Effectiveness of nanoemulsions of clove and lemongrass essential oils and their major components against Escherichia coli and Botrytis cinerea.

Nanoemulsions exhibit a number of advantages to carry and deliver lipophilic compounds such as essential oils (EOs) due to their good stability and high surface area per volume unit. The purpose of this work was to assess the long-term stability of nanoemulsions of clove and lemongrass (LG) EOs and their principal components eugenol and citral (CI), respectively, at 3 different concentrations (2, 5 or 10 times their respective minimum inhibitory concentrations) and at two storage temperatures (1 °C and 21 °C). The initial droplet size of LG and CI-loaded nanoemulsions was below 100 nm and most of them kept droplet sizes in the nano-range until the end of storage at both temperatures. The ζ-potential was lower than - 40 mV, but it increased through storage, indicating a weaker alginate adsorption at the oil surface at both temperatures. The antimicrobial activity increased with the EOs concentration and was negatively affected by the highest storage temperature. Nanoemulsions containing CI and LG were able to significantly decrease Escherichia coli counts during storage, particularly at 1 °C. Nanoemulsions containing 1.0 and 2.0% w/w CI and 2.5% w/w LG were the most efficient in reducing Botrytis cinerea growth through storage, mainly at 1 °C. The nanoemulsions containing 1.0 and 2.0% w/w CI, as well as, 1.25 and 2.5% w/w LG better maintained their stability and antimicrobial effect along 6-months storage mainly when at 1 °C, making those nanoemulsions suitable as edible coatings for food preservation. Future studies should be oriented to evaluate the impact of these nanoemulsions on the organoleptic properties of coated foods and their potential toxicity.

Elucidating the effect of additives on the alkyl chain packing of a double tail cationic surfactant.

HYPOTHESIS: Some low molecular weight additives can strongly influence the phase behavior of aqueous surfactant systems, and this offers an important handle to control the properties of surfactant solutions and thus to optimize the stability and performance of various formulations. EXPERIMENTS: The surfactant dioctadecyldimethylammonium chloride (DODAC) self-assembles into two lamellar phases in water, the gel phase (Lß) and the liquid crystalline phase (Lα). Here, we present approaches to tune the gel-to-liquid crystalline transition temperature (Tm) with the use of additives. The effects of urea, sodium butyrate and butyric acid on the packing behavior of DODAC were determined. The surfactant phases were characterized using polarized optical microscopy (POM), differential scanning calorimetry (DSC), and small/wide angle X-ray scattering (SWAXS). FINDINGS: All three additives - urea, sodium butyrate and butyric acid yield a single and stable lamellar phase. Urea and sodium butyrate have only minor effects on Tm, butyric acid gives a large decrease as it stabilizes the Lα phase with respect to the Lß phase. From the bilayer thickness of the gel phase an interdigitated or tilted packing of the surfactant molecules is suggested. The addition of sodium butyrate gives a highly interdigitated gel structure and resulted in the transition from lamellar liquid crystal to an isotropic L3 phase.

Development of carboxymethyl cellulose-chitosan hybrid micro- and macroparticles for encapsulation of probiotic bacteria.

Novel carboxymethyl cellulose-chitosan (CMC-Cht) hybrid micro- and macroparticles were successfully prepared in aqueous media either by drop-wise addition or via nozzle-spray methods. The systems were either physically or chemically crosslinked using genipin as the reticulation agent. The macroparticles (ca. 2mm) formed are found to be essentially of the core-shell type, while the microparticles (ca. 5µm) are apparently homogeneous. The crosslinked particles are robust, thermally resistant and less sensitive to pH changes. On the other hand, the physical systems are pH sensitive presenting a remarkable swelling at pH 7.4, while little swelling is observed at pH 2.4. Furthermore, model probiotic bacteria (Lactobacillus rhamnosus GG) was for the first time successfully encapsulated in the CMC-Cht based particles with acceptable viability count. Overall, the systems developed are highly promising for probiotic encapsulation and potential delivery in the intestinal tract with the purpose of modulating gut microbiota and improving human health.

