Antimicrobial activity of copper-humate-cellulose sheets

During our research, cellulose-based test sheets containing copper and potassium humate were prepared and tested against several bacterial and fungal strains. The energy dispersive X-ray spectrometry and attenuated total reflectance (ATR) Fourier-transform infrared spectroscopy revealed that humate contributed to increase the copper content of the test sheets. The antimicrobial impact of test sheets on various fungi (Penicillium expansum, Wallemia sebi, and Aspergillus brasiliensis) and bacteria (genera Micrococcus, Enterococcus, Bacillus, Salmonella, Staphylococcus, Listeria genera) was tested using a modified agar diffusion-based assay. On the basis of the disc diffusion results, the sheets effectively inhibited all the tested fungi and Gram-positive bacteria. The highest antimicrobial effect was exhibited by the copper-containing test sheet with 1:0.1 and 1:0.25 cellulose:humate weight ratio. Deterioration test with blackberries demonstrated the shelf life extending effect of the sheets.     

Physicochemical properties and cellular toxicity of (poly)aminoalkoxysilanes-functionalized ZnO quantum dots

Luminescent ZnO nanocrystals were synthesized by basic hydrolysis of Zn(OAc)(2) in the presence of oleic acid and then functionalized with (poly)aminotrimethoxysilanes in the presence of tetramethylammonium hydroxide to render the QDs water-dispersible. The highest photoluminescence quantum yield (17%) was achieved using N(1)-(2-aminoethyl)-N(2)-[3-(trimethoxysilyl)propyl]-1,2-ethanediamine as surface ligand. Transmission electron microscopy and powder x-ray diffraction showed highly crystalline materials with a ZnO nanoparticle diameter of about 4?nm. The cytotoxicity of the different siloxane-capped ZnO QDs towards growing Escherichia coli bacterial cells was evaluated in MOPS-minimal medium. Although concentrations of 5?mM in QDs caused a complete growth arrest in E.?coli, siloxane-capped ZnO QDs appeared weakly toxic at lower doses (0.5 or 1?mM). The concentration of bioavailable Zn (2+) ions leaked from ZnO QDs was evaluated using the biosensor bacteria Cupriavidus metallidurans AE1433. The results obtained clearly demonstrate that concentrations of bioavailable Zn(2+) are too low to explain the inhibitory effects of the ZnO QDs against bacteria cells at 1?mM and that the siloxane shell prevents ZnO QDs from dissolution contrary to uncapped ZnO nanoparticles. Because of their low cytotoxicity, good biocompatibility, low cost and large number of functional amine end groups, which makes them easy to tailor for end-user purposes, siloxane-capped ZnO QDs offer a high potential as fluorescent probes and as biosensors.     

Synthesis, characterization and antimicrobial activity of conjugates based on fluoroquinolon-type antibiotics and gelatin

Different fluoroquinolon-type antibiotics were conjugated to gelatin with the aim to synthesize biomacromolecules with antimicrobial properties. The covalent linkage of the antibiotic was performed by a radical process involving the residues in the side chains of gelatin able to undergo oxidative modifications. The conjugation of antibiotic moieties onto the protein structure was confirmed by FT-IR, UV–Vis, fluorescence, and calorimetric analyses. Biocompatibility tests were performed on human bone marrow mesenchymal stromal cells and the antibacterial properties of bioactive polymers were investigated by appropriate tests against Klebsiella pneumoniae and Escherichia coli. With regard to the tests conducted in the presence of E. coli, a minimum inhibitory concentration (MIC) ranging from 0.05 to 0.40 μg mL^?1 was recorded, while in the presence of K. pneumoniae this concentration varies from 0.10 to 1.60 μg mL^?1. In all the conjugates, the drug moieties retain their biological activity and the MIC values are lower than the resistance parameters of fluoroquinolon-type antibiotics versus Enterobacteriacae. The collected data suggest a broad range of applications, from biomedical to pharmaceutical and food science for all conjugates.     

