bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis of tetradentate metal Schiff base complexes towards the synthesis of novel antimicrobials
Authors: Guimaraes Guilherme J. , Wolyniak Michael J. , Deifel Nicholas P.
Published: June 2021

Schiff bases are molecules that carry a characteristic imine bond that are derived from carbonyl compounds and are widely studied due to the versatility of their steric and electronic properties. Schiff bases are significant because of their biologically active properties that can improve and develop agrochemicals, anti‐inflammatories, antivirals, and even radiopharmaceuticals for cancer targeting. This study is focused on synthesizing a family of novel metal Schiff base complexes that have potential to act as antimicrobials over a wide range of pathogenic species.

In a previous study, a family of metal Schiff base complexes with an NN'NN coordination sphere was synthesized. This study found that antimicrobial activity using Schiff base complexes with chromium, iron, or manganese as their center showed greater inhibition of Aspergillus sp. growth than a variety of known antimicrobial drugs (Kumar et al., 2012). In an attempt to expand the antimicrobial potential of these compounds, we changed the structure of these Schiff base ligands to utilize an NN'OS coordination sphere with the same set of metal centers. We hypothesized that the chemical activity of the sulfur atom would provide an improved range of antimicrobial function.

Our synthesis of NN'OS coordination sphere‐based antimicrobials began with the compound cyclopentanone. Cyclopentanone was condensed with carbon disulfide, recrystallized, protonated, and methylated with dimethyl sulfate. The resultant methylated product was aminated with two different diamines to form a set of novel primary amines. These primary amines were reacted with salicylaldehydes with different substitutions to form a family of Schiff base ligands. The last step in the synthesis process was to add a transition metal to the Schiff base ligands (chromium, iron, or manganese) through a reflux reaction. After synthesis, each metal Schiff base complex was characterized through 1H and 13C nuclear magnetic resonance, infrared, and gas chromatography‐mass spectroscopy.

With synthesis complete, current work focuses on the examination of these compounds as effective antimicrobials. The compounds are being tested for these properties against Aspergillus furmigatus as a representative of a broad range of Aspergillus species. Characterized Schiff base complexes are being compared for their level of antimicrobial activity against that shown by Miconazole, an established antimicrobial used to treat an array of fungal infections. The results of these tests may establish Schiff base ligands with NN'OS coordination spheres as a novel effective means of treating an array of dangerous microbial infections.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using a High Resolution Melt Curve Assay to Examine the Single Nucleotide Polymorphisms of the HFE Gene
Authors: Ehrke Mariah Kay , Zwiesler‐Vollick Julie
Published: June 2021

Pharmacogenetics is the study of how inherited genetic variations correspond to an individual's specific reaction to medication. Pharmacogenetic testing determines an individual's genotype. This is beneficial in the medical field because it informs doctors and patients about drug efficacy, drug dosage recommendations, and the risk of adverse side effects for their genotype. The purpose of this project is to determine how to use high resolution melting analysis to characterize the different alleles caused by single nucleotide polymorphisms (SNPs). The SNP being tested is in the HFE gene which codes for an iron‐sensing protein which also regulates iron uptake. This SNP has been shown to predict the effectiveness of treatment with Humira® (adalimumab) in patients with Crohn's disease. We are presently developing a protocol for high resolution melting analysis of this SNP using the Roche LightCycler 480. The high resolution melting analysis should be specific enough to differentiate between the different SNPs. We will verify our SNPs results with DNA sequencing. The results of this study will be presented.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Monomethyl Branched Chain Fatty Acids link Mitochondrial Amino Acid Metabolism and Adipose Tissue Lipogenesis to Fatty Acid Diversity
Authors: wallace martina , Green Courtney , Roberts Lindsay , Lee Michelle , Cabrales Pedro , Ayres Janelle , Nomura Daniel , Loomba Rohit , Metallo Christian
Published: June 2021

BCAA's are important substrates for protein synthesis and bioenergetics in mammals, and this metabolic pathway is dysregulated in various disease states. Fatty acid biosynthesis occurs in many tissues, but the balance of lipogenesis in liver and adipose tissue is disturbed in the context of obesity and metabolic syndrome. Here, using in vitro and in vivo stable isotope tracing we demonstrate that these pathways are linked within adipose tissue, where mammalian FAS produces monomethyl branched chain fatty acids (mmBCFAs). BCAA catabolic flux and short chain fatty acids from the gut microbiome initiate fatty acid synthesis in brown and beige adipocytes to influence the diversity of fatty acids present in the body. Diet‐induced obesity decreases BCAA catabolism and adipose tissue de novo lipogenesis such that mmBCFAs are the most reduced fatty acid species detected and mmBCFA abundance correlates negatively with markers of non‐alcoholic steatohepatitis. These results highlight the origin and regulation of mmBCFAs in mammals, which may play functional roles in metabolic homeostasis, nutrient sensing, and membrane function in human disease.

