bluuebunny/crossref_metadata_2025_split huggingface dataset:
Atrazine Inhibition of Murine Electron Transport Chain: A CURE Biochemistry Laboratory
Authors: Martin Debra J. 
Published: June 2021

For the majority of students entering Biochemistry laboratory, basic biochemical skills are a new concept for them. Previous courses, such as lower division Introductory Biology and Genetics may have introduced them to some of the techniques; but overall, the concept of combining the techniques to actually apply it to research is lacking. Additionally, we want students to learn research skills, like interpreting evidence, reasoning, thinking critically, and conveying ideas and to be able to write like a scientist. Using a research based inquiry laboratory, students explore how a common herbicide might inhibit the electron transport chain (ETC). Additionally, they investigate how the herbicide could affect ETC when animals are exposed. By the end of the semester long course, the students submit a final paper in the format for submission to the Proceedings of the National Academy of Science. Since the implementation of this laboratory research project, pre/post Biochemistry knowledge assessment exam scores have increased 30%.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Leigh Syndrome Cell Model Development
Authors: Alverado Joshua , DeCuzzi Nicholaus , Hsueh Jessica , Madira Sarah , Lanning Nathan 
Published: June 2021

Leigh Syndrome (LS) is a neurological disease that stems from mutations in the subunits of mitochondrial respiratory chain subunits. These mutations decrease electron transport chain (ETC) deficiency, leading to decreased cellular ATP levels and increased reactive oxygen species (ROS) levels. There is currently no therapeutic approach to combat this degenerative disorder, and patients typically die by 8–10 years of age. There are also very few models with which to study this disease. We have generated LS cell models using both a chemical and genetic approach. Chemically, we inhibit ETC activity with low levels of rotenone, an ETC poison, in order to suppress ETC‐dependent ATP generation to levels seen in LS patients. Genetically, we use siRNA to knock down the ETC subunits that are mutationally inactivated in LS patients, also disrupting ETC function. Through these approaches, we have constructed LS cell models that can now be used to investigate potential therapeutics.Support or Funding InformationNIH 1R15GM123382, CSUPERB New Investigator GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Systems Approach Toward Elucidating The Molecular Basis Of Tropane Alkaloid Metabolism
Authors: D'Auria John C. 
Published: June 2021

Tropane and granatane alkaloids are key mediators of interspecies plant communication and valued by society for their pharmacological properties. Beyond their biological significance, the biocatalytic steps needed to generate tropane and granatane chemical scaffolds is not completely understood. This lack of knowledge of biosynthetic steps as well as kinetic and thermodynamic properties diminishes the scientific community's understanding of the chemistry of life. Tropane and granatane alkaloid metabolism consists of three modules: a first ring closure that forms a reactive N‐heterocyclic intermediate, a second ring closure that generates the tropane chemical scaffold and lastly modifications of the core chemical scaffold. The overarching question guiding this research is, how are the critical ring forming enzymes leading to these bicyclic compounds functioning at the molecular level? While tropane alkaloid metabolism has historically been studied in the Solanaceae, recent evidence indicates that non‐solanaceous species have independently evolved tropane metabolism using alternative biosynthetic machinery. Moreover, granatane alkaloids are structurally similar to tropane alkaloids and are hypothesized to use biocatalytic steps similar to those used to produce tropane alkaloids. Collectively, the tropane and granatane alkaloids constitute a comparative system for studying the mechanistic basis for the emergence and diversification of specialized metabolic enzymes. An integrated approach using biochemical, biophysical and molecular genetic currently being used to discover the molecular determinants of the bicyclic ring in tropane and granatane biosynthesis. Results from both the early and the terminal steps in the respective tropane and granatane pathways have begun to fill‐in gaps in our knowledge of their respective biosynthetic pathways as well as discover novel enzymes and divergent evolutionary mechanisms. For example, current data supports the hypothesis that a Type III Polyketide synthase is responsible for the extension off the first ring and ultimate formation of the second ring in the bicyclic tropane or granatane nucleus. We have identified Type III PKS enzymes and the genes encoding them from members of the Erythroxylaceae, Solanaceae, and Lythraceae. These enzymes all produce their respective predicted non‐cylclized polyketide enzyme product while sharing closer identity with PKS enzymes from within their own lineages.Support or Funding InformationThis research is funded via the National Science Foundation of the United States of America award ID. 1714236This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of Daurichromenic Acid Synthase from Rhododendron dauricum
Authors: Taura Futoshi , Iijima Miu , Kurosaki Fumiya 
Published: June 2021

