Advanced Sunscreens: UV Absorbers Encapsulated in Sol-Gel Glass Microcapsules

Undesirable phototoxic and photoallergic reactions accompanying a justified increased use of sunscreen active ingredients within cosmetic products have encouraged the development of new products safer for human use. The sol-gel microencapsulation technology developed utilizes an interfacial polymerization process, allowing for the achievement of transparent silica glass microcapsules with sizes ranging between 0.3–3 microns and a characteristic core-shell structure. Within the sol-gel microcapsule structure a UV absorber core, constituting roughly 80% of the final product weight, is enclosed within a silica shell. These advanced sunscreen actives are then incorporated into a suitable cosmetic vehicle to achieve high Sun Protection Factors (SPF), while affording an improved safety profile, as the penetration of the UV absorbers is markedly reduced.     

Protein Stability in Reverse Micelles

The N‐terminal SH3 domain of the Drosophila signal transduction protein drk was encapsulated in reverse micelles. Both the temperature of maximum stability and the melting temperature decreased on encapsulation. Dissecting the temperature‐dependent stability into enthalpic and entropic contributions reveals a stabilizing enthalpic and a destabilizing entropic contribution. These results do not match the expectations of hard‐core excluded volume theory, nor can they be wholly explained by interactions between the head groups in the reverse micelle and the test protein. We suggest that geometric constraints imposed by the reverse micelles need to be considered.     

Denaturation and Leaching Study of Horseradish Peroxidase Encapsulated in Sol-Gel Matrices

Sol-gel matrices have been shown to be relatively inert while preserving the spectroscopic properties and biological activity of the encapsulated proteins. Horseradish peroxidase (HRP) is a hemeprotein widely used in the field of biosensors because of its high specificity for hydrogen peroxide. However, partial inactivation of the protein has been reported when incorporated in aged gels. Whether that inactivation comes from the unfolding of some of the encapsulated proteins or from the leaching of the heme non-covalent active site of HRP is evaluated by absorption and fluorescence spectroscopy. This study shows that the single Trytophan (Trp) fluorescence of HRP may be used to distinguish denaturation processes from leaching of the heme group, as well as to estimate the extent of the denaturation.     

Copper-Silica Sol-Gel Catalysts: Structural Changes of Cu Species upon Thermal Treatment

In this work the encapsulation of copper species in sol-gel silica catalysts thermally treated up to 1100°C was studied. XRD, TPR, BET data showed the occurrence of a vitrification/densification process upon treatment at temperatures higher than 900°C leading to the partial or complete encapsulation of the copper species by the silica matrix. As result of this process the copper catalyst particles become unavailable for the reaction with gas phase molecules and are not active for reactions such as carbon monoxide oxidation.     

ESR Study of a Nitroxide Radical in Sol-Gel Glasses

A spin probe TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidinyl-1-oxy) was dissolved in a tetraethyl orthosilicate sol-gel reaction system and measured by electron spin resonance spectroscopy at 295 K. The nitrogen hyperfine coupling constant was from 1.64–1.66 mT in the sol-gel solutions. The values were sensitive to the ethanol-to-water ratio of the solutions. The hyperfine coupling constant in the xerogels was 1.70 mT, which was almost the same as that in water, indicating that the probe molecules were trapped in silica pores with water adsorbed on the silica surfaces. The motion of TEMPOL in the xerogels was considerably slower than in the sol-gel solutions. The local viscosity estimated was from 70–90 cP. The ESR spectra of TEMPOL were altered during the sol-gel process, indicating that adsorbed water on the silicas surfaces has an important role for trapping organic molecules in sol-gel glasses.     

Proliferation and Insulin Secretion Function of Mouse Insulinoma Cells Encapsulated in Alginate/Sol-Gel Synthesized Aminopropyl-Silicate/Alginate Microcapsule

Alginate/aminopropyl-silicate/alginate microcapsules, ca. 15 m in membrane thickness and ca. 500 m in diameter, were prepared via sol-gel process. The aminopropyl-silicate membrane was derived from two silicone alkoxide precursors, tetramethoxysilane and 3-aminopropyl-trimethoxysilane on Ca-alginate micro gel beads. Pancreatic -cell line (MIN6) cells were encapsulated in the microcapsule. The encapsulated MIN6 cells proliferated and formed spheroidal tissues in vitro. The diameter of the MIN6 spheroids increased to approximately 250 m with an increase in the incubation period until the day 35. Storeptozotocin-induced diabetic mice became normoglycemia after implantation of the MIN6-enclosing microcapsules. The normoglycemic state remained until the retrieval of the implanted microcapsules for 1 month. These results indicate that the potential use of the alginate/aminopropyl-silicate/alginate microcapsule as a vehicle for a genetically engineered cell-enclosing therapeutic material delivery system.     