Honey as a Complementary Medicine.

The beneficial effects of honey on human health have long been recognized. Today, many of those positive effects have been studied to elucidate its mode of action. This review briefly summarizes the best studied features of honey, highlighting it as an appealing alternative medicine. In these reports, the health benefits of honey range from antioxidant, immunomodulatory, and anti-inflammatory activity to anticancer action, metabolic and cardiovascular benefits, prebiotic properties, human pathogen control, and antiviral activity. These studies also support that the honey's biological activity is mainly dependent on its floral or geographic origin. In addition, some promising synergies between honey and antibiotics have been found, as well as some antiviral properties that require further investigation. Altogether, these studies show that honey is effectively a nutraceutical foodstuff.

Identification and assessment of kefir yeast potential for sugar/ethanol-resistance.

Biochemical and molecular analysis was used for identification of different kefir yeasts species from Brazil, Canada and the United States of America. The sugar/ethanol-resistant activity of the yeasts was evaluated. Saccharomyces cerevisiae and Kluyveromyces marxianus had the highest growth rates, suggesting biotechnological applications possible for these strains.

Mixed protein-DNA gel particles for DNA delivery: role of protein composition and preparation method on biocompatibility.

Mixtures of two cationic proteins were used to prepare protein-DNA gel particles, employing associative phase separation and interfacial diffusion (Morán et al., 2009a). By mixing the two proteins, we have obtained particles that displayed higher loading efficiency and loading capacity values than those obtained in single-protein systems. However, nothing is known about the adverse effects on haemocompatibility and cytotoxicity of these protein-DNA gel particles. Here, we examined the interaction of protein-DNA gel particles obtained by two different preparation methods, and their components, with red blood cells and established cells. From a haemolytic point of view, these protein-DNA gel particles were demonstrated to be promising long-term blood-contacting medical devices. Safety evaluation with the established cell lines revealed that, in comparison with proteins in solution, the cytotoxicity was reduced when administered in the protein-DNA systems. In comparison with large-sized particles, the cytotoxic responses of small-sized protein-DNA gel particles showed to be strongly dependent of both the protein composition and the cell line being the tumour cell line HeLa more sensitive to the deleterious effects of the mixed protein-based particles. The observed trends in haemolysis and cell viabilities were in agreement with the degree of complexation values obtained for the protein-DNA gel particles prepared by both preparation methods.

Cyclodextrin-grafted cellulose: physico-chemical characterization.

Cyclodextrins (CDs) can form inclusion complexes with a wide variety of molecules making them very attractive in different areas, such as pharmaceutics, biochemistry, food chemistry and textile. In this communication we will report on the physico-chemical characterization of cellulose modified with CDs by means of infra-red spectroscopy (FTIR), cross polarization magic angle spinning solid state nuclear magnetic resonance (CP-MAS NMR), polarized optical microscopy (POM) and thermal gravimetric analysis (TGA). Both CP-MAS NMR and FTIR indicate that CDs are chemically attached to cellulose backbone through the formation of ester bonds. Furthermore, the CD-grafted cellulose was dissolved in a "superphosphoric" acid solution but, despite the increase of hydrophilicity due to the modification, POM revealed that grafted cellulose was less soluble when compared to the unmodified polymer. The formation of a complex CD-cellulose network is suggested.

Identification and assessment of kefir yeast potential for sugar/ethanol-resistance

Biochemical and molecular analysis was used for identification of different kefir yeasts species from Brazil, Canada and the United States of America. The sugar/ethanol-resistant activity of the yeasts was evaluated. Saccharomyces cerevisiae and Kluyveromyces marxianus had the highest growth rates, suggesting biotechnological applications possible for these strains.

Shear-induced defect formation in a nonionic lamellar phase.

