bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of Gambogic Acid on the Production of Reactive Oxygen Species and Neutrophil Extracellular Traps
Authors: Reinke Julia Grace , Gardner Cole , Jackson Alexandria , Kumar Shaili , Petreaca Melissa 
Published: June 2021

The innate immune system responds rapidly to the early signs of an infection or tissue damage by recruiting neutrophils to the affected area, where these inflammatory cells kill microbes through the production of reactive oxygen species (ROS) and neutrophil extracellular traps (NETs). Although these normal inflammatory responses are necessary for the elimination of potential pathogens before they proliferate and establish an infection, ROS and NETs are non‐specific mechanisms that can also damage host tissues. In chronic inflammation associated with various diseases, such as rheumatoid arthritis or Crohn's disease, prolonged or non‐resolving inflammation can cause severe pain and permanent tissue damage. We are currently investigating the potential of gambogic acid, a compound derived from the Garcinia hanburyi plant used in Ayurvedic and traditional Thai medicines, to reduce inflammation or promote its resolution. Previous studies have suggested that gambogic acid promotes apoptosis in multiple cell types, suggesting that this molecule could promote neutrophil apoptosis and thereby inhibit their inflammatory functions. In support of this hypothesis, we found that gambogic acid decreases PMA‐induced ROS and NETs by neutrophil‐like HL‐60 cells. Gambogic acid also decreases survival of neutrophil‐like HL‐60 cells in a WST‐8 survival assay, suggesting that it may reduce pro‐inflammatory neutrophil functions by inducing their apoptosis. We are currently investigating whether gambogic acid exerts similar effects on NET and ROS production in normal human polymorphonuclear cells (PMNs), and whether the gambogic acid impacts on neutrophil function are dependent upon its induction of neutrophil apoptosis. If so, this would suggest a therapeutic, inflammation‐resolving potential of gambogic acid in diseases characterized by excessive inflammation.Support or Funding InformationDePauw University, Science Research FellowsThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effect of TNF‐α on Mitochondrial Morphology in Model (NSC‐34) Motor Neurons
Authors: Brown Alyssa D. , Fogarty Matthew J. , Mantilla Carlos B. , Sieck Gary C. 
Published: June 2021

Mitochondrial dysfunction and fragmentation precede neuronal death in many neurodegenerative disorders, including those involving motor neurons. In other cell types, pro‐inflammatory agents such as TNF‐α cause marked mitochondrial fragmentation. These inflammatory insults may underlie the vulnerability of specific neurons during age‐associated conditions, such as sarcopenia. We hypothesize that motor neuron exposure to pro‐inflammatory cytokines induces mitochondrial fragmentation. Experiments were conducted using a mouse motor neuron‐like cell line, NSC‐34. These cells were differentiated by serum depletion until neurite structures (>50 um) were observed, ~24 h. The differentiated NSC‐34 cells were then treated with mouse TNF‐α at varying concentrations; control (0 ng), 10 ng/ml, and 20 ng/ml for 24 h. The cells were then loaded with MitoTracker Green to image mitochondria using a Nikon A1R confocal microscope (60x oil immersion 1.4 NA). Images were analyzed using MatLab macro to calculate form factor, aspect ratio and mitochondrial volume density. TNF‐α exposure caused mitochondrial fragmentation as reflected by decreased form factor (27% reduction) and aspect ratio (12 % reduction). These results are consistent with inflammatory cytokine exposure leading to mitochondrial fragmentation in motor neurons which may increase their vulnerability mitochondrial dysfunction and cell death.Support or Funding InformationNIH: RO1AG044615This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Oxidative Stress and Inflammation in Wound Repair: Molecular and Cellular Mechanisms
Authors: Pianissoli Lícia Gobeti , Silva Julia Paiva Rodrigues , Penteado Elis Arantes , Lopes Diana Ramlow Coelho , Mengal Vinicius Franskoviaky 
Published: June 2021