Encapsulation of Proteins in Bulk and Thin Film Sol-Gel Matrices

This paper considers the nature of the interactions between the sol-gel derived inorganic matrix and a specific biomolecule, cytochrome c. Optical absorption and impedance spectroscopies are used to characterize the influence of synthesis conditions on the protein’s stability and conformation within the silica matrix. In some instances, encapsulation within the sol-gel matrix provides stabilization. For example, protein denaturation is reversible and aggregation is prevented. Moreover, the drying process does not negatively affect the protein; it is possible to regenerate the aged gel state by rehydration. The flexibility of the sol-gel process enables high quality cytochrome c-doped SiO₂ thin films to be prepared. These films possess the characteristic reactivity and chemical function of cytochrome c in solution.     

荧光光谱仪在大分子研究中的应用

荧光光谱仪可定性、定量测定许多有机大分子和生物大分子,广泛应用于生物大分子、表面活性剂/大分子相互作用、高分子溶液相行为等方面的研究中,具有测定方法高效、灵敏、精准、简单易操作等优点,是大分子研究中重要的工具。     

Fast Biosynthesis of GFP Molecules: A Single‐Molecule Fluorescence Study

It's not easy being green : Real‐time visualization of labeled ribosomes and de novo synthesized green fluorescent protein molecules using single‐molecule‐sensitive fluorescence microscopy demonstrates that the mutant GFPem is produced with a characteristic time of five minutes. Fluorescence of the fastest GFP molecules appears within one minute (see picture).

     

Complexation and Fluorescence of Tricyclic Basic Dyes Encapsulated in Cucurbiturils

Tricyclic basic dyes (proflavine, acridine orange, pyronine, pyronine Y, oxonine, thionine and methylene blue) often form one‐to‐one or two‐to‐one complexes with CB[7] and CB[8], respectively. In the case of pyronine Y, the complexes with CB[7] and CB[8] have a one‐to‐one and three‐to‐one stoichiometry, respectively. The binding constants for CB[7] complexes range from 3.07×10⁶ to 1.70×10⁷ m ^?1. In the case of CB[8], the association constant varies between 3.24×10¹³ and 2.50×10¹⁶ m ^?2. Overall, these binding constants are four orders of magnitude higher than those reported for the same dyes in β and γ‐cyclodextrins. Formation of the host–guest complexes leads to an increase in the fluorescence quantum yields in the case of CB[7], while the dimeric or trimeric dye encapsulated in CB[8] are remarkably less fluorescent than the same dye in diluted solutions.     

Influence of Temperature and Time on the Stability of Silver in Silica Sol-Gel Glasses

The darkening of silica sol-gel glasses doped with 0.05 mol% silver was studied. Six sols were prepared from TEOS and silver nitrate. Different additives were used, to influence the chemical and physical states of silver: oxidizing or reducing agents (H₂O₂, As₂O₅), colloid stabilizer (sodium citrate) and network modifiers (Li₂O, CaO). Sols were gelified at 60°C and densified at 600°C. The samples without additives and those prepared with H₂O₂ at room temperature even if they were protected from light. With increased temperature, the darkening became samples were heated above, 400°C, reversible bleaching took place. This darkening-bleaching is of thermal nature (“thermochromic effect”) and seems to be determined by a reversible aggregation-disaggregation of tiny silver particles. The presence of sodium citrate, as an additive delayed the darkening effect and the presence of CaO delayed it even further. Lithium oxide inhibited it totally.     