(2)H NMR experiments on a nonionic oriented lamellar phase demonstrate that shear flow induces structural defects in the lamellar structure. These substantial structural changes give rise to a transition from a viscous to a solidlike behavior; the elastic modulus of presheared samples was found to increase, reversibly, with the applied preshear rate. A similar behavior was found when step-cycling the temperature toward the layer-to-multilamellar-vesicle transition and back at constant shear rate. However, while shear rate controls the defect density, the temperature is found to control the defect rigidity. The lamellar phase exhibits a shear-thinning behavior under steady shear conditions, following the power law eta approximately gamma(n), with n approximately -0.4. Both the shear thinning and the elastic behavior are in agreement with the available theoretical models. The observed shear-induced structural defects are reversible and can be regarded as a pretransition prior to the shear-induced formation of multilamellar vesicles.

Size determination of shear-induced multilamellar vesicles by rheo-NMR spectroscopy.

A model for analyzing the deuterium ((2)H) NMR line shapes of D(2)O in surfactant multilamellar vesicle (MLV, "onion") systems is proposed. The assumption of the slow exchange of water molecules between adjacent layers implies that the (2)H NMR line shape is simply given by a sum of Lorentzians if the condition of motional narrowing is also fulfilled. Using the classical two-step model for the NMR relaxation in structured fluids allows us to calculate how the NMR line shape depends on the MLV size. The model is tested on two different MLV systems for which the NMR line shapes are measured as a function of the applied shear rate using rheo-NMR. The MLV sizes obtained are in good agreement with previous data from rheo-small-angle light scattering.

Portuguese Thymbra and Thymus species volatiles: chemical composition and biological activities.

Thymbra capitata and Thymus species are commonly known in Portugal as thyme and they are currently used as culinary herbs, as well as for ornamental, aromatizing and traditional medicinal purposes. The present work reports on the state of the art on the information available on the taxonomy, ethnobotany, cell and molecular biology of the Portuguese representatives of these genera and on the chemotaxonomy and antibacterial, antifungal and antioxidant activities of their essential oils and other volatile-containing extracts.

Shear-induced transitions between a planar lamellar phase and multilamellar vesicles: continuous versus discontinuous transformation.

The shear-induced transitions between an oriented lamellar phase and shear-induced multilamellar vesicles (MLVs) in a nonionic surfactant system were studied by deuterium rheo-NMR spectroscopy as a function of time in start-up experiments at several temperatures and shear rates. By starting from an initial state of oriented lamellae and observing the transformation to the final steady state of MLVs and vice-versa, two different mechanisms were found, depending on the direction of the transition and the initial state. The transition is continuous when MLVs are formed, starting from the oriented lamellar phase. On the other hand, a discontinuous nucleation-and-growth process with a coexistence region is observed when transforming MLVs into an oriented lamellar phase.

Self-assembly of a hydrophobically modified naphthalene-labeled poly(acrylic acid) polyelectrolyte in water: organic solvent mixtures followed by steady-state and time-resolved fluorescence.

The solution properties of two water-soluble polymers, poly(acrylic acid) (PAA), covalently labeled with the fluorescent hydrophobic dye naphthalene (Np), have been investigated in water:organic solvent mixtures. The naphthalene chromophores have been randomly attached, onto the polymer, with two different degrees of labeling. Fluorescence measurements (steady-state and time-resolved) have been used to follow the photophysical behavior of the polymers and consequently report on the self-association of the polymers in the mixed organic (methanol or dioxane):aqueous solutions. The emission spectra of the high-labeled Np PAA reveal the presence of monomer and excimer bands whereas with the low-labeled polymer only monomer emission is observed. The excitation spectra collected at the monomer and excimer emission bands show significant differences, depending on the water content of the mixture, which indicate the simultaneous presence of preformed and dynamic dimers as routes to excimer formation. The time-resolved data decay profiles of the high-labeled polymer in the mixtures were always triple exponential whereas in pure methanol and dioxane they follow biexponential laws. The data in the mixtures are consistent with two types of monomers and one excimer. Both monomers are able to give rise to excimer in the excited state, one type involving the movement of long distant Np chromophores and the other involving a local reorientation of adjacent Np chromophores. These correspond to different decay times: (1) a long which corresponds to the long distant approach of non-neighboring Np chromophores forming an excimer and (2) a short corresponding to the fast adjustment of two neighboring Np chromophores in order to have the adequate parallel geometry. An additional decay time corresponding to the excimer decay was found to be present at all wavelengths. All the decay times were dependent on the water content of the mixture. An estimation of the two excimer forming rate constants values is made for the mixed media considered in this work. On the whole, using both steady-state and time-resolved fluorescence parameters, and by comparing data for a polymer with a small number of hydrophobes with a more highly modified one, it is possible in great detail to demonstrate how association is controlled by solvent quality for the hydrophobes and by the distance between hydrophobes.

