Spatial neurolipidomics—MALDI mass spectrometry imaging of lipids in brain pathologies

Given the complexity of nervous tissues, understanding neurochemical pathophysiology puts high demands on bioanalytical techniques with respect to specificity and sensitivity. Mass spectrometry imaging (MSI) has evolved to become an important, biochemical imaging technology for spatial biology in biological and translational research. The technique facilitates comprehensive, sensitive elucidation of the spatial distribution patterns of drugs, lipids, peptides, and small proteins in situ. Matrix-assisted laser desorption ionization (MALDI)-based MSI is the dominating modality due to its broad applicability and fair compromise of selectivity, sensitivity price, throughput, and ease of use. This is particularly relevant for the analysis of spatial lipid patterns, where no other comparable spatial profiling tools are available. Understanding spatial lipid biology in nervous tissue is therefore a key and emerging application area of MSI research. The aim of this review is to give a concise guide through the MSI workflow for lipid imaging in central nervous system (CNS) tissues and essential parameters to consider while developing and optimizing MSI assays. Further, this review provides a broad overview of key developments and applications of MALDI MSI-based spatial neurolipidomics to map lipid dynamics in neuronal structures, ultimately contributing to a better understanding of neurodegenerative disease pathology.     

New acetylcholinesterase inhibitors isolated from Delphinium uncinatum

The inhibition of acetylcholinesterase (AChE), the key enzyme in the breakdown of acetylcholine, is presently the most common pharmacological approach available for Alzheimer’s disease (AD). Despite research on the molecular bases of AD, potent therapeutic agent against its expansion is still needed. In searching for natural cholinesterase inhibitors, the present study was focused on the isolation of three new norditerpenoid alkaloids, uncinatine B-D together with known virescenine from Delphinium uncinatum. Chemical structures for all the isolated norditerpenoids (1–4) were established using latest spectroscopic techniques. The isolated undescribed compounds along with known virescenine were testified for their acetylcholinesterase inhibitory activity supported by docking analyses. Molecular docking simulation showed that the isolated compounds (1–4) were observed to adhered in the active site of AChE with docking scores ? 13.5322 (1), ?11.8173 (2), ?12.4240 (3) and ? 8.9352 (4) respectively. Overall results demonstrated that these natural norditerpenoids compounds were found as selective inhibitors of AChE. This is the first report regarding the use of bioactive ingredients of Delphinium uncinatum in testing against Alzheimer's disease.     

Characterizing the Memory Effect on the amylose tris(3,5-dimethylphenyl) carbamate stationary phase

Acid/base modifiers are sometimes used as additives in normal phase elution on columns packed with CHIRALPAK^® AD^®. They affect enantioseparations in ways that are not understood for the lack of systematic studies, which makes the scale-up of preparative separations difficult to predict. Once a column has been exposed to these modifiers, the selectivity of certain pairs of enantiomers may change, for the better or the worse. Numerous molecules that are affected by this phenomenon are listed in the literature. We selected five of them, the selectivity of which changes as more acidic or basic solutions are percolated through the column. The selectivity of ketoprofen, 4-chlorophenylalanine methyl and ethyl esters improves as a solution of Ethanesulfonic Acid is percolated through the column. The selectivity of Propranolol HCl and Tröger’s base increases as a solution of diisopropylamine is percolated through the column. Each one of these five compounds is inversely affected by the percolation of the opposite acid/base solution. We used trans-Stilbene Oxide (tso) as a “standard” to determine the columns stability because no Memory Effect is observed for it (its retention, enantioselectivity, and resolution remain constant). Karl Fisher titrations showed that only slight changes in the water content of the mobile phase occurred, and that a unique water to polymer moiety ratio is important. Analytical studies of the stationary phase suggest that slow protonation/deprotonation of water attached to the carbamate moiety may be responsible for the acid/base Memory Effect. Finally, we showed that the Memory Effect can be minimized by percolating through the column a sufficiently concentrated solution of the appropriate acid or base. Thus, columns that were unreliable for method development, due to the Memory Effect, can now be used. As a result, the need to store several CHIRALPAK AD columns, specific for each condition of the Memory Effect, is eliminated.     

Concentration Effect,Structural Properties,and Driving Force on Aβ28 Dimerization with and without Zn2+ Cooperation: Learning from Replica Exchange Sampling**

Zn²⁺ is a very important factor in promoting the formation of amyloid beta (Aβ) aggregates and amyloid plaques. The Zn²⁺-bound Aβ species generate amorphous or low molecular-weight oligomers. However, it is a lack of studies to approach the starting structural features (dimerization) in Aβ nucleation processes with and without Zn²⁺, which is the key point in understanding Zn²⁺-induced nucleation mechanisms. To better understand the effect of concentration, structural properties, and the driving force, 14 independent replica exchange molecular dynamics simulations were performed in Aβ₂₈ dimerization with and without Zn²⁺ (zAβ₂₈) cooperation. Our scanning results show that the aggregation propensity is easier in Aβ₂₈-Aβ₂₈ and Aβ₂₈-zAβ₂₈ systems than zAβ₂₈-zAβ₂₈ system. In binding property, the Aβ₂₈-Aβ₂₈ model (−61.5 kcal mol⁻¹) is stronger than zAβ₂₈-zAβ₂₈ (−26.6 kcal mol⁻¹) and Aβ₂₈-zAβ₂₈ (−7.24 kcal mol⁻¹) models. Further analysis confirmed that H13 and H14 residues play specific roles in the three systems. The key point is the orientation of N atom of the imidazole ring in histidine residues. Furthermore, we discovered different driving forces for each system. Our current study contributes to the understanding of how the Aβ₂₈ dimer interacts with Zn²⁺, which could lead to new insights into Zn²⁺-induced nucleation mechanisms.     

