Messenger RNA half-life measurements in mammalian cells.

The recognition of the importance of mRNA turnover in regulating eukaryotic gene expression has mandated the development of reliable, rigorous, and "user-friendly" methods to accurately measure changes in mRNA stability in mammalian cells. Frequently, mRNA stability is studied indirectly by analyzing the steady-state level of mRNA in the cytoplasm; in this case, changes in mRNA abundance are assumed to reflect only mRNA degradation, an assumption that is not always correct. Although direct measurements of mRNA decay rate can be performed with kinetic labeling techniques and transcriptional inhibitors, these techniques often introduce significant changes in cell physiology. Furthermore, many critical mechanistic issues as to deadenylation kinetics, decay intermediates, and precursor-product relationships cannot be readily addressed by these methods. In light of these concerns, we have previously reported transcriptional pulsing methods based on the c-fos serum-inducible promoter and the tetracycline-regulated (Tet-off) promoter systems to better explain mechanisms of mRNA turnover in mammalian cells. In this chapter, we describe and discuss in detail different protocols that use these two transcriptional pulsing methods. The information described here also provides guidelines to help develop optimal protocols for studying mammalian mRNA turnover in different cell types under a wide range of physiologic conditions.

Culture As a Process, the Process of the Lives of People and Their Behaviors

A Review of the 15th KUCES Lecture on Korean Culture entitled "Life as a Travel, Life as a Traum: Koreans' Way of Living in This Troubled World" presented by Professor Han Kyung-koo on November 20 [1999]

Cognitive Cities: An Application for Nairobi. Text Message Participation of Slum Inhabitants to Improve Sanitary Facilities

Population growth is always increasing, and thus the concept of smart and cognitive cities is becoming more important. Developed countries are aware of and working towards needed changes in city management. However, emerging countries require the optimization of their own city management. This chapter illustrates, based on a use case, how a city in an emerging country can quickly progress using the concept of smart and cognitive cities. Nairobi, the capital of Kenya, is chosen for the test case. More than half of the population of Nairobi lives in slums with poor sanitation, and many slum inhabitants often share a single toilet, so the proper functioning and reliable maintenance of toilets are crucial. For this purpose, an approach for processing text messages based on cognitive computing (using soft computing methods) is introduced. Slum inhabitants can inform the responsible center via text messages in cases when toilets are not functioning properly. Through cognitive computer systems, the responsible center can fix the problem in a quick and efficient way by sending repair workers to the area. Focusing on the slum of Kibera, an easy-to-handle approach for slum inhabitants is presented, which can make the city more efficient, sustainable and resilient (i.e., cognitive).