HPLC‐MS/MS method for quantitative determination of the novel dual inhibitor of FGF and VEGF receptors E‐3810 in tumor tissues from xenograft mice and human biopsies

We developed and validated a high‐performance liquid chromatography–tandem mass spectrometry analytical method to measure E‐3810, a novel dual inhibitor of fibroblast growth factor receptor 1 and vascular endothelial growth factor receptor 1–3 in tissues and determined the drug concentration in a biopsy of human breast cancer for the first time. The method is a modification of our previous one in plasma to study the clinical pharmacokinetics of the drug during the phase I/II trial. In view of the changes in matrix, we applied a partial validation protocol to determine recovery, sensitivity, range of linearity, precision, accuracy and stability of the method over three runs in a mouse tumor tissue and liver. The recovery of E‐3810 from liver or tumor homogenate was >69%, and the lower limit of quantification was 5 ng/ml. The method was linear in the concentration range 5.0–500.0 ng/ml, as demonstrated by a determination coefficient R² ≥ 0.9955. The range of the calibration curve was appropriate for the analysis, as demonstrated by the accuracy, which was between 91.4% and 106.7%. Interday precision and accuracy on quality control samples at 9, 30 and 300 ng/ml were 3.1‐11.2% and 98.3–111.4%, respectively. The assay was applied successfully to determine the intratumor concentration of E‐3810 in different mouse xenograft tumor models and in a biopsy of a patient with breast cancer included in the phase I/II trial of the drug. In mouse tumors, the concentrations of E‐3810 were higher than necessary to exert antitumor activity in vitro (1 µM). Even more of interest was the result obtained in a human biopsy of few milligrams, where E‐3810 reached 4.9 µg/g (11 µM). Copyright © 2014 John Wiley & Sons, Ltd.     

Inherently Chiral Macrocyclic Oligothiophenes: Easily Accessible Electrosensitive Cavities with Outstanding Enantioselection Performances

Linear conjugated oligothiophenes of variable length and different substitution pattern are ubiquitous in technologically advanced optoelectronic devices, though limitations in application derive from insolubility, scarce processability and chain‐end effects. This study describes an easy access to chiral cyclic oligothiophenes constituted by 12 and 18 fully conjugated thiophene units. Chemical oxidation of an “inherently chiral” sexithiophene monomer, synthesized in two steps from commercially available materials, induces the formation of an elliptical dimer and a triangular trimer endowed with electrosensitive cavities of different tunable sizes. Combination of chirality with electroactivity makes these molecules unique in the current oligothiophenes literature. These macrocycles, which are stable and soluble in most organic solvents, show outstanding chiroptical properties, high circularly polarized luminescence effects and an exceptional enantiorecognition ability.     

The Influence of Different Bleaching Protocols on Dentinal Enzymatic Activity: An In Vitro Study

This study aimed to investigate matrix metalloproteinase (MMP) activity in human dentin using in-situ and gelatin zymography, after at-home and in-office bleaching, related to their clinical exposure times. Dentin specimens (n = 5) were treated with 35% hydrogen peroxide (50 min per session/4 sessions), 10% carbamide peroxide (180 min/21 sessions), or no treatment. All were subjected to in-situ zymography. Dentin slices were, subsequently, obtained, covered with fluorescein-conjugated gelatin, and examined with confocal laser-scanning microscopy. The fluorescence intensity was quantified and statistically analyzed using one-way ANOVA and Bonferroni tests (α = 0.05). Furthermore, gelatin zymography was performed on protein extracts obtained from dentin powder (N = 8 teeth), treated with hydrogen peroxide or carbamide peroxide, with different exposure times (10/50 min for hydrogen peroxide; 252/1260 min for carbamide peroxide). The results of the in-situ zymography showed no statistical differences between the bleached specimens and the control group, with a medium level of gelatinolytic activity expressed in the dentin tubules. The results of gelatin zymography showed an increased expression of pro-MMP-9 in carbamide peroxide groups. The expression of pro-MMP-2 decreased in all the experimental groups. The bleaching treatments performed on the enamel of sound teeth do not influence dentinal enzymatic activity. However, when unprotected dentin tissue is bleached, matrix metalloproteinases are more expressed, particularly when carbamide peroxide is used, proportional to the exposure time.     

