bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cigarette Smoke Disrupts Calcium Homeostasis in Airway Smooth Muscle Cells
Authors: Lin Jinheng , Taggart Michael , Tarran Robert , Gray Michael 
Published: June 2021

Tobacco smoking is the largest risk factor for developing chronic obstructive pulmonary disease (COPD), and is associated with airway hyperresponsiveness. Hyperresponsiveness is linked to abnormal calcium metabolism, which regulates the contractility and hence the tone of airway smooth muscle (ASM). However, whether cigarette smoke (CS) directly alters or regulates calcium signaling in ASM cells (ASMC) is not fully known. In airway epithelial cells, acute CS exposure increases intracellular Ca2+ concentrations ([Ca2+]i). We hypothesise that CS impedes ASM function by disrupting calcium homeostasis in ASMC, and the present study aims to unravel the effects of CS on calcium handling in ASMC. Primary rat and human ASMC were isolated via enzymatic digestion, and maintained in culture up to 8 passages. ASMC were seeded onto glass coverslips, and deprived of serum for 24–48 hours prior to calcium imaging studies using fura‐2‐AM. ASMC were exposed to either whole CS or CS extract (CSE) from 3R4F reference cigarettes (University of Kentucky) with the particulate phase filtered out. We found that both CS and CSE elevated [Ca2+]i in rat and human ASMC, and this increase was not abolished following depletion of sarcoplasmic‐reticulum calcium stores with thapsigargin or cyclopiazonic acid. The increase in [Ca2+]i in response to CSE was concentration‐dependent. Removal of extracellular calcium attenuated the CSE‐induced elevation of [Ca2+]i in hASMC, suggesting that CSE may activate calcium influx pathways in cultured hASMC. We speculate that this elevation of [Ca2+]i would lead to exacerbated ASM contraction, especially under inflamed conditions in COPD, and hence narrowing of the airway lumen. Further investigations into the source of [Ca2+]i increase, identity of potential calcium influx channels activated, and the direct effects of CS/CSE on ASM contraction are on‐going.Support or Funding InformationThis work is supported by Newcastle University's JJ Hunter Bequest scholarship.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex differences in lung miRNA‐712 expression and inflammatory cytokines in a mouse model of asthma and air pollution exposure
Authors: Weaver Ashley Morgon , Fuentez Nathalie , Steckbeck Rachel , Riviera Lidys , Nicoleau Marvin , Silveyra Patricia 
Published: June 2021

BackgroundMicroRNAs (miRNAs) are short RNAs (18–22 nucleotides) that play a critical role in post‐transcriptional gene regulation. Specifically miRNA 712‐5p has been shown to play a major role in the regulation of inflammatory cytokine expression, which is one of the hallmarks of asthma. Asthma has been shown to afflict more adult females than males. Asthmatic females are also more susceptible to air pollution exposure, which leads to higher rates of respiratory disease. The hypothesis of our work is that differential expression of specific miRNAs that regulate inflammatory cytokine expression in the male and female lung upon exposure to an air pollutant could contribute to the mechanisms responsible for the higher susceptibility of females to respiratory disease.MethodsTo test this hypothesis, we developed an asthma model in which male and female C57BL/6 mice were sensitized with house dust mite (HDM) or phosphate buffer saline (PBS) for five‐weeks, followed by a three‐hour ozone or filtered air exposure challenge to induce inflammation. Following exposure, lungs were harvested and total RNA was extracted and converted to cDNA. Relative miRNA expression linked to inflammatory cytokine production was measured by real‐time PCR and normalized with U6. Cytokine gene expression was also determined by real time PCR.ResultsPreliminary results show that there are sex‐specific differences in miR‐712 expression as well as inflammatory cytokine gene expression in asthmatic mice. The data shows that both males and females express higher amount of miR‐712 upon sensitization to HDM compared to PBS and that ozone also increases the levels of miR‐712 expressed in both sexes. Upon exposure to ozone, females sensitized with PBS express higher amounts of miR‐712 than males. However, males sensitized with HDM and exposed to ozone express more miR‐712 than females.ConclusionMiR‐712 is differentially expressed between asthmatic males and females following exposure to a pollutant. Since, miR‐712 is known to regulate inflammation, the differential expression of this miRNA could account for the increased susceptibility of asthmatic adult females to pollution. The results of this study could help us to further understand the sex‐specific mechanisms of asthma, and develop novel therapies.Support or Funding InformationNIH (HL133520, HL141618)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Microglial Depletion Abolishes Compensatory Plasticity Induced by Prolonged Reductions in Respiratory Neural Activity
Authors: Mickelson Carly Rose , Gumnit Maia , Agbeh Abiye , Ewald Andrea , Meza Armand , Ouellette Jonathan , Braegelmann Kendra , Watters Jyoti , Baker Tracy 
Published: June 2021

