The role of deep traps in luminescence of anion-defective α-Al2O3: C crystals

This paper presents the results of a study of deep traps responsible for thermally stimulated luminescence in single crystals of the anion-defective corundum α-Al₂O₃: C observed at temperatures of about 703, 778, 830, and 903 K. The effect of the filling of these traps by carriers of both signs on the sensitivity to radiation of the main thermoluminescence peak at 450 K, optically stimulated luminescence, and phototransferred thermoluminescence has been investigated. The results obtained are treated in terms of a model involving interactive competition of traps of different depth.     

The role of basic amino acids in the molecular recognition of hydroxyapatite by statherin using solid state NMR

Organisms use proteins such as statherin to control the growth of hydroxyapatite (HAP), which is the principal component of teeth and bones. Though much emphasis has been placed on the acidic character of these proteins, the role of their basic amino acids is not well understood. In this work, solid state nuclear magnetic resonance was used to probe the interaction of the basic arginine side chains with the HAP surface. Statherin samples were individually labeled at each arginine site, and the distance to the surface was measured using the Rotational Echo DOuble Resonance (REDOR) technique. The results indicate a strong coupling between the R9 and R10 residues and the phosphorus atoms on the surface, with internuclear distances of 4.62 ± 0.29 Å and 4.53 ± 0.16 Å, respectively. Conversely, results also indicate weak coupling between R13 and the surface, suggesting this residue is more removed from the surface than R9 and R10. Combining these results with previous data, a new model for the molecular recognition of HAP by statherin is constructed.     

When is Mass Spectrometry Combined with Affinity Approaches Essential? A Case Study of Tyrosine Nitration in Proteins

Tyrosine nitration in proteins occurs under physiologic conditions and is increased at disease conditions associated with oxidative stress, such as inflammation and Alzheimer??s disease. Identification and quantification of tyrosine-nitrations are crucial for understanding nitration mechanism(s) and their functional consequences. Mass spectrometry (MS) is best suited to identify nitration sites, but is hampered by low stabilities and modification levels and possible structural changes induced by nitration. In this insight, we discuss methods for identifying and quantifying nitration sites by proteolytic affinity extraction using nitrotyrosine (NT)-specific antibodies, in combination with electrospray-MS. The efficiency of this approach is illustrated by identification of specific nitration sites in two proteins in eosinophil granules from several biological samples, eosinophil-cationic protein (ECP) and eosinophil-derived neurotoxin (EDN). Affinity extraction combined with Edman sequencing enabled the quantification of nitration levels, which were found to be 8?% and 15?% for ECP and EDN, respectively. Structure modeling utilizing available crystal structures and affinity studies using synthetic NT-peptides suggest a tyrosine nitration sequence motif comprising positively charged residues in the vicinity of the NT- residue, located at specific surface- accessible sites of the protein structure. Affinities of Tyr-nitrated peptides from ECP and EDN to NT-antibodies, determined by online bioaffinity- MS, provided nanomolar K_D values. In contrast, false-positive identifications of nitrations were obtained in proteins from cystic fibrosis patients upon using NT-specific antibodies, and were shown to be hydroxy-tyrosine modifications. These results demonstrate affinity- mass spectrometry approaches to be essential for unequivocal identification of biological tyrosine nitrations.     

Epitope Structure of the Carbohydrate Recognition Domain of Asialoglycoprotein Receptor to a Monoclonal Antibody Revealed by High-Resolution Proteolytic Excision Mass Spectrometry

Recent studies suggest that the H1 subunit of the carbohydrate recognition domain (H1CRD) of the asialoglycoprotein receptor is used as an entry site into hepatocytes by hepatitis A and B viruses and Marburg virus. Thus, molecules binding specifically to the CRD might exert inhibition towards these diseases by blocking the virus entry site. We report here the identification of the epitope structure of H1CRD to a monoclonal antibody by proteolytic epitope excision of the immune complex and high-resolution MALDI-FTICR mass spectrometry. As a prerequisite of the epitope determination, the primary structure of the H1CRD antigen was characterised by ESI-FTICR-MS of the intact protein and by LC-MS/MS of tryptic digest mixtures. Molecular mass determination and proteolytic fragments provided the identification of two intramolecular disulfide bridges (seven Cys residues), and a Cys-mercaptoethanol adduct formed by treatment with β-mercaptoethanol during protein extraction. The H1CRD antigen binds to the monoclonal antibody in both native and Cys-alkylated form. For identification of the epitope, the antibody was immobilized on N-hydroxysuccinimide (NHS)-activated Sepharose. Epitope excision and epitope extraction with trypsin and FTICR-MS of affinity-bound peptides provided the identification of two specific epitope peptides (5–16) and (17–23) that showed high affinity to the antibody. Affinity studies of the synthetic epitope peptides revealed independent binding of each peptide to the antibody.     

