Unravelling the Time Scale of Conformational Plasticity and Allostery in Glycan Recognition by Human Galectin-1

The interaction of human galectin-1 with a variety of oligosaccharides, from di-(N-acetyllactosamine) to tetra-saccharides (blood B type-II antigen) has been scrutinized by using a combined approach of different NMR experiments, molecular dynamics (MD) simulations, and isothermal titration calorimetry. Ligand- and receptor-based NMR experiments assisted by computational methods allowed proposing three-dimensional structures for the different complexes, which explained the lack of enthalpy gain when increasing the chemical complexity of the glycan. Interestingly, and independently of the glycan ligand, the entropy term does not oppose the binding event, a rather unusual feature for protein-sugar interactions. CLEANEX-PM and relaxation dispersion experiments revealed that sugar binding affected residues far from the binding site and described significant changes in the dynamics of the protein. In particular, motions in the microsecond-millisecond timescale in residues at the protein dimer interface were identified in the presence of high affinity ligands. The dynamic process was further explored by extensive MD simulations, which provided additional support for the existence of allostery in glycan recognition by human galectin-1.     

Validation of a HPLC/MS method for simultaneous quantification of clonidine,morphine and its metabolites in human plasma

A high‐performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of morphine, morphine's major metabolites morphine‐3‐glucuronide and morphine‐6‐glucuronide, and clonidine, to support the pharmacokinetic analysis of an ongoing double‐blinded randomized clinical trial that compares the use of morphine and clonidine in infants diagnosed with neonatal abstinence syndrome. Plasma samples were processed by solid‐phase extraction and separated on an Inertsil ODS‐3 (4 μm) column using an 0.1% formic acid in water–0.1% formic acid in methanol gradient. Detection of the analytes was conducted in the positive multiple reaction monitoring mode. The range of quantitation was 1–1000 ng/mL for morphine, morphine‐3‐glucuronide and morphine‐6‐glucuronide, and 0.25–100 ng/mL for clonidine. Intra‐day and inter‐day accuracy and precision were ≤15% for all analytes across the quantitation range. Extraction recovery rates were ≥94% for morphine, ≥90% for M3G, ≥87% for M6G and ≥ 79% for clonidine. Matrix effect ranged from 85–94% for clonidine to 101–106% for M3G. The method fulfilled all predetermined acceptance criteria and required only 100 μL of starting plasma volume. Furthermore, it was successfully applied to 30 clinical trial plasma samples.     

Time‐dependent finite element simulations of a shear‐thinning viscoelastic fluid with application to blood flow

A new finite element method is developed to simulate time‐dependent viscoelastic shear‐thinning flows characterized by the generalized Oldroyd‐B model. The focus of the algorithm is improved stability through a free‐energy dissipative scheme by using low‐order piecewise‐constant finite element approximations for stress. The algorithm is further modified by incorporating a pressure‐projection method, a DG‐upwinding scheme, a symmetric interior penalty DG method to solve the elliptic pressure‐update equation and a geometric multigrid preconditioner. The improved stability and cost to accuracy is compared when using higher order discontinuous bilinear approximation, where in addition, we consider the influence of a slope limiter for these elements. The algorithm is applied to the 2D start‐up‐driven cavity problem, and the stability of the free energy is illustrated and compared between element choices. An application of the model to modelling blood in small arterioles and channels is considered by simulating pulsatile blood flow through a stenotic arteriole. The individual influences of viscoelasticity and shear‐thinning within the generalized Oldroyd‐B model are investigated by comparing results to the Newtonian, generalized Newtonian and Oldroyd‐B models. Copyright © 2014 John Wiley & Sons, Ltd.     

The impact of blood on liver metabolite profiling – a combined metabolomic and proteomic approach

Metabolomics has entered the well‐established omic sciences as it is an indispensable information resource to achieve a global picture of biological systems. The aim of the present study was to estimate the influence of blood removal from mice liver as part of sample preparation for metabolomic and proteomic studies. For this purpose, perfused mice liver tissue (i.e. with blood removed) and unperfused mice liver tissue (i.e. containing blood) were compared by two‐dimensional gas chromatography time of flight mass spectrometry (GC × GC‐TOFMS) for the metabolomic part, and by liquid chromatography tandem mass spectrometry (LC‐MS/MS) for the proteomic part. Our data showed significant differences between the unperfused and perfused liver tissue samples. Furthermore, we also observed an overlap of blood and tissue metabolite profiles in our data, suggesting that the perfusion of liver tissue prior to analysis is beneficial for an accurate metabolic profile of this organ. Copyright © 2013 John Wiley & Sons, Ltd.     

不同抗凝比例对凝血四项检测结果的影响

目的观察并分析凝血四项检测结果与不同比例下的全血与抗凝剂之间的关系。方法研究对象取2015年6月来湖北省宜昌市第二人民医院参加体检的110例健康人员,抽取全血血液样本后按照不同比例与抗凝剂混匀,常规分离血浆并测定受试者凝血酶时间(TT)、凝血酶原时间(PT)、激活部分促凝血酶原激酶时间(APTT)以及纤维蛋白原(FIB),并对研究结果相关数据作统计学处理。结果当血液量与抗凝剂比例为1∶5时,标本TT、PT、APTT以及FIB各项指标较1∶9时差异显著而具有统计学意义(P0.05);抗凝比例1∶7的情况下,TT指标差异与1∶9抗凝比例标本相比差异具有统计学意义(P0.05),而PT、FIB以及APTT三项指标差异并无统计学意义(P0.05);抗凝比例为1∶11与1∶13的情况下,TT指标与1∶9抗凝比例标本相比差异具有统计学意义(P0.05),而PT、FIB以及APTT三项指标差异并无统计学意义(P0.05)。结论凝血四项检测工作中,标本质量检测控制最关键的环节在于准确采集血液量,倘若采集血量过多或过少,导致与抗凝剂比例失调往往会影响测定结果准确性,检验科工作人员应予以重视。     

