesis using sage leaves) size ranged from 20 to 50 nm and zeta potential was found to vary from -10 to 20 mV, with absorption maximum being 430 nm. The peroxidase-like activity was monitored in the presence of 5 mM 3,3',5,5'-tetramethylbenzidine (TMB) and hydrogen peroxide (30%). The nanoparticles showed about 80% activity of 1 mU horseradish peroxidase (HRP). In addition, AgNPs were applied to maize seedlings (5 days in hydroponics system) at concentrations of 1 mg/L, 50 mg/L, and 150 mg/L. Peroxidase activity in leaves after 48 hours was 59.5, 67.5, and 57.2 µg.mL-1 of HRP, respectively. Peroxidase activity of untreated plant leaves was 58.8 µg.mL-1 of HRP. Peroxidase activity in roots after 48 hours was 101.6, 109.0, and 156.7 µg.mL-1 of HRP, respectively. The peroxidase activity level of the control variant in root of treatment was 98.7 µg.mL-1 of HRP. The results indicate that the determination of peroxidase activity could be used to assess the phytotoxicity of nanoparticles in plants. © NANOCON 2019.All right reserved.

11th International Conference on Nanomaterials - Research and Application, NANOCON 2019 -- 16 October 2019 through 18 October 2019 -- -- 159862 2-s2.0-85097204138 The AgNPs have considerable industrial potential and are intensively studied with regard to their antibacterial properties. Using green synthesis, the nanoparticle surface can be coated with molecules that exhibit biologically significant properties. Increased use of nanoparticles enhances the risk of their release into the environment. However, there is still little known about the behaviour of AgNPs in the eco-environment. In this study, the effect of AgNPs prepared by green synthesis on germinated plants of maize was investigated. The effects on germination, basic growth and physiological parameters of the plant were monitored. AgNPs and silver nitrate were applied to maize seedlings (5 days in hydroponics system) at concentrations of 1 mg/L, 50 mg/L, and 150 mg/L. Chlorophyll a and b content was determined. We found that the growth inhibition of the above-ground parts of plants was about 40 % and AgNPs and silver nitrate had a significant effect on photosynthetic pigments. Chlorophyll a and b content changed with time and concentration. © NANOCON 2019.All right reserved.

Cancer represents one of the main causes of human death in developed countries. Most current therapies, unfortunately, carry a number of side effects, such as toxicity and damage to healthy cells, as well as the risk of resistance and recurrence. Therefore, cancer research is trying to develop therapeutic procedures with minimal negative consequences. The use of nanomaterial-based systems appears to be one of them. In recent years, great progress has been made in the field of possible use of nanomaterials with high potential in biomedical applications. Carbon nanomaterials, thanks to their unique physicochemical properties, are gaining more and more popularity in cancer therapy. They are valued especially for their ability to deliver drugs or small therapeutic molecules to these cells. Through surface functionalization, they can specifically target tumor tissues, increasing the therapeutic potential and significantly reducing the adverse effects of therapy. Their potential future use could, therefore, as vehicles for drug delivery. This review presents the latest findings of research studies using carbon nanomaterials in the treatment of various types of cancer. To carry out this study, different databases such as Web of Science, PubMed, MEDLINE and Google Scholar were employed. The findings of research studies chosen from more than 2000 viewed scientific publications from the last 15 years were compared.

The clinical use of doxorubicin (DOX) is limited by dose-related cardiomyopathy, which becomes more prevalent with increasing cumulative doses of the drug. Complexes of fullerene with DOX were designed and studied using biophysical methods. The ability of DOX to release from fullerene at different pHs was analyzed. It has been shown that the size of the fullerene-DOX complexes was ∼280 nm. The zeta potential for fullerene was -30 mV, for DOX -8 mV, and for fullerene-DOX conjugates -24 mV. Drug release was studied by CE with LIF detection. When fullerene-DOX conjugates were separated in a pH 7.5 buffer, 43% of all DOX signals were derived from free DOX, with the signal increasing as pH decreased. At pH 5.25, all DOX had been released and 100% of the signal was derived from free DOX. The release of DOX from complexes with fullerene at lower pH can be used in targeted antineoplastic therapy, resulting in lower toxicity for less acidic non-target tissue.

