bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification and Characterization of TNF Receptor Associated Factor 3 Interacting Protein 3 (Traf3ip3) in Skeletal Muscle
Authors: Golliher Ciara , DaSilva Anjelica , Waddell David 
Published: June 2021

Skeletal muscle atrophy is a physiological condition that results in decreased muscle size and strength and occurs in response to immobilization, denervation, spinal cord injury and aging. To better characterize the molecular genetic events of neurogenic atrophy, a previous study isolated gastrocnemius muscle from mice following 3 days and 14 days of sciatic nerve denervation. The gene expression profile in the denervated muscle tissue was then analyzed by microarray and compared to control muscle tissue in order to identify novel neurogenic atrophy‐induced genes. The microarray data revealed that TNF Receptor Associated Factor 3 Interacting Protein 3 (Traf3ip3) is expressed in skeletal muscle and is induced in response to denervation. To confirm that Traf3ip3 is expressed in muscle cells, we cloned the Traf3ip3 cDNA from cultured muscle cells and performed quantitative PCR (qPCR) to assess expression levels in both proliferating and differentiated muscle cells. The qPCR results demonstrated that Traf3ip3 expression levels are relatively low in proliferating myoblasts but show dramatically elevated expression in differentiated myotubes. To characterize the transcriptional regulation of Traf3ip3, fragments of the proximal promoter located immediately upstream of the start of transcription were cloned and fused to a reporter gene. The reporter plasmids were then transfected into C2C12 mouse muscle cells in combination with myogenic regulatory factor (MRF) expression plasmids, which resulted in differential effects on reporter gene activity. The discovery that Traf3ip3 is expressed in muscle combined with the observation that this gene is induced in response to neurogenic atrophy helps further our understanding of the molecular genetic events of skeletal muscle wasting.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Oral Ketone Supplementation Acutely Increases Markers of NLRP3 Inflammasome Activation in Human Monocytes
Authors: Neudorf Helena , Durrer Cody , Myette‐Cote Etienne , Makins Caitlyn , O'Malley Trevor , Little Jonathan P. 
Published: June 2021

Recent cell culture studies indicate that the ketone body β‐hydroxybutyrate (β‐OHB) directly inhibits the NLRP3 inflammasome, which plays an important role in regulating inflammation. However, there is limited direct evidence demonstrating this effect in humans.ObjectivesTo determine the effects of acutely raising blood β‐OHB on NLRP3 inflammasome activation in healthy humans.MethodsWe conducted two randomized double‐blind placebo‐controlled experiments wherein participants' blood β‐OHB levels were directly elevated by ingesting supplements containing ketone salts (0.3 g β‐OHB/kg; Study 1, N = 10 males) or ketone monoesters (0.482 g β‐OHB/kg; Study 2, N = 20, 50% females). Blood samples were obtained in the fasted state and 30‐minutes following ingestion of ketones or placebo. Markers of NLRP3 inflammasome activation included monocyte caspase‐1 (flow cytometry) and interleukin (IL)‐1β secretion (ELISA) in LPS‐stimulated whole blood cultures, and IL1B and NLRP3 mRNA expression (qPCR) from whole blood.ResultsBlood β‐OHB was significantly raised from baseline following both ketone salt (0.2 ± 0.1 mM to 0.9 ± 0.2 mM; P < 0.001) and ketone monoester (0.2 ± 0.1 mM to 3.2 ± 0.6 mM; P < 0.001) ingestion with no change in placebo. Caspase‐1 activation increased 30 minutes after consuming both ketone salt (condition X time interaction, P = 0.012) and ketone monoester (condition X time interaction, P = 0.016) supplements but not placebo. IL‐1β secretion from stimulated whole blood cultures was increased following ketone monoester consumption (9.1 ± 7.6 pg/mL to 10.9 ± 6.5 pg/mL; main effect of condition, P = 0.024) with no change in placebo. There were no significant effects of ketone supplementation on IL1B and NLRP3 mRNA expression.ConclusionsMeasures of NLRP3 activation increased when blood β‐OHB was acutely elevated using ketone supplements, suggesting that artificially increasing β‐OHB may have unintended metabolic effects that augment activation of circulating immune cells.Support or Funding InformationThis study was funded by a Natural Sciences and Engineering Research Council (NSERC) of Canada Discovery Grant (Grant number 435807). JPL is supported by a Canadian Institutes of Health Research (CIHR) New Investigator Salary Award (MSH‐141980) and a Michael Smith Foundation for Health Research (MSFHR) Scholar Award (16890). H.N. was supported by a NSERC Undergraduate Student Research Award.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
THE PRETERM INFANT GUT MICROBIOME, HUMAN MILK VOLUME AND IMMUNOBIOLOGY, AND GROWTH
Authors: Groer Maureen , Dutra Samia Valeria , Miller Elizabeth , Yee Alyson , Gilbert Jack 
Published: June 2021