Oxidative stress and inflammation are the critical factors attributed with delay in wound repairing process. Experimental studies established the dogma that inflammation is essential to the establishment of cutaneous homeostasis following injury, and in recent years information about specific subsets of inflammatory cell lineages and the cytokine network orchestrating inflammation associated with tissue repair has increased. Recently, this dogma has been challenged, and reports have raised questions on the validity of the essential prerequisite of inflammation for efficient tissue repair. Indeed, in experimental models of repair, inflammation has been shown to delay healing and to result in increased scarring. The main focus of present study was to explore the effect of oxidative stress and inflammation in the wound healing in patients with pressure injuries. The study was performed in patients in the medical clinic, from a general hospital, located in Espirito Santo‐Brazil. Weekly swab collection was performed in order to follow the healing process and therapeutic response. Molecular biology analyzes were performed to measure the presence increase, or time‐dependent reduction related to oxidative stress and inflammatory process. Our data show that pressure injuries in stages 2 and 3 of difficult healing present reduced of antioxidant enzymes, such as SOD and Catalase, as well as an increase in oxidant and inflammatory enzymes. We also evaluated the changes in inducible nitric oxide synthase (iNOS) and other inflammatory enzymes using western blot. Our results showed that, in pressure injuries, there is higher level of iNOS, inflammatory and oxidants enzymes. iNOS is linked to increased inflammation, leading to prolonged wound healing. Our results suggest that treatments that aim to reduce the local inflammatory process and consequently the oxidative stress can improve the healing process, as well as make the tissue repair process easier.Support or Funding InformationThis research was supported by a CNPq research grant. This study was funded by Fundação de Amparo a Pesquisa do Espirito Santo (FAPES‐Brazil) and Conselho Nacional de Desenvolvimento Científico e Tecnológico.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Nitric Oxide and Nitric Oxide Synthase isoforms are modulated in renal ischemia/reperfusion induced cardiac hypertrophy
Authors: Caio‐Silva Wellington , da Cruz Junho Carolina V , Panico Karine Cruz , Carneiro‐Ramos Marcela Sorelli 
Published: June 2021

IntroductionIt is known that kidney injury by ischemia/reperfusion (I/R) leads to cardiac hypertrophy mediated by immune system. Besides, the Nitric Oxide (NO) that can be produced by Nitric Oxide Synthase (NOS) plays a key role in cell signaling including heart and kidney. However, the role of NO and NOS, in pathophysiological cardiac hypertrophy caused by renal I/R has yet to be clarified. In this study, we begin to investigate the NOS expression and NO levels in kidney after I/R protocol.MethodsC57BL/6 male mice were subjected to surgical occlusion of left renal pedicle for 60 min followed for reperfusion (I/R) for 8 and 15 days. Morphometric analyses were performed using as parameters kidneys weight and body weight, and vimentin mRNA as molecular kidney injury marker. Real time PCR was performed to analyze gene expression. To assess the bioavailability of NO in the form of S‐Nitrosothiols we use an amperometric protocol for S‐nitrosothiol with an NO specific sensor.ResultsThe mRNA expression of all the three isoforms was analyzed in the left kidney. The expression of endothelial NOS and neuronal NOS were downregulated after 8 days of reperfusion (p<0.05) and returned to basal levels after 15 days. The induced NOS (iNOS) isoform followed the same pattern as the others at 8 days. However, the iNOS mRNA levels increased significantly after 15 days of reperfusion when compared to sham group (p<0.05). In relation to the bioavailability of NO in the form of S‐Nitrosothiols, the results showed that the left kidney 15 days I/R group presented a significant increase when compared to sham group. The right kidney presented no alteration on NO content.ConclusionThese results showed that renal I/R induced renal damage leads to NOS modulation remarkably to iNOS expression at 15 days of reperfusion. This modulation can promote an increase on NO bioavailability and contribute to oxidative stress dysregulation. Data herein suggest a possible mechanism that contributes to cardiac hypertrophy developmental.Support or Funding InformationFinancial Support: FAPESP 2015/19107‐5This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pendrin Mediates Bicarbonate Secretion and Enhances CFTR Function in Airway Surface Epithelia
Authors: Kim Dusik , Huang Junwei , Billet Arnaud , Abu‐Arish Asmahan , Goepp Julie , Matthes Elizabeth , Tewfik Marc A. , Frenkiel Saul , Hanrahan John W. 
Published: June 2021