Reduced respiratory neural activity elicits a compensatory enhancement in inspiratory motor output to the diaphragm, a form of plasticity known as inactivity‐induced inspiratory motor facilitation (iMF). Mechanisms that induce iMF differ depending on the pattern of reduced respiratory neural activity; spinal TNF‐α and subsequent activation of TNFR2 on phrenic motor neurons is necessary to induce iMF following prolonged reductions in respiratory neural activity, whereas spinal retinoic acid synthesis and subsequent activation of RARa on phrenic motor neurons is necessary to induce iMF following intermittent reductions in respiratory neural activity. Little is known regarding the cell subtypes required for iMF induced by either stimulus. Since microglia are an important source of TNF‐α in the CNS, we tested the hypothesis that microglia are necessary for iMF following prolonged, but not intermittent, reductions in respiratory neural activity. Male and female rats received vehicle or pexidartinib (PLX3397; 80 mg/kg, 7 days po), a colony stimulating factor 1 receptor (CSFR1) inhibitor that results in microglial apoptosis. PLX3397 significantly reduced Iba1+ and CD11b+ cells throughout the brain and spinal cord (−82±8%, n=4, p<0.05). The capacity for prolonged (30 min) and intermittent (5, ~1 min) reductions in respiratory neural activity to elicit iMF was then assessed in urethane‐anesthetized, mechanically ventilated rats. In vehicle‐treated rats, both intermittent and prolonged reductions in respiratory neural activity triggered significant increases in phrenic inspiratory output (75 ± 16% and 99 ± 32% baseline, n=6 each, p<0.05), indicating iMF. By contrast, in PLX‐treated rats, only intermittent (63 ± 13% baseline, n=6, p<0.05), but not prolonged (21 ± 6% baseline, n=6, p>0.05) reductions in respiratory neural activity increased phrenic inspiratory output, suggesting that microglial depletion selectively impairs iMF activated by prolonged, but not intermittent, reductions in respiratory neural activity. These data suggest that in response to prolonged reductions in inspiratory‐related synaptic inputs to phrenic motor neurons, microglia may release TNF‐α to trigger compensatory enhancement in phrenic inspiratory output. Studies are underway to investigate this idea further.Support or Funding InformationNIH R01 HL105511 and NS085226This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Female Mate Choice Influences on Monogamous Pair Bond Formation in Prairie Voles
Authors: Francis Amie , Curtis J. Thomas 
Published: June 2021

The prairie vole is an established model for social behavior, most notably, the monogamous pair bond formed between a mated pair. The formation of this bond hinges upon estrogen surges in the female shortly after being introduced to a potential mate. Our lab has correlated earlier estrogen surges with earlier achievement of pregnancy, which we have also found to be a determining factor in the formation of a monogamous pair bond for male prairie voles. Currently, our lab is examining how female mate choice influences the formation of a monogamous pair bond via both behavioral and endocrine measures.This study has three objectives: 1) To establish whether endocrine measures provide supporting data for behavioral measures of pair bond formation; 2) To examine whether female prairie voles allowed to pair with a mate of their choice will show an earlier estrogen surge and a shorter latency pregnancy; and 3) To determine if pairing and pregnancy is a greater stressor for animals not paired with their mate of choice.Sexually naïve female prairie voles were allowed to choose between two males and subsequently paired either with the male of their choice or the male they did not choose. An additional group of females was paired randomly with a male outside of the choice paradigm. Fecal samples were collected prior to and immediately after mate choice, and at select time points throughout the next 30 days or until the birth of a litter. Fecal samples were analyzed via ELISA for 17b‐estradiol and corticosterone content to examine sex hormone changes associated with pairing and pregnancy, as well as stress levels throughout pairing and pregnancy. These measures will be combined with partner preference testing on either the female or male subject to establish whether female mate choice can affect endocrine changes that may impact monogamous pair bond formation.Regardless of sex, animals in pairs that include the non‐chosen male tend to display social affiliation patterns that differ from those pairs that include the chosen male or a randomly assigned male. Despite differences in affiliative behavior, latency to birth in all groups was similar. These data suggest that female mate choice may influence pair bond formation, despite the lack of differences in latency to pregnancy.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Creatine‐Monohydrate on Cognitive Function in Subjects Who Differ in Dietary Meat Consumption
Authors: Smith Kaitlyn , Weickel Caroline , Petersen Jahkeema , Skelton Michele 
Published: June 2021

