Flow injection analysis of angiotensin I-converting enzyme inhibitory activity with enzymatic reactors

Assay of angiotensin I-converting enzyme (ACE) inhibitory activity always draws much attention because of diverse applications in the field of antihypertension and related pathogenesis. Recently, the use of a new synthetic substrate, 3-hydroxybutyrylglycyl-glycyl-glycine (3HB-GGG), for the assay of ACE inhibitory activity has been confirmed. To construct a rapid, economical, and automatic determination system of ACE inhibitory activity using 3HB-GGG, a flow injection analysis (FIA) system with enzymatic reactors was developed in this study. Enzyme reactors were composed of aminoacylase and 3-hydroxybutyrate dehydrogenase immobilized separately on CNBr-activated Sepharose 4B. The assay condition was optimized in terms of the conversion of 3HB-G into NADH by the enzymatic reactors when the reaction solution containing 3HB-G generated from 3HB-GGG (after the incubation with ACE) was repetitively injected into the FIA system. Under the optimized conditions, 3HB-G was converted to 3HB, and then 3HB was oxidized by NAD⁺ to form NADH. The developed system successfully detected practical ACE inhibitors with a great sensitivity, high sampling frequency (10 samples h⁻¹) and a durable stability of the enzymatic reactors.     

泰和乌骨鸡肉模拟胃肠道消化液中ACE抑制肽的分离纯化及结构鉴定

研究泰和乌骨鸡肉在胃肠道消化过程中的降血压活性,并鉴定出其中确定具有降血压活性的肽序列。采用体外模拟胃肠道消化法,测定了不同阶段消化液对血管紧张素I转化酶(ACE)的抑制作用,并选择其中的小肠道消化液,对其进行超滤、制备型反相高效液相色谱、凝胶柱色谱分离纯化,最后通     

Study of blood collection and sample preparation for analysis of vitamin D and its metabolites by liquid chromatography–tandem mass spectrometry

The analysis of vitamin D status, with special emphasis on 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D, is gaining interest in clinical studies due to the classical and non-classical effects attributed to this prohormone. In this research, the influence of the two steps preceding determination (viz. sample collection and preparation) on the quantitative analysis of vitamin D and its more important metabolites has been studied. Two preparation approaches, deproteination and solid-phase extraction (SPE), have been evaluated in terms of sensitivity to delimit their application, thus establishing that detection of 1,25-dihydroxyvitamin D cannot be addressed by protein precipitation. Concerning sample collection, serum and plasma reported high accuracy (above 83.3%) for vitamin D and metabolites, while precision, expressed as relative standard deviation, was below 12.9% for all analytes in both samples. Statistical analysis revealed that serum and plasma provided similar physiological levels for vitamin D₃, 24,25-dihydroxyvitamin D₃ and 25-hydroxyvitamin D₃, while significantly different levels were obtained for 1,25-dihydroxyvitamin D₃, always higher in plasma than in serum. Sample collection and treatment have proved to be significant in the analysis of vitamin D and its relevant metabolites.     

Fast evaluation of time domain fields in sub-wavelength source/observer distributions using accelerated Cartesian expansions (ACE)

Time domain integral equation solvers for transient scattering from electrically large objects have benefitted significantly from acceleration techniques like the plane wave time domain (PWTD) algorithm; these techniques reduce the asymptotic CPU and memory cost. However, PWTD breaks down when used in the analysis of structures that have subwavelength features or features whose length scales are orders of magnitude smaller than the smallest wavelength in the incident pulse. Instances of these occurring in electromagnetics range from antenna topologies, to feed structures, etc. In this regime, it is the geometric constraints that dictate the computational complexity, as opposed to the wavelength of interest. In this work, we present an approach for efficient analysis of such sub-wavelength source/observer distributions in time domain. The methodology that we seek to exploit is the recently developed algorithm based on Cartesian expansions for accelerating the computation of potentials of the form R^ν. In this paper, we present an efficient methodology for computing these polynomials for two different scenarios; where the size of the domain spans the distance travelled by light in (i) one time step and (ii) multiple time steps. These algorithms are cast within the framework of both uniform and non-uniform distributions. Results that demonstrate the efficiency and convergence of the proposed algorithm are presented.     