Antimicrobial activity of functionalised carbon nanotubes against pathogenic microorganisms

Carbon nanotubes represent one of the best examples of novel nanostructures, exhibit a range of extraordinary physical properties, strong antimicrobial activity and can pierce bacterial cell walls. This investigation handles the antimicrobial activity of functionalised multiwall carbon nanotubes (F‐MWNTs) as an alternative antimicrobial material compared to the commercial antibiotics. Antibacterial activities of F‐MWNTs are investigated through two different kinds of bacteria, E. coli and S. aureus. The results demonstrate that the best concentration of F‐MWNTs for the maximum inhibition and antibacterial functionality is 80 and 60 μg/ml for E. coli and S. aureus, respectively. The transmission electron microscope reveals the morphological changes damage mechanism for the cellular reliability on these microorganisms. F‐MWNTs are capable of biologically isolating the cell from their microenvironment, contributing to the development of toxic substances and placing the cell under oxidative stress leading to cellular death. The efficiency of F‐MWNTs is compared with the common antibiotics and shows an enhancement in the inhibitory effect with percentages reaches 85%. To account for the bactericidal performance of F‐MWNTs towards these pathogens, the dielectric conductivity and the bacterial growth measurements are conducted. The present study endeavour that F‐MWNTs could be exploited in biomedical devices and altering systems for hospital and industrial cleaning applications.Inspec keywords: antibacterial activity, biomedical materials, microorganisms, cellular biophysics, toxicology, nanomedicine, multi‐wall carbon nanotubes, transmission electron microscopy, electrical conductivity, permittivityOther keywords: F‐MWNTs, pathogenic microorganisms, antimicrobial activity, bacterial cell walls, functionalised multiwall carbon nanotubes, antibacterial activity, E. coli, S. aureus, antimicrobial material, physical properties, transmission electron microscopy, morphological changes, damage mechanism, cellular reliability, microenvironment, toxic substances, oxidative stress, cellular death, bactericidal performance, dielectric conductivity, bacterial growth measurements, biomedical devices, C     

Antimicrobial activity of chitosan attached to ethylene copolymer films

Ethylene copolymer film was coated with chitosan by attachment of the polymer to the corona‐treated surface of the film, and the composite film was analysed for antimicrobial activity. The film was active against bacteria in 0.625 mM phosphate buffer; it reduced colony counts of Escherichia coli 25922 and of Listeria monocytogenes Scott A by 5 and 2–3 log₁₀, respectively after 24 h exposure. The activity of the chitosan‐coated film against bacteria in buffer was increased when silver ions were incorporated into the films as demonstrated by complete killing of Escherichia coli O157:H7 DD3795 and the chicken exudate isolate Stenotrophomonas maltophilia within 2 h in buffer. The film was active against L. monocytogenes Scott A in 0.5% buffered peptone water up to a pH of about 7.0. Tests on beef and chicken meat exudates revealed antimicrobial activity of the film against Escherichia coli O157:H7 and L. monocytogenes Scott A of about 2 and 1–2 log₁₀ reductions in colony‐forming units, respectively. The antimicrobial activity of the film against L. monocytogenes Scott A was also tested on turkey breast, and a log₁₀ reduction of about 1.7 log₁₀ units after 10 days and 1.2 log₁₀ after 15 days at 4°C was achieved. Exposure to chitosan‐coated film and 350 MPa of pressure, 55°C or 1% sodium diacetate resulted in a synergistic effect. Copyright © 2008 John Wiley & Sons, Ltd     

Antimicrobial activity of UV-irradiated nylon film for packaging applications

Antimicrobial packaging could enhance food storage life and safety. An antimicrobial moiety that is permanently bound to the polymer surface and does not leach has particular appeal. The use of 193 nm UV irradiation to convert amide groups on the surface of nylon to amines having antimicrobial activity has been reported previously. We prepared materials accordingly and explored their mode of action and activity against pathogens. Three food related bacterial strains, Staphylococcus aureus, Pseudomonas fluorescens and Enterococcus faecalis were exposed to antimicrobial film in 0.2 M sodium phosphate buffer (pH 7.0). Samples were held shaken at 100 r.p.m. in a 25°C incubator. The antimicrobial film was effective in reduction of microbial concentration in the bulk fluid for all food-related bacteria tested. The effectiveness was dependent on the bacterial strain. Adsorption of bacterial cells diminished the effectiveness of amine groups. Experimental results indicate that the decrease in concentration of bacterial cells in bulk fluid is more likely to be the bactericidal action than adsorption of live cells. © 1998 John Wiley & Sons, Ltd.     

