Inorganic arsenic (iAs) is one of the most endemic toxicants worldwide and oxidative stress is a key cellular pathway underlying iAs toxicity. Other cellular stress response pathways, such as the unfolded protein response (UPR), are also impacted by iAs exposure, however it is not known how these pathways intersect to cause disease. We optimized the use of zebrafish larvae to identify the relationship between these cellular stress response pathways and arsenic toxicity. We found that the window ...[more]
Project description:This study was carried out to compare potential changes in the gene expression in 5dpf livers between untreated, 1.0 and 1.5mM iAs exposures. Overall design: Perform RNA-seq on 5 independent pools of 5 dpf livers collected after exposure to 1.0 and 1.5mM iAs from 4-5 dpf
Project description:This study was carried out to compare potential changes in the gene expression in 5dpf livers between untreated, 1.0 and 1.5mM iAs exposures. Overall design: Perform RNA-seq on 5 independent pools of 5 dpf livers collected after exposure to untreated, 1.0 and 1.5mMfrom 4-5 dpf
Project description:Ex-vivo liver perfusion (EVLP) is an ideal platform to study liver disease, therapeutic interventions, and pharmacokinetic properties of drugs without any patient risk. Rat livers are an ideal model for EVLP due to less organ quality variability, ease of hepatectomy, well-defined molecular pathways, and relatively low costs compared to large animal or human perfusions. However, the major limitation with rat liver normothermic machine perfusion (NMP) is maintaining physiologic liver function on an ex-vivo machine perfusion system. To address this need, our research demonstrates 24-hour EVLP in rats under normothermic conditions. Early (6 hour) perfusate transaminase levels and oxygen consumption of the liver graft are shown to be good markers of perfusion success and correlate with viable 24-hour post-perfusion histology. Finally, we address overcoming challenges in long-term rat liver perfusions such as rising intrahepatic pressures and contamination, and offer future directions necessary to build upon our work.
Project description:Heterogeneous Tg(hsp70-hRASG12V) embryos were obtained by mating the male homozygous transgenic fish to wildtype females. They were raised to 24 hour-post fertilization (hpf) stage and received heatshock at 37°C in waterbath for one hour, and kept in 28.5°C till 30hpf for RNA extraction. Wildtype embryos receiving the same heatshock treatment were used as controls. Each microarray sample was prepared by pooling 50 embryos and biological duplication was used. The zebrafish has been established as a powerful vertebrate model organism for large-scale genetic screens and chemical screens. Here, we seek to establish that zebrafish embryos can be utilized as an in vivo system to dissect crucial pathways for oncogenesis. A microarray analysis was performed by comparing transcription profile of heterogeneous Tg(hsp70-hRASG12V) embryos with heatshock to wildtype embryos with heatshock. Both groups of embryos received heatshock at 37°C for one hour at 24 hour-post fertilization (hpf) stage, and kept in 28.5°C till 30hpf for RNA extraction. Each microarray sample was prepared by pooling 50 embryos and biological triplicate was used. Ingenuity Pathway Analysis (IPA) was performed using the upregulated lists. “Cancer “ was the top diseases and disorders, with “Developmental disorder” and “Organismal Injury and Abnormalities” as the second and third diseases and disorders, validating that transient heatshock induction of oncogenic RAS in embryos activates major pathways in oncogenesis.