Solution NMR approaches for establishing specificity of weak heterodimerization of membrane proteins

Solution NMR provides a powerful approach for detecting complex formation involving weak to moderate intermolecular affinity. However, solution NMR has only rarely been used to detect complex formation between two membrane proteins in model membranes. The impact of specific binding on the NMR spectrum of a membrane protein can be difficult to distinguish from spectral changes that are induced by nonspecific binding and/or by changes that arise from forced cohabitation of the two proteins in a single model membrane assembly. This is particularly the case when solubility limits make it impossible to complete a titration to the point of near saturation of complex formation. In this work experiments are presented that provide the basis for establishing whether specific complex formation occurs between two membrane proteins under conditions where binding is not of high avidity. Application of these methods led to the conclusion that the membrane protein CD147 (also known as EMMPRIN or basigin) forms a specific heterodimeric complex in the membrane with the 99-residue transmembrane C-terminal fragment of the amyloid precursor protein (C99 or APP-βCTF), the latter being the immediate precursor of the amyloid-β polypeptides that are closely linked to the etiology of Alzheimer's disease.     

Dynamic ultrasound-assisted extraction of oleuropein and related biophenols from olive leaves

A continuous approach for the ultrasound-assisted extraction of olive biophenols (OBPs) from olive leaves is proposed. Multivariate methodology was used to carry out a detailed optimisation of extraction. Under the optimal working conditions, complete extraction of the target analytes (namely, oleuropein, verbacoside, apigenin-7-glucoside and luteolin-7-glucoside with LODs 11.04, 2.68, 1.49 and 3.91 mg/kg, respectively) was achieved in 25 min. The extract was injected into a chromatograph-photodiode array detector assembly (HPLC-DAD) for individual separation-quantification. No clean-up or preconcentration steps were required. Gas chromatography-mass spectrometry (without derivatization of the analytes) was used to identify OBPs at concentrations below the LODs obtained by HPLC-DAD. The efficacy of ethanol-water mixtures to extract OBPs from olive leaves has been demonstrated and compared with that of a conventional method which requires 24h for complete extraction; so these mixtures can substitute toxic extractants used to date.     

Multivariate optimisation of the microwave-assisted extraction of oleuropein and related biophenols from olive leaves

Microwave assistance is proposed for the first time in order to accelerate the extraction of biophenols from olive leaves. Under optimal working conditions, obtained using a multivariate methodology, complete extraction of the target analytes was achieved in 8 min. The extracts required no clean-up nor concentration prior to injection into a chromatograph–photodiode array detector assembly for individual separation–quantification. The optimal extractant (an 80:20 ethanol–water mixture) was also used in the development of a stirring-based extraction method which required around 24 h for complete extraction of the target compounds. These mixtures can be used as replacements for toxic extractants, with a view to exploiting olive leaves in order to obtain biophenols for human use.     

Hyperon production in pp collisions

We report on a study of the proton induced hyperon production reactions. We discuss the theoretical efforts made towards understanding the existing data and the uncertainties involved in the calculations. Our recent calculations of the missing mass spectra for the pp → K ⁺ Λp reaction which involve a proper coupled channel treatment of the final state Λp interaction are presented. Significant differences in the results using different models of the hyperon-nucleon interaction are found.     

Fast ethylamine gas sensing based on intermolecular charge-transfer complexation

We have investigated the fast ethylamine gas sensing of 2-chloro-3,5-dinitrobenzotrifluoride(CDBF) loaded poly(acrylonitrile) nanofiber based on an intermolecular charge-transfer complexation.Reversible response and recovery were achieved using alternating gas exposure.This system shows a fast ethylamine gas sensing within 0.4 s.     

