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On-Chip Lipid Extraction Using Superabsorbent Polymers for Mass Spectrometry
Bang, Geul,Kim, Young Hwan,Yoon, Junghyo,Yu, Yeong Jun,Chung, Seok,Kim, Jeong Ah American Chemical Society 2017 ANALYTICAL CHEMISTRY - Vol.89 No.24
<P>Pretreatment of samples is one of the most important steps in analytical methods for efficient and accurate results. Typically, an extraction method used for lipid analysis with mass spectrometry is accompanied by complex liquid–liquid extraction. We have devised a simple, rapid, and efficient lipid extraction method using superabsorbent polymers (SAPs) and developed a high-throughput lipid extraction platform based on a microfluidic system. Since SAPs can rapidly absorb an aqueous solution from a raw sample and convert it into the gel, the lipid extraction process can be remarkably simplified. The hydrophobic lipid components were captured into the fibrous SAP gel and then solubilized and eluted directly into the organic solvent without significant interference by this polymer. The small-scale lipid extraction process minimizes the liquid handling and unnecessary centrifugation steps, thereby enabling the implementation of a SAP-integrated microfluidic lipid extraction platform. The SAP method successfully induced reproducible extraction and high recovery rates (95–100%) compared to the conventional Folch method in several lipid classes. We also demonstrated the feasibility of the SAP method for the analysis of lipids in complex biological samples, such as the brain and liver, as well as <I>Escherichia coli</I>. This small-scale SAP method and its microfluidic platform will open up new possibilities in high-throughput lipidomic research for diagnosing diseases because this new technique saves time, labor, and cost.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/ancham/2017/ancham.2017.89.issue-24/acs.analchem.7b03547/production/images/medium/ac-2017-03547a_0007.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ac7b03547'>ACS Electronic Supporting Info</A></P>
Park Gaeun,Jang Wooyoung Eric,Kim Seoyeon,Gonzales Edson Luck,Ji Jungeun,Choi Seunghwan,Kim Yujin,Park Ji Hwan,Mohammad Hazara Begum,Bang Geul,Kang Minkyung,Kim Soobin,Jeon Se Jin,Kim Jin Young,Kim Kw 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-
Autism spectrum disorder (ASD) is a neurodevelopmental disorder associated with impaired social behavior and communication, repetitive behaviors, and restricted interests. In addition to genetic factors, environmental factors such as prenatal drug exposure contribute to the development of ASD. However, how those prenatal factors induce behavioral deficits in the adult stage is not clear. To elucidate ASD pathogenesis at the molecular level, we performed a high-resolution mass spectrometry-based quantitative proteomic analysis on the prefrontal cortex (PFC) of mice exposed to valproic acid (VPA) in utero, a widely used animal model of ASD. Differentially expressed proteins (DEPs) in VPA-exposed mice showed significant overlap with ASD risk genes, including differentially expressed genes from the postmortem cortex of ASD patients. Functional annotations of the DEPs revealed significant enrichment in the Wnt/β-catenin signaling pathway, which is dysregulated by the upregulation of Rnf146 in VPA-exposed mice. Consistently, overexpressing Rnf146 in the PFC impaired social behaviors and altered the Wnt signaling pathway in adult mice. Furthermore, Rnf146-overexpressing PFC neurons showed increased excitatory synaptic transmission, which may underlie impaired social behavior. These results demonstrate that Rnf146 is critical for social behavior and that dysregulation of Rnf146 underlies social deficits in VPA-exposed mice.
Effect of developmental exposure to bisphenol A on steroid hormone and vitamin D3 metabolism
Kim, Jae Kwan,Khan, Adnan,Cho, Seongha,Na, Jinhyuk,Lee, Yeseung,Bang, Geul,Yu, Wook-Joon,Jeong, Ji-Seong,Jee, Sun Ha,Park, Youngja H. Elsevier 2019 CHEMOSPHERE - Vol.237 No.-
<P><B>Abstract</B></P> <P>High exposure to bisphenol A (BPA) in children has been associated with the outcomes of several diseases, including those related to developmental problems. To elucidate the mechanism of BPA mediated developmental toxicity, plasma and urine from rats exposed to BPA was analyzed with high resolution metabolomics, beginning from post-natal day 9, for 91 days. Female and male rats were orally administered 5 different BPA doses to elucidate dose- and sex-specific BPA effects. Regarding dose-specific effects, multivariate statistical analysis showed that metabolic shifts were considerably altered between 5, 50 and 250 mg BPA/kg bw/day in treated rats. A nonmonotonicity and monotonicity between BPA dose and metabolic response were major trajectories, showing overall metabolic changes in plasma and urine, respectively. Metabolic perturbation in the steroid hormone biosynthesis pathway was significantly associated with dose- and sex-specific BPA effects. Intermediate metabolites in the rate-limiting step of steroid hormone biosynthesis down-regulated steroid hormones in the 250 mg treatment. Further, our study identified that BPA increased urinary excretion of vitamin D<SUB>3</SUB> and decreased its concentration in blood, suggesting that perturbation of vitamin D<SUB>3</SUB> metabolism may be mechanistically associated with neurodevelopmental disorders caused by BPA. Three metabolites showed a decrease in sex difference with high BPA dose because female rats were more affected than males, which can be related with early puberty onset in female. In brief, the results demonstrated that BPA induces dose- and sex-specific metabolic shifts and that perturbation of metabolism can explain developmental problems.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Nonmonotonicity and monotonicity between dose of BPA and metabolic responses. </LI> <LI> Early sexual maturation was only observed in females at 250 mg/kg bw/day. </LI> <LI> Female rats were more affected by bisphenol A than male, which leads to decreased sex difference at 250 mg/kg bw/day. </LI> <LI> Rate limiting step in steroid hormone biosynthesis pathway was down regulated, causing decreased steroid hormone.. </LI> <LI> Perturbation of vitamin D<SUB>3</SUB> metabolism by BPA can be associated with neurobehavioral problem. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>