Bicarbonate facilitates mucin unpacking and bacterial killing however its transport mechanisms in the airways are not well understood. cAMP stimulates anion efflux through the CFTR (ABCC7) anion channel and this is defective in CF. The anion exchanger pendrin (SLC26A4) also mediates HCO3− efflux and is upregulated by proinflammatory cytokines. Here we examined pendrin and CFTR expression and their contributions to HCO3− secretion by human nasal and bronchial epithelia. In native tissue, both proteins were most abundant at the apical pole of ciliated surface cells with little expression in submucosal glands. In well‐differentiated primary nasal and bronchial cell cultures, IL‐4 dramatically increased pendrin mRNA levels and apical immunostaining. Exposure to low‐Cl− apical solution caused intracellular alkalinization (ΔpHi) that was enhanced 4‐fold by IL‐4 pretreatment. ΔpHi was unaffected by DIDS or CFTRinh‐172 but was reduced by adenoviral shRNA targeting pendrin. Forskolin increased ΔpHi and this stimulation was prevented by CFTRinh‐172 implicating CFTR, yet forskolin only increased ΔpHi after pendrin expression had been induced by IL‐4. The dependence of ΔpHi on pendrin suggests there is minimal electrical coupling between Cl− and HCO3− fluxes and that CFTR activation increases anion exchange‐mediated HCO3− influx. Conversely, inducing pendrin expression increased forskolin‐stimulated, CFTRinh‐172‐sensitive current by ~2‐fold in epithelial and non‐epithelial cells. We conclude that pendrin mediates most HCO3− secretion across airway surface epithelium during inflammation and enhances electrogenic Cl− secretion via CFTR as described for other SLC26A transporters.Support or Funding InformationSupport: Cystic Fibrosis Canada, Canadian Institutes of Health Research, Canada Foundation for InnovationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Increased alcohol consumption and development of tolerance to alcohol‐induced sedation in NBCn1 knockout mice
Authors: Choi Inyeong , Schank Jesse R 
Published: June 2021

The Na/HCO3 cotransporter NBCn1 plays a role in regulating synaptic pH. We recently reported that a human SLC4A7 gene polymorphism identified as a marker for polysubstance abuse is responsible for the production of a transporter variant that has a defect in function and expression. This finding prompted us to test whether NBCn1 is involved in alcoholism. Here, we examined alcohol‐drinking behaviors, as well as locomotor/anxiety and spatial learning behaviors, in NBCn1 knockout (KO) mice. In the open filed test, NBCn1 KO mice displayed decreased locomotor activity compared to wild type (WT) littermates. However, the activity in the elevated plus maze test was similar between WT and KO mice, indicating negligible change in anxiety. In the Morris water maze test, both groups of mice had learning behaviors improving each day with similar latency to escape in repeated trainings and distance traveled. The two groups learned the platform location with similar time spent in the trained quadrant during the probe trials. Thus, hippocampal‐based spatial learning was not altered in KO mice. Next, alcohol‐related behaviors were tested in a two‐bottle choice drinking experiment, in which mice were given to 3–15% ethanol with 3% increments at 4 days and alcohol consumption was measured. KO mice consumed more alcohol than WT mice with higher alcohol preference (p < 0.05). The consumption was also increased after repeated alcohol withdrawals. In the loss of righting reflex test following alcohol administration (3.5 g/kg alcohol; intraperitoneal), KO mice had a longer duration of sedation, but blood alcohol concentration did not differ. The duration became shorter when KO mice were pretreated with alcohol for 7 days, indicating the development of tolerance to the hypnotic effects of alcohol. Whole cell current clamp of hippocampal neurons showed that neurons from KO mice had a higher action potential threshold, no recurrent action potential, and a more depolarized membrane potential. These results indicate that abnormalities in NBCn1 alter alcohol consumption and alcohol‐related behaviors by changing neuronal excitability. Increased voluntary alcohol consumption in KO mice is probably driven by decreased rewarding value of alcohol. Neurons from KO mice are less excitable, which would lead to decreased neurotransmitter release. Thus, the rewarding value of alcohol is reduced, requiring more alcohol to achieve the same level of hedonic response.Support or Funding InformationNIH and Emory URCThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Kölliker‐Fuse contains opioid‐sensitive inspiratory neurons and contributes to fentanyl‐induced apnea
Authors: Saunders Sandy E , Levitt Erica S 
Published: June 2021