We conducted a study of gut microbiome in very low birth weight (VLBW) infants along with measures of milk feedings (type and volume) and milk sIgA, cytokines and chemokines for six weeks after admission in 78 infants. Stools were collected weekly, and human milk collected daily and pooled weekly for a panel of immune markers. Mothers of these infants gave informed consent and the study was approved by the university IRB. Mean levels of sIgA, EGF, MIP1α, IP10, MCP1, IL‐8, IL‐4, IL‐6, IL‐10 and TNFα were measured each week for six weeks by MagPix.4.2. Stool DNA was extracted using MoBio kits, the V3–V4 region of the 16S rRNA gene was amplified and then sequenced on an Illumina MiSeq.These infants had a mean gestation age at birth of 28 weeks, 3 days, and a mean birth weight of 1078 Gms. The mean time to full enteral feeding was 12 days, and they were fed mothers own milk (MOM), donor milk or preterm infant formula in varying amounts. The infant stool demonstrated marked dysbosis over time, with a dominance of Gram negative Gammaproteobacteria. Mean levels of sIgA, EGF, MIP1α, IP10, MCP1, IL‐8, IL‐4, IL‐6, IL‐10 and TNFα were measured each week for six weeks. Data reported here are mean Shannon diversity, and mean number of operational taxonomic units (OTUs) over the first six weeks of admission to the NICU, the ratio of human milk volumes to total feed volumes, the mean concentrations of milk immune markers over the 6 weeks of measurement, and the relationship between growth and the microbiome in these infants.Shannon diversity significantly increased with increasing corrected gestational age (cGA) (Multiple R2: 0.09221, p= 1.28e‐08). Adjusting for cGA during the NICU stay, the alpha diversity also showed weak but significant correlations with overall rate of weight gain (Multiple R2: 0.144, p=0.00208) and total ratio of mother's own milk (Multiple R2: 0.1097, p = 0.024918).Over the 6 weeks of admission, the mean number of fecal OTUs was correlated with mean milk IL‐4 (pg/ml) (r=.25, p=.03), but no other milk immune markers. The time to enteral feeding was inversely correlated to the number of OTUs (r=‐.24, p=.036). Lower diversity was observed in infants with lower gestational age (t=.31, p=.006), sepsis (t=2.43, p=.017), and days on oxygen (r=.23, p=.04).This is the first study to show that growth of VLBWs may be influenced by the microbiome in these early weeks of life. There were few relationships between specific immune markers and gut microbiome, but the amount of MOM an infant receives may partially ameliorate the dysbiosis.Future studies are planned to investigate the growth/nutrition/microbiome relationship in VLBWs as they grow and develop into early school age.Support or Funding InformationNIH R01NR15546This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Resolving Subtle Treatment Effects through Pharyngeal Pumping Variability Assessment in the Nematode C. elegans
Authors: Harris Michael B , Barrientos Christopher , Bui Andy , Co Mary , Gill Sunny , Lim Shelby , Ly Lily , Ortiz Brianna , Parabo Ashley , Phillips Derek , Tran Adrian , Valdez Vanessa , Vu Tien , Taylor Barbara E , Berlemont Renaud 
Published: June 2021