Proteolytic activity monitored by fluorescence resonance energy transfer through quantum-dot-peptide conjugates

Proteases are enzymes that catalyse the breaking of specific peptide bonds in proteins and polypeptides. They are heavily involved in many normal biological processes as well as in diseases, including cancer, stroke and infection. In fact, proteolytic activity is sometimes used as a marker for some cancer types. Here we present luminescent quantum dot (QD) bioconjugates designed to detect proteolytic activity by fluorescence resonance energy transfer. To achieve this, we developed a modular peptide structure which allowed us to attach dye-labelled substrates for the proteases caspase-1, thrombin, collagenase and chymotrypsin to the QD surface. The fluorescence resonance energy transfer efficiency within these nanoassemblies is easily controlled, and proteolytic assays were carried out under both excess enzyme and excess substrate conditions. These assays provide quantitative data including enzymatic velocity, Michaelis-Menten kinetic parameters, and mechanisms of enzymatic inhibition. We also screened a number of inhibitory compounds against the QD-thrombin conjugate. This technology is not limited to sensing proteases, but may be amenable to monitoring other enzymatic modifications.     

Antimicrobial activity of parabens in submicron emulsions stabilized with lecithin

Antimicrobial efficacy of methyl and propylparaben combination as potential preservatives for submicron emulsions, and the effect of oil and lecithin concentration on the microbial growth were investigated. Parabens were ineffective in standard or doubled concentrations as per pharmacopoeial criteria. Poor growth inhibition and multiplication of reference strains point to protective and growth properties of submicron emulsions. No correlation was observed between oil/lecithin ratio and efficacy of parabens; partitioning of the latter into the oily phase and lipophilic domains could be the reason for such effect. Further studies are necessary to establish a stable and safe composition of such formulations.     

Size,shape and surface chemistry of nano-gold dictate its cellular interactions,uptake and toxicity

Colloidal gold is undoubtedly one of the most extensively studied nanomaterials, with 1000s of different protocols currently available to synthesise gold nanoparticles (AuNPs). While developments in the synthesis of AuNPs have progressed rapidly in recent years, our understanding of their biological impact, with particular respect to the effect of shape, size, surface characteristics and aggregation states, has struggled to keep pace. It is generally agreed that when AuNPs are exposed to biological systems, these parameters directly influence their pharmacokinetic and pharmacodynamic properties by influencing AuNPs distribution, circulation time, metabolism and excretion in biological systems. However, the rules governing these properties, and the science behind them, are poorly understood. Therefore, a systematic understanding of the implications of these variables at the nano–bio interface has recently become a topic of major interest. This Review Article attempts to ignite a discussion around the influence of different physico-chemical parameters on biological activity of AuNPs, while focussing on critical aspects of cellular interactions, uptake and cytotoxicity. The review also discusses emerging trends in AuNP uptake and toxicity that are leading to technological advances through AuNP-based therapy, diagnostics and imaging.     

In vitro antifungal activity and toxicity of itraconazole in DMSA-PLGA nanoparticles

Itraconazole (ITZ) is a drug used to treat various fungal infections and may cause side effects. The aim of this study was to develop and evaluate the in vitro activity of DMSA-PLGA nanoparticles loaded with ITZ against Paracoccidioides brasiliensis, as well as their cytotoxicity. Nanoparticles were prepared using the emulsification-evaporation technique and characterized by their encapsulation efficiency, morphology (TEM), size (Nanosight) and charge (zeta potential). Antifungal efficacy in P. brasiliensis was determined by minimal inhibition concentration (MIC), and cytotoxicity using MTT assay. ITZ was effectively incorporated in the PLGA-DMSA nanoparticles with a loading efficiency of 72.8 +/- 3.50%. The shape was round with a solid polymeric structure, and a size distribution of 174 +/- 86 nm (Average +/- SD). The particles were negatively charged. ITZ-NANO presented antifungal inhibition (MIC = 6.25 ug/mL) against P. brasiliensis and showed lower in vitro cytotoxicity than free drug (ITZ).     

