Effect of extreme heat processing on the Moroccan Zantaz' honey antioxidant activities.

The effect of an extreme heat processing on Zantaz honey samples was studied using a panel of physicochemical parameters, antioxidant activities and FTIR-ATR spectroscopy. Honey samples were heated at 121 °C for 30 min and the heat processing effect was confirmed indirectly through the assessment of hydroxymethylfurfural content, for which the values increased significantly (p < 0.01), and diastase activity, which was totally absent after the thermal processing. Besides, the effects of the heat on the antioxidant activities were diverse. Indeed, while the ability to scavenge 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid cation radicals (ABTS·+) and 2,2-diphenyl-1-picrylhydrazyl radicals was enhanced (p < 0.05 only for ABTS·+), after the heat processing, nitric oxide radicals scavenging activity was reduced drastically (p < 0.01). Regarding the chelating power, it was totally abolished following heating. Other activities showed no significant alteration. The initial values of antioxidant activities seem to be determinant in the changes occurring after the heat processing. Amongst the analysed parameters, following the heat processing, the honey colour was the variable where the influence of baseline values was the greatest. The spectral analyses confirmed that FTIR-ATR is a useful technique to discriminate the chemical differences occurring in honey after heat processing.

Insight on Propolis from Mediterranean Countries: Chemical Composition, Biological Activities and Application Fields.

This review updates the information upon the chemical composition of propolis from all Mediterranean countries as well as their biological properties and applications. The non-volatile fraction of propolis was characterized by the presence of phenolic acids and their esters and flavonoids. Nevertheless, in some countries, diterpenes were also present: Sicily (Italy), Croatia, Malta, Creta (Greece), Turkey, Cyprus, Egypt, Libya, Algeria and Morocco. The volatile fraction of propolis was characterized by the presence of benzoic acid and its esters, mono- and sesquiterpenes, being the oxygenated sesquiterpene ß-eudesmol characteristic of poplar propolis, whereas the hydrocarbon monoterpene α-pinene has been related with the presence of conifers. Regardless the chemical composition, there are common biological properties attributed to propolis. Owing to these attributes, propolis has been target of study for applications in diverse areas, such as food, medicine and livestock.

Impact of Biohybrid Magnetite Nanoparticles and Moroccan Propolis on Adherence of Methicillin Resistant Strains of Staphylococcus aureus.

Biofilm bacteria are more resistant to antibiotics than planktonic cells. Propolis possesses antimicrobial activity. Generally, nanoparticles containing heavy metals possess antimicrobial and antibiofilm properties. In this study, the ability of adherence of Methicillin Resistant Strains of Staphylococcus aureus (MRSA) to catheters treated with magnetite nanoparticles (MNPs), produced by three methods and functionalized with oleic acid and a hydro-alcoholic extract of propolis from Morocco, was evaluated. The chemical composition of propolis was established by gas chromatography mass spectrometry (GC-MS), and the fabricated nanostructures characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), Mossbauer spectroscopy and Fourrier transform infrared spectroscopy (FTIR). The capacity for impairing biofilm formation was dependent on the strain, as well as on the mode of production of MNPs. The co-precipitation method of MNPs fabrication using Fe(3+) and Na2SO3 solution and functionalized with oleic acid and propolis was the most effective in the impairment of adherence of all MRSA strains to catheters (p < 0.001). The adherence of the strain MRSA16 was also significantly lower (p < 0.001) when the catheters were treated with the hybrid MNPs with oleic acid produced by a hydrothermal method. The anti-MRSA observed can be attributed to the presence of benzyl caffeate, pinocembrin, galangin, and isocupressic acid in propolis extract, along with MNPs. However, for MRSA16, the impairment of its adherence on catheters may only be attributed to the hybrid MNPs with oleic acid, since very small amount, if any at all of propolis compounds were added to the MNPs.

Sodium triflate decreases interaggregate repulsion and induces phase separation in cationic micelles.