Support or Funding Information

Ajinomoto Innovation Alliance Program Grant.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Antioxidants Protect Diaphragm Function subjected to Ionizing Irradiation
Authors: Zuo Li , Lu Lanchun , Wu Shiyong , Zhou Tingyang
Published: June 2021

Diaphragm can be subjected to undesired radiotoxicity during radiotherapy when targeted tumors are located at the upper bodies such as in breast, stomach and lung cancers. This study tests the hypothesis that ionizing irradiation (IR) can induce reactive oxygen species (ROS) generation as well as functional decline in the diaphragm; while administration of antioxidants may attenuate muscle dysfunction associated with IR exposure. Male C57BL6 mice were irradiated with onetime IR of 10 Gy (6 MV energy, 600 cGy/min dose rate) on the diaphragmatic region. On days 1, 3, 5, 7 after IR, mice were sacrificed and the diaphragms were removed for experiments. Muscle force and extracellular ROS levels were measured during five‐min tetanic contraction. Control group followed the same protocol without IR exposure. Antioxidant‐treated mice received daily intraperitoneal injection of Tiron (200 mg/kg body wt) and N‐acetyl Cysteine (NAC, 100 mg/kg body wt) starting from one day before IR until sacrifice. These mice were sacrificed on days 5 and 7 after IR for muscle function and ROS assay. Our results show that IR exposure compromised diaphragmatic function compared to control on the 5th and 7th days after IR (n ≥ 6; p < 0.05). There was elevated ROS release associated with muscle contraction from days 3–7 after IR as compared to control muscles (n = 6; p < 0.05). In contrast, mice treated with antioxidants demonstrated no muscle functional decline on days 5 and 7 (n ≥ 5; p < 0.05). Further, antioxidants abolished IR‐induced oxidative stress during contraction on 5th and 7th days (n ≥ 10; p < 0.05). These data collectively suggest that 10‐Gy IR exposure can lead to decreased diaphragm function potentially via ROS formation. Antioxidant treatment may be regarded as a promising strategy for diaphragm protection among patients who receive radiotherapy on tissue/organs in proximity to diaphragm.

Support or Funding Information

This study was supported by American Physiology Society S&R Foundation Ryuji Ueno Award.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metabolic Control of Two Dynamic Pools of Diacylglycerol in Budding Yeast
Authors: Ganesan Suriakarthiga , Tavassoli Marjan , Sosa Maria Laura , Wagner Kelsey , Terebiznik Mauricio , Zaremberg Vanina
Published: June 2021

The location of lipids and their distributions across cellular membranes have critical biological consequences, particularly for lipids involved in cellular signaling. In this study, diacylglycerol pools were monitored in budding yeast under conditions where lipid homeostasis was altered. Two predominant pools of diacylglycerol were visualized using the C1 domain of mammalian PKCδ fused to GFP. One pool was associated with vacuolar membranes and the other localized to sites of polarized growth. Upon growth resumption, diacylglycerol pools appeared more dynamic than those of phosphatidic acid and phosphatidylserine. During this period, diacylglycerol enriched puncta and vacuolar rings experienced constant morphological changes with clusters of lipid droplets closely attached. Lack of conversion of diacylglycerol to phosphatidate by Dgk1 led to the accumulation of diacylglycerol in a vacuolar associated compartment. Diacylglycerol distribution was strongly affected in cells lacking the phosphatidylserine synthase Cho1/Pss1. Supplementation of lysophosphatidylserine to cho1 cells did not correct mislocalization of DAG, pointing to a role for phosphatidylserine synthesis and traffic in the establishment of cellular diacylglycerol pools.

Support or Funding Information

This work was supported by a National Science and Engineering Research Council of Canada to M.R. Terebiznik and V. Zaremberg.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Uncovering Clinically Relevant Mutations in Membrane Transporters by Genetic Analysis Linked to the Determination of Erythrocyte Membrane Protein Expression
Authors: Sarkadi Balazs , Zambo Boglarka , Bartos Zsuzsanna , Mozner Orsolya , Szabo Edit , Varady Gyorgy , Homolya Laszlo
Published: June 2021

The human red blood cell (RBC) membrane contains more than 300 integral membrane proteins, many with high relevance to disease conditions or pharmacological interventions. We have developed a flow cytometry method for the quantitative determination of the RBC membrane expression levels of selected membrane transporters, and examined their molecular genetic background, as well as their potential value in medical diagnostics as biomarkers. One example presented is the ABCG2 xeno‐ and endobiotic transporter, modulating the absorption and metabolism of pharmacological agents and causing multidrug resistance in cancer. ABCG2 is also involved in uric acid elimination and its impaired function is causative in gout. Analysis of ABCG2 expression in the erythrocyte membranes of healthy volunteers and gout patients showed an enrichment of lower expression levels in the gout patients, correlating with a genetic polymorphism, causing a Q141K variant protein. In addition, based on RBC expression levels, we found stop and frameshift mutations, as well as a relatively frequent, novel ABCG2 mutation (ABCG2‐M71V). This variant, according to cellular expression studies, causes reduced protein expression, although with preserved transporter capability. ABCG2‐M71V expression in vitro could be corrected by therapeutically relevant small molecules, with a potential in allele‐specific gout treatment. Another example is the plasma membrane calcium pump (PMCA4b) of the RBC membrane, which showed heterogeneous RBC expression levels in healthy volunteers. By genetic analysis we found a correlation of the low PMCA4b expression levels with a minor haplotype in the erythroid cell specific regulatory region of this protein. The respective regulatory regions in this minor haplotype have been characterized in detail. These results suggest that personalized medicine should consider the genetic analysis linked to RBC membrane protein expression, and these studies may provide a new tool to uncover clinically important variants of membrane proteins.