Traditional supplements of selenium generally have a low degree of absorption and increased toxicity. Therefore, it is imperative to develop innovative systems as transporters of selenium compounds, which would raise the bioavailability of this element and allow its controlled release in the organism. Nanoscale selenium has attracted a great interest as a food additive especially in individuals with selenium deficiency, but also as a therapeutic agent without significant side effects in medicine. This review is focused on the incorporation of nanotechnological applications, in particular exploring the possibilities of a more effective way of administration, especially in selenium-deficient organisms. In addition, this review summarizes the survey of knowledge on selenium nanoparticles, their biological effects in the organism, advantages, absorption mechanisms, and nanotechnological applications for peroral administration.

The theory of glucose-responsive composite membranes for the planar diffusion and reaction process is extended to a microsphere membrane. The theoretical model of glucose oxidation and hydrogen peroxide production in the chitosan-aliginate microsphere has been discussed in this manuscript for the first time. We have successfully reported an analytical derived methodology utilizing homotopy perturbation to perform the numerical simulation. The influence and sensitive analysis of various parameters on the concentrations of gluconic acid and hydrogen peroxide are also discussed. The theoretical results enable to predict and optimize the performance of enzyme kinetics.

Silver nanoparticles (AgNPs) are widely investigated with regard to their physical, chemical, but also biological properties. Antibacterial and antitumor properties of AgNPs have been intensively studied. In addition, the synthesis using a green approach brings further significant biological properties. However, it is also necessary to monitor the potential toxicity of such nanoparticles in different ecosystems. In this study, the effect of AgNO3 and AgNPs on germinated plants of Zea mays was studied. Effects on basic growth and physiological parameters were observed. There was a statistically significant difference between the variants tested.

The resistance of microorganisms to antibiotics is growing steadily. The development of new antibacterial agents is highly topical. Metal nanoparticles have shown significant antibacterial activity similar to the plant/animal materials used in traditional medicine. The study focuses on the synthesis of silver nanoparticles (AgNPs) modified with biomolecules from used plant extracts (T. serpyllum, S. officinalis, T. pratense). The obtained nanoparticles were studied in detail by physicochemical methods. In addition, they were deposited on acrylonitrile butadiene styrene (ABS). We created unique antibacterial material using 3D printing. 20-40% inhibition of S. aureus and E. coli was observed in the evaluation of their efficacy.

Preventing the spread of dangerous viral diseases such as flu, Ebola or HIV requires rapid and effective diagnostic approaches to detect these diseases at an early stage. Quantum dots (QDs) are nanocrystals that exhibit a variety of unique properties and are suitable for biomolecule labelling due to their high stability, ease of preparation, and biocompatibility. Modified QDs can be used to label nucleic acids or antibodies. Green synthesis method of QDs provides a platform for preparation of unique materials with new chemical or physical properties as compared to the original material. In this work, CdTe QDs were produced in the presence of plant extract which acted as a modifying agent. The Zea mays extract was added during the CdTe QDs synthesis at different time intervals and CdTe QDs showed a wide range of colors. The stability of the prepared QDs, including their application onto paper, was evaluated. The QDs were observed to show a remarkable electrochemical response for sensor applications and were also employed to label virus-specific antibody. The entire procedure was miniaturized and the viral particles were analyzed in a 3D-printed chip.