Daurichromenic acid (DCA) synthase catalyzes the oxidative cyclization of grifolic acid to produce DCA, an anti‐HIV meroterpenoid isolated from Rhododendron dauricum (Ericaceae). We identified a novel cDNA encoding DCA synthase by transcriptome‐based screening from young leaves of R. dauricum. The gene coded for a 533‐amino acid polypeptide with moderate homologies to flavin adenine dinucleotide oxidases from other plants. The primary structure contained an amino‐terminal signal peptide and conserved amino acid residues to form bicovalent linkage to the flavin adenine dinucleotide isoalloxazine ring at histidine‐112 and cysteine‐175. In addition, the recombinant DCA synthase, purified from the culture supernatant of transgenic Pichia pastoris, exhibited structural and functional properties as a flavoprotein. The reaction mechanism of DCA synthase characterized herein partly shares a similarity with those of cannabinoid synthases from Cannabis sativa, whereas DCA synthase catalyzes a novel cyclization reaction of the farnesyl moiety of a meroterpenoid natural product of plant origin.Moreover, in this study, we present evidence that DCA is biosynthesized and accumulated specifically in the glandular scales, on the surface of R. dauricum plants, based on various analytical studies at the chemical, biochemical, and molecular levels. The extracellular localization of DCA also was confirmed by a confocal microscopic analysis of its autofluorescence. These data highlight the unique feature of DCA: the final step of biosynthesis is completed in apoplastic space, and it is highly accumulated outside the scale cells. With respect to this unique extracellular biosynthesis and accumulation of DCA, we speculated the possibility that DCA would serve as a chemical defense component outside of the scale cells, the outermost layer of plants. In addition, we demonstrated that DCA and grifolic acid are phytotoxic meroterpenoids that induce cell death in the cell cultures of their producer plant R. dauricum, and thus have to be sequestered in the apoplast to avoid self‐poisoning.Support or Funding InformationThis work was supported by JSPS/MEXT KAKENHI (nos. 15K07994 and 17H05436) to FT, and a Grant‐in‐Aid for JSPS Fellows (no. 17J10178) to MI.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Glutathione Maintenance in Protection against Advanced Glycation End Product Induced Neurite Degeneration in SH‐SY5Y Cells
Authors: Stochelski Mateusz Andrzej , Wilmanski Tomasz Maciej , Burgess John R. 
Published: June 2021

Diabetes mellitus is one of the most common chronic diseases and peripheral neuropathy (PN) affects at least 50% of diabetic patients. Medications available for patients ameliorate symptoms (pain) but do not protect against cellular damage, and come with severe side effects leading to discontinued use. In our lab we use differentiated SH‐SY5Y cells treated with advanced glycation end products (AGE) as a model to mimic diabetic conditions and to study the mechanisms of oxidative stress (OS) mediated cell damage and antioxidant protection. Nacetylcysteine (NAC), a common supplement, was previously shown in our lab to fully protect against AGE‐induced damage. Here we report that NAC confers protection by maintaining intracellular glutathione (GSH), suggesting that GSH plays a critical role in neurite protection. Our lab has also shown that 3H‐1,2‐dithiole‐3‐thione (D3T), a cruciferous vegetable constituent and potent inducer of nuclear factor (erythroid‐derived 2)‐ like (Nrf2), can significantly increase cellular GSH concentrations and protect against H2O2–induced cell death. Paradoxically, D3T conferred no protection against AGE‐induced cell death or neurite degeneration. Furthermore, D3T increased reactive oxygen species (ROS) generation, depleted GSH, and increased glucose‐6‐phosphate dehydrogenase (G6PD) expression, suggesting its role as an NADPH generating agent may promote OS rather than provide protection. Inhibition of NADPH generation via G6PD, the protein product of a Nrf2‐responsive gene, with dehydroepiandrosterone was found to protect against AGE‐induced ROS generation, loss of viability, and neurite degeneration. These results suggest that using a strategy to protect against oxidative stress by upregulation of the endogenous antioxidant defense system via Nrf2 may backfire and promote further damage if the products of upregulation contribute to ROS generation.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Citrus polymethoxyflavones (PMFs) inhibit 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) induced mouse skin JB6 P+ cell transformation through the activation of Nrf2 pathway
Authors: Su Zheng‐Yuan , Pan Guan‐Wu , Tsai Chung‐Liang , Wu Hao‐Xuan , Wei Guor‐Jien 
Published: June 2021

