bluuebunny/crossref_metadata_2025 huggingface dataset:
Dissection of the Erv41‐Erv46 retrieval pathway suggests a redox‐regulated mechanism
Authors: Keiser Kristofer , Shibuya Aya , Barlowe Charles
Published: June 2021

The early secretory pathway consists of ER‐ and Golgi‐resident proteins involved in synthesis, folding, quality control, and post translational modifications of secretory proteins. Anterograde flux through this pathway can lead to the mislocalization of ER‐resident proteins as they passively leak to the Golgi. Fortunately there are several retrieval mechanisms to return mislocalized ER‐residents back to the ER. One such retrieval process identified by our lab relies on the Erv41‐Erv46 complex, which recognizes specific non‐HDEL ER‐residents in the Golgi and returns them to the ER in a COPI‐dependent manner. The focus of our work is to determine the mechanism by which the Erv41‐Erv46 retrograde receptor binds to escaped ER‐residents in the Golgi and releases this cargo in the ER.

Site‐directed mutagenesis to introduce Erv41 and Erv46 mutations in Saccharomyces cerevisiae revealed key residues involved in retrieval of non‐HDEL ER‐residents. One such series of mutations involves a cysteine rich region of Erv46 where Cys to Ser mutants result in a complete loss of function. This cysteine rich region contains two CXXC motifs typically found in redox proteins. As observed in other redox proteins, the residues between the vicinal cysteines in this region of Erv46 are important suggesting that the redox potential of these cysteines is finely tuned for activity. Furthermore, mutations of a four residue hydrophobic patch within the cysteine rich region as well as a distal basic residue in Erv46 leads to a failure in retrieval of ER‐residents. These residues, but not the cysteines, are conserved in Erv41, however concomitant mutations in Erv41 do not affect retrieval suggesting Erv46 is predominantly involved in binding while Erv41 may play a distinct role.

Based on our results, we propose the redox state of Erv46 is highly dynamic resulting in active and inactive disulfide configurations which can expose or mask a cargo binding hydrophobic patch. Defining the Erv41‐Erv46 binding mechanism may allow for a more complete understanding of ER homeostasis as well as providing a potential target to block pathogenic toxins that exploit Erv41‐Erv46 mediated retrieval for efficient delivery to the ER.

Support or Funding Information

NIH GM52549

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
mTOR‐dependent selective translation rapidly expands lysosome biogenesis, volume and retention capacity during phagocyte activatio
Authors: Botelho Roberto , Hipolito Victoria , Tandoc Kristofferson , Topisirovic Ivan
Published: June 2021

The molecular mechanisms that govern and adapt the number, size, morphology and activities of organelles to suit the needs of the many cell types and conditions that these cells may encounter are at the frontier of cell biology. Lysosomes are organelles that degrade cargo from various routes including endocytosis, phagocytosis and autophagy. Lysosomes have also emerged as a signalling platform that senses and integrates stress signals such as nutrient deprivation with regulatory kinase hubs like mTOR and AMPK to modulate metabolic activity. For phagocytes and antigen‐presenting cells like macrophages and dendritic cells, lysosomes are a kingpin organelle since they are essential to kill pathogens, and process and ultimately present antigens. During phagocyte activation with lipopolysaccharides, lysosomes undergo a striking reorganization, changing from dozens of globular objects to a tubular network, in a process that requires the phosphatidylinositol‐3‐kinase‐Akt‐mTOR signalling axis. Ultimately, lysosome tubulation is thought to promote pinocytic retention and antigen presentation. Here, we show that lysosome tubulation is accompanied by a rapid boost in lysosome volume and holding capacity of phagocyte activation. We reveal that lysosome expansion depends on biosynthesis of lysosomal proteins but is independent of TFEB and TFE3, transcription factors known to scale up lysosome biogenesis. Instead, we show that lysosome transcripts are selectively targeted for enhanced protein translation in an mTOR‐dependent manner. Indeed, lysosome transcripts were enriched in polyribosome fractions after lipopolysaccharide exposure in an mTOR‐dependent manner, while transcripts encoding non‐lysosomal proteins were unresponsive to LPS and/or mTOR inhibition. Collectively, we identified an example of rapid organelle expansion and remodelling driven by enhanced and selective protein translation during phagocyte activation.

Support or Funding Information

Canadian Institutes of Health Research, Canada Research Chair Program, and Ryerson University

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Deciphering the effect of Endoplasmic Reticulum (ER) stress on near‐mitochondrial localized translation
Authors: Maity Shuvadeep , Back Songhee , Rendleman Justin , Vogel Christine
Published: June 2021

Local synthesis of proteins near their active site is a critical cellular phenomenon. Recent studies have demonstrated that a subpopulation of cytoplasmic ribosomes is localized to organelles such as the endoplasmic reticulum and mitochondria. Due to the presence of different membranous compartments within eukaryotic cells, proper mRNA localization or targeted translation is required to focus the expensive process of translation to where it is needed most. During stress while global translation has been downregulated translation of stress responsive proteins as well other essential proteins remains active. During ER stress, substantial amounts of remodeling happen near ER localized translation populations. Due to its physical and functional relation with mitochondria it is also essential to regulate the translation process of mitochondrial destined proteins during the ER stress condition.

