Stabilization and activation of {alpha}-chymotrypsin in water-organic solvent systems by complex formation with oligoamines

Formation of enzyme–oligoamine complexes was suggestedas an approach to obtain biocatalysts with enhanced resistancetowards inactivation in water–organic media. Complex formationresults in broadening (by 20–40% v/v ethanol) of the rangeof cosolvent concentrations where the enzyme retains its catalyticactivity (stabilization effect). At moderate cosolvent concentrations(20–40% v/v) complex formation activates the enzyme (by3–6 times). The magnitude of activation and stabilizationeffects increases with the number of possible electrostaticcontacts between the protein surface and the molecules of oligoamines(OA). Circular dichroism spectra in the far-UV region show thatcomplex formation stabilizes protein conformation and preventsaggregation in water–organic solvent mixtures. Two populationsof the complexes with different thermodynamic stabilities werefound in -chymotrypsin (CT)–OA systems depending on theCT/OA ratio. The average dissociation constants and stoichiometriesof both low- and high-affinity populations of the complexeswere estimated. It appears that it is the low-affinity siteson the CT surface that are responsible for the activation effect.     

Design criteria and optical characteristics of one-dimensional photonic crystals based on periodically grooved silicon

Photonic bandgap (PBG) regions have been calculated for periodically grooved Si structures, acting as a one-dimensional photonic crystal. The wavelength range of the PBG as a function of the ratio (DSi/A) is presented, where DSi is the width of the Si walls and A is the grooved silicon lattice constant. The influence of the parameter DSi, the refractive index of the space between the Si walls and the number of structure periods, m, on the forming of PBG regions is discussed. A good correlation between the calculated and the experimentally observed PBG regions is obtained.     

Synergistic pseudo-hydroxide extraction: synergism and anion selectivity in sodium extraction using a crown ether and a series of weak lipophilic acids

The nature of the weak lipophilic acid used in synergistic combination with a model crown ether cation host was shown to have a strong effect on the strength and selectivity of sodium hydroxide separation from alkaline aqueous salt solutions. Sodium ion-pair extraction employing only cis-syn-cis-dicyclohexano-18-crown-6 (1) in nitrobenzene (NB) was correlated with the standard Gibbs energy (deltaG(p)o) of anion partitioning into NB and was notably weak and nonselective for the hydroxide ion, in accord with Hofmeister bias. The Hofmeister order can be selectively overcome for NaOH by utilization of acid-base chemistry coupled with complexation of sodium ion in the NB phase. Upon addition of a lipophilic organic acid into the solution of 1 in NB, sodium extraction was selectively enhanced due to the initiation of an exchange reaction between the aqueous sodium ion and the ionizable proton of the organic acid. A series of weak lipophilic hydroxy acids (HA) including fluorinated alcohols and phenols was tested. The resulting synergistic pseudo-hydroxide extraction correlates with the pKa of the employed HA; the most acidic cation exchangers provide the greatest synergism. The synergistic factor obtained using a fluorinated benzyl alcohol 7 was as high as 256. Ion-pair extraction of neutral sodium salts was not changed or only mildly enhanced by addition of HA into the NB solution of 1. This enhancement was explained by hydrogen bonding of HA with the anion as related to the hardness of the anion and the acidity of HA. In comparison with the synergism observed for NaOH, this enhancement was weak and unable to overcome the Hofmeister effect. Examination of extraction selectivity revealed that the combination of 1 and 7 preferentially extracted NaOH over all other sodium salts, including the normally preferred nitrate and perchlorate salts. Quantitative recovery of NaOH from the NB phase was demonstrated via hydrolysis of the organic acid upon a single contact of the loaded solvent with water.     

Production and characterization of pure and Cr<Superscript>3+</Superscript>-doped hydroxyapatite for biomedical applications as fluorescent probes

Pure and Cr³⁺-doped hydroxyapatite (HAP) were prepared via chemical precipitation route. The XRD measurements revealed that the typical HAP powder pattern was obtained. SEM analysis indicated that aggregates of nanoparticles were formed. EDX analysis indicates that the [Ca]/[P] concentration ratio was higher than the expected values but can be explained by the presence of carbonate groups as dopants. The optical absorption spectra of the doped samples presented absorption bands typical of Cr³⁺ occupying to different crystalline sites. From the position of the bands, it was possible to estimate the crystal field parameters for both sites of Cr³⁺ in the HAP matrix. The emission spectra of the Cr-doped samples were also investigated and typical transitions of the dopant ion, in trivalent state, were identified. The potential use of the Cr³⁺-HAP as fluorescent probes for medical applications was discussed.     