Quantification of Structural Integrity and Stability Using Nanograms of Protein by Flow-Induced Dispersion Analysis

In the development of therapeutic proteins, analytical assessment of structural stability and integrity constitutes an important activity, as protein stability and integrity influence drug efficacy, and ultimately patient safety. Existing analytical methodologies solely rely on relative changes in optical properties such as fluorescence or scattering upon thermal or chemical perturbation. Here, we present an absolute analytical method for assessing protein stability, structure, and unfolding utilizing Taylor dispersion analysis (TDA) and LED-UV fluorescence detection. The developed TDA method measures the change in size (hydrodynamic radius) and intrinsic fluorescence of a protein during in-line denaturation with guanidinium hydrochloride (GuHCl). The conformational stability of the therapeutic antibody adalimumab and human serum albumin were characterized as a function of pH. The simple workflow and low sample consumption (40 ng protein per data point) of the methodology make it ideal for assessing protein characteristics related to stability in early drug development or when having a scarce amount of sample available.     

化学气相沉积合成碳包裹镍纳米晶的初步研究

Nickel nanocrystals encapsulated in carbon shells were prepared by the large body knowledge developed for the growth of carbon nanotubes (CNTs), i.e. chemical vapor deposition. The products were characterized by transmission electron microscopy, XRD and FTIR. The results showed that the oxidization of CNT surface made it possible to interreact with nickel ions in solution. Ni-impregnated CNTs transformed into monocrystalline nickel nanoparticles supported on CNTs at 600 ℃ in nitrogen atmosphere. Subsequently, they would be covered with graphene layers during reaction with acetylene at 600 ℃. The formation mechanism has been preliminarily discussed on experimental results.     

Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions

The intracellular environment is overcrowded with a range of molecules (small and large), all of which influence protein conformation. As a result, understanding how proteins fold and stay functional in such crowded conditions is essential. Several in vitro experiments have looked into the effects of macromolecular crowding on different proteins. However, there are hardly any reports regarding small molecular crowders used alone and in mixtures to observe their effects on the structure and stability of the proteins, which mimics of the cellular conditions. Here we investigate the effect of different mixtures of crowders, ethylene glycol (EG) and its polymer polyethylene glycol (PEG 400 Da) on the structural and thermal stability of myoglobin (Mb). Our results show that monomer (EG) has no significant effect on the structure of Mb, while the polymer disrupts its structure and decreases its stability. Conversely, the additive effect of crowders showed structural refolding of the protein to some extent. Moreover, the calorimetric binding studies of the protein showed very weak interactions with the mixture of crowders. Usually, we can assume that soft interactions induce structural perturbations while exclusion volume effects stabilize the protein structure; therefore, we hypothesize that under in vivo crowded conditions, both phenomena occur and maintain the stability and function of proteins.     

How Post‐Translational Modifications Influence Amyloid Formation: A Systematic Study of Phosphorylation and Glycosylation in Model Peptides

A reciprocal relationship between phosphorylation and O‐glycosylation has been reported for many cellular processes and human diseases. The accumulated evidence points to the significant role these post‐translational modifications play in aggregation and fibril formation. Simplified peptide model systems provide a means for investigating the molecular changes associated with protein aggregation. In this study, by using an amyloid‐forming model peptide, we show that phosphorylation and glycosylation can affect folding and aggregation kinetics differently. Incorporation of phosphoserines, regardless of their quantity and position, turned out to be most efficient in preventing amyloid formation, whereas O‐glycosylation has a more subtle effect. The introduction of a single β‐galactose does not change the folding behavior of the model peptide, but does alter the aggregation kinetics in a site‐specific manner. The presence of multiple galactose residues has an effect similar to that of phosphorylation.     

Retention of Native Protein Structures in the Absence of Solvent: A Coupled Ion Mobility and Spectroscopic Study

Can the structures of small to medium‐sized proteins be conserved after transfer from the solution phase to the gas phase? A large number of studies have been devoted to this topic, however the answer has not been unambiguously determined to date. A clarification of this problem is important since it would allow very sensitive native mass spectrometry techniques to be used to address problems relevant to structural biology. A combination of ion‐mobility mass spectrometry with infrared spectroscopy was used to investigate the secondary and tertiary structure of proteins carefully transferred from solution to the gas phase. The two proteins investigated are myoglobin and β‐lactoglobulin, which are prototypical examples of helical and β‐sheet proteins, respectively. The results show that for low charge states under gentle conditions, aspects of the native secondary and tertiary structure can be conserved.