In the current study, a digitonin based extraction method has been successfully established to separate the free cytoplasmic ribosome and mitochondria bound ribosome from HEK293 cells and to perform ribosome profiling in order to capture the actively translating polysome populations of free and mitochondria bound translating ribosome populations. This method can successfully separate the two ribosomal populations and it is enriched with known near‐mitochondria localized ribosome. Previous studies identified a population of mRNAs which are translating near mitochondrial surface in yeast and most of them has been found to be conserved in higher organisms. Interestingly, a set of known near mitochondria localized RNA has been found to involve in active translation in our global riboseq data collected from Hela upon ER stress. It indicates a correlation between the near mitochondrial localized translation and ER stress regulation. Specifically, higher expression of different sub units of oxidative phosphorylation complexes has been found to be enriched upon ER stress. This motivates further detailed study to understand organeller cross talk at translation level during stress. Understanding the near mitochondria localized populations in presence and absence of ER stress in higher organisms would be helpful to understand the complex ER‐mitochondrial cross talk events.

Support or Funding Information

NIHR01

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of excessive mitochondrial fission protects the cardiomyocyte against palmitate‐induced lipotoxicity
Authors: Li Sin‐Jin , Chen Ching‐Yi
Published: June 2021

Mitochondria play the major role in maintaining the optimal cardiac function. Recently, mitochondrial dynamics have been connected with cardiovascular diseases. Cardiac oxidative stress and imbalanced mitochondrial dynamics (decreased fusion and excessive fission) are involved in lipotoxicity‐induced cardiomyopathy. Our aim was to investigate the role of mitochondrial dynamic in H9c2 cells under lipotoxicity. Treatment of H9c2 cells (cardiomyoblast) with 200 μM palmitate resulted in decreased cell viability and decreased ATP production. Palmitate treatment for 3 h elevated protein expression of the dynamin‐related protein‐1 (DRP1, a mitochondrial fission protein) and PTEN‐induced putative kinase 1 (PINK1, marker of mitophagy), but 24 h incubation with palmitate reduced expression of mitochondrial fusion‐ (mitofusin‐2, Mfn2), fission‐ (DRP1 and fission 1 protein, Fis1) and mitophagy‐ related proteins. To correct the mitochondrial imbalance on the initiation of lipotoxicity, we inhibited Drp1 to evaluate whether this action restores mitochondrial function. Knockdown of DRP1 resulted in decreased apoptosis in cells after palmitate treatment for 24 h. To further determine the functional effects of knockdown of DRP1 on palmitate stress in a short‐term or long‐term period (plamitate treatment for 1 h or 24 h), we measured cellular respiration using Seahorse analyzer. Treatment of H9c2 with plamitate for 1 h reduced the oxygen consumption rate and extracellular acidification rate during basal respiration, whereas inhibition of DRP1 maintained the mitochondrial functions. This result indicated that the energetic function was shifted to quiescence in palmitate‐treated cells whereas knockdown of DRP1 cell did not have this effect. Palmitate treatment resulted in a remarkable reduction ATP production while knockdown of DRP1 restored their reductions. Non‐mitochondrial oxygen consumption (e.g. ROS generation) was upregulated with palmitate treatment, but DPR1 suppression decreased ROS generation. However, there was lower basal respiration, ATP production and FCCP‐stimulated maximal respiration in both groups following 24 h‐palmitate treatment which indicated that DRP1 knockdown could not restore mitochondrial oxidative function. In conclusion, palmitate disrupted mitochondrial dynamic and induced cell death, whereas inhibition of mitochondrial fission (DRP1) at the onset of lipotoxicity maintained the mitochondrial function and cell survival; however, it did not restore mitochondrial function after prolonged palmitate stress, demonstrating its limitations.

Support or Funding Information

MOST‐104‐2313‐B‐002‐038‐MY3

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Molecular and structural recognition of Listeria cell‐wall glycopolymers by bacteriophage‐encoded SH3b domains
Authors: Shen Yang , Kalograiaki Ioanna , Prunotto Alessio , Dunne Matthew , Sumrall Eric , Cañada Francisco Javier , Loessner Martin
Published: June 2021