In C. elegans, feeding is achieved through a well characterized behavior, pharyngeal pumping. This pumping is controlled by two pairs of pharyngeal motor neurons (MC and M3). Each pump cycle corresponds to the propagation of a single pharyngeal muscle action potential, initiated by MC and terminated by M3. Many studies evaluate pharyngeal pumping to reveal experimental results and, traditionally, this has been quantified as an occurrence frequency by determining the number of pumps observed in a given time. Although rhythmic, pharyngeal pumping is both variable and periodic, and traditional pharyngeal pumping rate assessment does not consider underlying variability of pumping behavior or the sources of this variability. We use the recently developed ScreenChip system (NemaMetrix) to detect individual pump events, and have developed an algorithm (WormBeat) to normalize variation between subjects and distinguish variability associated with MC neuron pace and neuromuscular fidelity of the pharynx. The theory and practice of the WormBeat algorithm is described in a companion presentation. Here we use both traditional event‐frequency analysis and our algorithm to evaluate pharyngeal pumping. Studies are done in wild type (N2) C. elegans during pharmacological manipulations designed to alter the pace and regularity of MC and/or of the fidelity of pharyngeal neuromuscular junction. We also evaluate pharyngeal pumping in a series of mutant strains with known pumping defects. Our results illustrate that the WormBeat algorithm can identify subtle treatment effects and can identify and distinguish variation on motor neuron pacing from that produced by disruption of neuromuscular fidelity. These data support a hypothesis that underlying variability is a relevant and insightful index of functional decline in this model organismSupport or Funding InformationWork reported in this publication was supported by the National Heart Lung and Blood Institute and National Institute of General Medical Sciences of the National Institutes of Health under Award Numbers 1R15HL126105, 1SC2GM112570, UL1GM118979, TL4GM118980, and RL5GM118978. The work is solely the responsibility of the authors and does not necessarily represent the official view of the National Institutes of Health or any other funding body.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Primary Neurons from the Painted Turtle: A New Model for Comparative Neurobiology of Vertebrate Anoxia Tolerance
Authors: Riggs Claire L. , Xu Fenglian , Warren Daniel E. 
Published: June 2021

The painted turtle, Chrysemys picta, is the most anoxia‐tolerant tetrapod known, surviving over 170 days without oxygen at 3°C. A key component of C. picta's anoxia tolerance is metabolic depression: ATP use is dramatically reduced in the brain, in large part by suppression of nervous activity and channel arrest. This differs from mammals, which lack the ability to enter metabolic depression and withstand anoxia. Developing new tools to understand the neurobiology of the painted turtle has the potential to provide novel insight into the biology of anoxia‐tolerant vertebrates. While nearly all studies of anoxic turtle neuron function have been conducted in cortical sheets, the isolation and subsequent culture of primary neurons, to our knowledge, has not been conducted in this species. The goal of this study was to culture primary neurons from hatchling painted turtles and to develop a functional model for studying the neurobiology of anoxia tolerance. Primary neurons were dissociated from the cerebrocortex using enzymatic treatment and mechanical trituration and cultured in neurobasal medium supplemented with FBS, Penicillin/Streptomycin, B27, and L‐glutamine. Culturing the neurons on glass coverslips coated with laminin and poly‐D‐lysine at 37°C with 5% CO2 results in high neuronal survival and growth of neurites. Isolated neurons exhibit morphologies stereotypical of normal vertebrate neurons, with neuritic processes clearly identifiable. Importantly, the isolated cells showed functional responses characteristic of neurons, including calcium transients when excitatory factors, such as high concentration of KCl, were applied. This work represents the first in vitro primary culture of neurons from anoxia‐tolerant painted turtles that show normal vertebrate neuronal morphology and function.Support or Funding InformationThis work is supported by the NSF CAREER award 1253939 to Dr. Warren and the Saint Louis University Start‐Up Fund to Dr. Xu.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fluid‐structure interaction simulation of balance sensation in the avian lumbosacral organ
Authors: Wold Ethan , Daley Monica 
Published: June 2021

