The unique feature of riboswitches is to control selected gene expression by specific recognition of a cognate ligand. AdoCbl (adenosyl cobalamin, coenzyme B12) riboswitches regulate the expression of enzymes and transporters involved in bacterial AdoCbl biosynthesis or uptake on a transcriptional and/or translational level. The analysis of ligand recognition and the induced conformational changes requires a detailed knowledge of the kinetics and thermodynamics of ligand binding and of the secondary structure rearrangements. This chapter describes the investigation of coenzyme B12 binding to the btuB riboswitch from Escherichia coli by in-line probing assays and surface plasmon resonance spectroscopy. The experimental conditions, requirements, and performance of both methods are presented together with the evaluation of the experimental data to determine the associated conformational changes of the RNA and the kinetic and thermodynamic parameters of ligand binding. Owing to the light sensitivity of the cobalt(I)'carbon bond, these methods were specifically modified to ensure the chemical integrity of AdoCbl.
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http://dx.doi.org/10.1016/B978-0-12-801122-5.00020-9 | DOI Listing |
Mol Biol Rep
January 2025
Department of Biochemistry, All India Institute of Medical Sciences, Jodhpur, Rajasthan, 342005, India.
Background: Differential DNA methylation in the promoter region of tumour suppressor genes leads to gene function silencing.
Materials And Methods: In this study, we aimed to evaluate the salivary promoter methylation of EDNRB, MGMT and TIMP3 genes in H&NC patients (n = 100), premalignant lesions patients (n = 25) and healthy controls (n = 50). Blood and saliva samples were collected from all three groups and 20 concomitant tumour tissues were collected from the H&NC patients.
Biotechnol Bioeng
January 2025
Bioprocess Research and Development (BRD), WuXi Biologics, Shanghai, China.
Serving as a dedicated process analytical technology (PAT) tool for biomass monitoring and control, the capacitance probe, or dielectric spectroscopy, is showing great potential in robust pharmaceutical manufacturing, especially with the growing interest in integrated continuous bioprocessing. Despite its potential, challenges still exist in terms of its accuracy and applicability, particularly when it is used to monitor cells during stationary and decline phases. In this study, data pre-processing methods were first evaluated through cross-validation, where the first-order derivative emerged as the most effective method to diminish variability in prediction accuracy across different training datasets.
View Article and Find Full Text PDFJ Cardiothorac Surg
January 2025
The First Hospital of Lanzhou University, Lanzhou, China.
Background: This article aims to use high-throughput sequencing to identify miRNAs associated with ferroptosis in myocardial ischemia-reperfusion injury, select a target miRNA, and investigate its role in H9C2 cells hypoxia-reoxygenation injury.
Methods: SD rats and H9C2 cells were used as subjects. ELISA kits quantified MDA, SOD, GSH, LDH, and ferritin levels.
ACS Nano
January 2025
Department of Physics, University of Basel, Klingelbergstrasse 82, 4056 Basel, Switzerland.
High-precision molecular manipulation techniques are used to control the distance between radical molecules on superconductors. Our results show that the molecules can host single electrons with a spin 1/2. By changing the distance between tip and sample, a quantum phase transition from the singlet to doublet ground state can be induced.
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