Opioid overdose deaths have rapidly increased largely due to illicitly manufactured fentanyl. The cause of death from opioid overdose is respiratory depression. Opioid‐induced respiratory depression is characterized by decreases in respiratory rate, chemoreceptor reflexes and impairment of the upper airway. The Kölliker‐Fuse (KF) regulates upper airway coordination and modulates respiratory rate. Opioid agonists in the KF reduce respiratory rate and cause apneusis. The influence of the KF during systemic fentanyl‐induced respiratory depression was investigated using the in situ arterially perfused working heart‐brainstem preparation of rat. Application of fentanyl to the preparation resulted in dose‐dependent slowing of rate, apneusis consisting of low‐amplitude phrenic nerve bursts and loss of post‐inspiratory vagal output, and eventually apnea with occasional low amplitude phrenic bursts. Subsequent application of CTAP, a mu opioid receptor antagonist, directly into the KF increased the frequency of low amplitude phrenic nerve bursting to a rate similar to baseline, with no change in vagal output. A similar pattern emerged with pre‐application of the mu opioid antagonist CTAP into the KF prior to systemic fentanyl administration. In separate experiments, KF neurons were recorded extracellularly before and after fentanyl administration. Inspiratory neurons stopped firing upon fentanyl administration, whereas, expiratory neurons continued to fire in the presence of fentanyl. These results suggest that opioid‐sensitive inspiratory neurons in KF may initiate low amplitude phrenic bursts and reverse fentanyl‐induced apnea.Support or Funding InformationFunded by NIH Grant DA038069.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of a P2X2 Reporter Mouse Model
Authors: Kim Seol‐Hee , Taylor‐Clark Thomas 
Published: June 2021

P2X2 is a ligand gated ion channel and purinergic receptor that is stimulated by adenosine 5′‐triphosphate (ATP). P2X2 is expressed in subsets of peripheral nerves and mediates various cellular responses in peripheral sensory nerve system. In particular, functional experiments have shown that P2X2 is selectively expressed in vagal afferents derived from the placodes (nodose ganglion) but not from the neural crest (jugular ganglion).In this study, the Cre‐Lox reporter system was used to generate a mouse model enable us to target specific subtypes of sensory afferents. First, we generated a knockin P2X2‐Cre strain. Here a cre recombinase was incorporated into the endogenous P2rx2 gene, downstream of the last P2rx2 exon following a 2A sequence. Expression of Cre was linked to the expression of endogenous P2X2 via a 2A sequence. The P2X2‐Cre mouse was crossed with another strain carrying Floxed Ai9 ROSA tdTomato, which results in expression of tdTomato in only P2X2 expressing cells via Cre recombination. From these animals, sensory ganglia including vagal ganglia, trigeminal ganglia, superior cervical ganglia, carotid body and dorsal root ganglia were collected to visualize tdTomato‐expressing cells in combination with immunohistochemistry. Furthermore, brainstem was collected to map their central projections.tdTomato positive cells were observed only in the nodose and not in jugular of vagal ganglia. Some population of cells showed tdTomato expression in trigeminal ganglia, dorsal root ganglia and superior cervical ganglia. In addition, strong tdTomato expression was observed in carotid body, which appeared to surround anti‐tyrosine hydroxylase positive cells and nerves. These results imply that P2X2 positive afferent nerves exist in different ganglia and therefore, ATP‐mediated purinergic signaling occurs in various peripheral sensory nerve systems. Lastly, tdTomato expression in sequentially sectioned medulla showed distinct patterns. The main area that has tdTomato expression is commissural nucleus of solitary tract (NTS), gelatinous NTS and dorsolateral NTS, as well as area postrema. This data suggests that the nodose‐originated sensory afferents project to specific regions of the medulla. Overall, the transgenic mouse model generated using Cre‐Lox system successfully labeled P2X2 cells.Support or Funding InformationThis study is funded by National Institutes of Health Common Fund SPARC OT2 (2016–2019): “Functional mapping of peripheral and central circuits for airway protection and breathing”This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of Vagal Pathways in the Respiratory Depressant Effect of Codeine on Breathing in the Anesthetized Cat
Authors: Rose Melanie J , Shen Tabitha Y , Musselwhite M Nicholas , Davenport Paul W , Bolser Donald C 
Published: June 2021