Polyion adsorption onto catanionic surfaces. A Monte Carlo study.

The adsorption of a single and negatively charged polyion with varying flexibility onto a surface carrying both negative and positive charges representing a charged membrane surface has been investigated by using a simple model employing Monte Carlo simulations. The polyion was represented by a sequence of negatively charged hard spheres connected with harmonic bonds. The charged surface groups were also represented by charged hard spheres, and they were positioned on a hard surface slightly protruding into the solution. The surface charges were either frozen in a liquidlike structure or laterally mobile. With a large excess of positive surface charges, the classical picture of a strongly adsorbed polyion with an extended and flat configuration emerged. However, adsorption also appeared at a net neutral surface or at a weakly negatively charged surface, and at these conditions the adsorption was stronger with a flexible polyion as compared to a semiflexible one, two features not appearing in simpler models containing homogeneously charged surfaces. The presence of charged surface patches (frozen surface charges) and the ability of polarization of the surface charges (mobile surface charges) are the main reasons for the enhanced adsorption. The stronger adsorption with the flexible chain is caused by its greater ability to spatially correlate with the surface charges.

Analysis by gas chromatography-mass spectrometry of the volatile components of Teucrium lusitanicum and Teucrium algarbiensis.

The essential oils from four samples of Teucrium lusitanicum and one sample of Teucrium algarbiensis, grown in Algarve (southern Portugal) were analyzed by gas chromatography (GC) and gas chromatography-mass spectroscopy (GC-MS). Seventy-one volatile compounds were identified. Major compounds of T. algarbiensis oil were alpha-pinene (8.3%), sabinene (7.2%), beta-pinene (10.2%), limonene (11.8%) and germacrene D (7.6%). Concerning T. lusitanicum, some quantitative differences were found with regards to the major constituents of the oils from four populations: alpha-pinene (0.8-8.5%), sabinene (2.1-9.6%), beta-pinene (2.5-11.9%), limonene (1.2-11.5%) and elemol (2.6-12.0%).

Antimicrobial activity of essential oils isolated from Portuguese endemic species of Thymus.

AIMS: Thymus species are wild species mostly found in the arid lands of Portugal. Possible antimicrobial properties of Thymus essential oils have been investigated. The chemical composition of the essential oils and the antimicrobial activity of Thymus mastichina (L) L. subsp. mastichina, T. camphoratus and T. lotocephalus from different regions of Portugal were analysed. METHODS AND RESULTS: Hydrodistillation was used to isolate the essential oils and the chemical analyses were performed by gas chromatography (GC) and GC coupled to mass spectrometry. The antimicrobial activity was tested by the disc agar diffusion technique against Candida albicans, Escherichia coli, Listeria monocytogenes, Proteus mirabilis, Salmonella spp. and Staphylococcus aureus. Pure linalool, 1,8-cineole and a mixture (1 : 1) of these compounds were included. Linalool, 1,8-cineole or linalool/1,8-cineole and linalool/1,8-cineole/linalyl acetate were the major components of the essential oils, depending on the species or sampling place. The essential oils isolated from the Thymus species studied demonstrated antimicrobial activity but the micro-organisms tested had significantly different sensitivities. CONCLUSIONS: The antimicrobial activity of essential oils may be related to more than one component. SIGNIFICANCE AND IMPACT OF THE STUDY: Portuguese endemic species of Thymus can be used for essential oil production for food spoilage control, cosmetics and pharmaceutical use. Further studies will be required to elucidate the cell targets of the essential oil components.