Determination of sphingosine kinase activity for cellular signaling studies

Regulation of sphingosine and sphingosine-1-phosphate concentrations is of growing interest due to their importance in cellular signal transduction. Furthermore, new pharmaceutical agents moderating the intracellular and extracellular levels of sphingosine metabolites are showing promise in preclinical and clinical trials. In the present work, a quantitative assay relying on capillary electrophoresis with laser-induced fluorescence detection was developed to measure the interconversion of sphingosine and sphingosine-1-phosphate. The assay was demonstrated to be capable of determining the in vitro activity of both kinase and phosphatase using purified enzymes. The KM of sphingosine kinase for its fluorescently labeled substrate was 38 +/- 18 microM with a Vmax of 0.4 +/- 0.2 microM/min and a kcat of 3900 s-1. Pharmacologic inhibition of sphingosine kinase in a concentration-dependent manner was also demonstrated. Moreover, the fluorescent substrate was shown to be readily taken up by mammalian cells making it possible to study the endogenous activity of sphingosine kinase activity in living cells. The method was readily adaptable to the use of either bulk cell lysates or very small numbers of intact cells. This new methodology provides enhancements over standard methods in sensitivity, quantification, and manpower for both in vitro and cell-based assays.     

Antimicrobial activity of novel biocompatible wound dressings based on triblock copolymer hydrogels

Wound infection is a common complication often resulting in delayed healing with adverse clinical and financial consequences. Current antimicrobial treatments are far from ideal, side effects can include both bacterial resistance and toxicity. As a result, a great deal of effort over the last 20 years has been spent on investigating new forms of antimicrobial dressings. Here, we report the unexpected antimicrobial activity of a relatively new biocompatible thermo-responsive PHPMA–PMPC–PHPMA triblock copolymer gelator [where PHPMA denotes poly(2-hydroxypropyl methacrylate) and PMPC denotes poly(2-(methacryloyloxy)ethyl phosphorylcholine)]. In a radial diffusion assay, a 20% w/v copolymer gel produced an inhibitory zone up to six times greater than the corresponding control against Staphylococcus aureus. Similarly, in a broth inhibition assay the same copolymer reduced bacterial growth by 45% compared with control experiments conducted in the absence of any copolymer. Moreover, addition of the copolymer to a 3D-infected skin model reduced bacterial recovery by 38% compared to that of controls over 24–48 h. This is particularly relevant since these antimicrobial triblock copolymers were recently shown to be non-toxic when exposed to a tissue-engineered skin model. This antimicrobial activity was also successfully immobilised by grafting PMPC–PHPMA diblock copolymer brushes onto silicon wafers. Our results indicate that both PMPC–PHPMA diblock and PHPMA homopolymer brushes exhibit antimicrobial activity. Our hypothesis for the mode of action is that the moderately hydrophobic PHPMA chains penetrate the bacterial membrane, causing leakage of the cell contents. In summary, these gels and surfaces offer a promising new approach to antimicrobial dressings.     

Antimicrobial activity of composite nanoparticles consisting of titania photocatalytic shell and nickel ferrite magnetic core

Reverse micelle and hydrolysis have been combined to synthesize composite nanoparticles consisting of anatase–titania photocatalytic shell and nickel ferrite magnetic core. The average particle size of the composite nanoparticles was in the range of 10–15 nm. The photocatalytic shell of titania is responsible for the photocatalytic and anti-microbial activity and nickel ferrite magnetic core is responsible for the magnetic behavior, studied by superconducting quantum interference device. The anatase TiO2 coated NiFe₂O₄ nanoparticles retains the magnetic characteristics of uncoated nanocrystalline nickel ferrites, superparamagnetism (absence of hysteresis, remanence and coercivity at 300 K) and non-saturation of magnetic moments at high field. The magnetic measurements results encourage their application as removable anti-microbial photocatalysts. Bacterial inactivation with UV light in the presence of titania-coated NiFe₂O₄ nanoparticles is faster than the action with UV light alone.     