Support or Funding Information

Supported by OTKA K115375, K111678 and FIEK 16‐1‐2016‐0005, NKTH, grants.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phosphatidylinositol (4, 5)‐bisphosphate coordinates functional interactions in the dopamine transporter to promote amphetamine behaviors
Authors: Aguilar Jenny , Shekar Aparna , Matthies Heinrich , Galli Aurelio
Published: June 2021

The psychostimulant amphetamine (AMPH) mainly mediates its pharmacological and behavioral effects by increasing extracellular dopamine (DA) availability. DA homeostasis is maintained by the dopamine transporter (DAT), a presynaptic membrane protein that mediates the high‐affinity reuptake of released DA from the synaptic cleft. We have previously demonstrated that AMPH induces N‐term phosphorylation of the DAT, which leads to transport‐mediated efflux of DA. Furthermore, we have shown that phosphatidylinositol (4, 5)‐bisphosphate (PIP2) directly interacts with the DAT and facilitates AMPH‐induced DA efflux, but is not required for DA uptake. Specially, PIP2 binds DAT through electrostatic interactions with positively charged DAT N‐terminal residues. Disrupting the interactions between DAT and PIP2 or depleting PIP2 diminishes reverse transport (efflux) of DA. Previous studies on the human serotonin transporter show that non‐N‐terminal PIP2 binding sites (within the fourth intracellular loop) are essential for AMPH‐induced reverse transport of serotonin, but not substrate uptake. Together, these data led us to hypothesize that PIP2 coordinates functional interactions between the N‐terminus and the fourth intracellular loop to promote reverse transport of DA. Here, we demonstrate that depletion of PIP2 after DAT N‐terminus phosphorylation does not affect AMPH‐induced DA efflux, suggesting that N‐terminus phosphorylation is the limiting step of reverse transport of DA. Notably, we show that residue R443, which resides in the fourth intracellular loop, also modulates AMPH‐induced DA efflux. Thus, we focus on the role of the interaction between the DAT N‐terminus and R443 in mediating the reverse transport of DA. Furthermore, we translate our molecular discoveries in vivo by using a coordinated genetic and pharmacological approach to study AMPH‐induced behaviors in Drosophila melanogaster

Support or Funding Information

1F31MH114316 (JIA), AHA Predoctoral Award (AS), DA035263(AG)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Automated detection of GPCR exocytic events reveals distinct recycling populations
Authors: Weinberg Zara Yael , Phan Tiffany , Puthenveedu Manojkumar A.
Published: June 2021

G protein‐coupled receptors (GPCRs) are flexible scaffolds that allow cells to detect thousands of environmental signals. After binding ligand, receptor endocytosis enables cellular desensitization, after which the internalized receptor is sorted in the endolysosomal system for either degradation or recycling back to the plasma membrane. Throughout the postendocytic pathway, GPCRs continue to signal with different signals being attributed to differential endosomal localization of receptors during this process. Attempts to measure a given GPCR's recycling pathway have required perturbation of postendocytic dynamics through overexpression of specific marker proteins. We sought to understand receptor trafficking through the postendocytic pathway by looking for quantitative differences in individual recycling events that contained specific receptor cargos. Using three exemplar cargo –the GPCRs mu opioid (MOR) and B2‐adrenergic receptor (B2AR), and the classic endocytic cargo transferrin receptor (TfR) – we used total internal fluorescence microscopy to visualize individual exocytic events. We then analyzed these events by developing an automated analysis program which used supervised machine learning to discern and quantify discrete receptor exocytic events. We compared overall fluorescence intensity, event duration, and other quantitative characterizations of individual events. We discovered that TfR and B2AR recycle in a similar manner, while MOR recycled in a distinct manner with markedly less heterogeneity in its recycling population. These findings show that GPCR recycling offers another dimension through which receptors distinguish themselves, and suggest the value of measuring recycling as a potential means of dissecting a given receptor's signaling profile. Further, the tools developed in this study can be used to easily characterize the recycling phenotype of novel receptors, and can be extended to a variety of exocytic cargos.

Support or Funding Information

Supported by University of Michigan

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Myosin IIa is the most important effector of zyxin in cAMP‐mediated endothelial exocytosis
Authors: Wei Guoqin , Li Pin , Chen Liangyi , Luo Jincai
Published: June 2021

Endothelial exocytosis, one of the first lines of protection against vascular injury, is critical to maintaining the homeostasis of the blood vessels. Weibel‐Palade bodies (WPBs)1, endothelium‐specific secretory granules, deliver hemostatic and inflammatory mediators including von Willebrand factor (VWF) and P‐selectin in response to a variety of agonists2. Recently, using live‐cell super‐resolution microscopy3, we visualize the dynamic interaction between fine actin filaments and WPBs and found previously unappreciated reorganization of pre‐existing actin filaments around WPBs before fusion4. In addition, we show that focal adhesion protein zyxin mediates the formation of actin frameworks on exocytic WPBs4. However, the detailed mechanism how zyxin regulates the reorganization of actin filaments is still unknown.

In this report, we combined protein interactomics, genetic screening, high‐resolution imaging, and animal models, to show that the direct interaction between myosin IIa and zyxin is essential for cAMP‐mediated endothelial VWF secretion.