Association of surfactants and polymers studied by luminescence techniques.

Light emission spectroscopy has unique possibilities for the study of central issues of surfactants and associating polymers. With the help of luminescent probes, information may be obtained on matters such as molecular association, microstructure, and molecular dynamics; this constitutes an important contribution to the understanding and control of macroscopic properties, as well as biological function and technical applications. Important aspects of these systems considered in this review are: formation of micelles and hydrophobic microdomains; aggregation numbers of surfactants; shape of molecular aggregates; size of droplets in water or in oil in microemulsions; formation and stability of vesicles; intra- vs. intermolecular association in polymers; conformational changes in polymers; polymer-surfactant association; surfactant organization in adsorbed layers; kinetic aspects regarding the formation and disintegration of self-assembly structures; residence times of molecules in microdomains and migration of active molecules.

Applications and limitations of uranyl ion as a photophysical probe.

Uranyl ion has a long-lived luminescent excited state, which can be used as a probe for the aggregation behaviour in a variety of surfactant and polymeric systems. The general spectroscopy and photophysics of this species are discussed, and the applications to specific micellar, microemulsion, vesicular and liquid crystalline systems presented. It is shown that both dynamic and structural information can be obtained from spectral and kinetic data. Examples of applications of uranyl probe studies to technologically important mesoscopic and mesoporous systems involving metal oxides, or their salts, and to humic acids and soils will also be reviewed. Emphasis will be given to both the advantages and restrictions on the use of this useful photophysical probe.

DNA conformational dynamics in the presence of catanionic mixtures.

DNA conformational behavior in the presence of non-stoichiometric mixtures of two oppositely charged surfactants, cetyltrimethylammonium bromide and sodium octyl sulfate, was directly visualized in an aqueous solution with the use of a fluorescence microscopy technique. It was found that in the presence of cationic-rich catanionic mixtures, DNA molecules exhibit a conformational transition from elongated coil to compact globule states. Moreover, if the catanionic mixtures form positively charged vesicles, DNA is adsorbed onto the surface of the vesicles in a collapsed globular form. When anionic-rich catanionic mixtures are present in the solution, no change in the DNA conformational behavior was detected. Cryogenic transmission electron microscopy, as well as measurements of translational diffusion coefficients of individual DNA chains, supported our optical microscopy observations.

[Ovarian cystadenofibroma. 5-year experience at the "Dr. Manuel Gea González" General Hospital S.S.A. Report of 10 cases]. / Cistoadenofibroma de ovario. Experiencia de cinco años en el Hospital General "Dr. Manuel Gea González" S.S.A. Informe de 10 casos.

The ovarian cystoadenofibroma are tumors not very frequently of superficial epithelium. It can appears at all ages. It can form fibrotic nodules of strong consistency that initially appear with the papillae and it forms nodule when the time goes by; generally, they are unilateral. We studied 10 cases of cystoadenofibromas in a period of 5 years, the patients' age was from 18 to 68 years old being more frequent in the adult age; seven cases were unilateral, three were bilateral, two of these appeared clinically as acute abdomen due to rupture of the tumor the third one was a hysterectomy is finding. In seven cases there was association with uterine leiomiomatosis mainly in older patients. Macroscopically it was seen that in younger patients these tumors were cystic with small papillae, and in older patients they formed fibrotic nodules. The immunohystochemical study with Vimentin and Desmin showed that the Desmin was focally immunopositive in form of fine fibers at the union with superficial epithelium and greatly positive fibrous portion for Vimentina.