Chemoselective Hydrogenation of 6-Alkynyl-3-fluoro-2-pyridinaldoximes: Access to First-in-Class 6-Alkyl-3-Fluoro-2-pyridinaldoxime Scaffolds as New Reactivators of Sarin-Inhibited Human Acetylcholinesterase with Increased Blood–Brain Barrier Permeability

Novel 6-alkyl- and 6-alkenyl-3-fluoro-2-pyridinaldoximes have been synthesised by using a mild and efficient chemoselective hydrogenation of 6-alkynyl-3-fluoro-2-pyridinaldoxime scaffolds, without altering the reducible, unprotected, sensitive oxime functionality and the C−F bond. These novel 6-alkyl-3-fluoro-2-pyridinaldoximes may find medicinal application as antidotes to organophosphate poisoning. Indeed, one low-molecular-weight compound exhibited increased affinity for sarin-inhibited acetylcholinesterase (hAChE) and greater reactivation efficiency or resurrection for sarin-inhibited hAChE, compared with those of 2-pyridinaldoxime (2-PAM) and 1-({[4-(aminocarbonyl)pyridinio]methoxy}methyl)-2-[(hydroxyimino)methyl]pyridinium chloride (HI-6), two pyridinium salts currently used as antidote by several countries. In addition, the uncharged 3-fluorinated bifunctional hybrid showed increased in vitro blood–brain barrier permeability compared with those of 2-PAM, HI-6 and obidoxime. These promising features of novel low-molecular-weight alkylfluoropyridinaldoxime open up a new era for the design, synthesis and discovery of central non-quaternary broad spectrum reactivators for organophosphate-inhibited cholinesterases.     

Quantitative analysis of enasidenib in dried blood spots of mouse blood using an increased‐sensitivity LC–MS/MS method: Application to a pharmacokinetic study

A simple, sensitive and rapid assay method has been developed and validated as per regulatory guidelines for the estimation of enasidenib on mouse dried blood spots (DBS) using liquid chromatography coupled to tandem mass spectrometry with electrospray ionization in the positive‐ion mode. The method employs liquid extraction of enasidenib from DBS disks of mouse whole blood followed by chromatographic separation using 0.2% formic acid–acetonitrile (25:75, v/v) at a flow rate of 1.0 mL/min on an Atlantis dC₁₈ column with a total run time of 2.0 min. The MS/MS ion transitions monitored were m/z 474.0 → 267.1 for enasidenib and m/z 309.2 → 251.3 for the internal standard (warfarin). The assay was linear in the range of 1.01 – 3044 ng/mL. The within‐run and between‐run precisions were in the range of 3.18 – 9.06 and 4.66 – 8.69%, respectively. Stability studies showed that enasidenib was stable on DBS cards for 1 month. This novel method has been applied to analyze the DBS samples of enasidenib obtained from a pharmacokinetic study in mice.     

Numerical analysis of stress distribution in the upper arm tissues under an inflatable cuff: Implications for noninvasive blood pressure measurement

An inflatable cuff wrapped around the upper arm is widely used in noninvasive blood pressure measurement. However, the mechanical interaction between cuff and arm tissues, a factor that potentially affects the accuracy of noninvasive blood pressure measurement, remains rarely addressed. In the present study, finite element (FE) mod-els were constructed to quantify intra-arm stresses generated by cuff compression, aiming to provide some theoretical evi-dence for identifying factors of importance for blood pressure measurement or explaining clinical observations. Obtained results showed that the simulated tissue stresses were highly sensitive to the distribution of cuff pressure on the arm sur-face and the contact condition between muscle and bone. In contrast, the magnitude of cuff pressure and small variations in elastic properties of arm soft tissues had little influence on the efficiency of pressure transmission in arm tissues. In par-ticular, it was found that a thickened subcutaneous fat layer in obese subjects significantly reduced the effective pres-sure transmitted to the brachial artery, which may explain why blood pressure overestimation occurs more frequently in obese subjects in noninvasive blood pressure measurement.     

红细胞分布宽度与心房颤动负荷的关系研究

目的观察病态窦房结综合征患者心房颤动负荷与红细胞分布宽度（RDW）的关系。方法选择病态窦房结综合征植入永久性人工心脏起搏器的患者81例，回顾性分析患者入院时基本资料、血常规及生化指标。记录术后3个月起搏器记录的心房颤动负荷。根据患者心房颤动负荷分为高负荷组及低负荷组，比较分析两组患者RDW及相关临床资料。结果心房颤动高负荷组患者RDW（15.61±1.23）%，高血压比例71%，明显高于低负荷组的（13.50±1.03）%、46%（P＜0.01或0.05）。高负荷组左心房直径、左心室舒张末期内径、WBC、TG、TC均大于低负荷组（P＜0.01或0.05）。心房颤动负荷影响因素的logistic回归分析显示，RDW是心房颤动负荷高的预测因素（OR=10.32，P＜0.01）。高TG、高血压、高胆固醇也是心房颤动高发的预测因素。结论心房颤动负荷除与年龄，高血压有关外，还与RDW明显相关。RDW可成为病态窦房结综合征患者心房颤动负荷的预测因素之一。