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The present experiment describes a synthesis process of composites based on graphene oxide, which was tested as a carrier for composites of metal- or metalloid-based nanoparticles (Cu, Zn, Mn, Ag, AgP, Se) and subsequently examined as an antimicrobial agent for some bacterial strains (Staphylococcus aureus (S. aureus), methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli). The composites were first applied at a concentration of 300 M on all types of model organisms and their effect was observed by spectrophotometric analysis, which showed a decrease in absorbance values in comparison with the control, untreated strain. The most pronounced inhibition (87.4%) of S. aureus growth was observed after the application of graphene oxide composite with selenium nanoparticles compared to control. Moreover, the application of the composite with silver and silver phosphate nanoparticles showed the decrease of 68.8% and 56.8%, respectively. For all the tested composites, the observed antimicrobial effect was found in the range of 26% to 87.4%. Interestingly, the effects of the composites with selenium nanoparticles significantly differed in Gram-positive (G+) and Gram-negative (G) bacteria. Presentovaný experiment popisuje syntetizké procesy sloučenin založených na grafen oxidu, který byl testován jako nosič pro sloučeniny kovu - nebo podobných kovu založenýh na nanočásticích (Cu, Zn, Mn, Ag, AgP, Se) a zkumala, jako antimikrobiální prostředek pro některé bakteriální kmeny (Staphylococcus aureus (S. aureus), vůči rezistentnímu methicilinu Staphylococcus aureus (MRSA) a Escherichia coli (E. coli). Sloučeniny byly jako první aplikovány v koncentraci 300 uM na všechny typy modelových organismů a jejich účinek byl pozorován spektrofotometrickou analýzou, která ukázala pokles hodnoty absorbance ve srovnání s kontrolním neošetřeným kmenem. Nejvýraznější nárust zábrany růstu (87,4% )S. aureus byl pozorován po aplikaci oxidu grapheu sloučeného se selen nanočásticemi. Navíc, aplikace sloučenin stříbra a stříbrných fosfátových nanočástic ukázaly pokles o 68,8% a 56,8% . U všech testovaných sloučenin, byly pozorovánay antimikrobiální účinky účinky a byly nalezeny v rozsahu 26% na 87,4%. Je zajímavé, že účinky sloučenin selenových nanočástic se významně lišily v Gram-pozitivních (G +) a gram-negativních (G-) bakterií.

The present paper describes the synthesis, characterization, and utilization of multi-functional magnetic conjugates that integrate optical and magnetic properties in a single structure for use in many biomedical applications. Spontaneous interaction with eukaryotic cell membrane (HEK-239 cell culture) was determined using fluorescence microscopy, and fluorescence analyses. Both, differences in excitation, and emission wavelength were observed, caused by glutathione intake by cells, resulting in disintegration of core–shell structure of quantum dots, as well as adhesion of conjugate onto cell surface. When compared with quantum dots fluorescent properties, HEK-239 cells with incorporated nanoconjugate exhibited two excitation maxima (λ ex = 430 and 390 nm). Simultaneously, application of ideal λ ex for quantum dots (λ ex = 430 nm), resulted in two emission maxima (λ = 740 and 750 nm). This nanoconjugate fulfills the requirements of term theranostics, because it can be further functionalized with biomolecules as DNA, proteins, peptides or antibodies, and thus serves as a tool for therapy in combination with simultaneous treatment.

Doxorubicin (DOX) belongs to the group of anthracycline antibiotics with very effective anticancer properties. On the other hand, the cardiotoxic effects limit its application over the maximum cumulative dose. To overcome this obstacle, encapsulation of this drug into the protective nanotransporter such as apoferritin is beneficial. In this study, fluorescent behavior of DOX in various solvents was determined by fluorescence spectrometry, demonstrating the fluorescence quenching effect of water, which is often used as a solvent. It was found that by increasing the amount of the organic phase in the DOX solvent the dynamic quenching is significantly suppressed. Ethanol, acetonitrile and dimethyl sulfoxide were tested and the best linearity of the calibration curve was obtained when above 50 % of the solvent was present in the binary mixture with water. Moreover, pH influence on the DOX fluorescence was also observed within the range of 4–10. Two times higher fluorescence intensity was observed at pH 4 compared to pH 10. Further, the DOX behavior in capillary electrophoresis (CE) was investigated. Electrophoretic mobilities (CE) in various pH of the background electrolyte were determined within the range from 16.3 to −13.3 × 10 −9 m−2 V−1 s−1. Finally, CE was also used to monitor the encapsulation of DOX into the cavity of apoferritin as well as the pH-triggered release.

