China 
Africa 
Explore chapters and articles related to this topic
Current in vivo Models for Brain Disorders
Published in Carla Vitorino, Andreia Jorge, Alberto Pais, Nanoparticles for Brain Drug Delivery, 2021
Marta Guerra-Rebollo, Cristina Garrido
In the specific case of brain tumours, one of the most utilised techniques to visualise drug delivery into the tumour is using BLI. The firefly luciferase (FLuc) can be used as a reporter gene and d-luciferin as a substrate. FLuc catalyses d-luciferin oxidation in the presence of adenosine triphosphate (ATP) and coenzyme A (CoA) producing O2 and photons which can be detected in vivo. Recording of the emitted light by photodetectors, capable of linear response allows for real-time measurements [57, 58]. This technique is based on the stable expression of luciferase by the brain tumour cell line selected for the experiment. Once the cells are implanted into the mouse brain, d-luciferin could be injected intraperitoneally into the host mice, where it is distributed throughout the mouse body and crosses the BBB. The intensity of the emitted light correlates to the size of the tumour and allows comparisons of tumours size across different animals but also within the same animal in different time points (Fig. 13.1) [59].
*
Published in Chad A. Mirkin, Spherical Nucleic Acids, 2020
Liangliang Hao, Pinal C. Patel, All H. Alhasan, David A. Giljohann, Chad A. Mirkin
It is known that a single miRNA can regulate multiple targets through interactions with the 3’-UTRs of the target mRNA. To test whether the novel miRNA-AuNPs could function similarly, we cloned and inserted the complementary DNA (cDNA) fragment containing the 3’-UTR sequence of PRKCE downstream of firefly luciferase into the pMIRREPORT miRNA expression reporter vector (Applied Biosystems) and then transfected this plasmid into HeLa cells. When the ectopic miR-205 is successfully delivered to the cells by miRNA-AuNPs, it interacts with the miR-205 binding site in the PRKCE 3’-UTR fragment inserted into the reporter vector and causes translational repression of firefly luciferase. The pRL-SV40 renilla luciferase vector that did not contain a miR-205 binding site and could not be targeted by miR-205 was co-transfected as a control. Luciferase expressing HeLa cells were treated with 5 nM mimic miRNA-AuNPs targeting the 3’- UTR sequence of PRKCε. After 72 h, firefly luciferase signal was measured in cell lysates and normalized to the renilla luciferase signal using the DualGlo Luciferase Assay System (Promega).
Bioluminescence- and Chemiluminescence-Based Fiberoptic Sensors
Published in Loïc J. Blum, Pierre R. Coulet, Biosensor Principles and Applications, 2019
Loïc J. Blum, Sabine M. Gautier
The bioluminescence reaction of the American firefly Photinus pyralis has been extensively studied and well reviewed by DeLuca and McElroy (1–3). In vitro, firefly luciferase (EC 1.13.12.7) catalyzes the production of light in the presence of ATP, Mg2+, molecular oxygen, and a specific substrate, luciferin. The complex mechanism of the reaction is not yet completely understood, but the overall stoichiometry has been established: () ATP+luciferin+O2→Mg2+luciferaseAMP+oxyluciferin+PPi+CO2+hv
Adenosine triphosphate (ATP) bioluminescence-based strategies for monitoring atmospheric bioaerosols
Published in Journal of the Air & Waste Management Association, 2022
Yueqi Zhang, Bing Liu, Zhaoyang Tong
The use of adenylate kinase (ADK) and pyruvate kinase for ATP amplification has the potential to detect very low levels of ATP without the use of photon detectors. Lee et al. (2017) designed ATP amplification using (i) ADK as the first enzyme that converts AMP+ATP to two ADP molecules, and (ii) polyP kinase (PPK) as the second enzyme that converts ADP to ATP using polyP. In this reaction, excess AMP and polyP are added to the reaction mixture, which drives the ADK and PPK equilibriums to ADP and ATP formation, respectively. The amplified ATP is subjected to bioluminescence detection in a firefly luciferase reaction. The sensitivity of this method to ATP was about 10,000 times that of the bioluminescence method without ATP amplification. The ATP is amplified before the bioluminescence detection, and the luminescence amount can be improved without signal integration, thereby greatly improving the sensitivity of the bioluminescence detection to ATP.
Inhibitory effect of particulate matter on toll-like receptor 9 stimulated dendritic cells by downregulating mitogen-activated protein kinase and NF-κB pathway
Published in Journal of Toxicology and Environmental Health, Part A, 2020
Madeeha Arooj, Irshad Ali, Hee Kyoung Kang, Jin Won Hyun, Young-Sang Koh
Human embryonic kidney 293 T (HEK293 T) cells are well-established cell lineage extensively used as an expression tool for transfection (Thomas and Smart 2004). To examine whether PM2.5 exerted an inhibitory effect on CpG-DNA-stimulated activator protein-1 (AP-1) and NF-κB transcriptional activity, an AP-1 and NF-κB reporter gene assay was performed as previously described (Manzoor et al. 2013). Briefly, HEK293 T cells were plated in 24-well plate and grown overnight. HEK293 T cells were transfected with AP-1 or NF-κB reporter plasmid together with pRLnull, pcDNA3 (empty vector) or murine TLR9 expressing plasmid (pcDNA3-mTLR9) using Fugene 6. HEK293 T cells were pre-treated with PM2.5 for 1 hr and then stimulated with CpG-DNA (1 μM). After 18 hr, cells were lysed in passive lysis buffer and firefly luciferase activity was measured by using a dual-luciferase reporter assay system according to manufacturer’s instructions.
New approaches towards the discovery and evaluation of bioactive peptides from natural resources
Published in Critical Reviews in Environmental Science and Technology, 2020
Nam Joo Kang, Hyeon-Su Jin, Sung-Eun Lee, Hyun Jung Kim, Hong Koh, Dong-Woo Lee
To monitor the effect of chemical compounds on target pathway activity, a reporter frequently used for drug screening, the firefly luciferase (FLuc) reporter, is often employed (Xie et al., 2016). Reporter-based assays can be used to analyze gene expression and are highly sensitive, enabling accurate quantification of small changes in transcription. For instance, recently, the human H293-NF-κB-RE-luc2P reporter cell line was developed to identify anti-inflammatory bioactive compounds (Meijer, Vonk, Priebe, & Roelofsen, 2015). This system can be used to monitor a compound’s ability to inhibit TNF-α (Tumor Necrosis Factor-α)-induced NF-κB activity. The reporter line, derived from the human embryonic kidney cell line HEK293, contains NF-κB response elements five stably transfected upstream of the promoter of an optimized luciferase reporter gene (luc2P). Induction of the luc2P gene produces chemiluminescence when the substrate is added, and the amount of chemiluminescence generated is directly proportional to the changes in nuclear translocation of the NF-κB protein, and thus the assay can be used to measure inflammatory activity. The H293-NF-κB-RE-luc2P reporter cell line is a useful and sensitive tool for the rapid identification of BPs with anti-inflammatory activity.