Intra-Laboratory Validation of Alpha-Galactosidase Activity Measurement in Dietary Supplements

Introduction: Alpha-galactosidase (α-Gal) is an enzyme responsible for the hydrolyzation of glycolipids and glycoprotein commonly found in dietary sources. More than 20% of the general population suffers from abdominal pain or discomfort caused by intestinal gas and by indigested or partially digested food residuals. Therefore, α-Gal is used in dietary supplements to reduce intestinal gases and help complex food digestion. Marketed enzyme-containing dietary supplements must be produced in accordance with the Food and Drug Administration (FDA) regulations for Current Good Manufacturing Practice (cGMPs). Aim: in this work we illustrated the process used to develop and validate a spectrophotometric enzymatic assay for α-Gal activity quantification in dietary supplements. Methods: The validation workflow included an initial statistical-phase optimization of materials, reagents, and conditions, and subsequently a comparative study with another fluorimetric assay. A final validation of method performance in terms of specificity, linearity, accuracy, intermediate-precision repeatability, and system precision was then executed. Results and conclusions: The proven method achieved good performance in the quantitative determination of α-Gal activity in commercial food supplements in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals (ICH) guidelines and is suitable as a rapid in-house quality control test.     

An Assay for Screening Potential Drug Candidates for Alzheimer's Disease That Act as Chaperones of the Transthyretin and Amyloid-β Peptides Interaction

The protein transthyretin (TTR) modulates amyloid-β (Aβ) peptides deposition and processing and this physiological effect is further enhanced by treatment with iododiflunisal (IDIF), a small-molecule compound (SMC) with TTR tetramer stabilization properties, which behaves as chaperone of the complex. This knowledge has prompted us to design and optimize a rapid and simple high-throughput assay that relies on the ability of test compounds to form ternary soluble complexes TTR/Aβ/SMC that prevent Aβ aggregation. The method uses the shorter Aβ(12–28) sequence which is cheaper and simpler to use while retaining the aggregation properties of their parents Aβ(1–40) and Aβ(1–42). The test is carried out in 96-plate wells that are UV monitored for turbidity during 6 h. Given its reproducibility, we propose that this test can be a powerful tool for efficient screening of SMCs that act as chaperones of the TTR/Aβ interaction that may led to potential AD therapies.     

Synthesis of novel cytotoxic 3-azolylsteroids via Cu-catalyzed C–N coupling

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Antioxidant activity evaluation by physiologically relevant assays based on haemoglobin peroxidase activity and cytochrome c-induced oxidation of liposomes

Two new protocols for exploring antioxidant-related chemical composition and reactivity are described: one based on a chronometric variation of a haemoglobin ascorbate peroxidase assay and one based on cytochrome c-induced oxidation of lecithin liposomes. Detailed accounts are given on their design, application, critical correlations with established methods and mechanisms. These assays are proposed to be physiologically relevant and bring new information regarding a real sample, both qualitative and quantitative. The well-known assays used for evaluation of antioxidant (re)activity are revisited and compared with these new methods. Extracts of the Hedera helix L. are examined as test case, with focus on seasonal variation and on leaf, fruit and flower with respect to chromatographic, spectroscopic and reactivity properties. According to the set of assays performed, winter are the most antioxidant, followed by summer leaves, and then by flowers and fruits.     

综合性无机化学实验:保健药品中钙、铁、锌含量的测定*

在大学无机化学实验教学中,应用元素含量测定技术,对营养保健药品中钙、铁、锌的含量进行定量测定,详细介绍了实验条件、测定方法和计算过程,并重点讨论了应用常规仪器对待测样品进行分离、纯化的方法和对干扰元素有效的掩蔽方法。通过对比实验结果与产品说明书中相关元素的含量数据,表明该实验方法具有较好的准确性和可操作性。     

穿膜肽TAT-PTD的固相合成及其与DNA相互作用的研究

通过N-芴甲氧羰基(N-fluorenylmethoxycarbonyl,Fmoc)作为α-氨基的保护基,以逐个延伸的固相合成法合成TAT-PTD多肽,运用高效液相色谱和质谱鉴定合成多肽的纯度和相对分子质量。通过紫外吸收光谱和琼脂糖凝胶电泳阻滞实验研究合成的TAT-PTD多肽与质粒DNA相互作用,实验表明TAT-PTD多肽与质粒DNA可以通过静电作用相互吸引、靠近并结合,于是为下一步小单孢菌基因工程的细胞转导提供了依据。     

Establishment of a high-throughput screening system for universal anti-HIV targets

The process of identifying novel human immunodeficiency virus 1 (HIV-1) inhibitors presents a challenge for industrial and scientific research.Virus-cell-based screening approaches offer some advantages in the quest for novel inhibitors because they include multiple targets in a single screen and in some cases reveal targets not captured in biochemical assays.In this study,a high-throughput screening (HTS) system for HIV-1 inhibitors was developed,which allows the simultaneous screening of all the HIV-1 tar...     

潘婷强韧防掉发洗发露的Ames 试验研究

摘要: 目的应用Ames 试验探讨潘婷强韧防掉发洗发露是否具有致突变性。方法采用组氨酸缺陷型鼠伤寒沙门氏菌TA97,TA98,TA100 和TA102 进行潘婷强韧防掉发洗发露微生物回复突变试验。结果在1. 25 ～ 0. 156mg /皿剂量范围内,在加S9mix 和不加S9mix 平行条件下测试,试验结果为阴性。结论潘婷强韧防掉发洗发露在本实验中未见致突变性。     

硫黄素T对淀粉样β-蛋白质40聚集成核动力学的双重影响

荧光染料硫黄素T常用于淀粉样纤维聚集过程的定性定量检测。虽然有研究表明,某些抑制淀粉样蛋白质聚集的小分子抑制剂会与硫黄素T相互作用,影响其测试结果。但硫黄素T如何影响淀粉样蛋白质的聚集成核动力学尚不清晰。本文以淀粉样β-蛋白质40 (Aβ40)为模型,系统研究了硫黄素T对Aβ40聚集成核的影响。研究发现：硫黄素T能够显著改变Aβ40的聚集成核动力学,且影响程度与硫黄素T的浓度密切相关。即在低浓度硫黄素T存在下,Aβ40成核速率的延迟时间先随着硫黄素T浓度的升高而缩短,后随着硫黄素T浓度的升高延迟时间反而延长。但延伸的速率却随硫黄素T浓度的升高而缓慢增大。另外,硫黄素T基本不会影响Aβ40的二级结构和纤维形态。同时,等温滴定微量热实验结果表明,硫黄素T结合Aβ40之间的主要作用力为疏水相互作用。据此,本研究提出硫黄素T对Aβ40聚集成核动力学的双重影响机理。这些结果有助于进一步了解硫黄素T与淀粉样蛋白质的作用特点,为今后硫黄素T在Aβ40聚集成核动力学实验中的使用提供参考。