Targeting carbohydrate recognition plays a key role in multiple biological processes across all kingdoms. SH3b (or bacterial Src homology 3) domain is known as a conserved carbohydrate‐binding module that is responsible for carbohydrate recognition. However, the molecular and structural mechanisms underlying SH3b‐carborhydrate interaction remain elusive. Previously, we reported that three phage‐encoded SH3b domains (termed CBD500, CBDP35 and CBD025) exhibit distinct binding patterns that correlate with structural variations in Listeria wall teichoic acids (WTAs, or glycosylated polyribitol phosphate). Here, we use CBD500 to investigate how it recognizes specific carbohydrate ligand displayed on the bacterial surface. We unveil the contribution of glycosylation and O‐acetylation of WTA polymers to the binding by surface plasmon resonance analysis. Saturation Transfer Difference (STD)‐NMR further substantiates that disparity in GlcNAc to ribitol connectivity, O‐acteyl group, and glycosylated GlcNAc determine the binding specificity. Crystallography, epitope mapping and docking studies identify key residues that form a binding groove (including Phe175, Trp 198, Asn248, and Ser249) mediating CBD500‐WTA interactions, which are validated by site‐directed mutagenesis. This is the first report providing structural insights into the interactions between SH3b and native WTA polymer, thus guiding further studies on structure‐function relationship of SH3b‐carbohydrate complex, as well as support the potential use of endolysins as novel antimicrobials.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dolichol Phosphate Mannose Synthase: A Glycosyltransferase with Unity in Molecular Diversities
Authors: Baksi Krishna , Zhang Zhenbo , Serrano Negron Jesus E. , Banerjee Dipak K.
Published: June 2021

N‐glycans provide structural and functional stability to asparagine‐linked (N‐linked) glycoprotein, and add flexibility. Glycan biosynthesis is elaborative, multi‐compartmental and involves many glycosyltransferases. Failure to assemble N‐glycans leads to phenotypic changes developing infection, cancer, congenital disorders of glycosylation (CDGs) among others. Biosynthesis of N‐glycans begins at the endoplasmic reticulum (ER) with the assembly of dolichol‐linked tetra‐decasaccharide (Glc3Man9GlcNAc2‐PP‐Dol) where dolichol phosphate mannose synthase (DPMS) plays a central role. DPMS is also essential for GPI anchor biosynthesis as well as for O‐ and C‐mannosylation of proteins in yeast and in mammalian cells. DPMS has been purified from several sources and its gene has been cloned from 39 species (e.g., from protozoan parasite to human). It is an inverting GT‐A folded enzyme and classified as GT2 by CAZy (carbohydrate active enZyme; http://www.cazy.org). The sequence alignment detects the presence of a metal binding DAD signature in DPMS from all 39 species but finds cAMP‐dependent protein phosphorylation motif (PKA motif) in only 38 species. DPMS also has hydrophobic region(s). Hydropathy analysis of amino acid sequences from bovine, human, S. crevisiae and A. thaliana DPMS show PKA motif is present between the hydrophobic domains. The location of PKA motif as well as the hydrophobic domain(s) in the DPMS sequence vary from species to species. For example, the domain(s) could be located at the center or more towards the C‐terminus. Irrespective of their catalytic similarity, the DNA sequence, the amino acid identity, and the lack of a stretch of hydrophobic amino acid residues at the C‐terminus, DPMS has been classified as Type I and Type II enzyme. DPMS belonging to Type II sub‐class has been considered to be a multimeric protein requiring the DPM2 and DPM3 to anchor down the DPM1 to the ER membrane. Our results indicate that the recombinant DPMS from bovine adrenal medullary capillary endothelial cells can be expressed successfully in E. coli as an active enzyme. Furthermore, DPMS is regulated by extracellular signaling and microenvironment. So, we conclude that it is the hydrophobic domain that anchor down DPMS to the ER membrane.

Support or Funding Information

Supported in part by the grant NIH/NIMHD G12MD007583 (KB)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A novel FIJI workflow demonstrates dynamic changes in postnatal respiratory nuclei innervation by Nkx2.2‐ and Olig3‐derived neurons.
Authors: Liu Jillian , Fair Summer , Kaya Behiye , Zuniga Jessica , Mostafa Hasnaa , Alves Michele , Czeisler Catherine , Otero Jose
Published: June 2021

The autonomic nervous system undergoes rapid and significant change in the postnatal period of mammalian development. How the contributions of specific neural lineages to autonomic circuits change and develop during this phase remains to be determined. In this study, we utilized transgenic mouse modeling and immunofluorescent imaging to determine how Nkx2.2‐ and Olig3‐derived neural lineages develop postnatally and, more specifically, how their innervations into three hindbrain respiratory nuclei are altered during this epoch. We generated a novel FIJI workflow, which allowed the systematic and rapid analysis of Nkx2.2‐ and Olig3‐ derived synapses in regions of interest in each of the three hindbrain respiratory nuclei. This workflow provided more precise quantifications in only 1/25 of the time when compared to human quantifications. We demonstrated that Nkx2.2‐derived innervations into the PreBötzinger Complex significantly decreased in the first three weeks of postnatal life while Olig3‐derived afferents showed no such change during the same period. These data demonstrate utility in the application of this novel FIJI workflow to the investigation of neural circuit contributions to the postnatal development of the autonomic nervous system. Further, they indicate that the role of Nkx2.2‐derived neurons in autonomic function and respiration changes during the first three weeks of life.