Using affinity purification and co‐immunoprecipitation assay, we found that myosin IIa is the most abundant interacting protein of zyxin. Surprisingly, besides the zyxin‐related function on exocytosis, myosin IIa plays vital roles in the regulation of blood vessels homeostasis. Downregulation of myosin IIa by shRNA significantly suppresses both forskolin‐ and epinephrine‐induced VWF secretion. Using an inducible endothelial specific myosin IIa null mice model, we observed impaired epinephrine‐stimulated VWF release, prolonged bleeding time and thrombosis from those mice. Mechanistically, we found that in resting cells, the loss of myosin IIa, inhibition of myosin activity and blocking the phosphorylation of myosin IIa at S1943 decreased the peripheral localization of Rab27‐positive WPBs along stress fibers. Following stimulation with cAMP activating agents, myosin IIa interacted with zyxin and this required the phosphorylation of S1916 on myosin IIa mediated by CK2 to support the formation of functional actin framework, facilitating fusion and subsequent exocytosis.

In summary, our work revealed that myosin IIa is the most important interacting protein of zyxin, which couples the maintenance of mature WPB in peripheral region to the promotion of VWF secretion in synergy with zyxin via spatiotemporally distinctive actin remodelling using different phosphorylation sites (As shown in Figure 1). Our findings implicate myosin IIa as an important regulator of blood homeostasis under pathophysiological conditions.

Support or Funding Information

the National Science Funds(No. 91339111, No. 81470298, and Project 31521062), the Major State Basic Research Development Program of China (No. 2012CB945103), and the National Science and Technology Support Project (grant no. 2014BAI02B01).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of Bacterial Cell Wall Fragment Recognition by the Yeast Protein CYR1p
Authors: Scanlon Daniel J. , Burch Jason , Grimes Catherine
Published: June 2021

Within the human body are trillions of microorganisms which must maintain a proper balance with the human host as well as amongst each other. These organisms must be able to accurately identify and respond to the bacterial cells that surround them. Pattern recognition receptors (PRRs) can bind pathogen associated molecular patterns (PAMPs) and enable these microorganisms to recognize their surrounding microbiome. Similarly, in yeast species Candida albicans the adenylyl cyclase CYR1p is found to bind muramyl dipeptide (MDP) a fragment of the bacterial cell wall or peptidoglycan. Bacterial cell wall fragments like MDP are interestingly found to trigger a phenotypical change in C. albicans to enter pathogenic hyphal growth through a cascade initiated by the binding of MDP to CYR1p. Another member of the Candida family, Candida glabrata has been found to not experience the same phenotypical switch to hyphal growth observed in C. albicans in the presence of MDP despite both species being of the same family and both commonly involved in yeast infections. The Leucine Rich Repeat (LRR) domain specifically has been shown to be the site of MDP binding within the protein CYR1p in C. albicans. To understand the differences in the mechanism of peptidoglycan recognition by C. albicans and C. glabrata, the LRR domain of C. glabrata was expressed and purified. Analysis of the binding or lack thereof between MDP and the LRR domain of CYR1p from C. glabrata could help to elucidate why C. albicans has hyphae growth triggered by MDP but C. glabrata does not.

Support or Funding Information

University of Delaware Department of Chemistry and Biochemistry

This 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 Lipoprotein and Insulin Receptors in the Retina of Diabetic Mice
Authors: Sherman Josiah , El‐Desoky Dalia , Freter Robert , Adams Ryane , Mbony Hannah , Joseph Tiffany , Heron David , Hossain Ahamed , Bhattacharjee Partha
Published: June 2021

Diabetic retinopathy (DR) is a visual complication of diabetes mellitus and a leading cause of visual impairment across the world. Retinal neurodegeneration has been shown to precede the vascular abnormalities of the retina that are used as key clinical indicators of DR. An important feature of retinal neurodegeneration associated with DR is apoptosis of the retinal ganglion cells (RGCs). Our previous in vivo studies with genetically modified db/db mice (mimicking human type 2 diabetes mellitus) indicate a lipid metabolism‐regulated signaling pathway involving lipoprotein receptor‐related protein 1 (LRP‐1) protecting RGCs against apoptosis. Specifically, activation of LRP‐1 by ligand apoEdp (dimerized LRP‐1‐binding apolipoprotein E derived peptide) regulates the PI3K/Akt pathway in the retina, which appears to be involved in cross‐talk with insulin receptor and N‐methyl‐D‐aspartate receptor (NMDAR) signaling. Excess NMDAR activation by neurotransmitter glutamate in DR promotes glutamate excitotoxicity, leading to apoptosis of the cell and thinning of the neuroretina. Recent in vitro studies using cultured RGCs from neonatal mice were conducted and suggest the role of LRP‐1 in regulating glutamate‐induced excitotoxicity in RGCs. Our preliminary data from these in vitro studies suggest that LRP‐1 stimulation via apoEdp results in reduced glutamate‐induced excitotoxicity in RGCs.