The purpose of this study was to examine the effects of 4 weeks of creatine supplementation on cognitive function using the ImPACT test in vegetarians, subjects that consumed 1–10 servings of beef, chicken, pork, or fish per week, and subjects that consumed greater than 10 servings of beef, chicken, pork or fish per week. In a randomized, double‐blind placebo‐controlled study, twenty‐five subjects were divided into two groups. One group received creatine while the other group received a placebo. Subjects completed a 5‐day loading phase (20 grams per day) followed by a 24‐day maintenance phase (5 grams per day). Cognitive function was assessed prior to supplementation and immediately after a 4‐week supplementation phase. The ImPACT test was used and provides 6 measures of cognitive function. There were no significant changes in any of the cognitive measures between groups. However, vegetarians in the creatine group scored significantly higher than subjects who consumed greater than 10 servings of beef, chicken, pork or fish per week on the visual memory test. Preliminary analysis suggests that acute supplementation of creatine may enhance cognition in vegetarians but have no effect on those who consume meat regularly in their diet.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 Effect of Perinatal Blockade of Androgen Receptors on Adult Rat Behaviors and the Expression of Estrogen and Androgen Receptors in specific brain regions
Authors: Gopalan Chaya , Colley Zach , Niepoetter Paige , Waller Christopher , Cashikar Anil 
Published: June 2021

The brain, by default, differentiates into a female brain unless testosterone (T) is present during a critical window of time. In rats, there is a surge of T around the time of birth, more specifically four days prior to and four days after birth, referred to as the perinatal period. The purpose of this study was to examine the effects of blockade of androgen receptors (AR) during the perinatal period using flutamide (F) on sexual motivation (SM) and an open field (OF) test as well as the expression of AR and estrogen receptors (ER)‐alpha and ER‐beta in the hypothalamus, hippocampus, and amygdala. Four timed‐pregnant rats were divided into two groups: one group (N=2) received F during perinatal period whereas the other group (N=2) served as the control group receiving vehicle (95% Olive oil‐5% ethanol). Once the pups reached adulthood, the male rats underwent SM testing where they were exposed to either an estrus or an ovariectomized rat. Both F and control groups of male rats spent more time with the estrus rat compared to the ovariectomized rat suggesting that SM was not affected by their AR blockade during perinatal period. Open field testing recorded animal movements between the two zones, the center and the periphery, within an open arena. Male rats moved between the two zones more than the female rats did. At the end of the behavioral study, animals were sacrificed and the brains were harvested to isolate the hippocampus, hypothalamus, and amygdala. Real time PCR testing was carried out to quantify the levels of AR, ER alpha, and ER beta using B‐Actin as an internal control. Perinatal blockade of AR had no effect on the expression of any of the receptors in the three regions tested in the adult rats. Thus, the increased movement by the male rats in the OF testing is not associated with changes in the expression of AR, ER‐alpha, or ER‐beta.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sleep Restriction Worsens Daily Life Executive Functioning in Adolescents with ADHD
Authors: Fershtman Chaya E. M. , Kapadia Delna K. , Becker Stephen P. 
Published: June 2021

Attention‐deficit/hyperactivity disorder (ADHD) is a persistent neurodevelopmental disorder characterized by inattention and/or hyperactivity‐impulsivity. Adolescents with ADHD commonly experience deficits in areas of executive function (EF). These deficits are associated with academic, social, and emotional impairments. Experimental research shows shortened sleep duration to cause poorer daily life EF in typically developing adolescents. However, no previous research has looked at the casual relationship between inadequate sleep and executive functioning deficits in teenagers with ADHD. The objective of this study was to investigate whether sleep duration impacted daily life executive functioning. The hypothesis was that restricted sleep would negatively impact daily life executive functioning in adolescents with ADHD. A three‐week at home sleep manipulation protocol was implemented, using an experimental crossover design. Participants completed a phase stabilization week, followed by a sleep restriction (SR) week (6.5 hours) and sleep extension (SE) week (9.5). SR and SE conditions were counterbalanced across participants. Each week, participants attended laboratory visits at the medical center where executive functioning was measured with a validated rating scale, the Behavior Rating Inventory of Executive Function, Second Edition (BRIEF2). The BRIEF2 measured different areas of EF domains such as working memory, inhibition, planning/organization, emotional control, and initiation. Sleep duration was monitored with actigraphy and sleep diaries. Analyses looked at 48 participants who successfully adhered to the study protocol and who had complete actigraphy data. The results of a paired sample t‐test showed significantly (ps <.05) more deficits in most daily life EF domains during the sleep restriction phase compared to the sleep extension phase. This study provides the first evidence for shortened sleep duration as a causal contributor to worsened executive functioning in adolescents with ADHD. Increased sleep may significantly impact academic, social, and emotional functioning in adolescents with ADHD, and sleep may be an important future target for future intervention.Support or Funding InformationThis research was supported by the National Institute of Mental Health (NIMH; grant R03MH109787; Dr. Becker). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIMH.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Optogenetic Stimulation of Cholinergic Neurons in the Brainstem Induces Splenic Nerve Activity and Attenuates Systemic Inflammation
Authors: Kressel Adam M , Tsaava Téa , Chang Eric H , Addorisio Meghan E , Chang Qing , Pavlov Valentin A , Chavan Sangeeta S , Tracey Kevin J 
Published: June 2021