Dodecyltrimethylammonium triflate (DTATf) micelles possess lower degree of counterion dissociation (α), lower hydration, and higher packing of monomers than other micelles of similar structure. Addition of sodium triflate ([NaTf] > 0.05 M) to DTATf solutions promotes phase separation. This phenomenon is commonly observed in oppositely charged surfactant mixtures, but it is rare for ionic surfactants and relatively simple counterions. While the properties of DTATf have already been reported, the driving forces for the observed phase separation with added salt remain unclear. Thus, we propose an interpretation for the observed phase separation in cationic surfactant solutions. Addition of up to 0.03 M NaTf to micellar DTATf solutions led to a limited increase of the aggregation number, to interface dehydration, and to a progressive decrease in α. The viscosity of DTATf solutions of higher concentration ([DTATf] ≥ 0.06 M) reached a maximum with increasing [NaTf], though the aggregation number slightly increased, and no shape change occurred. We hypothesize that this maximum results from a decrease in interaggregate repulsion, as a consequence of increased ion binding. This reduction in micellar repulsion without simultaneous infinite micellar growth is, probably, the major driving force for phase separation at higher [NaTf].

Loading and protection of hydrophilic molecules into liposome-templated polyelectrolyte nanocapsules.

Compartmentalized systems produced via the layer-by-layer (LbL) self-assembly method have been produced by alternatively depositing alginate and chitosan layers onto cores of liposomes. The combination of dynamic light scattering (DLS), ζ potential, and transmission electron microscopy (TEM) techniques provides detailed information on the stability, dimensions, charge, and wall thickness of these polyelectrolyte globules. TEM microphotographs demonstrate the presence of nanocapsules with an average diameter of below 300 nm and with a polyelectrolyte wall thickness of about 20 nm. The possibility of encapsulating and releasing molecules from this type of nanocapsule was demonstrated by loading FITC-dextrans of different molecular weights in the liposome system. The release of the loaded molecules from the nanocapsule was demonstrated after liposome core dissolution. Even at low molecular weight (20 kDa), the nanocapsules appear to be appropriate for prolonged molecule compartmentalization and protection. By means of the Ritger-Peppas model, non-Fickian transport behavior was detected for the diffusion of dextran through the polyelectrolyte wall. Values of the diffusion coefficient were calculated and yield useful information regarding chitosan/alginate hollow nanocapsules as drug-delivery systems. The influence of the pH on the release properties was also considered. The results indicate that vesicle-templated hollow polyelectrolyte nanocapsules show great potential as novel controllable drug-delivery devices for biomedical and biotechnological applications.

Arbutus unedo L.: chemical and biological properties.

Arbutus unedo L. (strawberry tree) has a circum-Mediterranean distribution, being found in western, central and southern Europe, north-eastern Africa (excluding Egypt and Libya) and the Canary Islands and western Asia. Fruits of the strawberry tree are generally used for preparing alcoholic drinks (wines, liqueurs and brandies), jams, jellies and marmalades, and less frequently eaten as fresh fruit, despite their pleasing appearance. An overview of the chemical composition of different parts of the plant, strawberry tree honey and strawberry tree brandy will be presented. The biological properties of the different parts of A. unedo and strawberry tree honey will be also overviewed.

Encapsulation of DNA in macroscopic and nanosized calcium alginate gel particles.

Calcium alginate beads, which are biodegradable and biocompatible, have been widely employed as delivery matrices for biomacromolecules. In the present work, the feasibility of encapsulation of DNA (which is used as a model biomacromolecule) in calcium alginate nanobeads (sub-200 nm size), prepared using a recently developed protocol based on the phase inversion temperature (PIT) emulsification method [Machado et al. Langmuir 2012, 28, 4131-4141], was assessed. The properties of the nanobeads were compared to those of the corresponding macroscopic (millimeter sized) calcium alginate beads. It was found that DNA, representing a relatively stiff and highly charged polyanion (thus like-charged to alginate), could be efficiently encapsulated in both nanosized and macroscopic beads, with encapsulation yields in the range of 77-99%. Complete release of DNA from the beads could be accomplished on dissolution of the gel by addition of a calcium-chelating agent. Importantly, the DNA was not denatured or fragmented during the preparation and collection of the nanobeads, which are good indicators of the mildness of the preparation protocol used. The calcium alginate nanobeads prepared by the herein utilized protocol thus show good potential to be used as carriers of sensitive biomacromolecules.