Support or Funding Information

This work was sponsored by NIH/NHLBI grants R01HL132355, R01HL132355S1, and R01HL132355S2.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cardiac Conduction Disease in Huntington's Disease Mouse Model (BACHD)
Authors: Zhu Yujie , Shamblin Isaac , Ali Sameen , Gray Michelle , Huke Sabine
Published: June 2021

Objective

Huntington's disease (HD) is a progressive neurodegenerative disease caused by a CAG repeat expansion in the gene encoding the huntingtin protein. In addition to the central nervous system, recent evidence indicates cardiac involvement in HD patients. ECG screening in HD patients suggested an increased prevalence of conduction abnormalities and compromised sinoatrial node (SAN) function. Here we tested if the huntingtin gene defect would cause increased arrhythmia susceptibility.

Methods

We used a conditional human huntingtin (HTT) Bacterial Artificial Chromosome (BACHD) transgenic mouse model at 8 months (motor dysfunction present) and 18+ months of age (advanced disease). Fibrosis level was assessed histologically. Surface ECG was measured in mice anesthetized with isoflurane and arrhythmia susceptibility was tested in response to sympathetic and parasympathetic agonists and their combination (autonomic conflict).

Results

In 18+ month‐old mice overall cardiac structure was unremarkable. BACHD hearts were slightly larger, but HW/BW ratio was normal. Baseline ECG recordings showed prolonged QRS duration in BACHD mice. Stimulation with carbachol (parasympathetic agonist) resulted in increased premature atrial contractions (PACs), SAN exit block, tachy‐brady syndrome in BACHD mice. Stimulation with combined sympathetic and parasympathetic agonists (norepinephrine, epinephrine and carbachol) resulted in increased premature ventricular contractions, PACs, bigeminy and couplets in BACHD mice. Some of these events occurred also in younger BACHD mice, but to a lesser extent.

Conclusions/Broader Impacts

The BACHD mice showed signs of cardiac conduction system abnormalities and SA node dysfunction. This was observed in the absence of gross histological changes and during isoflurance anesthesia which dampened the influence of the autonomic nervous system. Stimulation with parasympathetic and sympathetic agonists uncovered increased atrial and ventricular arrhythmia susceptibility. Our study indicates that the huntingtin gene defect may cause progressive cardiac conduction system pathology that might increase the susceptibility to sudden cardiac death.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Alcoholic versus Non‐alcoholic Steatohepatitis: Levels of Expression of Some Proteins Involved in Tumorigenesis
Authors: Nguyen Luan , Masouminia Maryam , Mendoza Alejandro , Samadzadeh Sara , Tillman Brittany , French Barbara , French Samuel
Published: June 2021

Non‐alcoholic steatohepatitis (NASH) is commonly associated with obesity, type 2 diabetes, and/or hypertriglyceridemia, while alcoholic steatohepatitis (ASH) is associated with alcohol abuse. Both NASH and ASH patients can develop cirrhosis and hepatocellular carcinoma if left untreated. However, the rate of tumorigenesis in NASH and ASH appears to be different. Individuals with NASH progress to cirrhosis at a rate of 7–10% annually, when individuals with ASH progress to cirrhosis at a rate of 10–20% annually. Thus, the objective of our study is to determine if there are differences in NASH versus ASH in the levels of different proteins expressed involved in cancer development. The method used was measuring the proteins expressed in liver biopsied sections from NASH and ASH patients using immunohistochemical staining with fluorescent antibodies and then quantitating the fluorescence intensity morphometrically. Normal liver biopsies were used as controls. The 20 proteins tested were parts of the Ingenuity Canonical Pathway of Molecular Mechanisms of Cancer and included: RAP2B, NAIP, FYN, PAK6, SUV39H1, GNAI1, BAX, E2F3, CKDN2B, BAK1, BCL2, DIABLO, RASGRF2, GNA15, PIK3CB, BRCA1, MAP2K1, BIRC3, CDK2, and ATM. For the results, in ASH, the proteins which had upregulated levels of expression were SUV39H1, E2F3, BCL2, BAK1, BIRC3, and GNAI1. In NASH, the proteins which had upregulated levels of expression were BAK1 and GNAI1 and the one which had downregulated levels of expression was BCL2. Additionally, the levels of expression for SUV39H1, E2F3, BCL2, BAK1, BIRC3 and GNAI1 were significant upregulated in ASH compared to NASH. These results showed significant differences in ASH compared to normal liver, and significant differences in ASH compared to NASH. We conclude that there are more proteins involved in tumorigenesis in ASH compared to NASH and normal liver, which is consistent with the known tumor development rate in ASH and NASH.

Support or Funding Information

This study was funded by NIH:NIAAA grant # UO1‐021898‐05

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CXCR2 Regulates Hepatic Injury after Bile Duct Ligation by Altering Neutrophil Migration
Authors: Konishi Takanori , Schuster Rebecca , Lentsch Alex
Published: June 2021

Introduction

CXC chemokines and their receptor, CXCR2, are important components for progression of hepatic inflammation. However, the contribution of CXCR2 for cholestatic liver injury have not been elucidated. Neutrophil recruitment in the liver and their releasing reactive oxygen species during cholestasis contribute to liver injury. In the current study, we sought to determine the role of CXCR2 in hepatic injury after bile duct ligation (BDL).