Glucose/Ribitol Dehydrogenase and 16.9 kDa Class I Heat Shock Protein 1 as Novel Wheat Allergens in Baker’s Respiratory Allergy

Wheat allergens are responsible for symptoms in 60–70% of bakers with work-related allergy, and knowledge, at the molecular level, of this disorder is progressively accumulating. The aim of the present study is to investigate the panel of wheat IgE positivity in allergic Italian bakers, evaluating a possible contribution of novel wheat allergens included in the water/salt soluble fraction. The water/salt-soluble wheat flour proteins from the Italian wheat cultivar Bolero were separated by using 1-DE and 2-DE gel electrophoresis. IgE-binding proteins were detected using the pooled sera of 26 wheat allergic bakers by immunoblotting and directly recognized in Coomassie stained gel. After a preparative electrophoretic step, two enriched fractions were furtherly separated in 2-DE allowing for detection, by Coomassie, of three different proteins in the range of 21–27 kDa that were recognized by the pooled baker’s IgE. Recovered spots were analyzed by nanoHPLC Chip tandem mass spectrometry (MS/MS). The immunodetected spots in 2D were subjected to mass spectrometry (MS) analysis identifying two new allergenic proteins: a glucose/ribitol dehydrogenase and a 16.9 kDa class I heat shock protein 1. Mass spectrometer testing of flour proteins of the wheat cultivars utilized by allergic bakers improves the identification of until now unknown occupational wheat allergens.     

Synthesis and properties of new poly(ether sulfone)amides

High molecular-weight aromatic polyamides were obtained from 1,5- and 2,6-bis-(4′-carboxy-4-phenylenoxy-sulfonyl)naphthalene by direct polycondensation reaction in N-methyl-2-pyrrolidone with various aromatic diamines, using triphenyl phosphite and pyridine as condensing agents. The polymers were characterized by elemental analysis, thermogravimetric analysis, differential scanning calorimetry, and infrared analysis. The polyamides, obtained in quantitative yield, possessed inherent viscosities in the range 0.42–1.70 dL/g, glass transition temperatures between 245–310°C, and 10% weight loss temperatures in nitrogen and air above 435 and 424°C, respectively. Most of the polymers were soluble in aprotic solvents. The effect of the structure on properties, such as solubility, T_g, and thermal behavior, were also studied. © 1996 John Wiley & Sons, Inc.     

Flexural,impact, rheological and physical characterizations of POM reinforced by carbon nanotubes and paraffin oil

The paraffin oil dispersion technique innovated in the recent years to synthesize bulk polymer nanocomposite materials has a uniform dispersion. This research work aims to study the effect of added carbon nanotubes (CNTs) on flexural, impact and rheology behaviors of polyoxymethylene (POM) reinforced by 0–0.03 wt% of CNT using paraffin oil dispersion technique. The wettability and lamellar thickness were measured and rheological performance investigated using a parallel plate rheometer. The flexural and impact mechanical properties were also evaluated. The fracture surfaces were then examined by microscopy. The results showed that the energy to break, flexural strength and modulus increased proportionally with the addition of the amount of CNT in the matrix. For the rheology behavior, the viscosity decreased at the low percentage of CNT and then increased with increase in the percentage weight ratio of CNT in the POM matrix. It was also noted that the water contact angle rose with the increase the CNT percentages. Copyright © 2016 John Wiley & Sons, Ltd.     