The lumbosacral organ (LSO) is an enlargement of the avian spinal cord hypothesized to play a role in balance sensation. Several lines of anatomical and neurophysiological evidence suggest that the LSO is directly integrated with spinal motor circuits, independent of the vestibular system. However, no model currently exists to explain its physiological function. We hypothesized that mechanosensors may respond to differential strain in spinal ligaments. We segmented iodine‐enhanced CT scans of the quail LSO to visualize the 3D anatomy of the structure. We then utilized two‐way fluid structure interaction simulations to model possible balance sensing function. Our modeling suggests that the LSO spinal ligaments are able to sense the directionality of an applied balance perturbation in the form of differential strain patterns. Furthermore, given anatomical constraints, it is unlikely that fluid velocity alone is sufficient to cause deformation in the spinal tissue. Inertia of the fluid and soft tissue itself play significant roles in generating the ligament strain. This work is the first simulation study of the LSO in any species and provides a methodological framework for further fluid dynamics modeling of the system. Our 3D reconstruction suggests potential anatomical constraints on fluid flow and supports the ligament strain hypothesis of balance sensing function. Higher resolution simulation work, as well as experimental validation studies in live animals are needed to more precisely evaluate the hypotheses for balance sensation in the LSO.Support or Funding InformationThis work was made possible by support from the American Physiological Society and the National Science Foundation Integrative Organismal Systems Award (Award No. IOS‐1238831).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pharmacological identification of cholinergic receptor subtypes in modulation of neural circuits in Drosophila melanogaster
Authors: Bhutto Umair Fareed , Cooper Robin 
Published: June 2021

Acetylcholine (ACh) is an abundant neurotransmitter and neuromodulator in many species. In Drosophila melanogaster ACh is the neurotransmitter used in peripheral sensory neurons and is a primary excitatory neurotransmitter and neuromodulator within the central nervous system (CNS). The receptors that facilitate synaptic transmission at cholinergic synapses and beyond are divided into two broad subtypes: the ionotropic nicotinic acetylcholine receptors (nAChRs) and the metabotropic muscarinic acetylcholine receptors (mAChRs). This receptor classification is shared in both mammals and insects; however, both the pharmacological and functional characterization of these receptors within the Drosophila nervous system has lagged behind its mammalian model counterparts. In order to identify the impact of ACh receptor subtypes in regulating the performance of select neural circuits within the larval CNS, we have used a behavioral and electrophysiological approach to assess cholinergic modulation of locomotion, feeding, and sensory‐CNS‐motor (sensorimotor) circuit activity. We have exposed intact 3rd instar larvae to ACh agonists and antagonists to observe modulation of these behaviors and also expose an intact nervous system directly to solutions containing these compounds to address their influence on sensorimotor circuit efficacy. We reveal that chronic ACh exposure enhances locomotion but reduces feeding behavior and acute application excites a sensorimotor circuit. Nicotine exposure reduces activity through suspected rapid receptor desensitization. Moreover, chronic muscarine exposure reduces locomotion and feeding, but acute exposure enhances sensorimotor circuit activity. These results suggest a role for both nAChRs and mAChRs in modulating these select circuits and illuminates important pharmacological properties of cholinergic receptor subtypes in vivo.Support or Funding InformationFunding Dept. of Biology, Kentucky Science and Engineering Foundation (KSEF‐3712‐RDE‐019) at the Kentucky Science and Technology Corporation (RLC), Ribble Scholar funds, & personal funds (RLC).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of the Corticotropin‐releasing Factor (CRF) System in Atlantic Salmon (Salmo salar)
Authors: Lai Floriana , Royan Muhammad Rahmad , Gomes Ana Silva , Rønnestad Ivar 
Published: June 2021