Methods

Wild‐type (WT) and CXCR2−/− mice were subjected to BDL to induce cholestatic liver injury. Serum and liver samples were obtained 14 days after BDL. Serum levels of alanine amino transferase (ALT) and liver myeloperoxidase content (MPO) were measured. Liver sections were stained with hematoxylin and eosin for assessing the extent of liver necrosis. Neutrophils in liver tissue was determined by immunonhistochemical staining for Ly‐6G.

Results

After BDL, CXCR2−/− mice had significantly lower serum ALT level and less necrotic area than WT mice. There was no difference in MPO level in the whole liver, however, the distribution of recruited neutrophils was different between WT and CXCR2−/− mice. In WT mice, neutrophils selectively accumulated to necrotic areas in liver parenchyma and infiltrated into the peri‐biliary lesions. In contrast, in CXCR2−/− mice, neutrophils were randomly scattered in the liver parenchyma, with less concentration of neutrophils in necrotic areas and peri‐biliary lesions.

Conclusion

The data suggest that CXCR2 regulates liver injury after BDL by directing neutrophil migration to specific sites of hepatocyte and biliary cell injury.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Methodology to Determine Food Acquisition Strategies among Homeless Youth
Authors: Harris Cristen , Kloubec June
Published: June 2021

Background

Youth represent a growing, and particularly vulnerable, subgroup of the homeless population in urban areas. Social service institutions, health care centers, and voluntary organizations have responded to this growth by creating a variety of programs to assist homeless youth in obtaining food, shelter, and medical care. Still, many of these individuals struggle each day to get the food they need. Only a handful of studies have examined the nutritional quality of meals available to homeless youth and how they obtain food. Lack of data is primarily due to the difficulty in studying this group due to their transiency and distrust of establishment figures.

Objective

The aim of this study is to create a method to describe how homeless youth access food and the nutritional quality and adequacy of food that they obtain.

Methods

In this cross‐sectional study, investigators orally administer a ~50‐item survey, adapted from published research, to homeless youth ages 14 to 24 in the greater Seattle/Tacoma area. Data collection takes place at multiple settings and organizations in which homeless youth access food. Individuals who meet inclusion criteria are invited to participate at these settings and are offered $10 at the completion of the survey. The survey asks information on general demographics; participants' current housing situation; factors relating to their ability to access food; and the kinds of food they acquire. Anonymity of participants is maintained by obtaining verbal assent rather than signed consent. All survey data is entered by researchers on site using REDCap™ software.

Creating a method to gain a better understanding of how homeless youth obtain food and the nutritional quality of such food can help social services and nutrition programs improve the network of nutrition services and the quality of food available to this vulnerable population.

Support or Funding Information

This study was funded by a Bastyr University Faculty Student Research Grant (FSRG), award number H01A11.

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 Assessment of Cognitive Food Consumption Experience
Authors: Atakan S. Sinem , Wansink Brian
Published: June 2021

The goal of this research is to explore the cognitive experience of food consumption and develop a scale that can differentiate the various factors that individuals focus on while consuming food. Individuals differ in several aspects of the food consumption experience. Some pay more attention to the sensory aspects of the food whereas others focus on the quantity of the food or their surroundings. Currently, there are no reliable and valid instruments that assess the cognitive experience of food consumption.

In‐depth interviews with consumers highlighted that, during food consumption episodes, consumers pay attention to the sensory tasting experience of the food, quantity consumed, and the nutritional content of the food. Furthermore, self‐monitoring (expectations and norms set by others who are readily present) and trying to eat healthy seem to be crucial aspects of the experience. Finally, distracted (non‐food related) thinking also emerges as part of the cognitive experience during consumption.

Next, a survey study tested the dimensions that emerged from the in‐depth interviews. A pool of items for each dimension (attention to 1) the sensory experience, 2) quantity of food consumed, 3) nutritional content of the food, 4) healthy eating, 5) self‐monitoring, and 6) non‐food related thinking) was developed by the authors. Then, four professionals working on food consumption behavior were asked to review item content and question wording and to indicate any criteria that they believed were not adequately assessed. In addition, several consumers reviewed the scale questions for relevance and clarity. After making revisions based on feedback from both professionals and consumers, the questions were administered as part of a larger study that required consumers to consume several snack items. The participants (N = 56) were 18 years or older students at a large university. Exploratory factor analysis confirmed the six dimensions that emerged from the in‐depth interviews.