Support or Funding Information

NIH BUILD TL4GM118968‐04NIH BUILD 5RL5GM118966‐04NIH MARC 2T34GM007716‐38

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impaired Chemoreflex Response to Acute Hypoxia Correlates with Decreased Activation of the Medial Nucleus Tractus Solitarii in the STZ‐Induced Rat Model of Alzheimer's Disease
Authors: Brown Andrea G. , Thapa Mahima , Hooker John W. , Ostrowski Tim D.
Published: June 2021

Background

Alzheimer's disease (AD) is the most common cause of dementia and clinically diagnosed largely based on cognitive decline. Most AD patients also present with disorders of respiration, swallowing, and autonomic function. Current studies suggest that pathological changes in the brainstem manifest simultaneously or even before those in forebrain regions, and may account for non‐cognitive symptoms. Identifying the pathologic changes in brainstem centers may elucidate the mechanisms behind cardiorespiratory dysfunction in AD. This study concentrated on the respiratory deficits. Using the streptozotocin (STZ)‐induced model of AD, we determined cell activation in response to acute hypoxia in the nucleus tractus solitarii (nTS), a brainstem area that is integral for chemoreflex function.

Methods

Sporadic AD was induced in Sprague Dawley rats (280 ± 17g, n = 6 rats/group) by intracerebroventricular injections of streptozotocin (1.5 mg/kg STZ in citrate buffer; day 1 and 3). Vehicle injections served as control (CTL). Peripheral chemoreflex function was tested with two hours of acute hypoxia (10% O2) in a plethysmography chamber. Immediately following the hypoxic period, the brains were fixed with 4% paraformaldehyde. Four representative areas of the nTS were immunohistochemically analyzed for cell activation (c‐Fos).

Results

The chemoreflex‐mediated increase of minute ventilation in response to acute hypoxia was significantly blunted by ~20% in STZ rats when compared to CTL (p = 0.03). The diminished response continued over the entire 2 hours of hypoxia and was mainly due to alterations in respiratory rate. Immunohistochemical analysis revealed a ~25% decrease of hypoxia‐mediated cell activation in the medial nTS of STZ rats (−14.04 mm from bregma, p = 0.02). This decline in cell activation was not due to a general decrease in cell density as shown by DAPI staining. The level of activation in commissural and rostral nTS sections was not different from CTL.

Conclusion

We had previously found respiratory dysregulation in the STZ model of sporadic AD. This follow‐up study analyzed respiratory parameters over a two‐hour hypoxic period and the underlying pattern of cell activation in the nTS. Our data document a reduced activation of the medial nTS in the chemoreflex pathway of AD rats. This finding may help explain the respiratory symptoms observed in AD patients.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
pharmABCology: an initiative to create logical mnemonics systems for learning drug names
Authors: Liang Willmann
Published: June 2021

With a handful of comprehensive pharmacology textbooks on the market, students in any healthcare discipline are likely able to find at least one book that suits their individual needs. The more challenging part for students, however, is to become better acquainted with drug names. Textbooks seldom provide meaningful ways to help readers remember drug names and/or their mechanisms. Existing mnemonics, not widely used among students in general, may lack in providing the learner with an intuitive or logical approach to mentally connect with the drug names. For example, names of persons, objects or action verbs were chosen by random to represent a particular drug class or therapeutic drug use. The pharmABCology project was conceived with the final aim of creating logical mnemonics systems for drugs that students will commonly encounter in their studies. In this pilot presentation, two approaches, one based on the alphabet system and the other based on pharmacological mechanisms, were demonstrated in the mnemonics for four classes of drugs: 1) anti‐dyslipidaemics, 2) anti‐dysrhythmics, 3) anti‐hypertensives and 4) anti‐ischaemic drugs. For the alphabetical approach, the names, drug classes or mechanisms of anti‐dyslipidaemics and anti‐hypertensives are listed in alphabetical order. The other approach takes advantage of the spellings of anti‐dyslipidaemic and anti‐dysrhythmic drug names in formulating a meaningful sentence or phrase that is readily associated with the mechanisms of actions of these drugs. Some of the mnemonics systems above had been used in a sample (n = 30) of students in undergraduate MBChB tutorials, and the feedback was overwhelmingly satisfactory based on a 5‐point Likert scale. Of the 22 respondents (73 % response rate), 10 (45 %) and 9 (41 %), respectively, strongly agreed and somewhat agreed that pharmABCology mnemonics system helped with the learning of drug names. In light of the early promising results, similar approaches will be used to develop mnemonics for other drug classes with the intention of publicising this learning tool to the pharmacy, nursing and students in other healthcare‐related disciplines.

Support or Funding Information

Nil

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Will Use Of Cerego™ Spaced‐Learning And Self‐Test Sets Improve Medical Students' Performance On Summative Examination Pharmacology Questions?
Authors: Pham Grant , Quest Dale , Baatar Dolgor
Published: June 2021

PURPOSE

Spaced learning means studying at intervals, and in contrast to cramming, seems to enhance knowledge retention. Electronic software applications are available for medical students to utilize that approach. Our aim was to determine if spaced learning modules administered via the free‐to‐use Cerego™ learning platform might improve performance on pharmacology questions in the Scientific Principles of Medicine course unit summative exam.