Nanoparticles have gained increasing interest in medical and in vivo applications. Metallothionein (MT) is well known as a maintainer of metal ions balance in intracellular space. This is due to high affinity of this protein to any reactive species including metals and reactive oxygen species. The purpose of this study was to determine the metallothionein-quantum dots interactions that were investigated by spectral and electrochemical techniques. CuS, CdS, PbS, and CdTe quantum dots (QDs) were analysed. The highest intensity was shown for CdTe, than for CdS measured by fluorescence. These results were supported by statistical analysis and considered as significant. Further, these interactions were analysed using gel electrophoresis, where MT aggregates forming after interactions with QDs were detected. Using differential pulse voltammetry Brdicka reaction, QDs and MT were studied. This method allowed us to confirm spectral results and, moreover, to observe the changes in MT structure causing new voltammetric peaks called X and Y, which enhanced with the prolonged time of interaction up to 6 h. (C) 2014 Elsevier B.V. All rights reserved.

Carbon nanomaterials, including fullerenes, exhibit not only unique structure and electronic properties but also a significant potential to serve as radical scavengers and/or anti-oxidants. Their conjugation with anticancer drugs such as doxorubicin (DOX) may help to balance severe negative side effects of these cytostatics and also improve the delivery of the drug taking advantage of the enhanced cellular uptake, selectivity to cancer cells, and pH regulated release. In this study, the fullerene (C60) surface was oxidized by concentrated nitric acid, which enabled simple DOX‐fullerene conjugation based on ‐ stacking and hydrophilic interactions with carboxylic groups. The strength of this noncovalent binding is pH dependent. At a low pH, the amino group of DOX is protonated, however at a higher pH, the amino group is deprotonated, resulting in stronger hydrophobic interactions with the fullerene walls. CE and HPLC were employed for characterization of resulting complexes. The cell toxicity of the conjugates was evaluated using Staphylococcus aureusandfinally they were administered into the chicken embryo to assess the applicability for in vivo imaging.

Doxorubicin is a commonly used antineoplastic agent in the treatment of many types of cancer. Little is known about the interactions of doxorubicin with cardiac biomolecules. Serious cardiotoxicity including dilated cardiomyopathy often resulting in a fatal congestive heart failure may occur as a consequence of chemotherapy with doxorubicin. The purpose of this study was to determine the effect of exposure to doxorubicin on the changes in major amino acids in tissue of cardiac muscle (proline, taurine, glutamic acid, arginine, aspartic acid, leucine, glycine, valine, alanine, isoleucine, threonine, lysine and serine). An in vitro interaction study was performed as a comparison of amino acid profiles in heart tissue before and after application of doxorubicin. We found that doxorubicin directly influences myocardial amino acid representation even at low concentrations. In addition, we performed an interaction study that resulted in the determination of breaking points for each of analyzed amino acids. Lysine, arginine, β-alanine, valine and serine were determined as the most sensitive amino acids. Additionally we compared amino acid profiles of myocardium before and after exposure to doxorubicin. The amount of amino acids after interaction with doxorubicin was significantly reduced (p = 0.05). This fact points at an ability of doxorubicin to induce changes in quantitative composition of amino acids in myocardium. Moreover, this confirms that the interactions between doxorubicin and amino acids may act as another factor most likely responsible for adverse effects of doxorubicin on myocardium.

A diagnostics of infectious diseases can be done by the immunologic methods or by the amplification of nucleic acid specific to contagious agent using polymerase chain reaction. However, in transmissible spongiform encephalopathies, the infectious agent, prion protein (PrP(Sc)), has the same sequence of nucleic acids as a naturally occurring protein. The other issue with the diagnosing based on the PrP(Sc) detection is that the pathological form of prion protein is abundant only at late stages of the disease in a brain. Therefore, the diagnostics of prion protein caused diseases represent a sort of challenges as that hosts can incubate infectious prion proteins for many months or even years. Therefore, new in vivo assays for detection of prion proteins and for diagnosis of their relation to neurodegenerative diseases are summarized. Their applicability and future prospects in this field are discussed with particular aim at using quantum dots as fluorescent labels.