Background Ferritin is a globular intracellular protein that acts as the main reservoir for iron. Malignancies are associated with increased plasma ferritin concentrations. A number of studies show that tumor cells express high levels of transferrin receptors (TfR). Increased TfR expression was observed in prostate carcinoma. Apoferritin (APO) can be used as a protein nanotransporter into which a suitable medicinal substance can be encapsulated. Nanoparticles increase the permeability of tumor cells to nanotransporters and have a photothermal effect. The aim of this study was to encapsulate doxorubicin (DOX) into APO and to modify the resulting APO/DOX with gold (AuNPs) and silver nanoparticles prepared by green synthesis (AgNPsGS). Methods APO was characterized using 10% sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) - 120 V, 60 min, 24 mM Tris, 0.2 M glycine, 3 mM SDS. DOX fluorescence (Ex 480 nm; Em 650 nm) was observed, with a typical absorption maximum at 560 nm. Electrochemical measurement was performed in Brdicka solution (three-electrode setup). AgNPsGS were prepared by green synthesis using clover (Trifolium pratense L.). Results An electrophoretic study of APO and APO/DOX (5-100 μg/mL) was performed and the behavior of APO and APO/DOX (10 μM) as a function of pH was monitored. In an acidic environment, APO forms subunits of about 20 kDa; in an alkaline medium, it forms a globular protein of about 450 kDa. A change in APO/DOX mobility (about by 10%) was observed. A film of gold nanoparticles was applied to the APO/DOX surface. APO/DOX-AuNPs were washed with ultra-pure water. pH-dependent release of DOX a was monitored. The amount of DOX analyzed was increased by up to 50%. Furthermore, an AgNPsGS-DOX complex (1 mg AgNPsGS/100 μM DOX) was generated and prepared. Subsequently, the AgNPsGS-DOX complex was encapsulated into APO. To further improve therapeutic efficacy, the APO/AgNPsGS-DOX complex was coated with an Au layer. APO/AgNPsGS-DOX/AuNPs were stable and DOX was released from the complex after physical parameters had changed. Conclusion APO nanocomplexes were prepared and modified to increase therapeutic efficacy against tumors. Tumor cell targeting was achieved by binding to TfR and via increased tumor cell permeability and retention. Release of the drug was made possible due to a pH change and photothermal activation that will now be tested. This work was supported by COST European Cholangiocarcinoma Network CA18122 and International Collaboration Project of The European Technology Platform for Nanomedicine. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 21. 3. 2019 Accepted: 14. 5. 2019.

Cardiotoxicity is a serious complication of anticancer therapy by anthracycline antibiotics. Except for intercalation into DNA/RNA structure, inhibition of DNA-topoisomerase and histone eviction from chromatin, the main mechanism of their action is iron-mediated formation of various forms of free radicals, which leads to irreversible damage to cancer cells. The most serious adverse effect of anthracyclines is, thus, cardiomyopathy leading to congestive heart failure, which is caused by the same mechanisms. Here, we briefly summarize the basic types of free radicals formed by anthracyclines and the main processes how to scavenge them. From these, the main attention is paid to metallothioneins. These low-molecular cysteine-rich proteins are introduced and their functions and properties are reviewed. Further, their role in detoxification of metals and drugs is discussed. Based on these beneficial roles, their use as a new therapeutic agent against oxidative stress and for cardioprotection is critically evaluated with respect to their ability to increase chemoresistance against some types of commonly used cytostatics.

We present a utilization of carbon quantum dots (CQDs), passivated with polyethylene glycol as a fluorescent recognition probe for DNA damage. Synthesized CQDs were characterized in detail, using optical and electrochemical methods. Further, fluorescent behavior of CQDs was monitored in the presence of genomic DNA, isolated from Staphylococcus aureus. In laboratory conditions, after 30 min of exposure to UV irradiation (λ = 254 nm), the DNA/CQDs complex significantly increased its fluorescence. Further, stratospheric probe was suggested and crafted by using technology of 3D printing (acrylonitrile–butadiene–styrene as a material). CQDs were exploited to evaluate the DNA damage in stratospheric conditions (up to 20,000 m) by determination of fluorescence increase (λ exc = 245 nm, λ em = 400 nm), together with other parameters (temperature, humidity, altitude, pressure, UV intensity, and X-ray irradiation). The obtained data showed that the sensor utilizing the DNA/CQDs was able to identify the DNA damage, together with external conditions. It was shown the proposed concept is able to operate at temperatures lower than −70 °C. The proposed protocol may by applicable as a biosensor for long-term space missions, like international space station, missions to the Moon or Mars.

The goal of this study was to contribute to understanding of the mechanisms behind sensitivity differences between early and late instar larvae of Chironomus riparius and to address the influence of the differences in standard testing approaches on the toxicity evaluation. A 10-day contact sediment toxicity test was carried out to assess sensitivity to cadmium exposure in relation to different age and laboratory culture line origin of test organisms. Chironomid larvae of early (OECD 218 method) and late instar (US-EPA600/R-99/064 method) differed substantially in sensitivity of traditional endpoints (OECD: LOEC 50 and 10 μg Cd/g dry weight (dw); US-EPA: LOEC > 1000 and 100 μg Cd/g dw for survival and growth, respectively). Bioaccumulated cadmium and metallothioneins (MTs) concentrations were analyzed to investigate the role of MTs in reduced sensitivity to cadmium in late instar larvae. Metallothioneins were induced after treatment to greater Cd concentrations, but their levels in relation to cadmium body burdens did not fully explain low sensitivity of late instars to cadmium, which indicates some other effective way of detoxification in late instars. This study brings new information related to the role of MTs in age-dependent toxicant sensitivity and discusses the implications of divergence in data generated by chironomid sediment toxicity tests by standardized methods using different instars.

