Transcriptomics and Metabolomics for Co-Exposure to a Cocktail of Neonicotinoids and the Synergist Piperonyl Butoxide

Abstract

Here, the transcriptomics and metabolomics on a model of exposure to a cocktail of neonicotinoids (Neo) containing seven commercial compounds and a synergist piperonyl butoxide (PBO) were established. The results showed that Neo and PBO disrupted mRNA and metabolite levels in a dose-dependent manner. Neo caused tryptophan pathway-related neurotoxicity, reduced lipolysis, and promoted fat mass accumulation in the liver, while PBO induced an increase in inflammatory factors and damage to intercellular membranes. Co-exposure enhanced Neo-induced liver steatosis, focal necrosis, and oxidative stress by inhibiting oxidative phosphorylation (OXPHOS). Furthermore, diglycerides and metabolic biomarkers demonstrated that the activation of insulin signaling is associated with restricted OXPHOS, which commonly leads to a high risk of non-alcoholic fatty liver disease (NAFLD) and Alzheimer’s disease (AD) as the result of over-synthesis of lipids, low energy supply, and high thermogenesis. The study demonstrates that chronic disease can be induced by Neo and the synergist PBO at the molecular level.

CRISPR/Cas12a-Amplified Aptamer Switch Microplate Assay for Small Molecules

“…The activated Cas12a cleaves a fluorescent ssDNA reporter labeled with both a fluorophore and a quencher, producing high fluorescence signals. 20,22,23 When a small molecule target exists, the Apt-acDNA probe binds with the target and forms a stable conformation, inhibiting hybridization between the Apt-acDNA probe and cDNA on the microplate, which leads to a decrease in the amount of Apt-acDNA probes captured on the microplate. Thus, less Cas12a is activated and the fluorescence signal is reduced.…”

“…19−21 Particularly, the target DNA-activated Cas12a can indiscriminately cleave surrounding nonspecific ssDNA with high enzymatic efficiency (termed trans-cleavage activity). 20,22,23 This unique cleavage property of Cas12a has been widely deployed to construct a series of amplified detection platforms for the analysis of different targets. 19−21,24−28 For small molecule detection, aptamers are often used as affinity ligands, and a majority of methods utilize a structure-switchable aptamer to regulate the activity of Cas12a responsive to target information.…”

Innovative Approaches for Drug Discovery: Quantifying Drug Distribution and Response with Raman Imaging

“…Xu et al used deuterated essential amino acids to quantify single-cell growth rates as a ratio of the incorporated C−D bonds from newly synthesized labeled proteins and the total C−H bonds from unlabeled proteins. 179 With ratiometric C−D/C−H SRS, they observed variations in metabolic and morphological response to different drugs, including the TKIs gefitinib and lapatinib, as indicated in Figure 6b.i, as well as paclitaxel and etoposide. Using data from hundreds of single cells at each drug concentration, they were able to obtain a half-maximum inhibitory concentration (IC 50 ) that is consistent with the MTT assay.…”

Basic evaluation of the CRISPR/Cas system stability for application to paper-based analytical devices

“…Considering the application of CRISPR-related reagents for practical POCT use, most assays involve a nucleic acid preamplification step to achieve the required sensitivity. [29][30][31] Regardless of the pre-amplification method selected (e.g., PCR or isothermal), heating is generally involved. For the previously reported solution phase assays performed in microtubes, the processes of target amplification and CRISPR/Cas-based detection can be readily separated.…”