Magnetic particle mediated transport in combination with nanomaterial based drug carrier has a great potential for targeted cancer therapy. In this study, doxorubicin encapsulation into the apoferritin and its conjugation with magnetic particles was investigated by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). The quantification of encapsulated doxorubicin was performed by fluorescence spectroscopy and compared to CE-LIF. Moreover, the significant enhancement of the doxorubicin signal was observed by addition of methanol into the sample solution Magnetické částice zprostředkující transport v kombinaci s nanomateriály na bázi nosiče léčiv mají velký potenciál pro cílenou terapii nádorových onemocnění. V této studii byl doxorubicin zapouzdřeníý do apoferritinu a jeho konjugace s magnetickými částicemi byla zkoumána kapilární elektroforézou s laserem indukovanou fluorescenční detekcí (CE-LIF). Kvantifikace zapouzdřeného doxorubicinu byla provedena fluorescenční spektroskopií v porovnání s CE-LIF. Kromě toho, bylo pozorováno významné zlepšení signálu doxorubicinu přidáním methanolu do roztoku vzorku.

In this study a device for automatic electrochemical analysis was designed. A three electrodes detection system was attached to a positioning device, which enabled us to move the electrode system from one well to another of a microtitre plate. Disposable carbon tip electrodes were used for Cd(II), Cu(II) and Pb(II) ion quantification, while Zn(II) did not give signal in this electrode configuration. In order to detect all mentioned heavy metals simultaneously, thin-film mercury electrodes (TFME) were fabricated by electrodeposition of mercury on the surface of carbon tips. In comparison with bare electrodes the TMFEs had lower detection limits and better sensitivity. In addition to pure aqueous heavy metal solutions, the assay was also performed on mineralized rock samples, artificial blood plasma samples and samples of chicken embryo organs treated with cadmium. An artificial neural network was created to evaluate the concentrations of the mentioned heavy metals correctly in mixture samples and an excellent fit was observed (R2 = 0.9933). V této studii byla navržena automatická elektrochemická analýza. Detekční systém složený ze tří elektrod byl připojen k polohovacímu zařízení, které nám umožnilo manipulaci s elektrodou z jedné jamky na druhou mikrotitrační destičky. Jednorázový uhlíkový tip elektrody byl použit pro kvantifikaci Cd (II), Cu (II) a Pb (II) iontů, zatímco Zn (II) neměl signál v této konfiguraci elektrody. Aby bylo možné odhalit všechny uvedené těžké kovy současně pomocí tenkovrstvé rtuťové elektrody (TFME), byly vyrobeny podle elektrodepozicí rtuti na povrchu hrotů uhlíku. Ve srovnání s holýma elektrodami se TMFEs měli nižší detekční limity a lepší citlivost. Kromě čistého vodného roztoku těžkých kovů ....

The platinum group elements are significant compounds used in numerous fields of human life. Besides of often discussed toxic effect the platinum compounds show the therapeutic effects. Nowadays, platinum-based cytostatic are still the most frequently used drugs in oncology. Due to their proved medicinal purposes the behavior in the organism should to be intensively studied as well as their interactions with DNA and other important biological molecules. This review summarizes the recent results in the platinum drug field and discusses the behavior of platinum compounds in cells. The interaction of platinum and DNA with respect to the change of the DNA structure are also clarified.

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.