Toward bioinspired galectin mimetics: identification of ligand-contacting peptides by proteolytic-excision mass spectrometry

Clinically relevant bioactivities of human galectins (adhesion/growth-regulatory galactoside-specific lectins) inspired the design of peptides as new tools to elicit favorable effects (e.g., in growth control) or block harmful binding (e.g., in tissue invasion). To obtain the bioinspired lead compounds, we combined a proteolytic fragmentation approach without/with ligand contact (excision) with mass spectrometric identification of affinity-bound protein fragments, using galectin-1 and -3 as models. Two peptides from the carbohydrate recognition domains were obtained in each case in experimental series rigorously controlled for specificity, and the [157-162] peptide of galectin-3 proved to be active in blocking lectin binding to a neoglycoprotein and to tumor cell surfaces. This approach affords peptide sequences for structural optimization and intrafamily/phylogenetic galectin comparison at the binding-site level with a minimal requirement of protein quantity, and it is even amenable to mixtures.     

TD-GC-FID based approach for monitoring indoor borne styrene and optimization of irradiation conditions for radiopolymerization in consolidated artifacts

Styrene induces various adverse health impacts. Due to its wide applications, the quantification of personal exposure level is of great importance, especially in places that attract a large number of people daily, like museums. The aim of this study was to develop a TD-GC-FID based approach to assay the level of residual indoor borne styrene from artifacts consolidated through radio-polymerization. Optimization of the irradiation conditions and the minimum absorbed dose for total radio-polymerization were also investigated through the proposed methodology. Styrene’s assay was performed using a thermal desorption instrument coupled to a gas chromatograph equipped with a flame ionization detector (TD-GC-FID). Specific TD protocols were optimized for both declared objectives. Thermogravimetry/differential scanning calorimetry (TG/DSC) was used for optimization of the TD operational parameters. It was demonstrated that an adsorbed dose of 30 kGy ensures an almost complete radio-polymerization process. Less than 1.746?ppm of styrene is delivered from artifacts and may be inhaled by a person situated in its close proximity, a level placed within the accepted limits imposed by OSHA regulations. It was demonstrated that the TD-GC-FID approach might be useful to assess the occupational exposure to gaseous residual indoor borne styrene from museums exhibiting consolidated artifacts.     

An algorithm for solving the problem of convex programming with several objective functions

This work aims to establish an algorithm for solving the problem of convex programming with several objective-functions, with linear constraints. Starting from the idea of Rosen’s algorithm for solving the problem of convex programming with linear constraints, and taking into account the solution concept from multidimensional programming, represented by a program which reaches ”the best compromise”, we are extending this method in the case of multidimensional programming. The concept of direction of minimization is introduced, and a necessary and sufficient condition is given for as ∈R ⁿ direction to be a direction of minimization, according to the values of a criteria ensemble in a given point. The algorithm is interactive, and the intervention of the decident is minimal. The two numerical examples presented at the end validate the algorithm.     

Chemical Composition,Antioxidant and Antimicrobial Activity of Some Types of Honey from Banat Region,Romania

Honey is a natural product with multiple health benefits. The paper presents the chemical characterization and the antioxidant and antimicrobial potential of ten types of honey (knotweed, linden, wild cherry, acacia, honeydew, oilseed rape, sunflower, phacelia, plain polyflora and hill polyflora) from the Banat region, Romania. We studied the water content, dry matter, impurities, acidity and pH of honey. We also determined the content of reducing sugar, minerals and flavonoids and the total phenolic content. All honey samples analysed showed good nutritional characteristics according to the standard codex for honey. From the analysis of the mineral content of the honey samples, we observed a variability in the macro and microminerals, influenced by the botanical origin, ranging between 0.25% (wild cherry honey) and 0.54% (honeydew). The toxic metals’ (Cd and Pb) levels met the standard for almost all samples analysed except for knotweed. The flavonoid content of the samples ranged from 9.29 mg QE/100 g for wild cherry honey to 263.86 mg QE/100 g for linden honey, and for polyphenols between 177.6 mgGAE/100 g for acacia honey and 1159.3 mgGAE/100 g for honeydew. The best antioxidant capacity was registered in the case of linden honey (79.89%) and honeydew (79.20%) and the weakest in acacia (41.88%) and wild cherries (50.4%). All studied honey samples showed antimicrobial activity, depending on the type of honey, concentration and strain analysed. The novelty of this study is given by the complex approach of the study of honey quality, both from the perspective of chemical attributes and the evaluation of the antimicrobial potential on specific strains in correlation with the botanical and geographical origin of the analyzed area.     

Mechanism of formation of nonlinearity in the dose dependence of the thermoluminescence output for anion-defect α-Al2O3 crystals

The mechanism of formation of nonlinearity in the dose dependence of the thermoluminescence output was investigated for anion-defect α-Al₂O₃ single crystals. The experiments revealed a new specific feature: the nonlinearity of the dose dependence of the thermoluminescence output depends on the heating rate of crystals during recording of thermoluminescence. It was established that this effect and other thermoluminescence features studied earlier for anion-defect α-Al₂O₃ crystals are adequately described in the framework of a unified mechanism associated with the interactive interaction of dosimetric and deep traps. Recommendations are given for a decrease in the range of the superlinear portion in the dose dependence of the thermoluminescence output for the crystals under investigation.