Antimicrobial activity and biologic potential of silver-substituted calcium phosphate constructs produced with self-propagating high-temperature synthesis

There is significant demand for synthetic bone substitute materials that can decrease the incidence of implant-based bacterial infections. The intent of this research was to evaluate the antimicrobial activity and biologic potential of calcium phosphate (CaP) constructs substituted with silver (Ag) that were produced via self-propagating high-temperature synthesis (SHS). SHS is a combustion synthesis technique that has successfully generated porous CaP bioceramics intended for use in bone repair. SHS reactions are highly versatile; dopants can be added to the reactant powders to alter product chemistry and morphology. In this research, Ag powder was added to the reactants generating porous CaP constructs containing 0.5, 1, or 2 wt% Ag. Antibacterial performance of the constructs was assessed against Escherichia coli, a representative model for Gram-negative bacteria. Liquid solutions (1 μg/mL) of CaP–Ag particles to phosphate buffered saline were incubated with 10⁵ cells/mL. After 24 h, 10 μL of solution were spread on an LB agar plate and cultured for 24 h at 37 °C. Samples cultured with CaP–Ag showed complete bacterial inhibition while the controls (E. coli only and CaP without Ag) exhibited significant colony formation. The effects of Ag concentration on cytotoxicity and biocompatibility were tested in vitro. At 7 days, osteoblasts uniformly enveloped the CaP–Ag particles and displayed a healthy flattened morphology suggesting the concentrations of Ag incorporated into constructs were not cytotoxic. CaP–Ag constructs produced via SHS represent a source of synthetic bone substitute materials that could potentially inhibit, or reduce the incidence of post-operative bacterial infections.     

Improving anticancer activity and reducing systemic toxicity of doxorubicin by self-assembled polymeric micelles

In an attempt to improve anticancer activity and reduce systemic toxicity of doxorubicin (Dox), we encapsulated Dox in monomethoxy poly(ethylene glycol)-poly(ε-caprolactone) (MPEG-PCL) micelles by a novel self-assembly procedure without using surfactants, organic solvents or vigorous stirring. These Dox encapsulated MPEG-PCL (Dox/MPEG-PCL) micelles with drug loading of 4.2% were monodisperse and ～ 20 nm in diameter. The Dox can be released from the Dox/MPEG-PCL micelles; the Dox-release at pH 5.5 was faster than that at pH 7.0. Encapsulation of Dox in MPEG-PCL micelles enhanced the cellular uptake and cytotoxicity of Dox on the C-26 colon carcinoma cell in vitro, and slowed the extravasation of Dox in the transgenic zebrafish model. Compared to free Dox, Dox/MPEG-PCL micelles were more effective in inhibiting tumor growth in the subcutaneous C-26 colon carcinoma and Lewis lung carcinoma models, and prolonging survival of mice bearing these tumors. Dox/MPEG-PCL micelles also induced lower systemic toxicity than free Dox. In conclusion, incorporation of Dox in MPEG-PCL micelles enhanced the anticancer activity and decreased the systemic toxicity of Dox; these Dox/MPEG-PCL micelles are an interesting formulation of Dox and may have potential clinical applications in cancer therapy.     