A growing body of evidence indicates that systemic administration of centrally‐acting opioids can have actions on peripheral sensory receptors leading to complex effects on breathing. We speculated that the effects of systemically administered codeine on the breathing pattern would have a peripheral component. Experiments were conducted in anesthetized, spontaneously breathing cats (n=12). The animals were tracheotomized and electromyogram (EMG) electrodes were placed in the diaphragm to record inspiratory motor drive. The animals were allocated into two groups: vagal intact (VI) (n=5) or cervical vagotomy (CVx) (n=7). A dose‐response to intravenous codeine (0.1–10 mg/kg) was performed in each animal. Codeine elicited dose‐related decreases in respiratory frequency in both groups that were not significantly different from one another. This opioid also elicited dose‐dependent increases in diaphragm EMG magnitudes in both VI and CVx groups, but the change in EMG at the highest dose was significantly higher in the CVx group (change from control: CVx 189±43%; VI 40±21%, p<0.05). There was no significant difference in end‐tidal CO2 values suggesting that minute ventilation was similar between the two groups. These results support the hypothesis that codeine preferentially suppresses non‐diaphragmatic inspiratory motor drive in vagotomized animals, leading to larger compensatory increases in diaphragm activity than in vagal intact animals to maintain ventilation. An extension of this hypothesis is that vagal pathways maintain the excitability of intercostal and accessory inspiratory muscle motor drive with increasing doses of codeine.Support or Funding InformationSupported by OT2OD023854‐01, HL131716.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inspiratory Work, Power and Drive During Multiple Breath Extrinsic Resistive Loading in Healthy Adults
Authors: Davenport Paul Wesley , Davenport Matthew P. , Feinstein Justin , Khalsa Sahib , von Leupoldt Andreas 
Published: June 2021

An extrinsic resistive load (R) to breathing increases the work of breathing. The extrinsic airway resistive work (W) of breathing during inspiration is defined as: Waw = Paw * Vi.. The power (J) required to breathe with this load is defined as: Jaw = ∫ Waw*dt. With increasing R, the Waw and Jaw required to sustain minute ventilation (VE) increases. The respiratory control system in the brainstem provides an inspiratory drive over the inspiratory duration (Ti) to generate sufficient inspiratory pump force to move the air through the airways and into the alveoli. The element of inspiratory drive required to compensate for increased extrinsic resistance is a function of the inspiratory Paw generated over the Ti, hypothesized as the pressure‐time‐product: PtP = ∫ Paw*dt. Thus, we predicted the PtP must increase with increasing Jaw as a function of the magnitude of the R. In addition, we reasoned that sustained breathing against increased R's require increasing PtP to compensate for the Jaw required to sustain VE. Hence, we hypothesized during a 10‐breath R loading trial, the PtP for Breath 10 will be greater Breath 1 as a function of R. Finally, we hypothesized that the perception of breathing effort is directly proportional to the PtP and Jaw. Healthy participants were tested while breathing through a non‐rebreathing circuit with inspiratory flow‐resistive loads (R=0, 5, 10, 25, 50, 100 cmH2O/L/s). The subjects inspired for 10 consecutive breaths for each load magnitude in a randomized block order. Subjects estimated their perceived load intensity (MEInt) and unpleasantness (MEUn) for breaths 1, 5 and 10. VE was unchanged for all 10 breaths with all R loads. The Waw and Jaw increased with increasing R. The PtP significantly increased with increasing R and was directly proportional to the increase in Jaw. The PtP increased from breath 1 to 10 only for R=100 cmH2O/L/s. The MEInt and MEUn increased with increasing R directly proportional to the PtP and Jaw. The results of this study demonstrate the increase Waw associated with breathing against extrinsic R loads increases the Jaw required to sustain VE. The PtP (drive) increases proportionately to the Jaw. The MEInt and MEUn increased with the increased PtP and Jaw generating an increased sense of breathing effort and negative affective perception for loaded breathing.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.