Oxidative stress is one of the important factors to induce carcinogenesis, including skin cancers. Many bioactivities of polymethoxyflavones (PMFs), such as tangeretin and nobiletin, in citrus peels have been studied. In this study, the cancer chemopreventive potential potential and mechanism of tangeretin and nobiletin metabolites with low cytotoxicity were investigated. The results showed that PMF‐2, a tangertin derivate, at 50 μM showed the strongest activity to trigger Nrf2 pathway in human hepatoma HepG2‐C8 cells that were stably transfected with Nrf2‐antioxidant response element (ARE)‐luciferase plasmids. PMF‐2 also significantly suppressed 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA)‐induced anchorage independent growth of mouse skin JB6 P+ cells. Additionally, PMF‐2 increased mRNA and protein expression of Nrf2 and Nrf2 downstream genes such as heme oxygenase‐1 (HO‐1), NAD(P)H dehydrogenase quinone 1 (NQO1), and UDP glucuronosyltransferase 1A (UGT1A). Furthermore, PMF‐2 might regulate the expression of Nrf2 and Nrf2 downstream genes through regulating DNA methylation and Histone modification, which was elucidated by the determination of DNMTs, HDACs, and KDMs protein expressions, in vitro DNA methylation, methylation‐specific PCR (MSP) of Nrf2 promoter. The relationship between Nrf2 downstream genes and histone H3 acetylation (H3ac) was also analyzed by chromatin immunoprecipitation (ChIP)‐qPCR assay. Therefore, PMF‐2 has skin cancer chemopreventive potential through activating Nrf2‐related pathway that might be regulated by epigenetics modification.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Loss of sex hormones alters indices of prefrontal cortex and hippocampal insulin signaling and increases lipid content in a region‐specific manner independent of cardiac pressure overload in female aortic‐banded Yucatan miniature swine
Authors: Hayward Grant C. , Yang Alex J. , Leblanc Paul J. , Hiemstra Jessica A. , Edwards Jenna C. , Emter Craig A. , Olver T Dylan , MacPherson Rebecca E. 
Published: June 2021

Estimates indicate ~50% of patients with heart failure with preserved ejection fraction (HFpEF) experience dementia. Interestingly, postmenopausal women represent the largest cohort of patients with HFpEF. Impaired insulin signaling in the brain has been implicated in dementia, and may be related to regional changes in the neuronal lipid composition. How such processes are effected by the loss of female sex hormones remains to be elucidated. Using a translationally relevant large animal model of pressure overload‐induced heart failure, we tested the hypotheses that the loss of female sex hormones alters proteins involved in brain insulin signaling, and that such changes are associated with an altered lipid profile. Twenty‐eight female Yucatan miniature swine were divided into four groups (n=7): control (CON), aortic banding (AB; pressure overload model of heart failure), ovariectomy (OVX), and AB with OVX (AB‐OVX). Pigs underwent OVX and AB at 7 and 8 months of age, respectively, and prefrontal cortex and hippocampus tissues were collected at 14 months. Estrogen receptor‐α (p<0.05) content was reduced with OVX in both cortical and hippocampal brain regions. In the cortex, OVX reduced total Akt (p<0.05) and synaptophysin (p<0.10), and in the hippocampus OVX reduced phospho‐Akt Ser473 (p<0.05) and phospho‐ERK (p<0.05). Neuronal lipid composition in the prefrontal cortex and hippocampus revealed increases in total lipids (p<0.10), saturated (p<0.10), and monounsaturated fatty acids (p<0.05) with OVX. The hippocampus had increased total (p<0.10), monounsaturated (p<0.05), and polyunsaturated fats (p<0.10). No changes were observed in OVX for polyunsaturated fats. In conclusion, the loss of female sex hormones alters Akt and MAPK/ERK protein content/pathway activation predominantly in the hippocampus, independent of cardiac pressure overload. Impaired indices of hippocampal insulin signaling were associated with a local accumulation of saturated and monounsaturated fats. Overall, the loss of female sex hormones may contribute to the development of dementia in a translational swine model of pressure overload HF with potential relevance to human HFpEF through metabolic pathways influencing cognition.Support or Funding InformationNIH R01 HL112998, PI: CAEAmerican Heart Association #16POST2776005, PI: TDOAlzheimer Society of Brant, Haldimand, Norfolk, Hamilton, and Halton, PI: REKMThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cross‐talk Between the Gut and the Brain in Rats Subjected to LPS‐Induced Inflammation: Role of α‐Synuclein
Authors: Doursout Marie‐Francoise , Gaskey Gregory , Peng Zhanglong , Bick Diane L. , Smith Keri , Devalottai Ria , Kowada Minoru , Schiess Mya 
Published: June 2021