There is an arising and concerning issue in the field of bacterial resistance, which is confirmed by the number of deaths associated with drug-resistant bacterial infections. The aim of this study was to compare the effects of antibiotics on Staphylococcus aureus non-resistant strain and strains resistant to cadmium or lead ions. Metal resistant strains were created by the gradual addition of 2 mM solution of metal ions (cadmium or lead) to the S. aureus culture. An increasing antimicrobial effect of ampicillin, streptomycin, penicillin and tetracycline (0, 10, 25, 50, 75, 150, 225 and 300 mu M) on the resistant strains was observed using a method of growth curves. A significant growth inhibition (compared to control) of cadmium resistant cells was observed in the presence of all the four different antibiotics. On the other hand, the addition of streptomycin and ampicillin did not inhibit the growth of lead resistant strain. Other antibiotics were still toxic to the bacterial cells. Significant differences in the morphology of cell walls were indicated by changes in the cell shape. Our data show that the presence of metal ions in the urban environment may contribute to the development of bacterial strain resistance to other substances including antibiotics, which would have an impact on public health. K dispozici je vznikající a pokud jde o problém v oblasti bakteriální rezistence, což je potvrzeno počtu úmrtí souvisejících s drogami odolné proti bakteriálním infekcím. Cílem této studie bylo porovnat účinky antibiotik na kmen bez rezistentní Staphylococcus aureus a kmenů rezistentních k kadmia nebo olova ionty. Kmeny odolného kovu byly vytvořeny postupným přidáváním 2 mM roztoku kovových iontů (kadmium nebo olovo) na S. aureus kultury.Zvyšující antimikrobiální účinek ampicilin, streptomycin, penicilin a tetracyklin (0, 10, 25, 50, 75, 150, 225 a 300 um) na rezistentních kmenů byla pozorována za použití metody růstových křivek. Významná inhibice růstu (ve srovnání s kontrolou) buněk rezistentních kadmia byl pozorován v přítomnosti všech čtyř různých antibiotik. Na druhé straně, přidání streptomycinu a ampicilinu neinhibuje růst olova rezistentního kmene. Další antibiotika stále toxické pro bakteriálních buněk. Významné rozdíly v morfologii buněčných stěn byly označeny změny ve tvaru buňky. Naše data ukazují, že přítomnost kovových iontů v městském prostředí, může přispět k rozvoji bakterií kmene rezistence na jiné látky, včetně antibiotik, které by mohly mít vliv na veřejné zdraví. (přeloženo strojově, pravopis nekontrolován).

The aim of the study was the preparation of a liposome complex with encapsulated lead ions, which were electrochemically detected. In particular, experiments were focused on the potential of using an electrochemical method for the determination of free and liposome-encapsulated lead and determination of the encapsulation efficiency preventing the lead toxicity. Primarily, encapsulation of lead ions in liposomes and confirmation of successful encapsulation by electrochemical methods was done. Further, the reduction effect of the liposome matrix on the detected electrochemical signal was monitored. Besides encapsulation itself, comparison of toxicity of free lead ions and lead ions encapsulated in liposome was tested. The calculated IC50 values for evaluating the lead cytotoxicity showed significant differences between the lead enclosed in liposomes (28 mu M) and free lead ions (237 mu M). From the cytotoxicity studies on the bacterial strain of S. aureus it was observed that the free lead ions are less toxic in comparison with lead encapsulated in liposomes. Liposomes appear to be a suitable carrier of various substances through the inner cavity. Due to the liposome structure the lead enclosed in the liposome is more easily accepted into the cell structure and the toxicity of the enclosed lead is higher in comparison to free lead ions.

Carbon nanomaterials, including fullerenes, exhibit not only unique structure and electronic properties but also a significant potential to serve as radical scavengers and/or anti-oxidants. Their conjugation with anticancer drugs such as doxorubicin (DOX) may help to balance severe negative side effects of these cytostatics and also improve the delivery of the drug taking advantage of the enhanced cellular uptake, selectivity to cancer cells, and pH regulated release. In this study, the fullerene (C60) surface was oxidized by concentrated nitric acid, which enabled simple DOX-fullerene conjugation based on π-π stacking and hydrophilic interactions with carboxylic groups. The strength of this noncovalent binding is pH dependent. At a low pH, the amino group of DOX is protonated, however at a higher pH, the amino group is deprotonated, resulting in stronger hydrophobic interactions with the fullerene walls. CE and HPLC were employed for characterization of resulting complexes. The cell toxicity of the conjugates was evaluated using Staphylococcus aureus and finally they were administered into the chicken embryo to assess the applicability for in vivo imaging.

The technique described in this paper allows detection of quantum dots (QDs) specifically deposited on the polystyrene surface by laser-induced breakdown spectroscopy (LIBS). Using LIBS, the distribution of QDs or their conjugates with biomolecules deposited on the surface can be observed, regardless of the fact if they exhibit fluorescence or not. QDs deposited on the specific surface of polystyrene microplate in the form of spots are detected by determination of the metal included in the QDs structure. Cd-containing QDs (CdS, CdTe) stabilized with mercaptopropionic (MPA) or mercaptosuccinic (MSA) acid, respectively, alone or in the form of conjugates with metallothionein (MT) biomolecule are determined by using the 508.58 nm Cd emission line. The observed absolute detection limit for Cd in CdTe QDs conjugates with MT in one spot was 3 ng Cd. Due to the high sensitivity of this technique, the immunoanalysis in combination with LIBS was also investigated. Cd spatial distribution in sandwich immunoassay was detected.