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.

BACKGROUND Sarcosine (N-methylglycine) was previously delineated as a substantial oncometabolite of prostate cancer (PCa) and its metabolism seems to be significantly involved in PCa development and behavior. METHODS We focused on investigation whether the exposure of prostate cells (PNT1A, 22Rv1, and PC-3) to sarcosine-related amino acids (glycine, dimethylglycine, and sarcosine) affects their aggressiveness (cell mobility and division rates, using real-time cell based assay). The effect of supplementation on expression of glycine-N-methyltransferase (GNMT) mRNA was examined using qRT-PCR. Finally, post-treatment amino acids patterns were determined with consequent statistical processing using the Ward's method, factorial ANOVA and principal component analysis (P

The aim of the present study was to investigate the impact of copper-based pesticides (at concen - trations of copper of 20, 30, 40 and 70 µg/l) on one-year-old common carp (Cyprinus carpio L.) during 28 days of exposure. Abnormal behaviour was observed in fish exposed to 70 µg/l from Day 14. Histological alterations were noticed only in liver in the groups exposed to 40 and 70 µg/l. Significant changes (P ˂ 0.05) in almost all haema- tological indices were found, especially in the group exposed to the highest concentration of copper (70 µg/l).

The electrochemical behavior of adenine at a glassy carbon electrode (GCE) modified with a nanocomposite consisting of graphene oxide and polyaniline was investigated by cyclic voltammetry and differential pulse voltammetry. The nanocomposite was synthesized by polymerization and characterized by Raman and UV-vis spectroscopy, and its morphology was examined by scanning electron microscopy. Adenine is oxidized at the modified GCE at a working potential of 1.2 V (vs. Ag/AgCl) and gives a current density of approximately 2.64 nA.cm−2, which is distinctly increased compared to the 0.57 nA.cm−2 of a bare electrode. Peak current and adenine concentration are linearly related to each other in the range from 0.5 μM to 20 μM. The modified GCE exhibits acceptable analytical performance, with a detection limit of 72 nM and a limit of quantification of 240 nM. It is excellently reproducible, stable, and fabrication is simple.

The influence of anthocyanins on antioxidant activity and liver function was measured in three different animal species (rats, chickens and fish). Purple Konini wheat with the total anthocyanin content (TAC) of 41.70 mg/kg was fed to the experimental groups of animals (rats n = 32; chickens n = 32; fish n = 20). The same numbers of animals (control groups) were fed common wheat with TAC of 24.95 mg/kg. At the end of the experiment, the animals were killed and blood and liver tissues were taken for biochemical analyses. The liver enzyme activities were measured in blood. Antioxidant activity in the liver tissue was measured using the DPPH (2,2-diphenyl-1-picrylhydrazyl test), FR (Free Radicals method), FRAP (Ferric Reducing Antioxidant Power method), and ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) test. Significantly higher values (P< 0.05) measured by the DPPH and FR methods were determined in the liver from rats fed Konini wheat. The chickens fed Konini wheat showed significantly different values using DPPH, FR and ABTS methods. No significant differences were found in hepatopancreases of fish. Animals fed Konini wheat showed a tendency to lower liver enzyme activities in general, but a significant difference was found only in gamma-glutamyl transferase activity in the chicken’s blood. Our results suggest that feeding a higher content of anthocyanins can improve the antioxidant activity and function of the liver tissue.

In this study, enhancement of the electrochemical signals of etoposide (ETO) measured by differential pulse voltammetry (DPV) by modifying a glassy carbon electrode (GCE) with carbon quantum dots (CQDs) is demonstrated. In comparison with a bare GCE, the modified GCE exhibited a higher sensitivity towards electrochemical detection of ETO. The lowest limit of detection was observed to be 5 nM ETO. Furthermore, scanning electron microscopy (SEM), fluorescence microscopy (FM), and electrochemical impedance spectroscopy (EIS) were employed for the further study of the working electrode surface after the modification with CQDs. Finally, the GCE modified with CQDs under optimized conditions was used to analyse real samples of ETO in the prostate cancer cell line PC3. After different incubation times (1, 3, 6, 9, 12, 18 and 24 h), these samples were then prepared prior to electrochemical detection by the GCE modified with CQDs. High performance liquid chromatography with an electrochemical detection method was employed to verify the results from the GCE modified with CQDs.