The inflammatory reflex is a well‐defined neural circuit composed of afferent and efferent fibers that travel via the vagus nerve to regulate peripheral tumor necrosis factor (TNF) production. Electrical stimulation of the efferent fibers reduces splenic TNF output in an animal model of endotoxemia. However, the exact origin of these vagus nerve fibers in the brainstem and the means by which they innervate the spleen to modulate TNF levels is not yet understood. Using optogenetics, we selectively stimulated cholinergic neurons in the dorsal motor nucleus of the vagus nerve (DMV), the autonomic brainstem nucleus from which the efferent vagus nerve fibers originate. A fiber‐optic cannula was inserted using stereotactic guidance into the DMV of transgenic mice expressing channelrhodopsin under the choline acetyltransferase promoter (ChAT‐ChR2‐EYFP). Mice were subjected to either optogenetic or sham stimulation (n=20 per group) for five minutes (473nm laser, 20Hz, 25% duty cycle). After 24 hours, animals were subjected to endotoxemia by intraperitoneal administration of lipopolysaccharide (0.25mg/kg), blood was collected after 90 minutes, and serum TNF was quantitated. Optogenetic stimulation of the cholinergic neurons in the DMV of ChAT‐ChR2‐EYFP mice significantly decreased endotoxin‐induced serum TNF levels compared to sham controls (p=0.0004). Splenic nerve activity, recorded during DMV stimulation using a cuffed two‐channel electrode, was significantly increased over baseline (p=0.0002). Blocking the signals transmitted in the vagus nerve by adding bupivacaine, a sodium channel antagonist, eliminated this effect (p=0.0028), implicating a functional synapse between the vagus and splenic nerves. These studies provide the first direct evidence for the central origin of the efferent vagus nerve fibers regulating TNF production during endotoxemia and suggest novel anti‐inflammatory approaches based on targeting DMV cholinergic signaling.Support or Funding InformationThis study was funded by NIH/NIGMS/NIAID.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 baroreflex testing with fMRI reveals baroreflex mediated brainstem nuclei
Authors: Gerlach Darius Andreas , Manuel Jorge , Hoff Alex , Kronsbein Hendrik , Hoffmann Fabian , Heusser Karsten , Ehmke Heimo , Jordan Jens , Beissner Florian , Tank Jens 
Published: June 2021

BackgroundBrainstem nuclei mediate baroreflex adjustments in efferent sympathetic and parasympathetic traffic. Yet, human brainstem physiology is poorly understood given the lack of suitable methodology. We developed a novel approach combining pharmacological testing, beat‐by‐beat cardiovascular monitoring, and high‐resolution functional magnetic resonance imaging (fMRI) to assess human baroreflex regulation at the level of the brainstem.Method and materialsIn 11 healthy men (30.7 ±6.6 years; 24.1 ±1.9 kg/m2), we monitored continuous finger arterial blood pressure and ECG using customized hardware during multiband fMRI brain acquisitions. We applied repeated intravenous phenylephrine (PHE, 25 and 75 μg, n=8) and nitroprusside (NTP 25 and 75 μg, n=8) boluses using a remote controlled injector. Brainstem and hypothalamus fMRI images were analyzed to identify brainstem nuclei involved in baroreflex‐mediated blood pressure control using with masked general linear model (GLM). BP changes were correlated with the time‐courses of blood‐oxygen‐level dependent (BOLD) signals by mixed‐effects general linear model.ResultsPharmacological baroreflex testing yielded baroreflex sensitivity measurements with typical blood pressure and RR‐interval patterns. Correlation of the baroreflex‐mediated changes in systolic blood pressure with BOLD signals clearly revealed brainstem nuclei. The solitary tract (NTP: 7.3; PHE: 6.0 t‐value), the caudal ventrolateral medulla (PHE: 5.8 t‐value); the rostral ventrolateral medulla (NTP: 5.6; PHE: 7.1 t‐value), the nucleus ambiguus (PHE: 5.8 t‐value), the paraventricular nucleus (NTP:7.1; PHE: 4.5 t‐value), and raphe nuclei (NTP: 5.7; PHE: 6.0 t‐value), were identified with high sensitivity and corrected for multiple comparisons (p < 0.01).ConclusionWe developed novel approach testing baroreflex regulation at the level of the brainstem in humans. The methodology identified baroreflex‐mediated activation and deactivation patterns consistent with previous investigations in animal models. The methodology can be applied to elucidate human physiology and mechanisms of autonomic cardiovascular disease.Support or Funding InformationThis study was funded by the DLR internal cost object 2475023.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.