METHODS

Medical students were recruited from the Texas Tech University Health Sciences Center at El Paso, Paul L. Foster School of Medicine first‐year Class of 2020. The integumentary‐musculoskeletal‐peripheral nervous system (IMN) unit immediately preceded the cardiovascular‐respiratory (CVR) unit. No Cerego™ self‐test modules were created for IMN unit pharmacology sessions. Teaching materials for each of 7 CVR unit pharmacology sessions were used to generate 7 Cerego™ self‐test modules, each containing 10–45 questions approved by a faculty member, then made available to students on the Cerego™ learning platform. Cerego™ logs student attempts, time to complete a module, and an index of perceived difficulty. Subjects had unlimited access to Cerego™ while preparing for the CVR exam.

RESULTS

One‐way ANOVA showed students performed better on the ten pharmacology multiple‐choice questions on the CVR summative examination than they did on the eight multiple‐choice questions on the IMN summative examination. Linear regression analysis showed no significant dependence of students' performance on CVR summative exam pharmacology questions on scores averaged for all seven Cerego™ sets (R2: 0.0057).

CONCLUSION

The results of this study question the potential for use of Cerego™ sets to enhance retention of pharmacology content. Similar projects showed Cerego™ sets improved anatomy and micro summative scores. The disparate results of these studies provide impetus to see whether using Cerego™ helps students improve knowledge retention and exam performance in behavioral science, biochemistry, genetics, histology, immunology, neuroscience, pathology and physiology.

Support or Funding Information

All research was done on campus without financial assistance from outside sources. The authors declare no conflict of interest.
Lack of Evidence for Methylenedioxypyrovalerone (MDPV)‐Induced Persistent Dopaminergic or Serotonergic Deficits: Comparison with Methamphetamine and Mephedrone
Authors: Magee Charlotte P. , German Christopher L. , Curtis Peter S. , Hanson Glen R. , Fleckenstein Annette E.
Published: June 2021

β‐Ketoamphetamines such as methylenedioxypyrovalerone (MDPV) and mephedrone (MEPH) are synthetic cathinones with a structural similarity to methamphetamine (METH) that may account for some similarities in behavioral effects. However, the persistent effects of repeated high‐dose MDPV exposures on striatal dopaminergic and hippocampal serotonergic systems remain to be fully elucidated. Accordingly, the persistent effects of repeated high‐dose MPDV administrations were investigated. Results revealed that MDPV treatment did not alter striatal dopaminergic or hippocampal serotonergic systems as assessed 7 days later in synaptosomes prepared from treated rats. Further, MDPV did not alter hippocampal serotonin or 5‐hydroxyindoleacetic acid content at this time point. These data stand in contrast to our previous report that repeated high‐dose MEPH administrations cause persistent hippocampal serotonergic deficits. These data also stand in contrast to effects of repeated high‐dose METH administrations that include persistent striatal dopaminergic and hippocampal serotonergic deficits. Noteworthy, in these studies MDPV did not induce hyperthermia to the degree reported in previous studies of MEPH and METH. Further, the impact of MDPV on the dopamine transporter and vesicular monoamine transporter‐2 appears distinct from that of MEPH and METH. Taken together, these latter findings may contribute to the differential impact of MDPV, MEPH, and METH on dopaminergic and serotonergic systems.

Support or Funding Information

DA039145 and DA031883

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cannabidiol, a non‐psychoactive marijuana component, exhibits antioxidant and neuroprotective effects in neuronal Alzheimer's cell model SH‐5Y
Authors: Briggs Gwyneth Helen
Published: June 2021

Cannabidiol, a non‐psychoactive marijuana component, is a pleotropic PPAR‐gamma agonist which exhibits antioxidant and neuroprotective effects in several experimental models. Amyloid beta (Aβ), the precursor to extracellular senile plaques, has been implicated in the development of cognitive impairment and neuronal cell death. The effect of Cannabidiol against AD pathology has been linked to its ability to attenuate Aβ and tau aggregation in vitro. The present study evaluates the neuroprotective action of Cannabidiol on Aβ‐induced oxidative stress and memory loss. Differentiated SH‐SY5Y neuronal cell lines were pretreated with cannabidiol followed by Aβ treatment for 24 hours. SHSY5Y cells treated with Aβ exhibited increased reactive oxygen species and lipid peroxide levels. Enzymatic antioxidants including superoxide dismutase, catalase and glutathione reductase were decreased in the Aβ treated group when compared to the control group. Aβ treatment also increased the expression of total tau as well as phosphorylated forms of tau (CP13, S202/205; PHF1, S396/404). Expression of PSD‐95 and Arc proteins, essential for synaptic maturity and plasticity, was decreased by Aβ treatment. Cannabidiol treatment attenuated the accumulation of lipid peroxide levels, up‐regulated the antioxidant activities and improved the expression of memory‐associated proteins in Aβ treated SHSY5Y cells. Mechanistic study using SH‐SY5Y cell line transfected with APP695 (SH‐SY5Y‐APP) to investigate the Aβ ameliorating effects of cannabidiol showed no changes in protein levels of APP, α‐secretase or gamma secretase activity. In contrast, cannabidiol decreased β‐secretase levels and inhibited its activity thereby leading to reduced Aβ levels. These results suggest that Cannabidiol is capable of improving antioxidant activity, mitochondrial energy regulation, while decreasing Aβ, thereby indicating it to be a lead compound for AD drug development.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Co‐activation of adenosine and glutamate receptors modulates signaling in primary brain cell cultures
Authors: Gregory Karen J. , Hellyer Shane D. , Albold Sabine , Leach Katie , May Lauren T.
Published: June 2021

