Determination of Acid β-Galactosidase Activity: Methodology and Perspectives     

Radoslaw Kwapiszewski  Justyna Szczudlowska  Karina Kwapiszewska  Michal Chudy  Zbigniew Brzozka 《Indian journal of clinical biochemistry : IJCB》2014,29(1):57-62

Early, accurate diagnosis of lysosomal storage disorders is a major challenge, even for trained specialists. Finding innovative, accurate diagnostic methods, and high throughput, cost-effective tools are crucial to medical progress and will contribute to improved quality of life. The goal of this work was to improve currently used protocols to determine activity of acid β-galactosidase, and discuss the possibility analysing lysosomal enzymes with microfluidic systems. A principle of the determination of β-galactosidase activity was fluorometric measurement of a deprotonated form of 4-methylumbelliferone released in the enzymatic reaction. Measurements were performed using Jurkat T cells as a source of the enzyme. We observed the temperature-dependent substrate inhibition effect and determined the substrate (4-MU-β-d-galactopyranoside) concentration which should be used to determine acid β-galactosidase activity at 37 °C (0.8 mM) and at room temperature (0.6 mM). We proved that the sample incubation time may be significantly reduced to only a few minutes. We also showed that the amount of alkaline buffer used to stop the enzymatic reaction may be minimized and even, in some cases, eliminated. The presented results show how the sensitivity of the available methods to diagnose patients suffer from gangliosidosis GM1 or Morquio B disease can be improved. The proposed method may be easily implemented with microfluidic systems, which currently are promising tools for point-of-care applications.  相似文献   

Computing controversy: Formal model and algorithms for detecting controversy on Wikipedia and in search queries     

Kazimierz Zielinski  Radoslaw Nielek  Adam Wierzbicki  Adam Jatowt 《Information processing & management》2018,54(1):14-36

Controversy is a complex concept that has been attracting attention of scholars from diverse fields. In the era of Internet and social media, detecting controversy and controversial concepts by the means of automatic methods is especially important. Web searchers could be alerted when the contents they consume are controversial or when they attempt to acquire information on disputed topics. Presenting users with the indications and explanations of the controversy should offer them chance to see the “wider picture” rather than letting them obtain one-sided views. In this work we first introduce a formal model of controversy as the basis of computational approaches to detecting controversial concepts. Then we propose a classification based method for automatic detection of controversial articles and categories in Wikipedia. Next, we demonstrate how to use the obtained results for the estimation of the controversy level of search queries. The proposed method can be incorporated into search engines as a component responsible for detection of queries related to controversial topics. The method is independent of the search engine’s retrieval and search results recommendation algorithms, and is therefore unaffected by a possible filter bubble.Our approach can be also applied in Wikipedia or other knowledge bases for supporting the detection of controversy and content maintenance. Finally, we believe that our results could be useful for social science researchers for understanding the complex nature of controversy and in fostering their studies.  相似文献