Preparation of calcium alginate nanoparticles using water-in-oil (W/O) nanoemulsions.

A procedure for the preparation of calcium alginate nanoparticles in the aqueous phase of water-in-oil (W/O) nanoemulsions was developed. The emulsions were produced from mixtures of the nonionic surfactant tetraethylene glycol monododecyl ether (C(12)E(4)), decane, and aqueous solutions of up to 2 wt % sodium alginate by means of the phase inversion temperature (PIT) emulsification method. This method allows the preparation of finely dispersed emulsions without a large input of mechanical energy. With alginate concentrations of 1-2 wt % in the aqueous phase, emulsions showed good stability against Ostwald ripening and narrow, monomodal distributions of droplets with radii <100 nm. Gelation of the alginate was induced by the addition of aqueous CaCl(2) to the emulsions under stirring, and particles formed were collected using a simple procedure based on extraction of the surfactant on addition of excess oil. The final particles were characterized using cryo-transmission electron microscopy (cryo-TEM) and dynamic light scattering (DLS). They were found to be essentially spherical with a homogeneous interior, and their size was similar to that of the initial emulsion droplets. The herein presented "low-energy" method for preparation of biocompatible nanoparticles has the potential to be used in various applications, e.g., for the encapsulation of sensitive biomacromolecules.

Antioxidant and antiacetylcholinesterase activities of some commercial essential oils and their major compounds.

The commercial essential oils of Citrus aurantium L., Cupressus sempervirens L., Eucalyptus globulus Labill., Foeniculum vulgare Mill. and Thymus vulgaris L., isolated by steam distillation by a company of Morocco were evaluated in terms of in vitro antioxidant activity through several methods. In vitro acetylcholinesterase inhibitory activity was also determined. Citrus limon (L.) Burm. f. oil was also studied, but it was obtained by peel expression. The best antioxidant was T. vulgaris oil, independent of the method used, mainly due to the presence of the phenolic monoterpenes thymol and carvacrol, which when studied as single compounds also presented the best activities. Concerning the acetylcholinesterase inhibition activity, E. globulus was the most effective. Nevertheless its main components 1,8-cineole and limonene were not the most active, a feature that corresponded to d-3-carene.

Physicochemical characteristics and antiproliferative and antioxidant activities of Moroccan Zantaz honey rich in methyl syringate.

Zantaz honey is a monofloral variety produced from the melliferous plant Bupleurum spinosum (Apiaceae), a shrub that grows mainly in the Atlas Moroccan Mountains. Determination of the polyphenol composition revealed that methyl syringate accounts for more than 50% of total polyphenols, which represents a very useful parameter for the characterization of this monofloral honey. Epicatechin, syringic acid and catechin are also abundant. Caco-2 and THP-1 cells were used for determination of antioxidant and antiproliferative activities in Zantaz honey, respectively. All six commercial samples that were used for these studies exhibited antioxidant activity and inhibited cell proliferation. Interestingly, these activities had a positive correlation mainly with the content in methyl syringate and gallic acid. The recognition of health promoting activities in Zantaz honey should increase its commercial value, which would have a positive economic impact on the poor rural communities of Morocco where it is produced.

Antioxidant activity and enzyme inhibitory potential of Euphorbia resinifera and E. officinarum honeys from Morocco and plant aqueous extracts.

Natural products may be applied in a wide range of domains, from agriculture to food and pharmaceutical industries. In this study, the antioxidant properties and the capacity to inhibit some enzymatic activities of Euphorbia resinifera and Euphorbia officinarum aqueous extracts and honeys were assessed. The physicochemical characteristics were also evaluated. Higher amounts of iron, copper and aluminium were detected in E. officinarum honey, which may indicate environmental pollution around the beehives or inadequate storage of honey samples. This honey sample showed higher amounts of total phenols and better capacity for scavenging superoxide anion free radicals and DPPH free radicals as compared with E. resinifera honey, but poorer capacity for inhibiting lipoxygenase, acetylcholinesterase, tyrosinase and xanthine oxidase. The ratio plant mass:solvent volume (1:100) and extraction time (1 - 2 h) were associated with higher total phenols and better antioxidant activities and lipoxygenase, acetylcholinesterase and tyrosinase inhibitory activities, regardless of the plant species. The aqueous extracts had systematically higher in vitro activities than the respective honey samples.

