Characterization of the fruit proteolytic system of Bromelia serra Griseb. (Bromeliaceae) and its application in bioactive peptides release.

A crude extract with proteolytic activity was prepared from edible fruits of Bromelia serra, containing cysteine peptidases with molecular masses between 24.1 and 25.9 kDa. The extract presented an optimal pH range of 6.03-9.05, retained more than 80% of activity after thermal pre-treatments at 23, 37, and 45°C (120 min), but it was rapidly inactivated after 10 min at 75°C. These proteases were employed to hydrolyze soybean proteins, bovine casein and bovine whey, achieving degrees of hydrolysis of 18.3 ± 0.6, 29.1 ± 0.7, and 12.6 ± 0.9% (55°C, 180 min), respectively. The casein 180 min-hydrolysate (55°C) presented the maximum value of antioxidant activity (2.89 ± 0.12 mg/mL Trolox), and the whey protein 180 min-hydrolysate (55°C) showed the highest percentage of angiotensin-converting enzyme inhibition (91.9 ± 1.2%). This low-cost enzymatic preparation would be promising for the food industry because it requires mild working conditions and yields hydrolysates with biological activities useful as ingredients for functional food. PRACTICAL APPLICATION: Proteolytic enzymes are employed in the food industry in a wide variety of processes since they modify the properties of proteins causing beneficial effects such as improvement digestibility, diminution of allergenicity, and release of bioactive peptides. Fruits from Bromelia serra possess cysteine peptidases that could be used in food biotechnology because they are capable to hydrolyze soybean and milk proteins by mild working conditions and to provoke the release of bioactive peptides. These hydrolysates containing antioxidative and ACE-inhibitor activities would be useful as ingredients for functional foods or as nutraceuticals, which are nowadays two products highly required by consumers.

Bromide counterion as a spectroscopic sensor at the interface of cetyltrimethylammonium micelles.

The strong UV absorption of the bromide in aqueous solution undergoes a remarkable red shift of more than 10 nm induced by the addition of the salts that constitute a saline buffer. The maximum absorption wavelength of the bromide is displaced from approximately 194 nm in ultrapure water to wavelengths above 200 nm, depending on the composition of the solution. The bromide spectrum as counterion of the cetyltrimethylammonium in the surfactant CTAB also shows sensitivity to the aggregation behavior of the tensioactive, being able to detect intermolecular interactions even at concentrations lower than the critical micelle concentration. And, when the micelles are assembled, the bromide absorption detects the interfacial rearrangements caused by the incorporation of ions. To know more about those interfacial features, the pyrene molecular probe was used, taking advantage of the extensive knowledge of its spectroscopy. Pyrene verifies the existence of changes in the interfacial organization which confirm that the sensitivity of the bromide spectrum is based on the ability of the ion to detect its microenvironment, and therefore reaffirms that its absorption spectrum can be used as a local sensor. The present work encourages the use of bromide as a sensor ion in the UV region between 190 and 210 nm, which would avoid the introduction of external molecular probes that could disturb the system.

Host-guest molecular interactions in vanillin/amylose inclusion complexes.

The interaction of 4-hydroxy-3-methoxybenzaldehyde (vanillin) and Hylon VII due to the formation of an inclusion complex is studied using Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), and circular dichroism (CD). The results confirm the close interaction among the different functional groups of vanillin and its host. In addition, a second case study was carried out with an amylose from a different source (100% amylose [APT III]). As a result, remarkable differences were found in the vanillin complexation capability of this amylose, which is only shown in solution by circular dichroism spectroscopy studies through a clear Cotton effect. This finding confirms the value of using CD studies, which shows that, depending on the amylose source, inclusion complexes can be found in solution, or both in solution and the coexisting precipitates, as shown using other techniques, such as X-ray diffraction (XRD) or differential scanning calorimetry (DSC). Moreover, solubility assays and complexation of both starches with iodine and subsequent absorption spectroscopy studies gives more information regarding the possible source of the starch encapsulation capability. Thus, Hylon VII shows higher capacity as vanillin encapsulant than APT III, showing the formation of inclusion complexes both in solution and solid phase, whereas APT III complexes are only perceivable in solution.

Customized design of electronic noses placed on top of air-lift bioreactors for in situ monitoring the off-gas patterns.

A specially designed electronic nose was coupled to an air-lift bioreactor in order to perform on-line monitoring of released vapors. The sensor array was placed at the top of the bioreactor sensing the headspace in equilibrium with the evolving liquor at any time without the need of aspiration and pumping of gases into a separated sensor chamber. The device was applied to follow the off-gas of a bioreactor with Acidithiobacillus thiooxidans grown on beds of elemental sulfur under aerobic conditions. Evolution was monitored by acid titration, pH and optical density measurements. The electronic nose was capable to differentiate each day of reactor evolution since inoculation within periods marked off culture medium replacements using multivariate data analysis. Excellent discrimination was obtained indicating the potentiality for on-line monitoring in non-perturbed bioreactors. The prospects for electronic nose/bioreactor merging are valuable for whatever the bacterial strain or consortium used in terms of scent markers to monitor biochemical processes.

Violet ZnSe/ZnS as an alternative to green CdSe/ZnS in nanocrystal-fluorescent protein FRET systems.