Phenolic Compound Profile by UPLC-MS/MS and Encapsulation with Chitosan of Spondias mombin L. Fruit Peel Extract from Cerrado Hotspot—Brazil

Taperebá (Spondias mombin L.) is a native species of the Brazilian Cerrado that has shown important characteristics such as a significant phenolic compound content and biological activities. The present study aimed to characterize the phenolic compound profile and antioxidant activity in taperebá peel extract, as well as microencapsulating the extract with chitosan and evaluating the stability of the microparticles. The evaluation of the profile of phenolic compounds was carried out by UPLC-MS/MS. The in vitro antioxidant activity was evaluated by DPPH and ABTS methods. The microparticles were obtained by spray drying and were submitted to a stability study under different temperatures. In general, the results showed a significant content of polyphenols and antioxidant activity. The results of UPLC-MS/MS demonstrated a significant content of polyphenols in taperebá peel, highlighting the high content of ellagic acid and quercetin compounds. There was significant retention of phenolic compounds when microencapsulated, demonstrating high retention at all evaluated temperatures. This study is the first to microencapsulate the extract of taperebá peel, in addition to identifying and quantifying some compounds in this fruit.     

Effects of Regioisomerism on the Antiproliferative Activity of Hydroxystearic Acids on Human Cancer Cell Lines

A series of regioisomers of the hydroxystearic acid (HSA) was prepared, and the effect of the position of the hydroxyl group along the chain on a panel of human cancer cell lines was investigated. Among the various regioisomers, those carrying the hydroxyl at positions 5, 7, and 9 had growth inhibitor activity against various human tumor cell lines, including CaCo-2, HT29, HeLa, MCF7, PC3, and NLF cells. 10-HSA and 11-HSA showed a very weak effect. 8-HSA did not show inhibitory activity in all cell lines. The biological role of 7-HSA and 9-HSA is widely recognized, while little is known about the effects of 5-HSA. Therefore, the biological effects of 5-HSA in HeLa, HT29, MCF7, and NLF cell lines were investigated using the Livecyte’s ptychography technology, which allows correlating changes in proliferation, motility, and morphology as a function of treatment at the same time. 5-HSA not only reduces cell proliferation but also induces changes in cell displacement, directionality, and speed. It is important to characterize the biological effects of 5-HSA, this molecule being an important component of fatty acyl esters of hydroxy fatty acids (FAHFA), a class of endogenous mammalian lipids with noticeable anti-diabetic and anti-inflammatory effects.     

Metaheuristics for the team orienteering problem

The Team Orienteering Problem (TOP) is the generalization to the case of multiple tours of the Orienteering Problem, known also as Selective Traveling Salesman Problem. A set of potential customers is available and a profit is collected from the visit to each customer. A fleet of vehicles is available to visit the customers, within a given time limit. The profit of a customer can be collected by one vehicle at most. The objective is to identify the customers which maximize the total collected profit while satisfying the given time limit for each vehicle. We propose two variants of a generalized tabu search algorithm and a variable neighborhood search algorithm for the solution of the TOP and show that each of these algorithms beats the already known heuristics. Computational experiments are made on standard instances.     

Territorial origin of olive oil: representing georeferenced maps of olive oils by NMR profiling

ABSTRACT Proton NMR profiling is nowadays a consolidated technique for the identification of geographical origin of food samples. The common approach consists in correlating NMR spectra of food samples to their territorial origin by multivariate classification statistical algorithms. In the present work, we illustrate an alternative perspective to exploit territorial information, contained in the NMR spectra, which is based on the implementation of a geographic information system (GIS). Nuclear magnetic resonance spectra are used to build a GIS map permitting the identification of territorial regions having strong similarities in the chemical content of the produced food (terroir units). These terroir units can, in turn, be used as input for labeling samples to be analyzed by traditional classification methods. In this work, we describe the methods and the algorithms that permit to produce GIS maps from NMR profiles and apply the described method to the analysis of the geographical distribution of olive oils in an Italian region. In particular, we analyzed by ¹H NMR up to 98 georeferenced olive oil samples produced in the Abruzzo Italian region. By using the first principal component of the NMR variables selected according to the Moran test, we produced a GIS map, in which we identified two regions incidentally corresponding to the provinces of Teramo and Pescara. We then labeled the samples according to the province of provenience and built an LDA model that provides a classification ability up to 99% . A comparison between the variables selected in the geostatistics and classification steps is finally performed. Copyright © 2016 John Wiley & Sons, Ltd.