Vesicle-templated layer-by-layer assembly for the production of nanocapsules.

Hollow structures on the submicrometer scale (nm) are obtained with the assembly of polyelectrolytes according to the layer-by-layer (LbL) technique. Following the LbL procedure, polymers alginate and chitosan were alternatively adsorbed on a vesicular template made of didodecyldimethylammonium bromide (DDAB). Evidence for the removal of the vesicular template entrapped in the alginate/chitosan film is presented. The removal of the vesicular template was achieved through interactions between a nonionic surfactant (Triton X100) and the double-chained surfactant forming the vesicles. The application of this approach allowed the production of hollow nanospheres with a mild procedure, avoiding the use of strong acids or other extreme working conditions that can modify the shell integrity. The obtained nanostructures were characterized by means of dynamic light scattering (DLS), the zeta potential, and scanning electron microscopy (SEM). The SEM analysis demonstrated the presence, after the core removal process, of nanocapsules indistinguishable in size and shape from the parent core-shell system. The analysis of the surface charge of the hollow nanocapsules, after the core dissolution, by zeta potential measurements, indicates good aggregate stability. DLS measurements showed that the size of the nanocapsules is on the order of hundreds of nanometers. Moreover, the size of both the core-shell and the hollow particles did not appear to be perturbed by variations in temperature or ionic strength.

Interactions between DNA and nonionic ethylene oxide surfactants are predominantly repulsive.

In the present work, the interactions between double-stranded (ds) or single-stranded (ss) DNA and nonionic ethylene oxide (EO) surfactants, with special attention to the possible contributions from hydrophobic interactions, have been investigated using a multitechnique approach. It was found that the presence of ss as well as dsDNA induces a slight decrease of the cloud point of pentaethylene glycol monododecyl ether (C(12)E(5)). Assessment of the partitioning of DNA between the surfactant-rich and surfactant-poor phases formed above the cloud point showed that the polymer was preferably located in the surfactant-poor phase. Surface tensiometry experiments revealed that neither of the DNA forms induced surfactant micellization. Finally, it was shown by DNA melting measurements that another EO surfactant (C(12)E(8)) did not affect the relative stabilities of ss and dsDNA. To summarize, all experiments suggest that the net interaction between DNA and nonionic surfactants of the EO type is weakly repulsive, which can be attributed mainly to steric effects. In general, the results were practically identical for the ds and ss forms of DNA, except those from the cloud point experiments, where the decrease of the cloud point was less pronounced with ssDNA. This finding indicates the presence of an attractive component in the interaction, which can reasonably be ascribed to hydrophobic effects.

Facile control of surfactant lamellar phase transition and adsorption behavior.

This study sets out to investigate the effect of the presence of small water-soluble additives on the tunability of the surfactant gel-to-liquid crystalline (Lß-Lα) phase transition temperature (T m) for a bilayer-forming cationic surfactant and the phase behavior of such surfactant systems on dilution. This is strongly driven by the fact that this type of cationic surfactant has many interesting unanswered scientific questions and has found applications in various areas such as consumer care, the petrochemical industry, food science, etc. The underlying surfactant/additive interactions and the interfacial behavior of lamellar surfactant systems including the surfactant deposition on surfaces can provide new avenues to develop novel product formulations. We have examined dioctadecyldimethyl ammonium chloride (DODAC) in the presence of small polar additives, with respect to the phase behavior upon dilution and the deposition on silica. Differential scanning calorimetry (DSC) is used to track the transition temperature, T m, and synchrotron and laboratory-based small and wide-angle X-ray scattering (SAXS and WAXS) were used to determine the self-assembled surfactant structure below and above the T m. DSC scans showed that upon dilution the additives could be removed from the surfactant bilayer which in turn tuned the T m. A spontaneous transition from a liquid crystalline (Lα) phase to a gel (Lß) phase on dilution was demonstrated, which indicated that additives could be taken out from the Lα phase. By means of in situ null ellipsometry, the deposition of a diluted surfactant Lß phase upon replacement of bulk solution by deionized water was followed. This technique enables time-resolved monitoring of the deposited surfactant layer thickness and adsorbed amount, which allows us to understand the deposition on surfaces. Robust layers at least one bilayer-thick were deposited onto the surface and shown to be irreversibly adsorbed due to poor surfactant solvency in water. The thickest layer of surfactant deposited after dilution was found for mixtures with small amounts of additive since high amounts might lead to a phase-separated system.