This study is a preliminary exploration of the factors that shape the cognitive experience of food consumption. Future studies will focus on convergent, discriminant, and incremental validity of the newly devised scale.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Advocating the Responsible Use of Drugs through an Academic Service‐learning Activity in Higher Education.
Authors: Ignatova Tatiana , Yashchenko Oleksandre , Fernández Andrea , Sabaté Mar , Trullàs Eloi , Granados Victor , Morgó Judit , Solé Joel , Vacas Noemí , Vilaseca Jaume‐Elies , Garcia Maria Àngels , Cuffí Maria Laura
Published: June 2021

Even with the great availability of new drugs and the best clinicians, therapeutic results are not always as expected. Poor therapeutic adherence leads to a waste of scientific knowledge and resources of the public health system. Medical students from the University of Barcelona analyzed this situation in their local hospital via a survey oriented to patients, which indicated a lack of adherence to prescribed treatments as well as an excess of self‐medication.

A service‐learning activity begins with the identification of a real need and the preparation of an action plan (Kaye, 2004). The aim is to encourage best practices amongst the general public and health workers regarding drug consumption through an awareness campaign.

In order to improve awareness and understanding of the consequences of the problem described, an audiovisual campaign was chosen, focusing on promoting therapeutic compliance and avoiding the risks of self‐medication. Thanks to the collaboration between the Medicine and Audiovisual degrees, two video clips were produced: (https://drive.google.com/open?id=0B1aCcXVIkA_cNDFseTNPb3Z2TUE) (https://drive.google.com/open?id=0B1aCcXVIkA_cV2t3bHdhQUpxdWc)

This methodology improved the academic learning in Pharmacology and Audiovisual sciences as well as those related to other competencies required by our university.

The skills acquired were generally assessed according to their specific degree competencies. The outcomes for the participants were: 1) learning to communicate scientific subjects in an appropriate language for the subject and audience, 2) detecting some common mistakes in drug prescription and/or therapeutic adherence, 3) producing audiovisual and multimedia content and 4) networking with experts from another field of knowledge.

Kaye, C. B. The complete guide to service learning: Proven, practical ways to engage students in civic responsibility, academic curriculum, & social action. Free Spirit Publishing, 2004.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Clinical Potential of an Enzyme‐based Novel Therapy for Cocaine Overdose
Authors: Zheng Fang , Zhan Chang‐Guo
Published: June 2021

It is a grand challenge to develop a truly effective medication for treatment of cocaine overdose. The current available, practical emergence treatment for cocaine overdose includes administration of a benzodiazepine anticonvulsant agent (e.g. diazepam) and/or physical cooling with an aim to relieve the symptoms. The inherent difficulties of antagonizing physiological effects of drugs in the central nervous system have led to exploring protein‐based pharmacokinetic approaches using biologics like vaccines, monoclonal antibodies, and enzymes. In particular, we have rationally designed and discovered highly efficient cocaine‐metabolizing enzymes for therapeutic treatment of cocaine overdose and addiction. However, none of the pharmacokinetic agents has demonstrated convincing preclinical evidence of clinical potential for drug overdose treatment without a question mark on the timing used in the animal models. In this investigation, we have made use of animal models, including locomotor activity, protection, and rescue experiments in rats, of drug toxicity treatment with clinically relevant timing for the first time. It has been demonstrated that an efficient cocaine‐metabolizing enzyme developed in our previous studies can rapidly reverse the cocaine toxicity whenever the enzyme is given to a living rat, demonstrating promising clinical potential of an enzyme‐based novel therapy for cocaine overdose as a successful example in comparison with the commonly used diazepam.

Support or Funding Information

This work was supported by an NIDA Translational Avant‐Garde Award (grant UH2/UH3 DA041115) and NIDA R01 grants (DA035552, DA032910, DA013930, and DA025100).

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 Medial Prefrontal Cortex is Involved in Social Odor Recognition Memory
Authors: Granata Lauren M. , Robinson Siobhan , Hienz Robert D. , Davis Catherine M.
Published: June 2021

Objective

The medial prefrontal cortex (mPFC) is critical to mediating attention and social cognition, but its specific role in memory processing is unclear. In a rodent model of social odor recognition memory (SORM), we assessed the role of the prelimbic and infralimbic regions of the mPFC in social motivation and recognition memory at different stages of the memory process (encoding, consolidation, and retrieval). In the same animals, we also assessed the role of these subregions of the mPFC in sustained attention, using the rodent psychomotor vigilance test (rPVT).

Methods

The SORM task involves three phases (familiarization, habituation, and recognition), and requires olfactory‐based discrimination between familiar odors and novel odors from conspecifics, where failure to show preference for novel odors on the test phase indicates a memory deficit. The rPVT is a sustained attention test that measures accuracy, premature responding, and response times to the presence of a stimulus light that appears after a random waiting period between 3–10 seconds. Using pAAV‐hSyn‐hM4D(Gi)‐mCherry (“designer receptors,” DREADDs) injected bilaterally into the prelimbic or infralimbic regions of the mPFC, we remotely silenced these regions during behavioral testing via injections of the “designer drug,” clozapine‐N‐oxide (CNO).