相依结构数据的ACE拟合

利用ACE(alternating conditional expectations)方法,给出了寻找数据阿基米德copula相依结构的一种非参数算法.这种算法去掉了传统方法中对生成元需要事先假定结构类型的要求。拓宽了阿基米德copula的应用范围.模拟结果表明,ACE方法在寻找资产相依结构中是可以应用的,与传统方法相比也毫不逊色。     

Use of capillary electrophoresis as a versatile tool to measure interaction constants between a KDR‐binding PEGylated lipopeptide and pegylated phospholipid micelles

In the frame of our molecular imaging activities, a PEGylated lipopeptide has been developed as a specific ligand for the human vascular endothelial growth factor receptor 2, which is considered as one of the important molecular marker of angiogenesis. In this study, the potential of affinity capillary electrophoresis (ACE) is evaluated to measure the interactions of an active PEGylated lipopeptide, its hydrolysis product and its precursor consisting of a peptide structure with different micelles including Brij‐35, Tween‐20, and pegylated phospholipids. Given the amphiphilic structure of the PEGylated lipopeptide, a MEKC method allowing the simultaneous separation of the compounds of interest was set up, using low percentages of acetonitrile. Analytes were resolved using a BGE consisting of 100 mM borate buffer pH 9.0, 1 mM Brij, and 25% acetonitrile. Optimized conditions were then used to perform ACE experiments. The affinity constants of the analytes with the micelles were calculated on the basis of their mobility decrease when surfactant concentration increased in the electrolyte. The use of different linearization models to estimate affinity constants was discussed and comparison of different surfactants was reported. PEGylated lipopeptide interacted more strongly with pegylated phospholipid micelles than with Brij‐35 or Tween‐20. Moreover, it is likely that the chemical structure of the compounds, and particularly the lipidic part of the molecules, significantly affects the interaction with micelles. In conclusion, the ACE method can be readily applied to investigate interactions of our targeting lipopeptides with various micelles currently used for the preparation of pharmaceutical vehicles.     

具有智能评阅功能的在线有机化学习题平台:ACE: Organic简介及其在教学中的应用*

ACE: Organic(Achieving Chemistry Excellence: Organic Chemistry)是具有智能反馈功能的在线有机化学习题平台。学生利用系统中的画图软件在线完成每个练习题后,平台会判断学生的回答是否正确,即时给出评价与提示,引导学生找到解决问题的思路。对于复杂的机理与合成题,ACE可以接受多种合理的回答。教师可以通过查看平台的统计数据,随时了解学生对知识点的掌握情况。ACE设有论坛,便于教师和学生互动。教学实践证明,ACE的开放有效地提高了学生对有机化学的学习兴趣,培养了学生严谨的科学态度。     

On the Phan system of the Schur cover of SU(4, 32)

This article is part of the program described in Bennett et al. (in: Ivanov et al. (ed.) Groups, combinatorics, and geometry, 2003) [3]. We study the Phan amalgams and their universal completions that occur for q = 3 in rank n = 3 for the diagram A ₃ = D ₃, corresponding to SU(4, 3²) ≅ Spin ⁻(6, 3). We show that the associated geometries admit universal 9-fold coverings, by showing that the universal completion of the Phan amalgam is the central extension of SU(4, 3²) by its Schur multiplier. This information provides the last missing piece of information in the full classification of Phan amalgams and their universal completions for A _n and D _n .      

Virtual Screening in Search for a Chemical Probe for Angiotensin-Converting Enzyme 2 (ACE2)

We elaborate new models for ACE and ACE2 receptors with an excellent prediction power compared to previous models. We propose promising workflows for working with huge compound collections, thereby enabling us to discover optimized protocols for virtual screening management. The efficacy of elaborated roadmaps is demonstrated through the cost-effective molecular docking of 1.4 billion compounds. Savings of up to 10-fold in CPU time are demonstrated. These developments allowed us to evaluate ACE2/ACE selectivity in silico, which is a crucial checkpoint for developing chemical probes for ACE2.     

Ultrasound-assisted preparation of ACE inhibitory peptide from milk protein and establishment of its in-situ real-time infrared monitoring model

A scheme for preparing milk ACE inhibitory peptides by in vitro proteolysis and simulated gastrointestinal digestion was constructed. The ultrasonic assisted pretreatment was used to improve the enzymolysis of milk protein. The in-situ real-time infrared was used to establish monitoring model of enzymatic process. Results showed that under the conditions of single frequency 28 kHz, ultrasound time 40 min, ultrasound power density 20 W/L, milk protein concentration 34 g/L, batch ratio 2:4 and initial temperature 30 °C, the ACE inhibition rate of gastric digestion of enzymatic hydrolysate reached 67.20%, which was 22.87% higher than that of non-ultrasound samples. The results of secondary structure studies of proteins showed that after the ultrasonic treatment, the content of α-helix and β-corner reduced, and the content of β-folding and random coil increased. Compared with the control group, the ultrasonic treatment increased surface hydrophobicity and the content of SH while reduced the content of SS in milk protein, thus improving the ACE inhibitory activity of enzymatic hydrolysates. Furthermore, three quantitative prediction models of PLS, iPLS and Si-PLS for ACE inhibition rate of milk protease hydrolysates were established. And all these three different in-situ real-time prediction models had good predictive effect on the ACE inhibition rate of milk proteolysis products and gastrointestinal simulated digestion products.