Role of linker groups between hydrophilic and hydrophobic moieties of cationic surfactants on oligonucleotide-surfactant interactions.

The interaction between DNA and amino-acid-based surfactants with different linker groups was investigated by gel electrophoresis, ethidium bromide exclusion assays, circular dichroism, and melting temperature determinations. The studies showed that the strength of the interaction between the oligonucleotides and the surfactants is highly dependent on the linker of the surfactant. For ester surfactants, no significant interaction was observed for surfactant-to-DNA charge ratios up to 12. On the other hand, amide surfactants were shown to interact strongly with the oligonucleotides; these surfactants could displace up to 75% of the ethidium bromide molecules bound to the DNA and induced significant changes in the circular dichroism spectra. When comparing the headgroups of the surfactants, it was observed that surfactants with more hydrophobic headgroups (proline vs alanine) interacted more strongly with the DNA, in good agreement with previous studies.

Re-dissolution and de-compaction of DNA-cationic surfactant complexes using non-ionic surfactants.

Addition of a cationic surfactant to a solution of DNA causes the formation of compacted DNA-cationic surfactant complexes which precipitate from aqueous solution. It has been shown previously that addition of anionic surfactant will re-dissolve and de-compact the DNA-cationic surfactant complexes and we find that addition of non-ionic surfactants of the alkylpolyoxyethylene type can be used similarly. In principle, these de-compaction and re-dissolution processes could occur either by stripping of the cationic surfactant from the DNA into mixed micelles with the non-ionic surfactant or by solubilisation of the DNA-cationic surfactant complexes within the non-ionic micelles. Solubility phase-boundary measurements, fluorescence microscopy observations of the de-compaction process and light scattering results indicate that de-compaction and re-dissolution occur by the stripping mechanism, even for non-ionic surfactants where the favourable attractive electrostatic interaction between the two surfactants is absent. Using measurements of critical micelle concentrations and calculations based on regular solution mixed micelle theory, we show that re-dissolution and de-compaction of the DNA-cationic surfactant complexes occurs when the concentration of free monomeric cationic surfactant is reduced (by incorporation into mixed micelles) below a critical value.

Interaction between DNA and cationic surfactants: effect of DNA conformation and surfactant headgroup.

The interactions between DNA and a number of different cationic surfactants, differing in headgroup polarity, were investigated by electric conductivity measurements and fluorescence microscopy. It was observed that, the critical association concentration (cac), characterizing the onset of surfactant binding to DNA, does not vary significantly with the architecture of the headgroup. However, comparing with the critical micelle concentration (cmc) in the absence of DNA, it can be inferred that the micelles of a surfactant with a simple quaternary ammonium headgroup are much more stabilized by the presence of DNA than those of surfactants with hydroxylated head-groups. In line with previous studies of polymer-surfactant association, the cac does not vary significantly with either the DNA concentration or its chain length. On the other hand, a novel observation is that the cac is much lower when DNA is denaturated and in the single-stranded conformation, than for the double-helix DNA. This is contrary to expectation for a simple electrostatically driven association. Thus previous studies of polyelectrolyte-surfactant systems have shown that the cac decreases strongly with increasing linear charge density of the polyion. Since double-stranded DNA (dsDNA) has twice as large linear charge density as single-stranded DNA (ssDNA), the stronger binding in the latter case indicates an important role of nonelectrostatic effects. Both a higher flexibility of ssDNA and a higher hydrophobicity due to the exposed bases are found to play a role, with the hydrophobic interaction argued to be more important. The significance of hydrophobic DNA-surfactant interaction is in line with other observations. The significance of nonelectrostatic effects is also indicated in significant differences in cac between different surfactants for ssDNA but not for dsDNA. For lower concentrations of DNA, the conductivity measurements presented an "anomalous" feature, i.e., a second inflection point for surfactant concentrations below the cac; this feature was not displayed at higher concentrations of DNA. The effect is attributed to the presence of a mixture of ss- and dsDNA molecules. Thus the stability of dsDNA is dependent on a certain ion atmosphere; at lower ion concentrations the electrostatic repulsions between the DNA strands become too strong compared to the attractive interactions, and there is a dissociation into the individual strands. Fluorescence microscopy studies, performed at much lower DNA concentrations, demonstrated a transformation of dsDNA from an extended "coil" state to a compact "globule" condition, with a broad concentration region of coexistence of coils and globules. The onset of DNA compaction coincides roughly with the cac values obtained from conductivity measurements. This is in line with the observed independence of cac on the DNA concentration, together with the assumption that the onset of binding corresponds to an initiation of DNA compaction. No major changes in either the onset of compaction or complete compaction were observed as the surfactant headgroup was made more polar.