Variability of solar UV-B irradiance: in situ monitoring and model calculation of the vitamin D synthetic capacity of sunlight

The prediction of the biological effects of solar radiation on human health, in particular involving terrestrial level of solar ultraviolet (UV) radiation, requires the development of an adequate methodological strategy for remote sensing. To date, it is well understood that, in appropriate doses, UV radiation is beneficial for people, specifically due to the production of vitamin D₃ in the skin from its precursor 7-dehydrocholesterol. But as far as excessive UV exposure causes acute and chronic health effects, in most cases biological activity of solar UV radiation is calculated by weighting solar UV spectra with International Commission on Illumination (CIE) erythemal action spectrum. Yet the beneficial vitamin D synthetic capacity of sunlight cannot be correctly estimated in this way because of the significant difference between the erythemic and vitamin D synthesis action spectra. With due regard to the essential role of vitamin D₃ for human health, in this article we examine the possibility of simplified estimation in situ of provitamin D₃ photoconversion into previtamin D₃ from the UV absorption spectra of an in vitro model of vitamin D synthesis upon exposure to sunlight. A large-scale linear correlation (R = 0.99) was found on a clear summer day between the concentration of accumulated previtamin D₃ and maximum absorbance decline in the initial provitamin D₃ absorption spectrum. However, long-term observations showed a poorer (R = 0.77) correlation, and a source of ambiguity of such indirect estimation of previtamin D₃ concentration is discussed in detail. In addition, we propose a reliable algorithm for the direct calculation of previtamin D₃ accumulation using solar UV spectra as input data to the reaction model of previtamin D photosynthesis and demonstrate the critical dependence of previtamin D₃ accumulation on stratospheric ozone, season, latitude and cloudiness. The comparison of experimental and simulation data conforms to recent findings on Europe's darker atmosphere in the UV-B and implicates the practical certainty of the presented algorithm for the calculation of the vitamin D synthetic capacity from the remotely sensed solar spectra.     

Relaxation Properties of Pressure-sensitive Adhesives upon Withdrawal of Bonding Pressure

Relaxation properties of pressure-sensitive adhesives (PSA) have been studied with the squeeze-recoil tester used in the regime of parallel-plate dilatometer under conditions imitating the removal of compressive force in the course of adhesive bond formation. The relaxation properties of PSAs are compared with their adhesive behavior measured using the 180-Deg Peel Test. Two classes of PSAs are considered: 1) conventional rubbery adhesives based on the mixtures of styrene-isoprene-styrene (SIS) block copolymer with a tackifier resin and a plasticizer, and butyl rubber plasticized with low-molecular-weight polyisobutylene, and 2) hydrophilic PSAs composed of the blends of high-molecular-weight poly(N-vinyl pyrrolidone) (PVP) with oligomeric polyethylene glycol (PEG). By comparing the adhesive and relaxation behaviors of different PSAs, the relaxation criteria for pressure-sensitive adhesion have been stated. Relaxation behavior of the examined PSAs demonstrates two values of retardation time: the shorter retardation time of 10–70?sec and the longer time of 300–660?sec. These times can be associated, respectively, with small- and large-scale mechanisms of strain recovery. By comparing the relaxation and adhesive properties of PVP-PEG blend (which involves the formation of a hydrogen-bonded network through both terminal hydroxyl groups in PEG short chains) with the properties of covalently crosslinked copolymers of vinyl pyrrolidone (VP) with PEG-diacrylate and comb-like VP copolymers with PEG-monomethacrylate, the contributions of covalent crosslinking and H-bonding network have been characterized.     

Defensive Compounds and Male-Produced Sex Pheromone of the Stink Bug, Agroecus griseus

Agroecus griseus is a serious corn pest in Brazil. Contents of the dorsal abdominal glands (DAGs) in nymphs, and the metathoracic gland (MTG) in adults of this species were characterized and quantified. Compounds found were similar to those of other Pentatomidae species and included aliphatic hydrocarbons, aldehydes, oxo-alkenals, and esters. However, two compounds were found in the MTG that have not been described previously for this family. Mass spectrometry, infrared spectroscopy, and gas chromatographic analysis using coinjection with authentic standards confirmed the identities of the compounds as enantiopure (S)-2-methylbutyl acetate and 3-methyl-2-butenyl acetate. The five nymphal instars showed significantly different ratios of components, mainly between those of the first and later instars. No significant differences were detected in MTG contents between sexes. Gas chromatography (GC) analysis of aeration extracts of males and females showed the presence of a compound released exclusively by males. Gas chromatography – electroantennographic detection (GC-EAD) assays indicated that the male-specific compound is bioactive in females, suggesting the presence of an attractant pheromone. The mass spectrum and infrared data for this compound matched with methyl 2,6,10-trimethyltridecanoate, a sex pheromone component previously detected in the stink bugs, Euschistus heros and E. obscurus. The synthetic standard coeluted with the natural pheromone on two different GC stationary phases, confirming the proposed structure. Y-tube olfactometer assays showed that the synthetic standard was strongly attractive to females, and GC-EAD tests produced responses with antennae from females similar to those of the natural pheromone.