丰富环境改善阿尔茨海默病的基础研究进展

阿尔茨海默病(AD)是一种以进行性认知障碍和记忆能力损害为主的神经退行性疾病. 虽然针对AD发病机制和治疗的研究众多, 但目前仍缺乏十分有效的治疗手段. 一些研究显示, 通过改变生活环境和方式, 进行各种体育活动, 在预防及改善AD方面发挥重要作用. 丰富环境是指相对于标准实验室饲养条件而言可以增强动物感觉、认知和运动等各方面刺激的实验室饲养条件, 丰富环境可增加海马神经发生, 增强突触可塑性及提高认知行为等. 从最初发现丰富环境对神经退行性病变神经元丢失有改善作用之后, 关于丰富环境与AD的研究层出不穷, 从多角度如β淀粉样蛋白及Tau蛋白磷酸化等方面阐释丰富环境对AD的改善作用及相关机制. 本文就丰富环境对AD的改善作用及相关调节机制做一综述.     

采用移动极差统计合并技术来监控ICP-AES测量系统的不确定度

采用ICP AES(电感耦合等离子体 原子发射光谱法 )测定金属硅中铁杂质含量进行不确定度评估时 ,提出了平均移动极差 (MR)合并统计动态监控技术。研究认为 ,采用该技术来进行ICP AES测量系统的不确定度评估 ,其贡献量分别是由控制样品 (QC)和线性拟合两部分合成。这一结论的得出能最大限度合成各种变异因素的影响 ,避免了相关性的复杂计算 ,有利于ICP AES测量系统的不确定度评估     

Study of the enantiomeric separation of an acetamide intermediate by using supercritical fluid chromatography and several polysaccharide based chiral stationary phases

Four chiral stationary phases, based on the phenylcarbamate derivatives of amylose or cellulose: Chiralcel OD-H, Chiralpak AD, Lux Cellulose-2 and Lux Amylose-2, were evaluated for the enantiomeric separation of an acetamide chiral intermediate, the (4S-trans)-4-(ethylamino)-4-(N-acetamide)-5,6-dihydro-(6S)-methyl-4H-thieno-[2,3-b]thiopyran-7,7-dioxide, using SFC. The effect of the different modifiers and temperatures, on the separation, was also studied. The chiral separation could not be achieved using the Chiralpak AD column, nevertheless the other columns provided excellent results with analysis times close to 6 min and resolutions higher than 2. The highest enantioresolutions and retentions were obtained with the Lux Cellulose-2 column and 2-propanol as organic modifier. The isoelution temperatures were estimated from the van't Hoff plots, and in all the cases they were above the temperature range studied which means that the enantiomeric separation was enthalpy driven.     

All-atom molecular dynamics studies of the full-length β-amyloid peptides

β-Amyloid peptides are believed to play an essential role in Alzheimer’s disease (AD), due to their sedimentation in the form of β-amyloid aggregates in the brain of AD-patients, and the in vitro neurotoxicity of oligomeric aggregates. The monomeric peptides come in different lengths of 39–43 residues, of which the 42 alloform seems to be most strongly associated with AD-symptoms. Structural information on these peptides to date comes from NMR studies in acidic solutions, organic solvents, or on shorter fragments of the peptide. In addition X-ray and solid-state NMR investigations of amyloid fibrils yield insight into the structure of the final aggregate and therefore define the endpoint of any conformational change of an Aβ-monomer along the aggregation process. The conformational changes necessary to connect the experimentally known conformations are not yet understood and this process is an active field of research.     

Enantiomeric impurity determination of levetiracetam using capillary electrochromatography

CEC was used to develop a method for the enantiomeric excess determination of levetiracetam, an antiepileptic drug. Different types of calibration curve were evaluated for use in the range between 0.01 and 1 mg/mL when aniracetam was used as an internal standard. The method gave comparable results when only the areas of the impurity were used in the calibration curve. The predicted detection and quantification limits from the S/N were 1.1 and 3.6 microg/mL, respectively. However, experimental results showed that LOD and LOQ were underestimated. Repeatability of injection was demonstrated by the RSD values obtained for retention time, resolution, ratios of the areas impurity/internal standard, and areas of impurity and internal standard individually, which were below or equal to 9.30%. The between-days variability experiments indicated that it is better to make a calibration curve daily. The finally selected calibration curves were used to test the accuracy of the developed method on bulk samples and Keppra tablets containing 250 mg levetiracetam. Both selected calibration curves performed similarly. The one using the internal standard information gave overall recoveries between 88 and 118%, while the one using areas gave results between 84 and 118%.