Effect of the head-group geometry of amino acid-based cationic surfactants on interaction with plasmid DNA.

The interaction between DNA and different types of amino acid-based cationic surfactants was investigated. Particular attention was directed to determine the extent of influence of surfactant head-group geometry toward tuning the interaction behavior of these surfactants with DNA. An overview is obtained by gel retardation assay, isothermal titration calorimetry, fluorescence spectroscopy, and circular dichroism at different mole ratios of surfactant/DNA; also, cell viability was assessed. The studies show that the surfactants with more complex/bulkier hydrophobic head group interact more strongly with DNA but exclude ethidium bromide less efficiently; thus, the accessibility of DNA to small molecules is preserved to a certain extent. The presence of more hydrophobic groups surrounding the positive amino charge also gave rise to a significantly lower cytotoxicity. The surfactant self-assembly pattern is quite different without and with DNA, illustrating the roles of electrostatic and steric effects in determining the effective shape of a surfactant molecule.

Exploring the prebiotic effect of cyclodextrins on probiotic bacteria entrapped in carboxymetyl cellulose-chitosan particles.

In this work the prebiotic effect of different cyclodextrins, CDs, on the viability of model probiotic bacteria (Lactobacillus rhamnosus GG) encapsulated in carboxymethyl cellulose-chitosan (CMC-Cht) hybrid particles was studied. All the CDs tested were observed to considerably improve the viability (quantitatively like common prebiotics, such as corn starch) and encapsulation efficiency when compared to the CD-free particles, as inferred by plate counting method and fluorescence microscopy. The SEM data suggests that the morphology of the particles, the roughness of the surface and porosity, are dependent on the type of CD and may reflect different interactions between the CDs and the matrix components. The aging and stability of the samples with and without ß-CD were further evaluated. Remarkably, the viability count of the CD-doped samples was still reasonably high after one month storage at room temperature with acceptable values for practical uses. Moreover, when the CMC-Cht particles were exposed to in vitro simulated digestion fluids, the cell survival was much enhanced when the particles contained ß-CD.

Effect of headgroup on DNA-cationic surfactant interactions.

The interaction behavior of DNA with different types of hydroxylated cationic surfactants has been studied. Attention was directed to how the introduction of hydroxyl substituents at the headgroup of the cationic surfactants affects the compaction of DNA. The DNA-cationic surfactant interaction was investigated at different charge ratios by several methods like UV melting, ethidium bromide exclusion, and gel electrophoresis. Studies show that there is a discrete transition in the DNA chain from extended coils (free chain) to a compact form and that this transition does not depend substantially on the architecture of the headgroup. However, the accessibility of DNA to ethidium bromide is preserved to a significantly larger extent for the more hydrophilic surfactants. This was discussed in terms of surfactant packing. Observations are interpreted to reflect that the surfactants with more substituents have a larger headgroup and therefore form smaller micellar aggregates; these higher curvature aggregates lead to a less efficient, "patch-like" coverage of DNA. The more hydrophilic surfactants also presented a significantly lower cytotoxicity, which is important for biotechnological applications.