Static–dynamic sequential superheated liquid extraction of phenols and fatty acids from alperujo

Superheated liquids of different polarity have been used for sequential extraction of fatty acids and phenols from alperujo. Multivariate methodology has been used to optimise the static–dynamic extraction. Forty-two minutes are required to complete extraction (20 mg/kg of fatty acids and up to 2,200 mg/kg of hydroxytyrosol in the raw material used). The efficacy of the extraction has been demonstrated and compared with that of conventional methods (Folch and stirring-based methods for fatty acids and phenols, respectively), which needed 4.5 and 24 h for the extraction of fatty acids and phenols, respectively. The non-polar and polar extracts were injected into GC–MS and HPLC–MS–MS equipment, respectively, for individual separation–quantification of the target compounds. The simplicity of the experimental setup and the low costs of the raw material make the proposed method advisable when extraction of both fractions is required.     

Liquid chromatography/triple quadrupole tandem mass spectrometry with multiple reaction monitoring for optimal selection of transitions to evaluate nutraceuticals from olive-tree materials

Optimal transitions have been selected for the identification and quantitation of the most interesting hydrophilic biophenols in extracts from olive-tree materials, which are of interest because of their nutraceutical properties. The tested materials were extra virgin olive oil, waste from oil production (known as alperujo), and olive-tree materials such as leaves, small branches and fruit stones. The identification and determination steps of the target biophenols are based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) with a triple quadrupole (QQQ) mass detector. The interface between the chromatograph and the QQQ was an electrospray ionization source operated in the negative ion mode. Highly selective identification of the biophenols was confirmed by multiple reaction monitoring (MRM) using the most representative transitions from the precursor ion to the different product ions. Quantitative MS/MS analysis was carried out by optimization and selection of the most sensitive transition for each analyte, which resulted in estimated detection limits of 5.10 to 11.65 ng/mL for the extracts. The biophenols were extracted from the tested samples by different methods: liquid-liquid extraction for virgin olive oil, microwave-assisted leaching for olive leaves, branches and stones, and pressurized liquid leaching for alperujo. This study provides valuable information about the most suitable source for the isolation of each nutraceutical biophenol and enables us to obtain a complete profile of them in Olea Europaea.     

Multifragmentation and the phase transition: A systematic study of the multifragmentation of 1A GeV Au, La and Kr

A systematic analysis of the multifragmentation (MF) in fully reconstructed events from 1A GeV Au, La and Kr collisions with C has been performed. Detailed comparisons of the various fragment properties are presented as a function of excitation energy, E*_th. The charged particle multiplicity from MF stage shows a saturation beyond E*_th ∼ 8 MeV/nucleon for Kr. The universal behavior of intermediate mass fragment yields and of the size of the largest fragment is observed only for Au and La when scaled with size of the system. The Kr data are found to lack this property. Moments of the fragment size distribution show that the Kr MF is different than the MF of Au and La. A power law behavior is observed for Au and La with exponent τ>2, while for Kr τ<2. The results are compared with the statistical multifragmentation model (SMM). A single value of all the parameters of the model fits the data for all the three systems. The breakup of Au and La is consistent with a continuous phase transition. The data indicate that both E*_th and the isotope ratio temperature T _Hc-DT decrease with increase in system size at the critical point. The breakup temperature obtained from SMM also shows the same trend as seen in data. This trend is attributed primarily to the increasing Coulomb energy with finite size effects playing a smaller role. The percolation and Ising model studies for finite size neutral matter show behavior which is opposite to the one seen in the present work. EOS Collaboration     

Correction to: Robust optimal classification trees under noisy labels

Advances in Data Analysis and Classification -     

Relación entre el parámetro flecha-luz y la carga crítica de pandeo lateral para arcos parabólicos biempotrados bajo diferentes hipótesis de cálculo

Curved graphs that relate the f/L ratio and the λ = qL³/EI_y parameter for clamped arches with the different calculation hypothesis are going to be proposed in this article.It has been necessary to determine the real critical lateral strength of a series of clamped arches considering different calculation hypothesis respect to geometrical linearity and a steel material characteristic.The elastic-plastic material hypothesis with non geometrical linearity considers the existence of residual stresses and initial lateral deflection.