Comparative cytotoxicity of gold-doxorubicin and InP-doxorubicin conjugates

Direct comparisons of different types of nanoparticles for drug delivery have seldom been performed. In this study we compare the physical properties and cellular activity of doxorubicin (Dox) conjugates to gold nanoparticles (Au) and InP quantum dots of comparable diameter. Although the Au particles alone are non-toxic and InP is moderately toxic, Au-Dox is more effective than InP-Dox against the Dox-resistant B16 melanoma cell line. Light exposure does not augment the efficacy of InP-Dox, suggesting that conjugates are breaking down. Electron and confocal microscopy and atomic absorption spectroscopy reveal that over 60% of the Au-Dox conjugates reach the cell nucleus. In contrast, InP-Dox enters cell nuclei to a very limited extent, although liberated Dox from the conjugates does eventually reach the nucleus. These observations are attributed to faster Dox release from Au conjugates under endosomal conditions, greater aggregation of InP-Dox with cytoplasmic proteins, and adherence of InP to membranes. These findings have important implications for design of active drug-nanoparticle conjugates.     

抗菌涂层和抗菌塑料在食品包装中的研究发展

目的从可食用抗菌涂层和食品抗菌塑料包装的角度出发提升食品的品质安全,满足消费者对安全优质食品的追求。方法根据食物的腐烂周期和保质方式的特点,系统性地总结目前可食用抗菌涂层的几种主要覆盖技术,以及非食用性抗菌塑料的载体和制备方法,并将其运用到食品的包装设计中。结论针对保鲜周期短的易腐食品和保质期较长的常规食品提出了可行的包装方案,对可食用抗菌涂层和非可食用性抗菌塑料在未来食品包装领域的发展和研究做了指向性分析。     

Comparison of the abilities of ambient and manufactured nanoparticles to induce cellular toxicity according to an oxidative stress paradigm

Nanomaterial properties differ from those bulk materials of the same composition, allowing them to execute novel activities. A possible downside of these capabilities is harmful interactions with biological systems, with the potential to generate toxicity. An approach to assess the safety of nanomaterials is urgently required. We compared the cellular effects of ambient ultrafine particles with manufactured titanium dioxide (TiO2), carbon black, fullerol, and polystyrene (PS) nanoparticles (NPs). The study was conducted in a phagocytic cell line (RAW 264.7) that is representative of a lung target for NPs. Physicochemical characterization of the NPs showed a dramatic change in their state of aggregation, dispersibility, and charge during transfer from a buffered aqueous solution to cell culture medium. Particles differed with respect to cellular uptake, subcellular localization, and ability to catalyze the production of reactive oxygen species (ROS) under biotic and abiotic conditions. Spontaneous ROS production was compared by using an ROS quencher (furfuryl alcohol) as well as an NADPH peroxidase bioelectrode platform. Among the particles tested, ambient ultrafine particles (UFPs) and cationic PS nanospheres were capable of inducing cellular ROS production, GSH depletion, and toxic oxidative stress. This toxicity involves mitochondrial injury through increased calcium uptake and structural organellar damage. Although active under abiotic conditions, TiO2 and fullerol did not induce toxic oxidative stress. While increased TNF-alpha production could be seen to accompany UFP-induced oxidant injury, cationic PS nanospheres induced mitochondrial damage and cell death without inflammation. In summary, we demonstrate that ROS generation and oxidative stress are a valid test paradigm to compare NP toxicity. Although not all materials have electronic configurations or surface properties to allow spontaneous ROS generation, particle interactions with cellular components are capable of generating oxidative stress.     

Antimicrobial activity of microencapsulated lemongrass essential oil and the effect of experimental parameters on microcapsules size and morphology

Lemongrass (Cymbopogom citratus) essential oil, known due to its broad-spectrum antimicrobial activity, was microencapsulated by simple coacervation. Poly(vinyl alcohol) (PVA, 78,000 Da and 88 mol% degree of hydrolysis) crosslinked with glutaraldehyde was used as wall-forming polymer. The influence of stirring rate and oil volume fraction on the microcapsule size distribution were evaluated. Sodium dodecil sulphate (SDS) and Poly(vinyl pirrolidone) were tested in order to avoid microcapsules agglomeration during the process. Depending on the experimental conditions, microcapsules in the range of 10 μm to 250 μm were obtained. Microcapsules presenting no agglomeration were obtained when SDS at 0.03 wt.% was used. The composition and the antimicrobial properties of the encapsulated oil were determined, demonstrating that the process of microencapsulation did not deteriorate the encapsulated essential oil.