The irradiance of ultraviolet (UV) radiation is a physical parameter that significantly influences biological molecules by affecting their molecular structure. The influence of UV radiation on nanoparticles has not been investigated much. In this work, the ability of cadmium telluride quantum dots (CdTe QDs) to respond to natural UV radiation was examined. The average size of the yellow QDs was 4 nm, and the sizes of green, red and orange QDs were 2 nm. Quantum yield of green CdTe QDs-MSA (mercaptosuccinic acid)-A, yellow CdTe QDs-MSA-B, orange CdTe QDs-MSA-C and red CdTe QDs-MSA-D were 23.0%, 16.0%, 18.0% and 7.0%, respectively. Green, yellow, orange and red CdTe QDs were replaced every day and exposed to daily UV radiation for 12 h for seven consecutive days in summer with UV index signal integration ranging from 1894 to 2970. The rising dose of UV radiation led to the release of cadmium ions and the change in the size of individual QDs. The shifts were evident in absorption signals (shifts of the absorbance maxima of individual CdTe QDs-MSA were in the range of 6&ndash, 79 nm), sulfhydryl (SH)-group signals (after UV exposure, the largest changes in the differential signal of the SH groups were observed in the orange, green, and yellow QDs, while in red QDs, there were almost no changes), fluorescence, and electrochemical signals. Yellow, orange and green QDs showed a stronger response to UV radiation than red ones.

Background: Sarcosine is an amino acid that is formed by methylation of glycine and is present in trace amounts in the body. Increased sarcosine concentrations in blood plasma and urine are manifested in sarcosinemia and in some other diseases such as prostate cancer. For this purpose, sarcosine detection using the nanomedicine approach was proposed. In this study, we have prepared superparamagnetic iron oxide nanoparticles (SPIONs) with different modified surface area. Nanoparticles (NPs) were modified by chitosan (CS), and sarcosine oxidase (SOX). SPIONs without any modification were taken as controls. Methods and Results: The obtained NPs were characterized by physicochemical methods. The size of the NPs determined by the dynamic light scattering method was as follows: SPIONs/Au/NPs (100⁻300 nm), SPIONs/Au/CS/NPs (300⁻700 nm), and SPIONs/Au/CS/SOX/NPs (600⁻1500 nm). The amount of CS deposited on the NP surface was found to be 48 mg/mL for SPIONs/Au/CS/NPs and 39 mg/mL for SPIONs/Au/CS/SOX/NPs, and repeatability varied around 10%. Pseudo-peroxidase activity of NPs was verified using sarcosine, horseradish peroxidase (HRP) and 3,3′,5,5′-tetramethylbenzidine (TMB) as a substrate. For TMB, all NPs tested evinced substantial pseudo-peroxidase activity at 650 nm. The concentration of SPIONs/Au/CS/SOX/NPs in the reaction mixture was optimized to 0⁻40 mg/mL. Trinder reaction for sarcosine detection was set up at 510 nm at an optimal reaction temperature of 37 °C and pH 8.0. The course of the reaction was linear for 150 min. The smallest amount of NPs that was able to detect sarcosine was 0.2 mg/well (200 µL of total volume) with the linear dependence y = 0.0011x − 0.0001 and the correlation coefficient r = 0.9992, relative standard deviation (RSD) 6.35%, limit of detection (LOD) 5 µM. The suggested method was further validated for artificial urine analysis (r = 0.99, RSD 21.35%, LOD 18 µM). The calculation between the detected and applied concentrations showed a high correlation coefficient (r = 0.99). NPs were tested for toxicity and no significant growth inhibition was observed in any model system (S. cerevisiae, S. aureus, E. coli). The hemolytic activity of the prepared NPs was similar to that of the phosphate buffered saline (PBS) control. The reaction system was further tested on real urine specimens. Conclusion: The proposed detection system allows the analysis of sarcosine at micromolar concentrations and to monitor changes in its levels as a potential prostate cancer marker. The whole system is suitable for low-cost miniaturization and point-of-care testing technology and diagnostic systems. This system is simple, inexpensive, and convenient for screening tests and telemedicine applications.