Results

For the SORM test, injecting CNO (1 mg/kg, i.p.) 30 minutes prior to the habituation phase did not impair novel odor discrimination on this phase, but did result in impaired memory for a novel odor on the recognition test day 24‐hrs later. These effects were not a function of CNO administration alone because the impairment was found on the drug‐free test day 24‐hrs after the previous CNO injection. In a second study, injections of CNO (0–5 mg/kg, i.p.) were administered 30‐min prior to different rPVT sessions. CNO at 1 mg/kg had effects on premature responding in the rPVT, without impairing response times.

Conclusions

Silencing the prelimbic and infralimbic subregions of the mPFC impaired memory performance the recognition test phase of the SORM test, without inducing a motor impairment in the psychomotor vigilance test, indicating that the mPFC could be involved in encoding, and possibly consolidating, information about social odors acquired during habituation.

Support or Funding Information

This work was supported by a grant from NASA (NNX15AC71G) to CMD.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining Key Carbonic Anhydrase Isozymes Involved in Learning and Memory via Mouse Memory Assays
Authors: Lemon Nicole , Sanku Rajesh Kishore Kumar , Chong Anthony M. , Ilies Marc A. , Walker Ellen A.
Published: June 2021

Carbonic anhydrase (CA) is a zinc enzyme that catalyzes the reversible reaction between carbon dioxide and bicarbonate. This reaction is important in many tissues and its functions include recycling and secretion of cerebrospinal fluid and pH homeostasis. In mammals, 16 isozymes have been described to date, with different catalytic activity, sub‐cellular localization, and tissue distribution. CA isozymes located in the CNS are believed to be involved in neuronal signaling and synaptic strengthening suggesting that activation of these isozymes may enhance learning. The role of CA in learning and memory processes is not fully elucidated, predominantly due the fact that most of the carbonic anhydrase activators (CAA) available today are either inefficient activators or may lack appropriate physicochemical properties to cross blood brain barrier. The purpose of the current study is to profile a set of lipophilic CA activators and to determine the important isozymes of CAs involved in learning and memory. Therefore, we designed and synthesized series of novel lipophilic CAAs with discreet activation profiles for different CA isozymes in the CNS. Our strategy is to selectively activate one, two, and/or three different isozymes with our discrete set of CA activators. In the current study, a series of novel CAAs were profiled using a mouse memory assay known as the Novel Object Recognition Test (NORT). Solubility and lipophilicity for each compound were also determined and compared along with the activation profiles (KA) of multiple isozymes for all compounds. Representatives with different activation profiles and optimal physicochemical properties were assessed for learning and memory in mice to identify the main CA isozymes likely involved in learning and memory. When tested in NORT, CAAs protected mice from scopolamine‐, ketamine‐, and acetazolamide‐induced recognition deficits when tested using doses of 0.03 mg/kg CAA, i.p. and a 30 min pretreatment. Considering both the physicochemical and activation profiles, preliminary results gathered by testing CAAs in NORT revealed that several CA isozymes present in the CNS are responsible for learning and memory. Moreover, our data suggest that CAAs protect retention from drug‐induced memory disruption without affecting the motor activity.

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 Effects of HIV‐1 Gp120 on the Puncta Count and Expression of α5‐Containing GABAA Receptors in Cultured Rat Hippocampal Neurons
Authors: Jo Adrienne , Green Matthew , Thayer Stanley
Published: June 2021

Approximately 50% of the >30 million people worldwide affected by human immunodeficiency virus‐1 (HIV‐1) suffer from cognitive impairments known as HIV‐1‐associated neurocognitive disorders (HAND). HIV‐1 can infect microglia, leading to the release of toxic proteins, such as envelope glycoprotein 120 (gp120IIIB), which causes neurotoxicity and synaptodendritic damage. Gp120 is known to induce the release of interleukin‐1β (IL‐1β), a cytokine released by microglia. Previous work shows that IL‐1β increases the function of α5‐containing GABAA receptors (α5‐GABAA‐Rs), which lowers cell excitability and may contribute to cognitive impairments. Thus, we tested whether gp120 would increase the expression of protein clusters of α5‐GABAA‐Rs in primary hippocampal cultures derived from embryonic day 17 Sprague Dawley® rats. Immunocytochemistry was performed to examine the localization of α5‐GABAA‐Rs relative to microtubule‐associated protein 2 (MAP2) immunoreactivity. Based on previous electrophysiology experiments on α5‐GABAA‐Rs, we hypothesize that gp120 increases α5 puncta. Preliminary data demonstrates that gp120 leads to a trending increase in α5 puncta, which might explain the increased neuronal inhibition and decreased cell excitability seen in vitro. Altered excitability may lead to the cognitive impairments found in HAND patients. Thus, inhibition of α5‐GABAA‐Rs may be a novel target for the treatment of HAND.

Support or Funding Information

This investigation was supported by DA07304 and the National Institute on Drug Abuse of the National Institutes of Health under Award Number 1R25DA039074‐01A1 through the Summer Research Program for Diversity Students in PharmacoNeuroImmunology (PNI) under Life Sciences Summer Undergraduate Research Programs.