G protein‐coupled receptors (GPCRs) represent the largest class of targets for current therapeutics. Two distinct modulatory GPCRs within the CNS, the adenosine A1 receptor (A1AR) and the metabotropic glutamate receptor subtype 5 (mGlu5), have been implicated in Alzheimer's disease (AD) pathology and as potential targets to treat cognitive impairments and disease progression. Both GPCRs are present on the same CNS‐resident cell types, e.g. neurons and astrocytes, and are expressed in brain regions implicated in AD pathology. GPCR discovery programs generally only consider GPCR activity in isolation, without factoring in the influence of other GPCRs or stimuli present. Of note, both glutamate and adenosine are often present within culture medium and/or released from cultured cells. GPCR cross‐talk and/or heteromerization can introduce pharmacological heterogeneity and offer new avenues for targeted drug development. We sought to test the hypothesis that coincident activation of co‐located GPCRs modulates signaling in primary cultures of CNS resident cells. Primary cultures were derived from striatum and cortices of E16 mice and cultured for 6–8 days. High‐throughput signaling assays (iCa2+ mobilization, inositol phosphate (IP1) and cAMP accumulation) and radioligand binding studies were performed to confirm expression and function of mGlu5 and A1AR in the cultures by assessing the ability of: 1) A1AR selective (MeCCPA) and non‐selective (NECA) agonists to inhibit forskolin stimulation of cAMP accumulation; and 2) mGlu5 selective allosteric agonist (VU0424465) and group I mGlu selective orthosteric agonist (DHPG) to stimulate iCa2+ and IP1 accumulation. Experiments performed in the presence and absence of adenosine deaminase revealed that ambient adenosine was not influencing mGlu5 signalling. Coincident activation of A1AR enhanced mGlu5‐mediated iCa2+ mobilization in response to both orthosteric and allosteric agonists in primary striatal and cortical neuronal cultures. However, A1AR activation had no effect on mGlu5‐mediated IP1 accumulation in cortical neurons. Conversely, coincident activation of mGlu5 had little influence on A1AR‐mediated inhibition of cAMP accumulation, but inhibited AR‐stimulation of ERK1/2 phosphorylation. Collectively, our data demonstrate that coincident activation of mGlu5 and A1AR differentially modulates intracellular signaling pathways. Future work exploring the underlying mechanisms may reveal new strategies for targeting these GPCRs to treat AD and cognitive disorders.

Support or Funding Information

This work was supported by National Health and Medical Research Council of Australia (APP1123722 to KJG and LTM) and Australian Research Council Future Fellowships to KJG and KL.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Seizure‐protective Receptors GPR37L1 and GPR37 Regulate Calcium‐binding Protein S100A5 Expression and Secretion
Authors: Nguyen TrangKimberly Thu , Giddens Michelle , Duong Duc , Hall Randy
Published: June 2021

Objective

The objective of this study is to investigate the mechanisms through which S100A5 expression and secretion is regulated by the receptors, GPR37L1 and GPR37.

Methods

Proteomic data obtained via mass spectrometry of mouse brain lysates was performed through the Emory Proteomics Core and follow‐up studies were performed studying cultured cells.

Summary of results

A mutation in the G protein‐coupled receptor GPR37L1 was recently found in a large family to be associated with a progressive form of epilepsy in which seizures start around puberty and increase in severity with age, ultimately resulting in death in the late teens to early twenties. Studies from our lab have revealed that knockout mice lacking GPR37L1 (and/or the closely related receptor GPR37) exhibit greatly increased susceptibility to seizures. To shed light on the potential mechanistic underpinnings of this phenotype, we performed proteomic analyses of whole mouse brain tissue from wild‐type vs. GPR37L1/GPR37 double knockout mice to identify proteins regulated by the absence versus presence of these receptors. The most dramatically downregulated protein in the knockout versus wild‐type brain tissue was S100A5, a brain‐specific calcium‐binding protein. The reduction of S100A5 expression in the knockout brain tissue was confirmed via Western blot. Furthermore, when S100A5 is co‐expressed with either GPR37L1 or GPR37 in HEK293T cells, a robust increase in secretion of S100A5 was observed. This secretion was receptor‐dependent, as over‐expression of S100A5 alone did not result in detectable S100A5 in the media. To dissect the G protein signaling pathway that mediates this secretion, inhibitors of the different G protein signaling pathways were used to treat cells that co‐expressed S100A5 with either receptor. These studies suggested that GPR37 and GPR37L1 signal through Gs to mediate S100A5 secretion, as treatment with H89, a protein kinase A (PKA) inhibitor, sharply reduces S100A5 secretion from transfected HEK293T cells. These findings may be of general interest because other members of the S100A protein family are known to be secreted and have neuroprotective actions within the brain, but little is known about the mechanisms controlling their secretion.

Conclusions

S100A5 expression and secretion are both regulated by GPR37 and GPR37L1. Our studies reveal that these receptors signal in a PKA‐dependent manner to regulate S100A5 secretion.