Wine consumption has been popular worldwide for many centuries. Based on in vitro and in vivo studies, a certain amount of everyday wine consumption may prevent various chronic diseases. This is due, in part, to the presence and amount of important antioxidants in red wine, and, therefore, research has focused on them. Wine polyphenols, especially resveratrol, anthocyanins, and catechins, are the most effective wine antioxidants. Resveratrol is active in the prevention of cardiovascular diseases by neutralizing free oxygen radicals and reactive nitrogenous radicals; it penetrates the blood-brain barrier and, thus, protects the brain and nerve cells. It also reduces platelet aggregation and so counteracts the formation of blood clots or thrombi. The main aim of this review is to summarize the current findings about the positive influence of wine consumption on human organ function, chronic diseases, and the reduction of damage to the cardiovascular system. © 2018 by the authors., [DG16P02R017]

Approximately 90 % of head and neck cancers are squamous cell carcinomas (HNSCC), and the overall 5-year survival rate is not higher than 50 %. There is much evidence that human papillomavirus (HPV) infection may influence the expression of commonly studied HNSCC markers. Our study was focused on the possible HPV-specificity of molecular markers that could be key players in important steps of cancerogenesis (MKI67, EGF, EGFR, BCL-2, BAX, FOS, JUN, TP53, MT1A, MT2A, VEGFA, FLT1, MMP2, MMP9, and POU5F). qRT-PCR analysis of these selected genes was performed on 74 biopsy samples of tumors from patients with histologically verified HNSCC (22 HPV-, 52 HPV+). Kaplan-Meier analysis was done to determine the relevance of these selected markers for HNSCC prognosis. In conclusion, our study confirms the impact of HPV infection on commonly studied HNSCC markers MT2A, MMP9, FLT1, VEGFA, and POU5F that were more highly expressed in HPV-negative HNSCC patients and also shows the relevance of studied markers in HPV-positive and HPV-negative HNSCC patients.

Fluorescence resonance energy transfer (FRET) in combination with quantum dots (QDs) and their superior properties has enabled designing of the new and improved sensors. In this review, the latest novelties in development and application of FRET nanosensors employing QDs are presented. QDs offer several advantages over organic dyes - broad excitation spectra, narrow defined tunable emission peak, longer fluorescence lifetime, resistance to photobleaching and 10-100 times higher molar extinction coefficient. These properties of QDs allow multicolor QDs to be excited from one source by common fluorescent dyes without emission signal overlap and results in brighter probes comparing to conventional fluorophores. Due to these benefits, QD-FRET-based nanosensors gained a wide spread popularity in a variety of scientific areas. These sensors are most frequently applied in the domain of the nucleic acid and enzyme activity detection. Other applications are detection of peptides and low-molecular compounds, environmental pollutants, viruses, microorganisms and their toxins, QD-FRET-based immunoassays, and pH sensors.

Background: Infections, mostly those associated with colonization of wound by different pathogenic microorganisms, are one of the most serious health complications during a medical treatment. Therefore, this study is focused on the isolation, characterization, and identification of microorganisms prevalent in superficial wounds of patients (n = 50) presenting with bacterial infection. Methods: After successful cultivation, bacteria were processed and analyzed. Initially the identification of the strains was performed through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry based on comparison of protein profiles (2–30 kDa) with database. Subsequently, bacterial strains from infected wounds were identified by both matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and sequencing of 16S rRNA gene 108. Results: The most prevalent species was Staphylococcus aureus (70%), and out of those 11% turned out to be methicillin-resistant (mecA positive). Identified strains were compared with patients’ diagnoses using the method of artificial neuronal network to assess the association between severity of infection and wound microbiome species composition. Artificial neuronal network was subsequently used to predict patients’ prognosis (n = 9) with 85% success. Conclusions: In all of 50 patients tested bacterial infections were identified. Based on the proposed artificial neuronal network we were able to predict the severity of the infection and length of the treatment. Keywords: Bacterial strains, MALDI-TOF, Sequencing, Superficial wounds