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 Monoamines on MPP+ and H2O2 Induced Oxidative Stress Response
Authors: Cobb Shelley , Shepherd Kennie
Published: June 2021

Parkinson's Disease is a neurodegenerative disorder characterized by the depletion of monoamines, degeneration of dopaminergic neurons, and subsequent symptoms of bradykinesia, muscle rigidity, and other movement deficiencies. The production of reactive oxygen species has also been implicated in PD. It was previously demonstrated in vivo, that monoamines are depleted in the brain prior to the degeneration of dopamine neurons induced by the Parkinson's inducing agent MPTP. The current study seeks to examine the effects of monoamines such as dopamine, norepinephrine, and serotonin in buffering the effects oxidative stress and cell death mediated by hydrogen peroxide and the Parkinson's inducing agent MPTP/MPP+. The study also aims to elucidate how the monoamines provide this neuroprotective effect. The data has shown differing effects between the different monoamines which may be due to their differing structures. It is hypothesized that they interact with the Nrf2/KEAP1 pathway which regulates the antioxidant enzyme system. The data from these studies will provide potential insight into the role that monoamines play in the neurodegenerative process.

Support or Funding Information

This work was supported by NIH Grant U54NS083932 and NIMHD Grant #5S21MD000101. The investigation was conducted in a facility constructed with support from Research Facilities Improvement Grant #C06 RR‐07571 from the National Center for Research Resources, NIH.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Methylene Blue Protects The Dopaminergic Neurons through Upregulation of BDNF
Authors: Bhurtel Sunil , Choi Dong‐Young , Katila Nikita
Published: June 2021

Parkinson's disease is a relatively common neurodegenerative disease without known causes. There are no effective therapies to halt or heal the neurodegeneration. Methylene blue (MB) possesses neuroprotective property, and has been tested in a clinical trial for Alzheimer's disease treatment. However, few studies have elucidated the effects of MB in Parkinson's disease. In this investigation, we evaluated effects of MB in the 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) mouse model of Parkinson's disease. Mice received MB (20 mg/kg/day, i.p.) for four consecutive days, and then MPTP (30 mg/kg/day, i.p.) for the next four days. Immunohistochemical stainings displayed that MB significantly attenuated MPTP‐induced loss of the dopaminergic neurons in the nigrostriatal region. Activation of glial cells was observed in the substantia nigra and striatum challenged with MPTP, and pretreatment of MB markedly decreased the glial reaction. Moreover, MB restored MPTP‐induced depletion of dopamine in the striatum. To illuminate the underlying mechanism for the neuroprotection of MB, we analyzed the level of neurotrophic factors such as brain derived neurotrophic factor (BDNF). The western blotting showed MB as a strong inducer of BDNF and its downstream signaling pathways, suggesting BDNF might be a critical contributor to the beneficial effects. In study with SH‐SY5Y cells, inhibition of the receptor tyrosine kinase B, a receptor for BDNF reversed MB‐mediated protection against MPP+ toxicity implying that BDNF was responsible for the preventive effect. These results indicate that MB protects neurons against MPTP neurotoxicity through induction of BDNF, representing MB as a potential modulating agent for Parkinson's disease.

Support or Funding Information

Korea Foundation for the Advancement of Science & Creativity (KOFAC)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of early life sleep deprivation on later life behaviors in rats
Authors: Atrooz Fatin , Liu Hesong , Kochi Camila , Salim Samina
Published: June 2021

Increasing body of epidemiological studies suggest that sleep deprivation at early life stages (childhood and adolescence) is associated with behavioral and cognitive impairment. While these studies are important, yet, the relationship between early life sleep deprivation and development of later life psychiatric symptoms is not clearly understood. While examining the link between early life sleep deprivation and later life behavioral and cognitive well‐being is important, conducting such clinical studies in children are difficult. Therefore, animal models are valuable in studying the behavioral and biochemical consequences of early life sleep deprivation across the developmental course. Extended wakefulness increases cellular metabolism and induces reactive oxygen species (ROS) formation leading to oxidative stress. Hence, we examined the role of oxidative stress on behavioral impairment caused by early life SD. We employed two groups of male Sprague Dawley rats; control (CON) and sleep deprivation (SD) groups. Rats, at postnatal day (PND) 19 were subjected to SD for 14 days, (6–8 hours/day) using Pinnacle automated sleep deprivation system. Behavioral and cognitive tests were performed at three stages; early adolescent (PND32), late adolescent (PND60) and adult stage (PND90). Rats in the SD group exhibited anxiety‐like behavior at PND32 and 60 but not at PND90. Interestingly, SD rats did not exhibit depression‐like behavior at PND32 or at PND60 but developed depression‐like behavior at PND90, as indicated by increased immobility time in forced swim test compared to control group. Plasma levels of corticosterone (indicator of stress) were significantly increased in SD rats compared to control rats. We conclude that early life sleep deprivation during postnatal brain development, by engaging oxidative stress cascades, might alter the normal synapse formation and pruning causing anxiety‐like behavior early in life (at PND32 and 60), which in later life transforms into depression‐like behavior (PND 90).

Support or Funding Information

2R15MH093918‐02

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