Fluorescent proteins from the green fluorescent protein family strongly interact with CdSe/ZnS and ZnSe/ZnS nanocrystals at neutral pH. Green emitting CdSe/ZnS nanocrystals and red emitting fluorescent protein dTomato constitute a 72% efficiency FRET system with the largest alteration of the overall photoluminescence profile, following complex formation, observed so far. The substitution of ZnSe/ZnS for CdSe/ZnS nanocrystals as energy donors enabled the use of a green fluorescent protein, GFP5, as energy acceptor. Violet emitting ZnSe/ZnS nanocrystals and green GFP5 constitute a system with 43% FRET efficiency and an unusually strong sensitized emission. ZnSe/ZnS-GFP5 provides a cadmium-free, high-contrast FRET system that covers only the high-energy part of the visible spectrum, leaving room for simultaneous use of the yellow and red color channels. Anisotropic fluorescence measurements confirmed the depolarization of GFP5 sensitized emission.

Evaluation of the molecular polarizability using the IPPP-CLOPPA-INDO/S method. Application to molecules of biological interest.

The IPPP-CLOPPA-INDO/S method is introduced to investigate the static molecular polarizability in macromolecules. As an example of application, the polarizability of phospholipidic compounds, with and without the presence of water molecules has been estimated. The IPPP technique was employed to calculate the polarizability of the polar head and the hydrocarbon chains separately to analyze the feasibility of evaluating the total polarizability of the molecule by addition of these two projected results. INDO/S dipole moments of different fragments of the complex molecule were obtained by means of localized molecular orbitals in order to evaluate the charge transfer in the system.

Silicon based materials for drug delivery devices and implants.

This patent review focuses on silicon based materials for drug delivery systems and implant devices devoted to medical applications. The article describes some representative examples of the most depictive silicon based compounds associated with drug release formulations and tissue engineering biomaterials. Ranging from inorganic to organic and hybrid inorganic-organic silicon compounds, the paper referrers to patents describing inventions which make use of the best properties of silicon dioxide, silica aerogel and xerogel, silicon bioactive materials, silicones and ormosils, pointing out the usefulness of each kind of compound within the invention embodiment.

Electronic nose screening of ethanol release during sol-gel encapsulation. A novel non-invasive method to test silica polymerisation.

Porous silica matrices prepared by sol-gel process yield biocompatible materials adequate for encapsulation of biomolecules or drugs. The procedure is simple and fast, but when alkoxyde precursors like tetraethoxysilane (TEOS) are used the polymerisation reaction leads to the formation of alcohol as a by-product, which can produce undesirable effects on the activity of entrapped enzymes or modify a drug release kinetic. Therefore, it is critical to determine that no remnant ethanol is left prior using or storing the obtained biomaterial. In this regard, the technique used in the alcohol determination should be non-invasive and non-destructive to preserve the encapsulation device intact and ready to use. In this work we have successfully used a portable electronic nose (e-nose) for the screening of silica polymerisation process during theophylline encapsulation. TEOS reaction was "smelt" since precursor pre-hydrolysis until the end of ethanol release, sensed directly at the headspace of matrices slabs. Measurements showed that ethanol was negligible since 10th day in polymeric slabs of 10 mm width and 2 cm diameter. This first use of e-nose following a polymerisation reaction opens a wide number of putative applications in pharmaceutical and biochemical fields.

Electronic nose screening of limonene release from multicomponent essential oils encapsulated in pectin gels.

Multicomponent essential oils Tagetes Minuta and Poleo as well as pure limonene were encapsulated in Tween doped-high methoxylated pectin gels. Optical microscopy reveals that the obtained gels containing limonene consisted in a highly heterogeneous oil-in-water emulsion stabilised by the gelled medium. The influence of limonene encapsulation in pectin gelation kinetics and the gel structural properties were followed by dynamic rheological measurements. An electronic nose device developed in our laboratory was used to follow the flavour release of the three systems in order to discriminate the samples according to the main components released to the headspace. PCA and Neural Network Analysis allowed us to discriminate Tagetes Minuta from Poleo due to the difference in their limonene content. It is remarkable that the fingerprints of encapsulated complex mixtures differ from those obtained for the non-encapsulated oils, showing a preferential release of some components. In the case of limonene, the effect of the encapsulated concentration on the detected odour was also studied.

Dynamic rheological measurements and drug release kinetics in swollen scleroglucan matrices.

The structure of scleroglucan gel matrices was characterized by dynamic rheological studies. The results were compared with the release kinetics of theophylline in analogous samples using a Franz diffusion cell, fitting the drug release data with a semi-empirical power law. Dynamic rheology gave information about the viscous and elastic components (loss and storage moduli, respectively) of the gel which could influence the drug-release profiles. Scleroglucan gels showed two structural transitions within the gel regime that coincided with changes in the release pattern. It was found that the introduction of 0.4% (w/w) of theophylline decreased the loss and storage moduli in the 2% (w/w) scleroglucan gels by 50%. The influence of the same wt.% theophylline in other gels was strongly dependent on the gel concentration. These results demonstrated the value of rheological studies to detect matrix structural changes produced by the inclusion of drugs which may modify the drug-release profile.

Correlation between gel structural properties and drug release pattern in scleroglucan matrices.

The drug delivery behavior of scleroglucan hydrogels was correlated with the structural features found by means of steady-state fluorescence studies. Theophylline at 0.4% in weight was used as the model drug and its release was measured using a Franz cell device. The results were fitted with a semi-empirical power law equation, finding significant differences between the 0.5% and higher than 0.8% scleroglucan concentrations (w/w). Four molecular fluorescent probes were used to test microenvironmental properties of the gel cavities, which will play a key role on the release of the pharmaceutical drugs. The results found by the fluorescence analysis are in good agreement with the ones obtained by release kinetics.