It is now well established that the involvement of the gastrointestinal tract is of great interest as a contributing factor to the development and progression of neurodegenerative diseases. The dysregulation of the Gut‐Brain‐Axis (GBA) may be associated with gastrointestinal manifestations preceding disorders of cognitive and/or memory and learning process, supporting the hypothesis that the pathological process is spread from the gut to the brain (Mayer et al. 2014). The extensive involvement of the gut in neurodegenerative diseases such as Parkinson's disease, even in its early stages, has led to the evaluation of enteric α‐synuclein as a possible biomarker (Forsyth et al. 2011). Therefore, the overall goal of the proposed study is to assess the time course of the compromised intestinal barrier capable of influencing brain function in rats challenged with LPS. Specifically, we hypothesize that alterations of the intestinal barrier (or leaky guts) enhances inflammation as well as the production of a unique protein α‐synuclein which has characteristic of degenerative diseases. To induce inflammation, Lipopolysaccharide (LPS) was administered intravenously (IV) in rats at a dose of 20 mg/kg (Doursout et al. 2013). A single dose LPS was elected based on previously reported results from a mouse model (Qin et al. 2007). Animals were divided into 2 groups (saline and LPS). Animals were divided into 2 sub‐groups (short term; 1 week and long term 4 weeks). Following sacrifice at each time point, tissues (gut and brain e.g. hippocampus and olfactory bulb) were harvested. We measured edema formation in the gut in rats subjected to LPS as compared to saline by wet/dry ratio. Alpha‐synuclein was determined using ELISA (R&D Systems) in small and large intestines as well as in the brain at each time point. Data are expressed as Mean ± SEM. Our data show that LPS induced edema in the gut, suggesting inflammation. Our data demonstrate that increases in α‐synuclein protein concentrations were time‐dependent in the hippocampus and in the olfactory bulb. We postulate that LPS induce an increase in α‐synuclein concentrations in the gut and spread to the brain at a later time point.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Enhancement of adiponectin ameliorates non‐alcoholic fatty liver disease via inhibition of FoxO1 in type I diabetic rats
Authors: Xie Xiang , Xia Zhengyuan 
Published: June 2021

Non‐alcoholic fatty liver disease (NAFLD) is a common liver disease which is previously associated with Type 2 Diabetes Mellitus (T2DM). Recent research has indicated Type 1 Diabetes Mellitus (T1DM) is also involved in the development of non‐alcoholic fatty liver disease, whereas the underlying mechanisms are not utterly understood. Forkhead box O1 (FoxO1) and adiponectin (APN) have been proposed to play an important role in the processes in NAFLD in T1DM. We herein investigated the effects of FoxO1 and APN on the NAFLD and the underlying mechanism in T1DM. Serum AST and ALT was determined by commercially available kits. Blood glucose levels were measured by One Touch Ultra Glucose Meter. Relevant protein expression was tested by Western blot analysis. We found that serum AST and ALT were both increased in T1DM rats, which was ameliorated by application of APN or block of FoxO1. Moreover, APN and FoxO1 inhibitor both decreased the expression of FoxO1 which was significantly increased in diabetic rats. However, the inhibition of FoxO1 didn't alter the expression of APN and its receptors. We also found that Akt1 expression was significantly declined in diabetic rat which was restored by APN. Together, these results suggest that APN ameliorates NAFLD via inhibition of FoxO1 through Akt1/FoxO1 signaling pathway. The present study demonstrated the effort of T1DM on NAFLD and presented a novel role of APN in the underlying mechanism.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sustained crosstalk between TLR2 and IL‐1R1 exacerbates neurodegeneration in Parkinson's Disease
Authors: Johnson Marguerite Elizabeth , Kortagere Sandhya 
Published: June 2021

Current evidence suggests the causal role of inflammation in the pathogenesis of Parkinson's Disease (PD). PD is a neurodegenerative disorder that predominantly affects dopaminergic neurons within the substantia nigra (SNc) region of the brain and is typically accompanied by the accumulation of alpha‐synuclein containing fibrils called Lewy Bodies (LB). Studies have demonstrated that alpha‐synuclein fibrils function as damage‐associated molecular patterns (DAMPs) to induce an inflammatory response via Toll‐like Receptor 2 (TLR2) activation on both neurons and microglia. The role of TLR2‐mediated inflammation in neurodegeneration is further supported by increased microglial activation, pro‐inflammatory cytokines (e.g. TNF‐a and IL‐1B), and TLR2 protein expression in LBs. Two recent reports have shown that IL‐1R1 antagonism attenuates inflammation and behavioral deficits in PD animal models. Despite these studies, the mechanism by which both TLR2 and IL‐1R1 contribute to neurodegeneration in PD remains elusive. Recently, our lab has demonstrated that activation of TLR2 by PAM3CSK4 induces an increase in IL‐1R1 expression in SH‐SY5Y cells. Further, activation of TLR2 or IL‐1R1 leads to an increase in phosphorylation of p38‐MAPK and ERK1/2 and an increase in cytokine levels. These results suggest a crosstalk between TLR2 and IL‐1R1 may be essential to produce degeneration of the dopaminergic neurons in the substantia nigra. Sustained crosstalk between TLR2 and IL‐1R1 could lead to exacerbation of this proinflammatory response in PD.Support or Funding InformationDrexel UniversityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.