In contrast to most other teleosts studied to date, salmonids has undergone four whole genome duplications (WGD), commonly referred to as the Ss4R (salmonids‐specific 4th vertebrate WGD) event. Large genomic rearrangements and the restoring of the disomic chromosome inheritance in the ancient polyploidy have been crucial for the post Ss4R re‐diplodization process, leading to possible divergent functions of duplicated genes. This evolutionary genome duplication, together with the available genome sequence information for Atlantic salmon (Salmo salar), provide a unique opportunity to investigate the pivotal roles of gene diversity and functionality.When facing variable external or internal adverse effects beyond their homeostatic state, teleosts engage a stress response through the Hypothalamic‐Pituitary–Interrenal gland (HPI) axis to restore the compromised physiological steady state and enable the animal to overcome the threat. Corticotropin‐releasing factor (CRF), CRF‐binding proteins (CRFBPs) and CRF receptors (CRFRs) are commonly used as markers for stress responsiveness. This system has been poorly explored in Atlantic salmon, thus, in this study, we aim to investigate all the CRF system players.As for other salmonids, the Ss4R event induced the tetradiploidization in the Atlantic salmon genome leading to the several duplicated genes (paralogues) in these species. We currently investigate the crf, crfbp and crfr genes to explore the roles of the different paralogues. In silico analysis show that the Atlantic salmon genome has four crf paralogue genes distributed among chromosome (Chr)Ssa03, ChrSsa14, ChrSsa19 and ChrSsa29, two crfbp genes located in the ChrSsa01 and ChrSs11, two crfr1 paralogue genes placed in the ChrSsa03 and ChrSsa06 and two crfr2 paralogue genes in ChrSsa03 and ChrSsa14. To evaluate their function in the HPI stress response axis, we have analysed the mRNA expression of the different transcripts in different brain regions of Atlantic salmon post‐smolts. Our initial data shows that all crf transcripts are present in the hypothalamus and pituitary. However, further analysis are needed and particularly the characterization of these several transcripts under control and stressed conditions.Support or Funding InformationFunded by the Research Council of Norway (NFR 267626 LeuSENSE).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gestational Intermittent Hypoxia Reduces Basal Levels of Interleukin‐1 alpha Gene Expression in Neonatal Brainstem Microglia: Implications for Respiratory Control
Authors: Knutson Andrew Owen , Ewald Andrea , Kiernan Elizabeth , Baker Tracy , Watters Jyoti 
Published: June 2021

The in utero environment strongly impacts the developing fetus, and perturbations during this critical developmental window can have long lasting, negative consequences on offspring physiology. Intermittent hypoxia, a hallmark of obstructive sleep apnea, causes profound inflammation, is common during the third trimester of pregnancy, and is rising in parallel with the obesity epidemic. Although maternal immune activation is well‐known to influence fetal brain development and microglial immune activities, the consequences of gestational intermittent hypoxia (GIH) on microglia in CNS regions essential for respiratory neural control have not been studied. Given the important role of microglia in brain development during the first 2 postnatal weeks of life, and the importance of the critical period in postnatal respiratory development, we evaluated inflammatory gene expression in brainstem microglia in 14 day old (P14) rat pups whose dams were exposed to intermittent hypoxia (2 minutes alternating between 21% O‐2/10.5% O‐2) or normoxia (alternating between 21% O2/21% O2; GNX) for 8hrs/day during gestational days 10–21. Microglia were immunomagnetically isolated from the brainstems of GNX and GIH P14 pups, and inflammatory gene expression was analyzed by qRT‐PCR. While there were sex‐dependent differences in the basal expression of several key inflammatory cytokine genes in brainstem microglia, we focus here on interleukin‐1 alpha (IL‐1α). The expression of IL‐1α was 3.5‐fold higher in GNX female microglia than in male (p=0.014; N=5 each), indicating a strong sexual dimorphism in the expression of this cytokine in normal brainstem microglia. Very interestingly however, IL‐1α mRNA levels were significantly reduced by GIH only in female microglia (p=0.03; N=5 each), although they approximated those in GNX male microglia. IL‐1α is a unique cytokine in that it is an alarmin‐ a molecule that is released early after insult that signals tissue damage and activates the immune system. It has not been specifically studied in the respiratory neural control system to our knowledge, but the downregulation of IL‐1α by GIH in female brainstem microglia may reflect a compensatory response that is absent in males to combat the inflammatory uterine environment caused by maternal intermittent hypoxia. Studies are currently underway to further investigate the role of IL‐1α in respiratory neural control.Support or Funding InformationSupported by NS085226, HL105511, and Wisconsin Alumni Research Foundation UW2020 grantThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.