Support or Funding Information

Pharmacological Sciences T32 Predoctoral Training Grant (4T32GM008602‐20), Pharmacological Sciences T32 Predoctoral Training Grant (4T32GM008602‐21), NIH Grants R21‐NS91986 and R01‐NS088413

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EET‐agonist Prevents and Reverses Heart Failure in Obesity Induced Diabetic Cardiomyopathy
Authors: Singh Shailendra P. , McClung John A. , Bellner Lars , Schragenheim Joseph , Abraham Nader G.
Published: June 2021

Previously we demonstrated that an EET‐agonist has pleiotropic effects including inhibition of Soluble Epoxide Hydrolase expression and reversed cardiomyopathy by decreasing inflammatory molecules and increasing antioxidant signaling. We hypothesized that an EET agonist would increase PGC‐1α, which controls mitochondrial function and HO‐1 induction and negatively regulates the expression of the proinflammatory adipokine NOV in both cardiac and pericardial tissues. This pathway would be expected to further improve left ventricular systolic function as well as increase insulin receptor phosphorylation. The effect of an EET agonist was measured in cardiac and pericardial adipose tissues in db/db mice on oxygen consumption, fractional shortening, blood glucose levels, thermogenic and mitochondrial signaling proteins. Control db/db mice developed signs of metabolic syndrome including insulin resistance, hypertension, inflammation, LV dysfunction, and increased NOV expression in pericardial adipose tissue. EET agonist decreased pericardial adipose tissue expression of NOV, normalized FS, increased PGC‐1α, HO‐1 levels, insulin receptor phosphorylation and improved mitochondrial function. HO‐1‐ derived carbon monoxide (CO) levels, mitochondrial function (p<0.01), insulin receptor phosphorylation (p<0.05), ATP levels and VO2 (p<0.05). Deletion of PGC‐1α reversed all of the above beneficial effects, increasing the expression of NOV and inflammatory cytokines in pericardial adipose tissue. These studies demonstrate that an EET agonist increases insulin receptor phosphorylation, mitochondrial and thermogenic gene expression, decreases cardiac and pericardial tissue NOV levels, and ameliorates cardiomyopathy in an obese mouse model of the metabolic syndrome. Further, this study demonstrated that EET‐mediated inhibition of NOV is a potential therapeutic target for the prevention of heart failure and the development of metabolic syndrome.

Support or Funding Information

National Institutes of Health grant (HL34300 to NGA)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Podocyte‐Specific Deletion of Asah1 Gene Produced Podocytopathy without Sclerotic Pathology: A Potential Mouse Model for Steroid‐Insensitive Minimal Change Disease
Authors: Li Guangbi , Lohner Hannah , Dempsey Sara , Yuan Xinxu , Zhang Qinghua , Gehr Todd , Ritter Joseph , Li Pin‐Lan
Published: June 2021

Lysosomal acid ceramidase (AC) has been shown to be critical for the ceramide hydrolysis and thereby regulates lysosome function and related cell function such as autophagy and vesicle/molecular trafficking. Despite that ceramide is involved in the regulation of glomerular function, it remains unknown whether AC activity contributes to the control of podocyte function and whether its deficiency causes glomerular disease. In the present study, we generated a mouse line with podocyte‐specific deletion of α subunit in Asah1 (AC gene code in mouse) gene, a main subunit for its activity in lysosomes using Cre‐Lox recombination technology. This Asah1fl/fl/PodoCre mouse colony was characterized by several genetic, molecular and biochemical approaches. AC α subunit was not detectable in podocytes in glomeruli of Asah1fl/fl/PodoCre mice, compared with their control littermates (Asah1fl/fl/WT and WT/WT mice). Using LC‐MS/MS, remarkable ceramide accumulation was detected in isolated glomeruli of Asah1fl/fl/PodoCre mice, but not in their control littermates (increased from 42.99 pg/glomerulus to 237.19 pg/glomerulus). Although Periodic acid‐Schiff staining showed no significant increase in glomerular damage index in Asah1fl/fl/PodoCre mice compared with other two types of mice, severe proteinuria and albuminuria were found in Asah1fl/fl/PodoCre mice started at 8 weeks old (increased from 28.84 μg/24h/gbw to 181.41 μg/24h/gbw of urinary protein excretion, increased from 1.22 μg/24h/gbw to 10.39 μg/24h/gbw of urinary albumin excretion). Correspondingly, glomerular permeability to albumin was markedly increased by podocyte‐specific deletion of AC α subunit. Moreover, hypoalbuminemia and edema were observed in Asah1fl/fl/PodoCre mice when they grew to 12 weeks old. Under transmission electron microscope, Asah1fl/fl/PodoCre mice were found to have foot process effacement, vacuolation, and microvillus formation in podocytes. Treatment with dexamethasone (DEX) for 4 weeks failed to reverse podocyte injury and proteinuria in Asah1fl/fl/PodoCre mice. Immunohistochemistry showed no changes in α‐dystroglycan, but decreases in β‐dystroglycan in glomeruli of Asah1fl/fl/PodoCre mice, compared with their control littermates. These results suggest that lysosomal AC and associated ceramide hydrolysis is essential for the maintenance of podocyte structural and functional integrity and that the defect of AC expression and its function deficiency may result in podocytopathy and related glomerular disease such as steroid‐resistant minimal change disease.

Support or Funding Information

This study was supported by NIH grants DK54927 to Pin‐Lan Li and DK102539 to Joseph K. Ritter.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.